COMMISSION OF THE
EUROPEAN
COMMUNITIES
    UNITED STATES
   ENVIRONMENTAL
PROTECTION AGENCY
                       WORLD HEALTH
                       ORGANIZATION
            INTERNATIONAL  SYMPOSIUM


                     PROCEEDINGS


                    Recent Advances
          in the Assessment of the Health Effects
                of Environmental Pollution
                        Volume IV


                   '• '



   24 to 28 June 1974
      EUR 5360

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LEGAL NOTICE

The Commission of the European Communities
and its departments decline at)
responsibility with regard to the use of
the information contained herein.

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                           TAGUNGSBERICHTE
                      INTERNATIONALES  SYMPOSIUM

              NEUESTE ERKENNTNISSE  IN DER BEURTEILUNG
        DER GESUNDHEITLICHEN FOLGEN DER UMWELTVERSCHMUTZUNG
                             PROCEEDINGS
                       INTERNATIONAL SYMPOSIUM

                  RECENT ADVANCES  IN THE ASSESSMENT
         OF THE HEALTH EFFECTS OF  ENVIRONMENTAL POLLUTION
                                ACTES
                       SYMPOSIUM  INTERNATIONAL

                PROGRES REGENTS DANS L1EVALUATION DES
       EFFETS DE  LA POLLUTION DE L'ENVIRONNEMENT SUR LA SANTE
                                 ATT I
                       SIMPOSIO  INTERNAZIONALE

                 RECENTI PROGRESSI  NELLA VALUTAZIONE
    DEGLI EFFETTI  DELL'INQUINAMENTO DELL' AKSIENTE SULLA SALUTE
                                VORSLAG
                      INTERNATIONAAL SYMPOSIUM

            RECENTE VORDERINGENBIJ DE VASTSTELLING VAN
     DE GEVOLGEN  VAN MILIEUVERONTREINIGING  VOOR DE GEZONDHEID
                  Paris  (France)f  June 24-28, 1974
                          organized jointly by

               CEC - Commission of the European Communities
     Directorate General for Social Affairs - Health Protection Directorate

             EPA - United States Environmental Protection Agency

                                 and

                     WHO - World Health Organization
          Published by the Commission of the European Communities
Directorate General Scientific and Technical Information and Information Management

                            Luxembourg, 1975

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                                 XCVII
ORGANISATIONSAUSSCHUSS / ORGANIZING COMMITTEE / COMIT£  ORGANISATEUR
             COMITATO ORGANIZZATIVO / ORGANISEREND
         Vorsitzend / Chairmen / Presidents / Presidents / Voorsitters:
                         B.H. DIETERICH  (W.H.O.)
                          S. GREENFIELD (E .P .A .)
                            P.  RECHT  (C.E.C.)
        Stellvertretende Vorsizende / Vice Chairmen / Vice-presidents /
                      Vicepresidenti / Viae-Voorzitters:

                            D.  EARTH  (E.P.A.)
                         M. CARPENTIER  (C.E.C.)
                           J.  KUMPF (W.H.O.)


           Generalsekretar / Secretary General / Secretaire general /
                  Segretario generate / Algemeen Secretaria:

                           J.  SHEETS  (C.E.C.)
           Vieeenschoftliche Sekretoriat / Scientific Secretariat /
             Secretariat scientifique / Segretariato scientific*) /
                        Wetenschappelijk Secretariaat:

                           A.  BERLIN  (C.E.C.)
                            R. ENGEL  (E.P.A.)
                           V.B.  VOOK  (W.H.O.)
         Assistentin / Assistant / Aseistante / Assistente / Assistent

                         G.  TREU-RICCO'  (C.E.C.)
               ifiesenschaftliche Berater / Scientific advisors /
              Conseillers scientifiques / Consiglieri scientifici /
                         Wetenschappelijke Adoiseups:

             Ph.  BOURDEAU  (C.E.C.), G.C.  BUTLER  (Canada)
          M. COLOMBINI  (Italy) ,  G. DEAN (Republic  of Ireland) ,
               J.F.  FINKLEA  (U.S.A.), L. FRIBERG  (Sweden),
               H.E.  GRIFFIN  (U.S.A.), W.W. HOLLAND  (U.K.),
               M. KEY  (U.S.A.),  A. LAFONTAINE  (Belgium),
           D.  RALL (U.S.A.),  H.-W. SCHLIPKOTER  (F.R. Germany),
            R. SENAULT  (France), J. SPAANDER  (Netherlands),
               L. TEPPER  (U.S.A.), N.K. WEAVER  (U.S.A.)

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                        XCIX


                  TAGUNGSBERICHTE
               INTERNATIONALES SYMPOSIUM
        NEUESTE ERKENNTNISSE IN DER BEURTEILUNG
  DER GESUNDHEITLICHEN FOL6EN DER UMWELTVERSCHMUTZUNG
                        Band I
                   ERO'FFNUNGSSITZUNG
               POPULATIONSUNTERSUCHUNGEN
  UNTERSUCHUNG DER WIRKUNGEN AUF DEN MENSCHEN
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                   PROCEEDINGS
             INTERNATIONAL SYMPOSIUM
   RECENT ADVANCES IN THE ASSESSMENT OF  THE HEALTH
        EFFECTS OF ENVIRONMENTAL POLLUTION

                     Volume I
                 OPENING SESSION
               POPULATION STUDIES
           HUMAN  EFFECTS  STUDIES  (Panel)
              TOXICOLOGICAL STUDIES
            EXPOSURE  MONITORING  (Panel)

                     Volume II
              HUMAN  EFFECTS STUDIES
                  ANIMAL  STUDIES
                   METABOLISM
                   INTERACTIONS
              INDICATORS  OF EXPOSURE

                   Volume III
               TISSUE MEASUREMENTS
                 MONITORING NEEDS
            ENVIRONMENTAL MEASUREMENTS
                EXPOSURE  MONITORING
                      MODELS
               SUPPLEMENTARY  PAPERS

                     Volume IV
THE SCIENTIFIC DATA BASE  REQUIRED FOR DECISIONS TO
  PROTECT HUMAN HEALTH   (Plenary discussion group)
            EXPERIMENTAL INVESTIGATIONS
             MONITORING NEEDS (Panel)
            TISSUE MEASUREMENTS  (Panel)
              HEALTH EFFECTS STUDIES
                  CLOSING SESSION
               LIST OF PARTICIPANTS

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                             CI


                          ACTES
                  SYMPOSIUM  INTERNATIONAL
           PROGRES REGENTS DANS L'EVALUATION  DES
  EFFETS DE LA  POLLUTION  DE  L*ENVIRONNEMENT SUR  LA SANTE

                         Volume I
                    SEANCE D'OUVERTURE
                 ETUDES SUR  LA POPULATION
           ETUDES DES EFFETS SUR  L*HOMME (Panel)
                   ETUDES TOXICOLOGIQUES
               MESURE DE  L'EXPOSITION (Panel)

                         Volume II
               ETUDES DES EFFETS  SUR L'HOMME
                  ETUDES  SUR LES  ANIMAUX
                       METABOLISMS
                       INTERACTIONS
                 INDICATEURS D'EXPOSITION

                       Volume III
         MESURES RELATIVES  AUX TISSUS BIOLOGIQUES
        BESOINS EN MATIERE  DE MESURE DE  L'EXPOSITION
               MESURES DANS  L'ENVIRONNEMENT
                  MESURE  DE  L'EXPOSITION
                         MODELES
                 RAPPORTS SUPPLEMENTAIRES

                         Volume IV
          LES DONNEES SCIENTIFIQUES REQUISES  A
LA PRISE DE DECISION POUR LA PROTECTION DE LA SANTE HUMAINE
        (Groupe de discussion en  assembles plSniere)
                 RECHERCHES EXPERIMENTALES
    BESOINS EN MATIERE DE MESURE  DE L'EXPOSITION  (Panel)
      MESURES RELATIVES AUX TISSUS BIOLOGIQUES  (Panel)
              ETUDES DES EFFETS SUR LA SANTE
                     SEANCE DE CLOTURE
                  LISTE DES PARTICIPANTS

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                             CII

                           ATT I
                  SIMPOSIO INTERNAZIONALE
            RECENTI PROGRESS I NELLA VALUTAZIONE
DEGLI EFFETTI DELL'INQUINAMENTO DELL'AMBIENTE SULLA SALUTE

                         Volume I
                     SESSIONS  INAUGURALE
                  STUDI SULLA POPOLAZIONE
           STUDI DEGLI EFFETTI SULL'UOMO (Panel)
                   STUDI TOSSICOLOGICI
              MISURA DELL'ESPOSIZIONE (Panel)

                         Volume II
               STUDI DEGLI EFFETTI SULL'UOMO
                    STUDI SUGLI ANIMAL!
                        METABOLISMO
                        INTERAZIONE
                 INDICATORI DI ESPOSIZIONE

                        Volume III
               MISURE NEI TESSUTI BIOLOGICI
      NECESSITA" RELATIVE ALLA MISURA DELL1 ESPOSIZIONE
                     MISURE AMBIENTALI
                  MISURA DELL'ESPOSIZIONE
                          MODELLI
                  RAPPORTI SUPPLEMENTARI

                         Volume IV
         I DATI SCIENTIFICI DI BASE RICHIESTI PER
 PRENDERE DECISIONI AL FINE DI PROTEGGERE LA SALUTE UMANA
                (Gruppo di discussione plenaria)
                   INDAGINE SPERIMENTALE
NECESSITA' RELATIVE ALLA MISURA DELL'ESPOSIZIONE  (Panel)
           MISURE NEI TESSUTI BIOLOGICI  (Panel)
             STUDI DEGLI EFFETTI SULLA SALUTE
                     SESSIONS  FINALE
                  ELENCO DEI PARTECIPANTI

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                              cm

                            VORSLAG
                    INTERNATIONAAL SYMPOSIUM
            RECENTE VORDERINGEN BIJ DE VASTSTELLING
 VAN DE GEVOLGEN VAN MILIEUVERONTREINIGING VOOR DE GEZONDHEID

                              Band I
                        OPENINGSZITTING
                       BEVOLKINGSSTUDIES
        ONDERZOEKINGEN NAAR EFFECTEN BIJ DE MENS  (Panel)
                 TOXICOLOGISCHE ONDERZOEKINGEN
                   METING VAN EXPOS IE  (Panel)

                             Band II
           ONDERZOEKINGEN NAAR EFFECTEN BIJ DE MENS
                   ONDERZOEKINGEN BIJ DIEREN
                          METABOLISME
                          INTERACT IES
                    INDICATOREN VAN EXPOS IE

                           Band III
                METINGEN VAN BIOLOGISCH WEEFSEL
   El SEN VOOR NET TOT STAND BRENGEN VAN TOEZICHT OP EXPOS IE
                    METINGEN  IN HET MILIEU
                     METINGEN VAN EXPOS IE
                           MODELLEN
                   SUPPLEMENTAIRE DOCUMENTEN

                            Band IV
  WETENSCHAPPELIJKE GEGEVENS NOODZAKELIJK VOOR HET NEMEN VAN
  BESLISSINGEN TER BESCHERMING VAN DE GEZONDHEID VAN DE MENS
                    (Plenaire discussiegroepj
                    EXDERIMENTEEL ONDERZOFK
El SEN VOOR HET TOT STAND BRENGEN VAN TOEZICHT OP EXPOS IE  (Panel
            METINGEN VAN BIOLOGISCH WEEFSEL (Panel)
        ONDERZOEKINGEN NAAR GEVOLGEN VOOR DE GEZONDHEID
                          SLOTZITTING
                        DEELNEMERSLIJST

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INHALTSVERZEICHNIS




     CONTENTS





TABLE DES MATIERES





      INDICE





   INHOUDSOPGAVE

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                              CVII
DIE WISSENSCHAFTLICHEN  DATENGRUNDLAGEN,  DIE FUER DIE
ENTSCHEIDUNG UEBER DEN  GESUNDHEITSSCHUTZ BENOETIGT WERDEN
(Aussprache im P.ahmen des Gesamtausschusses)

THE SCIENTIFIC DATA BASE  REQUIRED FOR DECISIONS TO PROTECT
HUMAN  HEALTH (Plenary discussion group)

LES DONNEES SCIENTIFIQUES REQUISES A LA  PRISE DE DECISION
POUR LA  PROTECTION DE LA  SANTE HUMAINE  (Groupe de discus-
sion en  assemblee pleniere)

I DATI SCIENTIFICI DI BASE RICHIESTI PER PRENDRE DECISIONI
AL FINE  DI  PROTEGGERE LA  SALUTE UMANA   (Gruppo di
discussione plenaria)

WETENSCHAPPELIJKE GEGEVENS NOODZAKELIJK  VOOR  MET NEMEN
VAN BESLISSINGEN TER BESCHERMING VAN DE  GEZONDHEID VAN DE
MENS (Plenaire discussiegroep)	1873
EXPERIMENTELLE FORSCHUNG
EXPERIMENTAL INVESTIGATIONS
RECHERCHES  EXPERIMENTALES
INDAGINE  SPERIP.ENTALE
EXPERIKENTEEL ONDERZOEK

     The state of the  art regarding the experimental investigations
     of the  effects of pollutants	1967
     D.V. BATES

     The effects of low levels of S0_  and ozone in the same
     atmosphere on human pulmonary function  	   1979
     D.V. BATES, M.  HAZUCHA

     Effects of sulfur dioxide on healthy and peripheral airway
     impaired subjects	1989
     F.W. WEIR,  P.A. BROMBERG

     Emergency population exposure:  a methodological approach. .   2005
     P.E. JOOSTING,  M.M. VERBERK

     L'influence du bruit sur le sommeil de travailleurs de nuit et
     des travailleurs par equipes alternantes 	   2031
     P. KNAUTH,  J. RUTENFRANZ

     Alteration in urinary D-glucaric  acid excretion as an indication
     of exposition to xenobiotics  	   2047
     W.R.F.  NOTTEN,  P.Th. HENDERSON

     The assessment of environmental carcinogen risks in terms of
     life shortening  	2061
     R.E. ALBERT, B. ALTSHULER

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                               CVUI
 ERFORDERNISSE IM HINBLICK AUF MESSUNGEN (Panel)
 MONITORING  NEEDS (Panel)
 BESOINS EN  MATIERE DE  MESURE DE L  EXPOSITION  (Panel)
 NECESSITY RELATIVE ALLA  MISURA DELL'ESPOSIZIONE  (Panel)
 El SEN VOOR  NET TOT STAND BRENGEN VAN TOEZICHT  OP EXPOS IE  (Panel

      Pollutant-oriented integrated monitoring systems  and lead
      exposure  assessment     	    2073
      E.A. SCHUCK, G.B. MORGAN, D.S.  BAPTJI

      Integrated assessment of health effects of air pollution  . . 2083
      L.R. BABCOCK jr,  N.L.  KAGDA,  A.T.  RCSSANO

     Mexico city's pindex level	2091
      H.BRAVO  A,  M.E.  ESPINOSA,  A.C.  LOWE,  S.  SALAZAR

      A method for  simulating the true human exposure of critical
      population groups to air pollutants	2097
      W.R. OTT, D.T.  MAGE

      Personal and  indoor exposure meters 	 2109
      A.P. ALTSHULLER,  A.E. O'KEEFFE

      Die automatische Ueberwachung der Kraftfahrzeugabgas-Immission
      im Arbeits- und Wohnbereich   	2115
      D.  JOST, W.  RUDOLF

      Incorporation of population exposure  concepts in air quality
      criteria documents	2143
      A.C. STERN,  Y. HORIE

           Panel  Discussion	2153


GEWEBSMESSUNGEN   (Panel)
TISSUE MEASUREMENTS  (Panel)
MESURES  RELATIVES AUX  TISSUS  BIOLOGIQUES  (Panel)
MISURE NEI TESSUTI BIOLOGICI  (Panel)
METING VAN BIOLOGISCH  WEEFSEL   (Panel)

      Intercomparison programme on the analysis of lead, cadmium
      and mercury in biological fluids	2185
     A.  BERLIN,  R. LAUWERYS,  J.P.  BUCKET,  H.  ROELS,
     P.  Del  CASTILHO,  J.  SHEETS

     A comparison  of methods for analysis of cadmium in food and
     biological material.  A co-operative study between Sweden,
     Japan and U.S.A	2197
     T.  KJELLSTROM, K. TSUCHIYA,  E. TOMPKIKS,  F. TAKAJJATAKE
     B.  LIND, L.  LINNMAN

     The determination of  lead and cadmium in blood by atomic
     absorption spectroscopy 	  2215
     H.T.  DELVES

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                                CIX
     Mesure du plomb dans les os humains provenant de la region
     parisienne	          2225
     L. JEANMAIRE,  F.  PATTI, N.  CIALELLA/n!  GROS"

     Zur Blei- und Cadmiumanalytik  in biologischen Matrices  .  .  .  2231
     M. STOEPPLER,  F.  BACKHAUS,  R.  DAHL, M. DUMONT,
     H. HAGEDORN-GOETZ,  K. HILPERT, P. KLAHRE, K. RtfTZEL,
     P. VALENTA, H.W.  NUERNBERG

           Panel Discussion	2247

GEWEBSMESSUNGEN  (Panel)
TISSUE  MEASUREMENTS (Panel)
MESURES RELATIVES  AUX TISSUS BIOLOGIQUES   (Panel)
MISURE  NEI TESSUTI  BIOLOGICI   (Panel)
METING  VAN BIOLOGISCH WEEFSEL  (Panel)

     The concentrations of  copper,  iron,manganese, zinc and  cadmium
     in human hair as a possible indicator of their tissue concen-
     trations    	,_	2263
     E.  VUORI, A.  HUUNAN-SEPPALA,  J. KILPIO

     Trace elements in lung and hair	,	2271
     G.W.  GIBBS, E. BOGDANOVIC

     Trace metal content of hair and  the levels of 5-HIAA in the
     urine of population exposed to lead	2285
     B.  STANKOVIC,  M. STANKOVIC, S. MILIC,  Z. KORICANAC,
     Lj.  MILOVANOVIC, M. DUGANDZIC

           Panel Discussion	2293


UNTERSUCHUNG DER GESUNDHEITLICHEN WIRKUNGEN
HEALTH  EFFECTS STUDIES
ETUDES  DES EFFETS  SUR LA SANTE
STUDI DEGLI EFFETTI SULLA SALUTE
ONDERZOEKINGEN NAAR GEVOLGEN VOOR DE GEZONDHEID

     Health effects studies - now and in the future	2307
     L.  FRIBERG

     The carcinogenic risk  for man  of environmental chemicals  .  .  2317
     L.  TOMATIS

     Interaction of environmental chemical,enzymatic  changes and
     their possible effects on health 	  2331
     H.L.  FALK

     Perspectives in the environmental toxicology  of  trace metals  2351
     K.  TSUCHIYA

     Assessment of the influences of  environmental pollutants on
     cancer and other chronic diseases   	  2367
     S.S.  EPSTEIN

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                                 ex
      The use of quantitative  EE3 for detecting low-level prolonged
      exposure to pesticides  	  2387
      J.A.  SANTOLUCITO

      Internal pollution - Our first priority. A review of the studies
      of the special commission of internal pollution	2397
      P.  BEACONSFIELD, N.  BORLAUG, A. CAPRI DI RISMINI,
      H.  KREBS,  R. PETERS,  R.  RAINSBURY

      A comprepensive toxicological limitation of toxic substances in
      the environment.   Functional and metabolic criteria of noxious
      action	,	,.	2409
      N.  IZMEROV, I.V.  SANOTSKY

      Behavioral methods for investigating environmental  health
      effects	    2415
      B.  WEISS
SCHLUSSITZUNG
CLOSING SESSION
SEANCE DE  CLOTURE
SESSIONE FINALE
SLOTZITTING

     Conclusions  et reflections  des conseillers  scientifiques.  . . .  2437
     A.  LAFONTAINE

     B.H.  DIETERICH	2451
      Director Division of Environmental Health, World Health
     Organization

      A.C.  TRAKOWSKI	2455
      Assistant Administrator for Research and Development - Ihited
      States Environmental Protection Agency

      P.  RECHT	2461
      Directeur de la Protection Sanitaire - Commission des
      Conanunautes Europeennes

TEILNEHMERLISTE - LIST  OF PARTICIPANTS  - LISTE DES
PARTICIPANTS  -  LISTA DEI PARTECIPANTI - LUST VAN
DE DEELNEMERS	2469

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                            1873
Aussprache im Kahmen des Gesamtausschusses - Plenary Discussion
Group - Groupe de Discussion en Assemblee pleniere - Gruppo
     di Discussione Plenaria - Plenaire Discussiegroep
   DIE WISSENSCHAFTLICHEN DATENGRUNDLAGEN, DIE FUER DIE
 ENTSCHEIDUNG UEBER DEN GESUNDHEITSSCHUTZ BENOETIGT WERDEN

    THE SCIENTIFIC DATA BASE REQUIRED FOR DECISIONS TO
                   PROTECT HUMAN HEALTH

 LES DONNEES SCIENTIFIQUES REQUISES A LA PRISE DE DECISION
          POUR LA PROTECTION DE LA SANTE HUMAINE

 I DATI SCIENTIFICI DI BASE RICHIESTI PER PRENDERE DECISIONI
           AL FINE DI PROTEGGERE LA SALUTE UMANA

 WETENSCHAPPELIJKE GEGEVENS NOODZAKELIJK VOOR HET NEMEN VAN
  BESLISSINGEN TER BESCHERMING VAN DE GEZONDHEID VAN DE MENS
Chairman :
D.S. EARTH
Members :
D. BENINSON

K. EIERSTEKER
G.C. BUTLER

J.R. GOLDSMITH

P. RECHT

J. TREMOLIERES

V.B. VOUK
U.S. Environmental Protection Agency, National
Environmental Research Centre, Las Vegas,
Nevada, U.S.A.
Gerencia de Proteccion, Radiologica y Seguridad,
Buenos Aires, Argentina
Municipal Health Department, Rotterdam, Nederland
Division of Biological Sciences, National
Research Council of Canada, Ottawa, Ontario,
Canada
National Cancer Institute and California State
Health Dept., Bethesda, Maryland, U.S.A.
Commission des Communautes Europeeennes,
Direction Protection Sanitaire, Luxembourg
Laboratoire de Nutrition Humaine, Institut
National de la Sante et de la Recherche
Medicale, Hopital Bichat, Paris, France
World Health Organization, Division of  Environment^
Health, Geneva, Helvetia

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                              1875
    THE  SCIENTIFIC DATA BASE REQUIRED FOR DECISIONS
                TO PROTECT HUMAN  HEALTH
                    (Chairman's Note)

                      D. S.  EARTH

The purpose  of  the Plenary Discussion Group was to provide
a forum for  discussion of the interface between scientists
doing research  to  assess health effects of environmental poll-
utants and decision makers who must use the resulting infor-
mation to develop  adequate environmental protection plans,
Each member  of  the Plenary Discussion Group was selected on
the basis of his scientific expertise and was asked to repre-
sent his  personal  views and convictions in responding to quest-
ions and  in  discussing issues. Thus, the answers provided
to questions represented the personal views of the individual
responders and  not the collective views of the entire Plenary
Discussion Group.

Each member  of  the Group was given an opportunity for an open-
ing statement to address those issues which he considered
to be of  the highest priority. They were requested to consid-
er the following topics as part of their opening statement:
    o  What is the contribution  and significance of epidemic-
       logical, clinical and toxicological studies in deter-
       mining  protection guides?
    o  In determining exposure-effect relationships, what
       is the  relative importance of parameters such as age,
       race, state of health, nutritional status and eating
       habits, climate, occupation, smoking history, socio-
       economic status, etc?
    o  How  important are biological responses, such as increa-
       ses  in  pollutant burden or physiological effects of
       uncertain  significance, in setting protection guides?
    o  In the  development of protection guides, how does
       one  account for the difference between acutely toxic

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                               1876
         and  cumulatively  toxic  environmental  pollutants?
      o   How  should  exposure-effect relationships  be  expressed
         and  analyzed  to be most helpful  in  developing protect-
         ion  guides?   Is a "threshold"  or a  "non-threshold"
         concept preferable?
      o   Is it possible to account for, or correct for,  concurr-
         ent  exposure  to multiple pollutants which may be syn-
         ergistic or antagonistic?
      o   What factors  go into determining an adequate margin
         of safety for a protection guide?
      o   What factors  go into determining meaningful  relation-
         ships between public health protection guides and
         occupational  health protection guides?
      o   should a fixed value be set as a protection  guide
         or should a range of values be given?
      o   What are the  population groups (aged, children,  diseas-
         ed,  etc.) to  be considered in proposing protection
         guides for different pollutants?
      o   What is the importance  of indoor  measurements vs.
         outdoor measurements in determining exposure?
      o   Is it possible to relate one study  to another when
         different measurement methods were used to assess
         either exposure or effects?
Following these presentations, questions  which had been  pre-
viously  submitted in  writing by the participants  were answered
by one or more members of the Group.   The original questions
asked in writing by the participants at  this Symposium are
reproduced at the end of this session.  The Panel Members and
the Scientific Secretariat met prior to  the plenary  session
to summarize and combine the original questions into  a fewer
number so all subject areas germane to this discussion could
be covered within the time allotted.  During the course of answer-
ing the panel questions, it became apparent that  there would
not be sufficient time to allow oral questions and discussions
from  the floor.   The Chairman, feeling that oral participat-
ion by Symposium participants was desirable, deferred answering

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                               1877
some of the panel questions with the promise that all questions
not answered during the session would be answered in the Pro-
ceedings.  Thus,  the last part of the session was devoted to
oral interchanges between participants and members of the Group.
At the conclusion of the session, there were many participants
still asking for recognition to bring up additional questions.
To accommodate these additional questions, the Group agreed
to accept  pertinent written questions after the session with
the promise that all such questions would be answered in the
Proceedings.
           OPENING OF THE PLENARY GROUP DISCUSSION

BJVRTH
As Chairman, I would like to call this session to order please.
This is the Plenary Discussion Group on the subject  'the  Scien-
tific Data Base Required for the Decisions to Protect  Human
Health1.   First I would like to read from a document which
was prepared by the Scientific Advisors, to advise the organiz-
ing committee with regard to the purpose of such a session
as this.   Specifically the words are as follows:   'as  stated
in the announcement the symposium is of particular interest
for all those concerned with the public health and research
aspects of environmental pollution as well as those  concerned
with its control.  The purpose of this panel session is to
develop strategies for organising the scientific knowledge
of the exposure-effect relations for the development of criteria.
This discussion should therefore address itself to three  aspects
of the scientific data required by administrators.   First is
the kind of data required.  Ideally the administrator  should
be provided with estimates of the probability of the effects
produced in a variety of receptors by each pollutant.  How
the data are presented will determine his course of  action.
Second is the quantitative description of the damage.  The
description should be given as an estimate of absolute risk.

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                               1878
This can then be converted  to a  relative  risk  and  compared
with other risks to the  same population.   Finally,  crucial
lacks in the data base must be identified,  ordered  in  priority
and considered in planned research.

In a letter that I sent  to  the members  of the  P'lenary  Discuss-
ion Group, I wrote the following paragraph,  which  is just an
extension of what we have stated.   "The purpose  of  this  Plenary
Discussion Group will be to provide  an  interchange  between
the assessment of details,  scientific data and the  decisions
required for the protection of human health  from environmental
pollutants."  Today our  discussion must come to  grips  with
the kinds of questions facing decision  makers  and  how  best
to furnish technical information in  terms which  will be  under-
standable to them.  Philosophical concepts  are necessary, but
wherever possible the discussion should refer  to practical
guidelines for real situations.

Each member of the group will have an opportunity  to make
some introductory remarks and each member is asked  to  limit
those remarks to less than  five  minutes.   Following the state-
ment from each one of the members, we will  then  proceed  to
answer written questions which have  been  submitted  to  us from
the floor.  Because of the  number of questions and  the large
area of subject matter,  I will read  a condensed  paraphrased
version of the submissions.  I think that those  of  you who
wrote the questions will recognize your subject.

BENINSON
The basic objectives of protection are  to prevent  the  occurr-
ence of deterministic effects (acute or late)  and  to limit
the probability of occurrence of stochastic  effects to levels
deemed to be acceptable.  The first  of  these objectives  is
easily met due to the existence  of thresholds  and probably
exposure rate effects.  The second objective relates to  much
more complicated problems,  mainly due to  the absence of  human
data, particularly at the levels of risk  which would be  considered

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                              1879
safe.  As it has been the normal practice in radiation protect-
ion, it is possible here to adopt a conservative assumption,
namely, that a non-threshold linear relationship exists between
the exposure and the probability of such late effects as  the
induction of malignancies and deleterious hereditary effects.
Furthermore it can be assumed that the risk per unit exposure
which might be deduced from observations at high exposures
apply to the low exposure range relevant for protection.

An implicit consequence of these assumptions is that no expos-
ure is absolutely safe.  The main issue is therefore the  accept-
ability of the implied risks, in relation to both the accept-
ability of other risks by society and the benefits  expected
from the operations causing the exposure.

At the low risk levels relevant for protection, the following
basic information appears to be required for setting limits:
     o  adequate exposure parameters  (probably time integrals
        of concentration of the pollutant in the critical organs)
     o  risk-exposure relationships (at least for high expos-
        ures, which could be conservatively assumed to apply
        at low exposures)
     o  a selection of an "acceptable" level of risk for  mem-
        bers of the public.

As many possible effects can be due to a given pollutant,
a parameter quantifying the total impact may be required.

Using the approach developed for radiation protection, the
total impact could be represented by the "detriment" as origin-
ally defined by the ICRP.  The "detriment" in a population
is the expectation of harm incurred from an exposure taking
into account not only the probabilities of each type of deleter-
ious effect but the severity of the effects as well.  Thus
if P.^ is the probability of suffering the effect i, the sever-
ity of which is expressed by a weighting factor g^f then  the

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                                1880
 detriment G in a group composed of P persons is = PZ(p. g.).
 Under the assumption of linearity of each p± with exposure
 pi = riE(where r± is a risk coefficient), and further assum-
 ing that the severity of stochastic effects is independent of
 their frequency, the detriment can be expressed as G = PEE(r.g.),
 where the sum is a constant for each pollutant.

 In addition to limits of exposure (Primary Protection Standards)
 it is necessary to derive limits for discharge of pollutants  •
 and for levels in the environment.   The use of environmental
 models allows the establishment of  relationship between dis-
 charges,  environmental levels and exposures.   The basic data
 required  are the transfer parameters relating compartments
 of the model.

 As in the case of radiation protection,  when  more quantitative
 toxicological  and ecological data become available,  further
 requirements of  protection  will have to  be  taken  into account,
 namely the  "justification"  of  a given practice  and the  "optimiz-
 ation"  of protection.

 Assessing the  justification of  a given source  involves  cost-
 benefit analysis  considering its  total positive and  negative
 effects as well  as the availability  of alternative procedures.
 The optimization  of protection,  on the other hand, is assessed
 by differential  cost-benefit analysis, with the purpose of
 finding the  lowest reasonably achievable exposure.  This is
 obtained  when the costs of  further reduction outweigh the  achie-
 ved reduction of detriment.  Both types of cost-benefit analysis
 require the  assignment of a  monetary  cost per unit exposure.
This is a very complex problem, but some values have been  pub-
 lished at least in the field of radiation protection and similar
values could be derived for  other pollutants if the exposure-
risk relationships become available.

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                               1881
 BIERSTEKER
 Environmental  health can be defined as the collection of scien-
 tifically  sound  data that can help man in establishing positive
 relations  with his  environment,  leading to better health and
 welfare, now and in future generations.

 The first  part of this  definition is a means,  the second a
 goal or objective.   About the objective exists little differ-
 ence in public health circles, though health is harder to define
 than in the past, when  indicators like the infant mortality
 rate, life expectancy and incidence of communicable diseases
 were simple yardsticks.   Nowadays the content  of life seems
 to gain in importance and it is  not unusual that large numbers
 of people  are  willing to trade in a few months or even one
 or two years of  life expectancy  for a beautification of the
 remaining  years.

 To study the means  of our definition,  we have  to collect data
 that demonstrate  qualitative and  quantitative  relations between
 man and his environment.   So far  this  has been mainly the task
 of epidemiologists.   Thanks  to the  work of a relatively small
 number of  scientists, the role of  the  environment in the trans-
 mission of communicable  diseases  has  been successfully document-
 ed and communicable  disease  control possesses  enough scientif-
 ically sound data to control  diseases  like poliomyelitis,  mea-
 sles, diphtheria, rubella, smallpox, whooping  cough,  malaria,
 schistosomiasis,  cholera,  typhoid fever,  etc.

When we look at these victories,  insiders  realize,  however,
 that in the list of  diseases  which  I mentioned,  the environ-
ment often plays  a minor  role, at least  from the  standpoint
of control.  In many  cases the environment is  left  untouched,
 for instance when vaccination is used  as  the main tool to
prevent communicable diseases.  In  other  control  programs  the
focus is on only  a very  small part  of  the  environment.  Malaria
control, for example, does not aim  at Anopheles eradication
but at  elimination of those members of  the malaria  parasites

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                               1882
transmitting species, which have just taken a blood meal from
a potential parasite carrier.

The fact that malaria eradication and schistosomiasis eradic-
ation are so much harder to achieve than smallpox and diphtheria
eradication, is due to the fact that the the environment plays
a more important role in the transmission of the first two
diseases.  The difference is also due to the fact that man
himself plays an active role in malaria and schistosomiasis
eradication. If he does not participate fully due to lack of
understanding or simply unwillingness, the eradication becomes
a doubtful goal.  Where this cooperation means a change in
his way of life, in his habits, the risk of failure increases
enormously.

The relation between man and his chemical environment is much
less understood than the relationship between man and his
environmental microbes.  This should caution us against a too
optimistic view of the contribution that medicine can make
in this non-communicable sector of environmental health.  Even
where we succeed in establishing true cause and effect relations,
control may be difficult because control means a change in
his way of life, in his philosophy, and in the values he places
on things.  To achieve an environment safe for health, there
will be no easy victories as were obtained by vaccination,
chlorination of water supplies, indoor spraying of DDT, etc.
It will take scientific data about man at risk from the envir-
onment as well as about the environment at risk from man.
And it will take data about means to make man change his way
of life.

The contribution this meeting makes is mainly in the first
of these three fields of action; i.e., data acquisition about
the effect of chemical pollutants in the environment on mans1
health.  A large number of epidemiological studies has been
necessary to elucidate such simple questions as:  where do

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                               1883
 populations  begin to show an increase in respiratory disease
 prevalence or  incidence if we measure sulfur oxides and smoke
 as  indicators  of  pollution.   From the standpoint of public
 health,  smoke  and sulfur oxides can never do good to a human
 lung.  The fact that we study the relationships is already
 a concession.  Society expects data on which it can base an
 ethical  judgment.   We do not know at what prevalence or incid-
 ence society wants action, but we all know that there is a
 sentimental  feeling about the physically weak in a society.
 The standards  for air pollution that society wants seem in
 the first place to aim at the protection of people with exist-
 ing cardio-respiratory impairments.   For the long term exposure
 standards we even lack proof whether effects are due to the
 short increases in pollution or to the chronic load of small
 particles that exists in such environments. Surprisingly enough,
 the same lack  of  insight exists about oxidants.   Whether the
 intermittent exposure has long term effects is still a matter
 of doubt.  We  are  somewhat better off with specific pollutants
 like carbon  monoxide and lead,  where biological  responses can
 be reproduced  in  the laboratory,  though the public health
 significance of the  responses that have been used for standard
 setting may  in the  long run  not impress society  sufficiently
 to act on these data alone.

 The problem  illustrated by these  few examples is that we are
 lacking data in three fields:
     o  what standards  does  society  exactly want to set to
        protect environmental  health,
     o  what effects  does  man  even after setting such standard
        s possibly  still  have  on  the environment,
     o  what means do we  have  to  make man cooperate in keeping
        the  standards.

 The possibility exists  that man's effect on the  environment
 takes place at levels of  exposure which  are not  directly detri-
mental to his health  as expressed in terms  of  life expectancy
 or prevalence of known diseases.   The  beautification  of life,

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                               1884
 or  the  contrast  of  it "and  the  absence of  beauty due  to  malodors
 or  hazes  or needless noise for example, so  far has not  been
 expressed in  standards,  though we know that more people in
 polluted  environments are  aware of a problem.  It is, based
 on  my Rotterdam  experience, possible that the movement  for
 better  environmental health finds its real  roots in  a welfare
 society.   This would mean  that standards  for direct  physical
 health  risks  are needed.   But also data to  set standards for
 beautification of the environment. Every  general practitioner
 nowadays  deals more and more with patients  who are not  ill
 in  the  classical way but who are considered ill from deficien-
 cies in their social and physical environment.  In this meeting
 we  have talked about the physical environment mainly, and it
 will probably be an easier field to remedy  than the  social
 field.

 For the ever changing human behavior, we  may well need  more
 help from  the social disciplines than we  realize at  this moment.
 I fear, however, that their data collection for the  successful
 implementation of environmental health projects has  hardly
 begun.
BUTLER
The purpose of this Plenary Discussion Group should be to
initiate a dialogue between scientists and administrators
involved in environmental affairs.  To begin with they should
use the same vocabulary and I propose to use that recommended
for the Stockholm Conference.  Since there is at present no
standard usage, a number of terms have been defined by the
Preparatory Committee for the discussion of international co-
operation for pollution control at Stockholm:
     o  exposure;  the amount of a particular physical or chem-
        ical agent that reaches the target;

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                       1885
target  (or receptor):   the  organism,  population or
resource to be protected from  specified  risks;
risk;   the expected  frequency  of undesirable  effects
arising from a given exposure  to a  pollutant;
criteria  (or exposure-effect relationships):the quantit-
ative relations between the exposure  to'  a  pollutant
and the risk or magnitude of an undesirable effect
under specified circumstances  defined by environmental
variables and target variables;
primary; protection standard;   an accepted  maximum
level of a pollutant  (or its indicator)  in the  target,
or some part thereof, or an accepted  maximum  intake
of a pollutant or nuisance  into the target under specif-
ied circumstances;
derived jgorking levels(or limits);  maximum acceptable
levels of pollutants in specified media  other than
the target designed  to ensure  that  under specified
circumstances a primary protection  standard is  not
exceeded;
       derived working  levels  are known  by a variety
       of names, including  environmental or ambient
       quality standards, maximum permissible limits
        and maximum  allowable  concentrations.   When
        derived working levels apply  to  products such
       as food or detergents,  they  may be  known as
       product standards;
the maximum acceptable release of a pollutant from
a given source to a  specified  medium  under specified
circumstances may be termed a  discharge  (or effluent
or emj.sjsi.on) standard or a release  limit.   Effluent
charges levied on the release  of pollutants and mater-
ials taxes or prices adjustments levied  on materials
which may become pollutants may also  be  used  to limit
the release of pollutants;
       in order to meet discharge standards or  release
       limits, it may be necessary  to set  various types

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                                1886
                of technological standards or codes of practice
                concerned with the performance and design of
                those technologies or operations leading to
                the release of pollutants? or
      o  derived working levels and the various means used to
         meet them are collectively termed derived standards
         and other controls;
      0  action  level; the level of a pollutant at which specif-
         ied emergency countermeasures, such as the seizure
         and destruction of contaminated materials, evacuation
         of  the  local population or closing down the sources
         of  pollution, are to be taken.
 It  is  perhaps also necessary to define "critical organ" as
 "the  organ  in which the entry of a given pollutant into the
 body results  in the greatest body injury.   This is often, but
 not always,  assumed to be the body organ in which there is
 the greatest  concentration of pollutant."   This will depend
 on  the nature and chemical form of the pollutant and its route
 of  entry into the body.   It  is also necessary  to define "thres-
 hold", which means  that below some finite  level of dose there
 is  no  observable  effect.   Certain other terms  that I use will
 reflect  my  background in radiation protection  and will  be differ-
 ent from those  used by many  toxicologists  and  environmental
 protection  officers.

 In  organizing his knowledge  of  the consequences,  for human
 health,  of  environmental contamination the  scientist thinks
 first  of exposures  and effects  in the  critical  organ, where
 this can be identified,  and  how  these  vary  with time.   Where
 the critical organ  cannot be  identified  the scientist may be
 reduced  to  observing  symptoms displayed  by  the  whole animal.
 Thus exposures may  be  brief  or protracted  (sometimes chronic)
depending on: the residence  of the  subject  in the  polluted
 environment, the  persistence  of  the  contaminant  in  the  environ-
ment and the chemical  form and route of  entry of  the pollutant.

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                              1887
Effects can be classified into three types depending on the
time course of pollutant level and its effects in the critical
organ:
     o   Early (sometimes called "acute") effects result from
        a transient peak of pollutant concentration in the
        critical organ followed immediately by a peak of effect
        which, if it is not fatal, subsides rapidly.  The plots
        of concentration vs time and effect vs time can be
        made if one knows the retention equation for the agent
        in the critical organ and the persistence of the effect
        as a function of time.  Short-lived effects of short-
        lived agents usually result from the reduction of the
        rate of some vital process.  The dose-effect curves
        usually display a marked threshold.  Examples of such
        agents are: cyanide, parathion and caffeine.  The kind
        of protection guide required for this effect should
        ensure that the instantaneous concentration in the
        critical organ never rises above the threshold which
        could be provided by a limit on concentration in the
        environment.
     o  Late effects may result from the build-up of the
        pollutant concentration in the critical organ to a
        maintained level which produces a commensurate damage
        in a proportion of the cells.  Under constant intake
        the tissue concentration increases with time until
        it becomes constant at some equilibrium level.  When
        the level of pollutant concentration and proportionate
        damage are high enough the symptoms appear.  From a
        knowledge of the retention equation for the agent
        in the critical organ and the rate of uptake one can
        calculate the tissue concentration for any given time
        or what level of uptake will lead to what equilibrium
        level.  Examples of agents that behave in this fashion
        are methyl mercury and warfarin.  Environmental protect-
        ion guides for such agents should limit the product
        of concentration and time of uptake so as to prevent
        the equilibrium level in the critical organ exceeding

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                               1888
        the threshold for damage. For constant uptake  there
        should be a limit on environmental concentration while
        for intermittent uptakes there should be a  limit on
        the total concentration X time for all the  episodes.
     °  "Stochastic" effects is the name sometimes  given to
        effects that accumulate with little repair  throughout
        the life of the receptor and are produced in proportion
        to the amount of the agent in the critical  organ,
        however small.  Here the dose-effect curve  shows no
        threshold and the effect will be proportional  to the
        prior time integral of agent concentration  in  the
        critical organ, whether it resulted from a  single,
        multiple or chronic exposure.  This independent variable
        can be calculated from the retention equation  for the
        agent in the critical organ.  The environmental protect-
        ion guide for this type of effect should ensure that
        the subject does not receive during his lifetime an
        accumulated exposure that will result in an unaccept-
        able risk of stochastic effect, usually cancer, mutat-
        ion or foetal damage.  Examples of such agents are:
        ionizing radiations or radioactive nuclides and carcin-
        ogenic hydrocarbons which produce cancer.   Another
        type is organic phosphorus compounds such as trior-
        thocresyl phosphate which produce permanent paralysis.

There are other examples of different combinations  of persis-
tence of agent and its effects; e.g., alcohol with  a short
persistence but the effects of which accumulate and DDT with
a long persistence and short-lived effects.

It is apparent that of primary interest to the scientist is
the time pattern of concentration of the agent (pollutant)
in the critical organ and the primary protection standards
to limit this to acceptable levels commensurate with accept-
able risks of damage.  The administrator has a greater need
for derived working limits which for industrial workers could

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                               1889
be the amount of agent excreted per day and  for  the  populat-
I6n, ambient quality standards to  limit intake.

Ideally, criteria reports from the scientist to  the  adminis-
trator should give the probability of effects  for  a  range  of
levels of pollutant, with confidence limits.   The  administrator
also needs to know how these vary with:

     o  the state of the subject; e.g., age,  health  and  nutrit-
        ion,
     o  the presence of other co-acting pollutants,
     o  other environmental factors such as  temperature, sun-
        light, humidity.
This type of information, among others, is required  for  decid-
ing what safety factors to use in extrapolating  from the ex-
perimental results to acceptable levels for  protection guides.

So far I have mentioned only the scientific  information  requir-
ed for primary protection standards, derived working limits
and ambieat quality standards.  In controlling pollution it
may be easier to impose emission standards instead of the  above
protection guides, for which the administrator must  have in
addition quantitative information on the relation  between  the
rates of emission and the probabilities of their occurrence
on the one hand and the uptake by the receptor on  the other.

The kind of information described above for  pollutants and
their effects as functions of time is also needed  for current
and candidate pesticides in order to make intelligent decisions
about replacing "dangerous" or too persistent  agents such  as
DDT with "less dangerous" or less persistent pesticides.

GOLDSMITH
The main purpose of assessment of carcinogenic effects of  pol-
lutants is to provide a basis for intervention.  This requires
evaluation of epidemiological and experimental evidence  in
response to a variety of findings.  The major  types  of findings

-------
                              1890
may be excess cancer occurrence among a population,  (epidemio-
logical evidence) or the finding of a known carcinogen to which
exposure by one or more routes is occurring.  While  the paper
does not evaluate the carcinogenic effects of occupational
exposures, occupational exposures both interact with non-occu-
pational exposures, as well as provide many of 'the findings
of the first type.

The procedures used in evaluation of possible relationships
of air pollutants and respiratory cancer and in asbestos in-
gestion and gastro-intestinal cancer are used to illustrate
the principles of evaluation.  The procedures require the app-
lication of judgment, and hence different conclusions from
such procedures can be expected from different individuals
and groups.

Environmental exposures, including occupational ones, have
been epidemiologically associated with cancer of sinuses, pleura
and peritoneum, mouth, skin, bladder, lung, larynx,  liver,
scrotum and with leukemia,  (1).  The IARC is publishing a series
of monographs on the Evaluation of Carcinogenic Risk of Chemicals
to Man (2). The experts who evaluate the human effects data
usually do not find such data adequate.  In most examples to
date, the evaluation of human health effects refers  to occupat-
ional exposures.

Several decades of human exposure to environmental agents are
usually required before cancer manifests itself, and this
property of human cancer is one of the reasons why it is
hazardous to draw quantitative or even qualitative inferences
for man from experimental research.  Reliance on epidemiological
evaluation is therefore an  important basis  for conclusions.
However, the epidemiologically detectable increases  in cancer
are often not recognized and responded to sufficiently early.

Along with environmental exposures, inborn  factors,  nutrition,

-------
                              1891
other diseases or injury, socio-economic status  and  age-sex
dependent factors are major variable clusters  in human  carcin-
ogenesis. For many forms of cancer, smoking adds another  criti-
cal variable. Epidemiological evaluation requires consideration
of a sufficiently complete set of variables and  in the  appro-
priate form.  Path diagrams are shown to be a  useful device
to compare completeness of variable sets.

Path analysis originally proposed by Sewall Wright for  statisti-
cal genetics (Li) and recently applied to sociology  (3) , (4) ,
has been proposed for application to chronic disease epidemiol-
ogy.  Path analysis describes an adaptation of multivariate
analysis in which, either on an a priori basis,  or as a result
of partial correlation analysis, the roles of  "independent"
(measured) and exogenous (unmeasured) variables  are  structurally
ordered in relationship to dependent variables,  in this case,
incidence rates of cancer.  Path analysis methods are currently
being tested for their appropriateness in environmental cancer
epidemiology. They appear to have the desirable  feature of
being able to partial out the contributory role  of a multiplicity
of factors, but the assumptions needed may restrict  their
applicability.

Evaluation of the role of air pollution in respiratory  cancer
is based on four types of findings:
     o  that inhaled materials of various types  can  and do
        lead to increased lung cancer rates,
     o  that carcinogenic agents are found in  polluted  air,
     o  that there is an urban excess of lung  cancer,
     o  differential effects of migration of populations on
        lung cancer rates.
The conclusion  hinges on judgment concerning the principal
arguments supporting and opposing the causal role of air pollut-
ion in the urban excess of lung cancer.  While the weight
of evidence now available does not support the conclusion that
air pollution is an important causal factor, neither can the
possibility be  dismissed.

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                               1892
In the case of ingested asbestos and the possible role  it may
have in cancer of the gastro-intestinal tract, the  following
findings need to be considered:
     o  there is epidemiological evidence that gastro-intestinal
        cancer rates are elevated in groups occupationally
        exposed to asbestos.
     o  asbestos fibers and fibrils can be found in water
        and some foods.
     o  although asbestos minerals can enter water  both from
        natural and technogenic sources, the associations be-
        tween asbestos levels in water  (or probable differences
        in such levels) and gastro-intestinal cancer  rates
        have not yet been systematically examined;  neither
        is uptake from ingested asbestos well understood.
     o  a few orally ingested agents have been shown  to be
        carcinogenic for man.

Other types of evaluation are that of arsenic in water  (6)
or air (7) based on estimates of body burden, and that  of nitrate
in water (8) based on an association with increased methemo-
globin in infants. These agents, for which a carcinogenic potent-
ial is suspected, are being assessed and controlled on  the
basis of non-malignant responses.

Among the factors to be considered in evaluating human  carcino-
gens are:
     o  distribution of exposures in time and place
     o  number of people at risk
     o  available information and its validity concerning ex-
        posures, responses  (carcinogenic or not) and  other*
        relevant variables.
     o  adequacy of model and data with respect to  completeness
        of variable set and variable representation.
     o  adequacy of epidemiological strategy and of statistical
        analysis.
     o  ease of preventing hazardous exposure
     o  potential availability of additional data

-------
                              1893
     o  the nature of other health reactions to the
        agent(s).

A satisfactory strategy for environmen-tal cancer control
depends on:
     o  realistic models of exposure including evaluation
        of effects of non-environmental variables,
     o  suitable population studies, with special attention
        to occupationally exposed groups
     o  study and inferences based on non-malignant
        responses to carcinogenic exposures, such as
        body burden estimates, cytological changes,
        and metabolic responses,
     o  utilization of a consistent set of principles
        for evaluation and
     o  appropriate applications of the results of evaluation.

In light of experience with asbestos, vinyl chloride,
benzo(a)pyrene, arsenic, and beryllium, as community
pollutants, the study of non-malignant responses to possible
carcinogens should have a prominent place in such a strategy.
1.   J. Higginson and L. Tomatis, "Industrialization,
     Cancer Incidence and Possible Prevention", Proc.
     XVI, Int. Cong, occup. Health, Tokyo, 246-25O, 1971
2.   International Agency for Research on Cancer.  IARC
     Monographs on the Evaluation of the Carcinogenic
     Risk of Chemicals to Man, vol. 1  (1972), vol. 2,
     (1973), vol. 3  (1973), vol. 4  (1974) and vol. 5  (1974)
3.   D. R. Heise, "Problems in Path Analysis and Causal
     Inference" in Sociological Methodology, (1969), E.
     F. Bergetta and G. W. Bohrnstedt  (Ed) Jossey-Bass,
     San Francisco.
4.   K. C. Land, "Principles of Path Analysis", op.cit.
5.   J. R. Goldsmith and K. Berglund,  "Epidemiological
     Approach to Multiple Factor Interactions in Pulmonary
     Disease: The Potential Usefulness of Path Analysis",
     Proc. N.Z. Acad. Science, 221, 361-375, 1974.

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                            1894
 6.   J. R. Goldsmith, M. Deane, J. Thorn, and G. Gentry
      "Evaluation of Health Implications of Elevated
      Arsenic in Well Water",  Water Resch., 6, 1133-
      1136, 1972.
 7.   S. Milham, Jr. and T. Strong, "Human Arsenic
      Exposure in Relation to  a Copper Smelter", Environ.
      Res., 7_, 176-182,  1974.
 8.   L. A. Shearer, J.  R. Goldsmith,  C.  Young,  O.A.
      Kearns, and B. R.  Tamplin, "Methemoglobin  Levels
      in Infants in an Area with High  Nitrate Water
      Supply", Am.  J.  Public Health, 62,  1174-118O,
      1972.                         —

 RECHT
 Dans  le domaine de la protection de 1'homme  et  de
 1'environnement,  les  criteres  sont definis habituellement
 comme les  relations entre 1'exposition d'une cible
 a  une pollution ou nuisance et le  risque et/ou  1'ampleur
 de 1'effet defavorable  ou indesirable  qui en resulterait
 dans  des circonstances  donnees.  II est  souhaitable
 que cette  relation soit quantitative et  gue,  notamment,
 1'exposition  soit  exprimee sous  forme  de valeurs  numeriques,
 de concentration,  d'intensite,  de  duree  ou de frequence.
 Le risque  est  lui-meme  defini  comme une  probabilite
 d1apparition d'effets defavorables ou  indesirables  resultant
 d'une exposition donnee a un ou plusieurs polluants ou
 nuisances  pris  isolement  ou en  combinaison.  Le risque
 et  1'effet defavorable  ou  indesirable doivent, autant
 que possible, etre  exprimes d'une maniere precise.

La recherche et 1'etablissement de ces criteres representent
pour  les autorites  responsables de decisions reglementaires
et administratives  un element de jugement et une base
 scientifique de reflexion  et de decision.  Idealement,
 les criteres devraient  etre proposes avant qu'une reglementation
ne soit mise en oeuvre  pour reduire les pollutions et
ou les nuisances.  En raison meme de son objectif, 1'etablisse-
ment de criteres est une entreprise etendue et souvent

-------
                            1895
malaisee; elle doit affronter de nombreuses difficultes dont
les unes sont liees aux premiers termes de la relation, c'est-
a-dire I1exposition, et dont les autres concernent le risque
ou 1'effet defavorable.  II y a d'abord le grand nombre de pol-
lluants a considerer, la variete des facteurs physico-chimiques
intervenant dans 1'exposition, la diversitfi  des voies par les-
quelles 1'homme peut etre touche, I1absence de donnees chiffrees
et comparables sur 1'etat de la pollution du milieu, la pres-
ence simultanee dans 1'air inhale ou dans 1'eau et les aliments
ingeres, de nombreux polluants ayant des actions synergiques
ou antagonistes. II y a, en outre, la complexite des processus
metaboliques et biochimiques intervenant dans la contamination
ou dans 1'atteinte humaine, la definition du caractere defavor-
able ou indesirable de 1'effet ou la nature du risque a consid-
erer, 1'interpretation des donnees.  Deux elements importants
doivent retenir I1attention et concernent d'une maniere directe
un des buts essentiels de la protection centre le danger des
pollutions et nuisances; ils constituent d'ailleurs des impe
ratifs majeurs dans la recherche des criteres.   II y a d'abord
le fait que les concentrations caracterisant 1'exposition sont,
en general, relativement basses et variables dans le temps et
selon les regions.  II s'agit par ailleurs d'une exposition
chronique a laquelle ne s'appliquent pas necessairement les ex-
periences acquises lors d1accidents aigus de pollution ou les
donnees de I1hygiene industrielle.  Si pour certains polluants,
on peut retenir une relation lineaire dose/effet, elle n'est
pas demontree, ni meme demontrable pour un grand nombre d'autres
polluants en considerant des effets determines.  Le caracte
re de la population dans son ensemble doit etre pris en conside-
ration; la population comprend des groupes d'individus non homo-
genes, dont I1age et les conditions sanitaires sont differents
et parmi lesquels nous trouvons des enfants, des vieillards,
des femmes enceintes et des malades.

Frequement, dans 1'interpretation des resultats des facteurs
concomitants apparaissent, qui n'ont rien a voir avec le

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                            1896
polluant considere et creent certaines perturbations  (par exe-
mple: tabac, medicaments, alimentation, habitudes  sociales,
etc...),  un des problemes fondamentaux de  la pathologic spe-
ciale de 1'environnement est la difficulte  de qualifier d'inde-
sirable ou de defavorable un effet sur la sante  humaine.  En
raison du niveau relativement bas des expositions,  les effets
a prendre en consideration appartiennent au domaine des effets
"subcliniques"; ils ne sont pas nettement pathologiques. Les
signes indiquant 1'atteinte sont rarement pathognomoniques et
beaucoup d'entre eux ne sont reveles que par des etudes stati-
stiques.   La radioprotection et 1'hygirene industrielle nous
avaient habitues a une certaine signification dans  I1applica-
tion de modules metaboliques etablis selon  les caracteristiques
de 1'homme standard.

Faut-il un ou plusieurs crite"res est une question  qui peut
Stre posee et constitue certainement un probleme interessant
a discuter.  Pour les autorites responsables des decisions
en matie"re d1 environnement, il est plus commode de  n1 avoir a
considerer qu'un seul crite"re.  On peut, dans ce cas, fixer le
niveau £ partir duquel le premier effet defavorable apparalt;
un exemple est 1'oxyde de carbone. On sait  qu'a partir de 4%
de carboxyhemoglobine il y a un effet defavorable  certain sur
1'appareil cardio-vasculaire du a 1'hypoxie des tissus:  a
partir de 2% il y a une interference avec les performances
d'exercices physiques chez les malades cardio-vasculaires.
On sait aussi que 1'intensite de I1effet augmente avec I1expo-
sition et qu'en outre d'autres effets defavorables  (systeme ner-
veux artherosclerose, foetus)  peuvent intervenir.   Pour le dio-
xyde de soufre et la fumee, on sait par les etudes  epidemiolo-
giques qu'a partir d'une concentration de 25O ug/m3 de SO2
associee 1 des concentrations plus faibles  de fumee, il y a un
effet defavorable et significatif du point  de vue sanitaire
sur 1'appareil respiratoire; mais vu la difficulte  de fixer a
1'heure actuelle des limites precises, on a ete amene a pre-
senter d'autres relations dose/effet a des  seuils inferieurs

-------
                            1897
ou superieurs d'exposition et lies a des atteintes ou a des
risques d1atteintes exprimes en general d'une maniere assez va-
gue.  Dans le cas particulier de ces polluants, une serie de
criteres se justifie et laisse aux autorites sanitaires le soin
de decider en tenant compte des considerations sociales ou meme
economiques, quel est le critere a considerer dans I1etablissement
de normes.

Un troisieme exemple qui peut illustrer les difficultes est ce-
lui du plomb et de 1'impregnation saturnine des populations.
Les voies par lesquelles I1 nomine se contamine sont nombreuses
(inhalation, ingestion, peau). Les sources de contamination va-
rient notamment selon les regions et les habitudes alimentaires.
Neanmoins, on peut reconnaitre que la plombemie ou la mesure
de 1'ALAD representent des indicateurs precieux de I1impregnation
saturnine globale humaine.  A partir de certains taux de plom-
bemie, il apparalt une atteinte de 1'enzyme intervenant dans
la synthese de 1'heme.  Une atteinte significative de cette en-
zyme est generalement consideree comme indesirable et peut etre
liee a un taux de plombemie relativement precis.   Le groupe
le plus sensible est celui des enfants; des lors, dans le cri-
tere, la plombemie des enfants devient le facteur determinant.
En partant de ce critere, que I1on pourrait qualifier de fon-
damental, on peut "deriver" des normes pratiques concernant
certaines voies de contamination de 1'homme.  S'il est prouve
que 1'appareil respiratoire peut representer 1'organe critique
dans le cas d'une contamination humaine par inhalation, on peut
etablir un deuxieme critere, qui est la relation entre une con-
centration du plomb dans I1air et un effet determinesur 1'appa-
reil respiratoire ou certains de ses mecanismes de defense
(macrophages).

D'autres exemples pourraient etre apportes conformant qu'on ne
peut eriger en regie absolue la fixation d'un seul critere et,
qu'au contraire, un ensemble de relations correspond mieux a
1'etat actuel des connaissances scientifiques.  II est evident

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                            1898
 que,  dans  ce  cas,  chaque  critere  doit etre  assort!  de convmen-
 taires  explicatifs qui  puissent permettre aux  autorites respon-
 sables  des decisions  d'en comprendre  la portee et la signifi-
 cation  reelle.

 Quant au probleme  des groupes de  population a  proteger, 1'ana-
 lyse  precedente  revele  qu'en raison de la non  homogeneite de
 la population il faut prendre en  consideration un groupe de pop-
 ulation sensible mais significatif et representatif.  Dans le
 cas des enfants  et de la  contamination par  le  plomb, le prob-
 leme  est simple  car il  s'agit d'un groupe important de la popu-
 lation  et  constitue d'individus bien  portants.

 Quand le groupe  identifie comme sensible est composed1individus
 maladesf le probleme  est  plus complexe et plus delicat; par
 exemple, le monoxyde  de carbone a partir de 4% de saturation
 de 1'hemoglobine a une  action certaine sur  1'appareil cardio-
 vasculaire  surtout chez les individus  presentant  deja des ano-
 malies  ou  consideres  comme individus  "a risque".   II ne s'agit
 pas en  fait de malades  hospitalises mais d'individus relativement
 nombreux dans la population actuelle  et qui, a partir d'un
 certain age,  sont  susceptibles de presenter un accident cardio-
 vasculaire.   II  est raisonnable dans  ce cas de baser une deci-
 sion normative eventuelle  sur un  groupe representatif et signi-
 ficatif du point du vue demographique.  Mais on sait egalement
 que le monoxyde  de carbone peut avoir,  meme en  dessous  de 2%
 de saturation, une action  sur des malades dont  1'etat de sante
 est particulierement  deficient au point de  vue  cardio-vasculaire,
 ce sont les malades "in extremis" dont il a ete fait reference
 dans le rapport  technique  5O6 de  1'OMS.  Vu la  tres grande
 fragilite de  ces malades, on pourrait  arriver  a la  conclusion
                                                       »
 que meme une  legere augmentation  du COHb par rapport aux niveaux
 d'origine endogene constituerait  un risque.  Est-il  raisonnable,
dans un programme  de  reduction des nuisances, de  baser  une

-------
                            1899
decision sur un critere aussi severe?  Le probleme reste ouvert
et peut, sans aucun doute, constituer une question importante
a discuter.
Criteria for the protection of man and the environment are nor-
mally defined as the relation between the exposure of a target
to a pollutant or nuisance and the risk or magnitude of a harm-
ful or undesirable effect which would result under specified
circumstances.  This relationship should preferably be quantit-
ative t and in particular the exposure level should be expressed
as a numerical value, in terms of concentration, intensity,
duration or frequency.  The risk itself is defined as the expect-
ed frequency of harmful or undesirable effects arising from a
given exposure to one or more pollutants or nuisances, consider-
ed alone or in combination.  The risk and the harmful or undesir-
able effect must be expressed in as precise a manner as possible.

For the authorities responsible for legislative and administra-
tive decisions, the search for and establishment of such cri-
teria provide a means of evaluation and a scientific basis for
consideration and decision.  Ideally, these criteria should be
put forward before standards are instituted for the reduction
of pollution of nuisances. By reason of its aim, the establish-
ment of criteria is a far-reaching and often difficult task;
there are many difficult points to be considered, some of which
are connected with the first part of the relationship, namely,
exposure, and others with the risk or harmful effect.  First,
there is the enormous number of pollutants to be considered,
the diversity of physico-chemical factors involved in exposure,
the many different ways in which man may be affected, the absence
of comparative numerical data on the state of environmental pol-
lution and the presence at one and the same time in air inhaled
or food and water ingested of numerous pollutants having syner-
gistic or antagonistic effects.  There is in addition the prob-
lem of the complex nature of the metabolic and biochemical pro-
cesses involved in contamination and in the effect on man, the
definition of the harmful or undesirable nature of the effect
or the character of the risk under consideration and the inter-
pretation of the data.  Two significant factors merit attention
and are directly concerned with one of the essential aims of
protection against the dangers of pollutants and nuisances;
what is more, they amount to major essentials in the search for
criteria.  First, there is the fact that concentrations involved
in exposure are, in general, relatively low and vary according
to weather and district. We are dealing, moreover, with chronic
exposure, to which experience obtained on the occasion of acute
accidents arising from pollution, or the findings of industrial
hygiene, do not necessarily apply.  Although for certain pollu-
tants a linear dose/effect relationship may be accepted, it has
not been proved, and indeed cannot be proved  for a large num-
ber of other pollutants, as regards given effects.  The charac-
teristics of the population as a whole must be taken into

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                            1900
 consideration,  and it  is  composed of heterogeneous groups of
 individuals  of  different  ages  and states  of health,  amongst whom
 we  find  children,  the  aged,  pregnant women and diseased persons.

 In  interpreting results we  frequently come across concomitant
 factors  which are  unconnected  with the pollutant under consid-
 eration  and  which  cause certain  complications  (for example:
 tobacco, medicines,  food, social habits,  etc....).  One of the
 fundamental  problems of special  environmental  pathology is the
 difficulty of classifying an effect on human health as undesir-
 able or  harmful.   Because of the relatively low exposure rate
 the effects  to  be  considered are 'subclinical ' - that is, they
 are not  clearly pathological.  The signs  indicative of an injur-
 ious effect  are rarely pathognomic and many of them become evid-
 ent only through statistical studies.   Radiation protection and
 industrial hygiene  had previously accustomed us to attributing
 a certain significance in practice to metabolic models based
 on  the characteristics of the  average man.

 Wt, may ask whether  we need  one or several  criteria,  and the pro-
 blem is  certainly  an interesting one.Fortheauthorities res-
 ponsible for making  decisions  in environmental matters it is
 more convenient to  have only one criterion to  consider.  Thus
 one may  establish  the  level  at which the  first harmful effect
 appears;  carbon monoxide   provides one example.   He know that,
 starting at a carboxyhaemoglobin level of 4%,  there is a defin-
 /"•'<* harmful effect  on the cardiovascular  system,  owing to hypo-
 -,*a of the tissues.  We know  also that the  intensity of this ef-
 fect increases  with  exposure and that in  addition other harmful
 effects may occur  (affecting the nervous  system or the foetus
 or in the form  of  arteriosclerosis).   For  sulphur dioxide and
 smoke	 we know  through epidemiological  surveys that above
 a concentration of  250 vg/m* of  S0~ associated with ISO vg/m*
 of smoke a significant injurious  effect develops  threatening
 the health of the  respiratory  organs;  but  owing to the difficul-
 ty of determining precise limits  at the present time,  other
 dose/effect relationships have been prepared involving higher
 or  lower exposure  limits  and connected with injurious  effects,
 or the risk of  such  effects, expressed generally  speaking in
 rather a vague  way.  In the  individual case of this pollutant,
 the adoption of a  series  of  criteria is justified,  the health
 authorities being  left to decide,  having  regard to social or
 even economic considerations, what  criterion is to be  taken
 into account in  laying down  standards.

A third example which may illustrate the  difficulties  is that
 of lead and its concentrations in  the  population.  The  ways in
which man may become contaminated  are  numerous  (inhalation,
 ingestion,  skin).  The sources of contamination vary in particu-
 lar according to district and feeding  habits.. Nevertheless,
 it is true that  the  blood-lead level or the ALAD  level are valu-
able indicators of the aggregate  lead-contamination  level in
man.  Above certain  blood-lead levels,  the  enzyme  responsible
for haerr, synthesis is damaged.    A  significant  effect is gener-
ally considered  to be undesirable  and  may  be linked  with a

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                             1901

 relatively precise level.  Children constitute the most sensit-
 ive group.  For this reason the blood-lead level in children
 is the determining factor in the criterion.  On the basis of
 this parameter, which may be described as fundamental, it is
 possible to work out practical criteria or norms with regard
 to certain ways in which man becomes contaminated. If it is
 proved that the respiratory system may represent the critical
 organ in the case of human contamination by inhalation, a second
 criterion may be elaborated,  namely the relationship between
 a given concentration of lead in the atmosphere and the measured
 effect on the respiratory system or on certain of its defence
 mechanisms (macrophages).

 Other examples might be cited showing that it is impossible
 to lay down a single criterion in the form of an absolute
 rule,  but that on the contrary a group of relationships corres-
 ponds more accurately to the  present state of scientific know-
 ledge.   It is evident that in such circumstances each criterion
 must  be ^accompanied by explanatory notes allowing the authorities
 responsible for taking decisions to understand its true scope
 and meaning.

 On the question of the groups of persons to be protected,  the
 foregoing analysis shows that in view of the  heterogeneous
 nature  of the population it is necessary to consider groups
 which  are not only at risk but also significant and representat-
 ive.   In  the ^case  of children and contamination by lead,  the
 problem is  simple,  since we are  concerned with a significant
 group  of the  population composed of healthy individuals.   If
 the group identified as being at risk is composed of diseased
 persons,  the  problem is both  more complex and more delicate:
 for example  a saturation level of carbon monoxide  in haemoglo-
 bin exceeding 4%  has  a definite  effect on the cardiovascular
 system,  especially  in individuals already displaying anomalies
 or  viewed as  being  'at risk'.   We are not concerned with  patients
 in  hospitals  but  with the fairly numerous individuals  in  the
 present-day population who, above a certain age,  have  a tendency
 to  cardiovascular  accidents.   In such a case  it is reasonable
 to  base any  legislative  decision on a group which  is demograph-
 ically  representative and significant.   But we  also know  that
 carbon  monoxide,  even below 4%  saturation,  may  have an effect
 on  diseased persons  whose  health is particularly  deficient  as
 regards the cardiovascular  system.   These are the  patients  'in
 extremis' referred  to in  No.  506 WHO Technical  Report.  In  view
 of  the  very delicate  condition  of such  patients,  one might  argue
 that even a slight  increase in  the  COHb above their original
 endogenous  level would constitute  a risk.  Is  it reasonable,  in
a programme dealing^ with  the  reduction  of nuisances  to  base  a
decision  on so  strict  a  criterion?  The  question still  remains
 unanswered, and can without doubt  provide  an  important  subject
for discussion.

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                            1902
 TREMOLIERES
 Pour juger des effets de la sante de la pollution de 1'environ-
 nement, un grand vide subsists:  le flou, voire 1'absence
 d'une methodologie permettant de transposer a I'homme les
 travaux experimentaux sur 1'animal; ceci aussi bien pour clas-
 ser qualitativement les polluants, que pour fixer quantitati-
 vement la dose journaliere acceptable.

 Le Comite "Contamination de la Chaine Biologique" du Ministere
 de 1'Environnement francais a etabli, sous le nom de "toxi-
 cologie metabolique",  une methodologie  dont 1'objet est d'e
 tablir pour  les substances etrangeres les processus de meta-
 bolisation maintenant  etablis pour les  nutriments, suivant
 les especes,  les  etats de nutrition,  les developpements, les
 associations  de toxiques.

 Sous-jacente  aux  etudes  de toxicologie  aiguefsubaigue ou chro-
 nique,  des toxicites de  relais,  des pouvoirs  cancerigenes et
 teratogenes,  qui  restent des guides indispensables,  la toxi-
 cologie  metabolique s'efforce  d'expliquer  les  mecanismes des
 faits  observes.   Elle  comporte:
    o   la physiologic de  la  diffusion  et de  la  transformation
       par  les divers tissus  suivant les doses  et  les
       associations;
    o   les  systemes enzymatiques  detoxiquants  (oxydases,
      peroxydases, etc.	);
    o   les repercussions  de  ces systemes sur les metabolismes
      de la bioenergetique et des biosyntheses nucleiques,
      proteiques et membranaires;
    o  les effets physiologiques  et pharmacodynamiques  sur
      les divers organes du produit et  de ses metabolites
      suivant 1'etat nutritionnel et metabolique du recepteur.

ABSORPTION ~ DIFFUSION - CONCENTRATION  - ELIMINATION
    II faut etudier suivant les condition d'ingestion:
      o  le sort du compose dans le tube digestif et son
         absorption.  On devra approfondir le role:

-------
                          1903
          oo  de la flore digestive (exemple:   les cyclamates
              sont complement metabolises par  la flore
              intestinale de certains  individus seulement,
              aussi bien dans 1'espece humaine que chez le
              lapin New Zealand),
          oo  des  mucus digestifs,  les modes de transport a
              travers  1'enterocyte, 1'intervention des
              tissus lymphoides de  la  sous-muqueuse
              intestinale,  de cellules de Kupffer, des
              macrophages,  comme barrieres successives
              possibles,
      o   la repartition et les possibilites d'accumulation
          dans  les  divers tissus:   ceci pose le probleme  des
          techniques d'identification, de detection et de
          dosage du compose et de ses  eventuels derives
          dans  les  substances biologiques comportant les
          chelates  proteines  metaux.
    II faut prevoir un travail de  chimie organique pour
    la preparation de  substances etalons,
      o   1'excretion dans  la bile,  les feces,  1'urine,
          1'air expire.

METABOLISATION du  TOXIQUE  a  L'ECHELON CELLULAIRE
    Suivre  le passage  d'un compose  a travers  1'organisme
    implique done  la reconnaissance de son  metabolisme
    par les divers  tissus.   Quels  derives   sont produits?
    Duns quels tissus?

    «•   SijSt-em.es enz_yr;gfiqi
-------
                           1904
    Enfin, le compose peut etre excrete inchange.   Ou
    bien encore, les divers processus peuvent jouer
    simultanement.

    On salt que les transformations metaboliques des
    composes etrangers ont lieu dans les reins,  le poumon
    et surtout dans le foie; mais le tissu adipeux
    pourrait aussi jouer un r<5le ainsi aue la muqueuse
    intestinale.  Les reactions ont lieu en deux phases:

        o  phase I     :oxydations, reductions,  hydrolyses
        o  phase II    :syntheses, reactions de conjugaison
    Les enzymes de la phase I sont localisflees dans les
    microsomes.

On les appele en anglais "processing enzymes" ou "drug
metabolizing enzymes".  Ces processus n'aboutissent pas
forcement a la detoxification de la molecule "traitee".
         On s'attachera :a reconnaitre:
         o   quelles sont ces enzymes;
         o   dans quelles conditions physico-chimiques
             agissent-elles?  Ce qui  revient a explorer
             leur environnement: entre autres, ces enzymes
             necessitent la presence de plusieurs co-facteurs,
             notamment NADPH, Cytochrome P45O, etc.  Les
             mecanismes de transmission des electrons du
             substrat a 1'oxygene posent d'ailleurs de
             difficiles problemes, etant donne la rr.ulti-
             plicite des co-facteurs  intermediaires dont le
             role est mal connu;
         o   dans quelles conditions  physiologiques agissent-
             elles?  En d'autres termes, peut-on mettre
             en evidence un induction ou une repression de
             la synthese de ces enzymes dans differentes
             conditions experimentales ou physio-patholo-
             giques.

-------
                           1905
   B.  Les  effets du compose et de sa m&tabol-isation suv
       les  metabol-ismes ceHula-ires
       Le comporteraent physiologique et biochimique de
       I1animal  soumis a un traiteraent a  long terme
       devra  etre examine le plus finement possible sous
       son  double aspect:  comportement habituel, compor-
       tement en reponse a une stimulation  (jeune, autre
       nuisance  ou desequilibre nutritionnel, etc...).
   o   On presentera un profil descriptif de I1animal:
       oo etude  de la croissance, des ingesta et du
          rendement alimentaire;
       oo echanges respiratoires, depenses d'energie
          et  d'azote;
       oo etat hormonal  {insulinemie, corticosteronemie,
          etc.)
       oo poids  des differents organes et composition
         corporelle, etudes histologiques, exploration
         fonctionnelle, etc...).
   o   On evaluera  les  techniques habituelles comportant
       1'emploi  de  traceurs radioactifs:   les vitesses
       de renouvellement des acides  nucleiques,  des
       proteines, des phospholipides dans les divers
       tissus.
   o   On determinera  les. activites  enzymatiques
       potentielles de  certaines  voies metaboliques.
   o   L'etude cinetique des concentrations  des
       metabolites  et des  co-facteurs dans un  tissu,  soit
        sur  I1animal  entier,  soit  sur des  organes,
        tissus ou organites,  est  particulierement
        interessante pour reveler  les differences de
       comportement metabolique  et  proposer  des
       hypotheses sur  les  mecanismes d1action  des
        composes dans  un tissu.

LES CONDITIONS EXPERIMENTALES
    La plupart des experimentations  reposent sur  des
    animaux  bien nourris et toujours soumis a un  seul

-------
                           1906
    facteur:  1'agent ou'on veut etudier.  Or les
    resultats concernant des sujets sains ne s'appliquent
    pas forcement a ceux dont le metabolisme est deja
    "biaise".  II y aurait interet a reprendre les
    etudes decrites chez des animaux en etat d'obesite
    nutritionelle, diabetiques, etc., ou recevant un regime
    carence en proteines, en diverses vitamines, ou riche
    en alcool ethylique ou en saccharose.  De telles
    manipulations rapprocheraient les modeles experimentaux
    de certaines conditions qui sont realisees dans 1'espe-
    ce humaine par des conditions de vie particulieres.   On
    sait par exemple que les "processing enzymes" sont
    inhibes par 1'intoxication chloroformique, la castration,
    la denutrition, la thyroxine, la morphine; induits
    par les barbituriques, les hydrocarbures polycycliques
    et diverses classes de steroides comme les androgenes,
    les glucocorticoides, les anabolisants, les organoch-
    lores, etc.

    Les problemes de toxicologie metsbolique se posent en
    termes particuliers lors du developpement,
    o   Au cours de la vie foetale:
        oo action du toxique administre experimentalenient
           directement au foetus sur son developpement
           general et sur certains aspects non "recuperables"
           ulterieureroent du developpement:  differenciation
           sexuelle, differenciation du systeme nerveux
           central;
        oo passage transplacentaire du toxique.
    o   Au cours de 1'allaitement:
        oo passage de la substance toxique dans* le lait.
    o   Au cours de developpement.

CONCLUSION
    II s'agit:
    o   d'affermir les bases d'une methodologie de

-------
                           1907
        1'evaluation toxicologique;

    o   d'elaborer pas a pas une nomenclature permettant de

        classer les polluants suivant leurs effets

        metaboliques;
    o   de developper une approche pluridisciplinaire,

        analogue a celle qui a elabore la physiologie des

        metabolismes en nutrition.


There is a great gap in our knowledge for assessing the
effects of environmental pollution on health, namely,
the vagueness or indeed the absence of a methodology for
extrapolating animal experiments to man, both for the
qualitative classification of a given element in pollutants
and for the quantitative assessment of the acceptable
daily dose.

The French Ministry of the Environment's Committee on
the Contamination of the Biological Chain has elaborated
a methodology under the name of  'Metabolic Toxicology',
which aims to determine, for foreign substances, the processes
of metabolization which have now been determined for 'nut-
rients, according to species, state of nutrition, growth
and associations of toxins.

Underlying the studies of acute, sub-acute or chronic
toxicology, relayed toxicity, carcinogenicity and terato-
genicity, which remain invaluable  guides, metabolic  toxi-
cology attempts to explain the workings of observed facts.
It involves:
        I.  The physiology of diffusion and  transformation
            by various tissues, according  to  dose  and
            associations;
        2.  -  detoxicating  enyzme  systems  (oxidases,
               peroxidases,  etc.);
            -  the  effects of these systems  on  the mata-
               bolisms of bioenergetics and  of  nucleic,
               proteinic and membrane  biosynthesis;
        3.  The physiological and pharmacodynamic  effects
            of the  product and  its  metabolites  on  the
            various organs in relation to  the nutritional
            and metabolic state of  the subject.

j.  -  ABSORBTION - DIFFUSION - CONCENTRATION - ELIMINATION

The following must  be considered, according  to  the circum-
stances of  ingestion:

        o   what happens  to the  compound in the  alimentary

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                           1908



        canal, and its absorption.

Ve must further -investigate the role of:

    o   digestive flora (for example:  cyclamates are
        completely metabolized by the intestinal flora
        of certain individuals only, both in man and
        in the New Zealand rabbit),

    o   digestive mucus, the manner of passage through
        the enterocyte, the action of the lymphoid tissues
        of the intestinal submucosoe, Kupffer cells and
        macrophages as possible successive barriers.

    o   distribution and the chances of accumulation
        in the various tissues:  this poses the problem
        of techniques for identifying, detecting and
        measuring concentrations of the compound and of
        any derivatives in biological substances containing
        metallic protein chelates.

A research project in organic chemistry will have to be
undertaken to establish standard substances:

    o   excretion in the bile, faeces, urine and exhaled
        air.

II.  -  METABOLIZAT20N OF TOXIN AT CELL LEVEL

Following the progress of a compound  through the organism
implies ascertaining its metabolization by  the various
tissues.  Which derivatives are produced, in which  tissues?

    A.  Enzymatic detoxication or  transformation systems

Ve thus come  quite naturally to studying and carrying out
research into the enzyme processes responsible for  the
transforming  of  the compound.  The results  of such  research
might of course  be negative; for example, thalidomide
undergoes spontaneous  change without  the action  of  enzymes.
Enzymes in the intestinal flora must  be considered  separately,
Finally, the  compound  may be excreted unchanged  or,  again,
the various processes  may take place  simultaneously.

Ve know that  metabolic  transformation of foreign compounds
takes place in the kidneys,  the  lungs,  and  above all the
livery but adipose tissue and  the  intestinal mucosa might
also be involved.  Reactions  take  place  in  two  stages:

        stage I           :     oxidation, reduction,  hydro-
                                lysis
        stage II         :     synthesis, conjugation re-
                               actions.

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                           1909
The enzymes involved in stage I aye localized in the micro-
somes. They are called 'processing enzymes' or  'drug-meta-
bolizing enzymes'.  These processes do not necessarily
result in the detoxification of the molecule subjected
to processing.

We shall try to discover:

        o  which these enzymes are;
        o  under what physico-chemical conditions they
           operate.  This amounts to investigating their
           environment; amongst other things* these
           enzymes require the presence of several other
           cofactors - in particular NADPH, cytochrome P 450,
           etc.  Moreover, mechamisms by which  electrons are
           transferred from substrate to oxygen present
           difficult problems in view of the many intermed-
           iate cofactors, whose role is not well under-
           stood;
        o  under what physiological conditions  they operate.
           In other words, can we show that the sythesis
           of these enzymes is either stimulated or
           inhibited by different experimental  or physio-
           pathological conditions?


5.  The effects of  the compound and its metabolization
    on cell metabolism*

The physiological and biochemical reactions of  an ammal
subjected  to  long-term treatment must be observed as  closely
as possible both  for normal reactions and  for reactions
in response to a  stimulus  (starvation, other dietary  dis-
turbances  or  imbalances,  etc).

        a)  an outline of the animal's condition will be
        given, covering:
         - study  of growth,  ingesta,  nutritional efficiency;
         - respiratory exchange, expenditure of energy
           and nitrogen;
         - hormone  level  (insulinaemia,  corticosteronaemia,
           etc..);
         - weight  of  the  various organs  and  physical
           composition  (histological  study,  functional
           examination,  etc).

         b) An evaluation  will  be made of the usual  techniques
         using radioactive tracers,ie.  rate of  renewal
         of nucleic  acids,  proteins  and phosphol^pids
         in the various  tissues.

         c)   The  potential enzyme  activity  of certain  meta-
             bolic processes  will  be determined.

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                           1910
        d) The kinetic study of metabolite concentrations
         and cofactors in a tissue, either1 in the whole
         animal or in certain organs, tissues or organs
         is of particular value for detecting differences
         in metabolic action and for putting forward
         hypotheses on the ways in which compounds act
         in a tissue.

III. - EXPERIMENTAL CONDITIONS

The majority of experiments are carried out on well-nourish-
ed animals which are subjected to one factor only, namely,
the factor to be investigated.  Results obtained for  healthy
subjects are not necessarily applicable to those whose
metabolism is already in a state of imbalance.  It would
be interesting to repeat the experiments described in the
case of animals which are obese due to diet, diabetic,
etc., receiving a low-protein diet or a diet deficient
in certain vitamins, or one with a high ethyl alcohol
or saccharose content.  Such variable conditions would
approximate to experimental models for certain conditions
induced in man by certain conditions of life.  Ve know,
for instance, that  'processing enzymes' are inhibited
by barbiturates, polycyclic hydrocarbons and various  types
of steroids such as androgens, glucocorticoids, anabolising
agents, crganochlorinated substances, etc.  The problems
of metabolic toxicology are particularly apparent during
development.

        a)  During foetal life:
            effect of the toxin experimentally administered
             directly to the foetus on its general development
            and on certain irreversible aspects of development
            such as sexual differentiation, differentiation
            of the central nervous system;

         -  transplacental transfer of toxins;

        b)  During  lactation:
            transfer of the toxic  substances  in milk.

        c)  During further growth.

CONCLUSION

I.  The foundations of a methodology for  toxicological
evaluation must be firmly laid.

2.  A nomenclature  should be worked out step  by step  making
it possible to classify pollutants according  to their effects
upon metabolism.

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                           1911
3.   A multidisciplinary approach should be  used  similar
to  that adopted in charting the physiology  of  nutritional
metabolisms.

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                           1912
VOUK/FALK
An  expanded  and  accelerated  WHO  programme  on  environmental
health criteria,  stressing the need for  an integrated
approach, was developed  in 1972.   In response to a  number
of  World Health  Assembly Resolutions (2) and  recommendations
of  the United Nations Conference  on the  Human Environment  (3),
a WHO Scientific Group was convened in April  1973 to examine
the scientific basis of  the  proposed programme (4).  The
programme is being  implemented in close  collaboration
with national institutions and WHO collaborating centres,
and has the  following objectives:
    o  establishment of health criteria for environmental
      quality, and  provision of guidelines for the  setting
      up of  exposure limits  to protect man's  health,
    o  promotion  of  research  relevant to  the development
      of environmental health criteria,
    o  identification of  new  environmental  hazards to public
      health.
The following topics are  briefly  discussed in  this  contribut-
ion to the plenary  discussion group on the scientific data
base required for decisions  to protect human  health:  o
definition of environmental  health  criteria as  used  in
the WHO programme,  o  shortcoming of  the existing knowledge
on  exposure/health  effect relationship, o   the  concept
of a threshold for  biological effects of environmental
hazards, o  the progress made in  the  preparation  of  the WHO
criteria document and the identification of new or potential
hazards.

Definition of environmental  health  criteria
Ideally, environmental health criteria may, be defined as
complete sets of quantitative exposure/response  relationships
for all environmental factors, involving different effects
and different population groups,  and  covering the whole
range of anticipated exposure levels  (1).  This relationship
has not been totally defined for any  single agent and less
for combinations of agents and factors in  the environment,

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                             1913
 although studies on ioniying radiation have come close to
 meeting the  situation for practical purposes.
 Health  effects  of environmental  agents
 Linking environmental exposure to prompt  effects (i.e.  those
 appearing relatively soon — within days  or weeks after expos-
 ure)  such as acute disease or death,  is usually much less
 difficult than  associating the exposure with chronic disease,
 which may be a  very subtle effect and often not detectable
 except  with  statistical methods  applied to  comparatively large
 human populations.   In addition  to the characteristics  of ex-
 posure  (intensity,  frequency, variability,  rate,  duration),
 the biological  response depends  on a  variety of host factors.
 Some  population groups such as the young, the old,  the  sick
 or debilitated  may be  particularly susceptible  to environmental
 factors.   An exposure  that would have no  effect on  a normal
 adult individual  could aggravate the  course of  a  chronic ill-
 ness, e.g. cardio-vascular or respiratory disorders.  This
 susceptibility  may be  temporary  or permanent, inherited
 or acquired.  The  presence of factors other than  those  under
 consideration and  their interactions  may  significantly  and
 profoundly modify  the  response to a given environmental agent.

 It is obviously unsatisfactory to await the occurrence  of epi-
 sodes of high or accidental  pollution, with their associated
 adverse health  effects, as  a means of identifying environ-
mental hazards; a  detailed  study of such occurrences  is,  how-
 ever, of great  value particularly when an exposure-effect
relationship can be  established.   The  primary basis  for  envir-
 onmental health criteria  and exposure  limits will have  to be
 the experimental work  done  in the  laboratory before  human in-
 jury has occurred.  Nevertheless  such  experimental work  must
be supplemented by epidemiclogical  clinical and toxicological
programs that seek to  identify indices of adverse biological
effects in man, for there  is wide  variability in  human  res-
ponse.  Man does not necessarily  respond in a manner  predicted
by animal experimentation, where  variability in the  response
of different species and  strains  also  exists.

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                            1914
Exposure limits
An important concept for the establishment of exposure
limits is the threshold of response, i.e. the exposure
level below which no demonstrable effects occur.  It may be
difficult to prove or disprove experimentally the existence
of a true threshold for certain environmental agents  and
certain effects.  Effects which appear not to have a thres-
hold occur in independent cells, in cell cultures, or in cir-
cumstances in which affected cells are able to react autono-
mously or when repair processes are not effective.

There are some experimental data indicating that there may
not be a demonstrable threshold for mutagenesis but for other
effects on reproductive processes it is assumed that thres-
hold levels exist.  Since it has not been possible to reach
agreement as to whether there is a threshold for chemical
carcinogens and, if it exists, how to determine it  (5),some
authorities consider it prudent to extrapolate exposure-res-
ponse curves to zero by a straight line.

For some environmental factors  (e.g. temperature, essential
elements) there is an optimal level of exposure, and departure
from this level, in either direction, positive or negative,
may cause unfavorable biological effects.

The general principle to be followed in  setting-up the  exposure
limits for effects other than mutagenesis or carcinogenesis
is that the limit should be set in all cases below the  "no-
adverse effect  level" i.e. below the exposure  level at  which
no pathological effects or physiological or biochemical impair-
ment has been demonstrated  (4)  and that  the population  group
in which the effects are likely to occur be kept  as  small
as possible.  The practical application  of  this principle
is difficult because of disagreement as  to what constitutes
physiological or biochemical  impairment.  This  brings  us to
the problem of  the sensitivity  of  the  tests  used  to  identify
such  impairment, and to the meaning  to be attached  to  a

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                             1915
statistically  significant departure  from  the  "normal"  state
of the organism.
Interactions
A most important  and difficult consideration  in establishing
exposure limits is the possibility of  interactions  among
various environmental factors.  Of these,  synergism-the exagg-
eration of the effect considerably beyond  the additive— pre-
sents the greatest problem in determining  safe exposure levels.
The interactions  may have important effects on the  exposure/
response relationships, though they may be observed only at
certain exposure  levels, while at very low exposures  there
may be no evidence of synergism or antagonism.

Interactions among physical, chemical  and biological  factors
take place against a background of social, cultural and econo-
mic factors in the community, which may influence the pattern
and intensity of  environmental interactions,  so that  consid-
eration of the entire complex situation is fundamental to
an understanding  of events and to achievement of control.
Only by considering man in the context of his total, multi-
factorial environment can we hope to attain that state of
health that is not merely the absence  of disease but complete
physical, mental  and social well-being.

Risk/benefit evaluation
To permit wise steps to be taken towards legislative control
of hazardous substances in man's environment, it is essential
to make a very careful evaluation of risks and benefits, but
this may be extremely difficult.

The expertise needed for the evaluation of risk is  different
from that needed  for benefit evaluation. When evaluating the
risk,  concern is  focused on adverse health effects  on man,
damage to the environment, and misuse of natural resources.
On the benefit side, the emphasis is on value to the consumer

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                             1916
and the country, i.e. improvement in the health of the popul-
ation and the availability of cheaper and better products,
particularly those that modern man considers indispensable.
Economic considerations, such as the improvement of productiv-
ity, the development of resources, or a better balance of
trade are also important. It should be noted that very often
the population group which is exposed to the risk is not the
same as the population group which may have the benefits from
a given environmental situation.

In both types of evaluation consideration must be given to
the consequences that could arise from a regulatory action.
It is possible that the remedy may prove to have worse con-
sequences than the evil it was intended to cure (6).

WHO environmental health criteria programme
The priority environmental pollutants and hazards for which WHO
criteria documents or preliminary reviews will be prepared
have been selected taking Into account the following consid-
erations (4) :
   "o  Severity and frequency of observed or suspected
       adverse effects on human health.  Of importance
       are irreversible or chronic effects, such as
       genetic, neurotoxic, carcinogenic, and embryotoxic
       effects including teratogenicity.  Continuous or
       repeated exposures generally merit a higher
       priority than isolated or accidental exposures.
    o  Ubiquity and abundance of the agent in man's
       environment.  Of concern are inadvertently
       produced agents, the levels of which may be
       expected to increase rapidly, and agents that
       add to a natural hazard.
    o  Persistence in the environment.  Pollutants that
       resist environmental degradation and accumulate,
       in man, in the environment, or in food chains
       deserve attention.

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                             1917
    o  Environmental transformations or metabolic alterations.
       Since these alterations may lead to the production
       of chemicals that have greater toxic potential,
       it may be more important to ascertain the distribution
       of the derivatives than that of the original pollutant.
    o  Population exposed.  Attention should be paid to
       exposures involving a large portion of the general
       population, or occupational groups, and to selective
       exposures of highly vulnerable groups represented
       by pregnant women, the newborn, children, the infirm,
       or the aged".

Using these considerations, a priority list of some seventy
chemical, biological and physical agents was prepared  (4).
Environmental agents for which criteria documents or prelimin-
ary reviews (Preliminary reviews deal with substances for
which toxicological information s limited) have been initiated
in 1973 or will be initiated in 1974 are given in Table I.

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                             1918
                       TABLE I

   PRIORITIES FOR THE PREPARATION OF WHO CRITERIA
            DOCUMENTS AND PRELIMINARY REPORTS
                        1974
           to be initiated in
                          1975
Criteria documents
Preliminary reviews
manganese
nitrates, nitrites
nitrosamines
PCBs
mycotoxins
cadmium
mercury
lead
oxides of nitrogen
asbestos
noise
Sb, Bi
Se, Mo, Te
Ti, Ge, Sn
organic dusts
selected products
of petroleum indus-
try
nickel
vanadium
sulfates, H_SO.
aerosol
fluorides
chlorinated biocides
 and chlorine
arsenic
beryllium
chromium
SO- and suspended
 particulate matter
carbon monoxide
ozone and oxidants
polycyclic hydrocarbons
carbon disulfide
Li
Ba
La, Al, Ga,  Zn
Fe, Ni, Co, Pd, Pt
inert dusts
selected plastics and
plasticizers

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                             1919
The principle which has been used in the preparation of the
criteria documents and preliminary reviews is that of an integ-
rated approach, i.e. the exposure of man through various path-
ways (air, water, food, home and working environment) is ex-
amined simultaneously to arrive at a total exposure.  When-
ever possible, guidelines will be given for primary protection
standards, i.e., "an accepted maximum level for pollutants  (or
its indicator) in the target or some part thereof or an accep-
ted maximum intake of a pollutant or nuisance into the target
under specified circumstances"(7).

Based on these primary protection standards procedures will
have to be developed for deriving 'working limits' i.e. "maxi-
mum acceptable levels of pollutants in specified media other
than the target designed to ensure that under specified cir-
cumstances a primary protection standard is not exceeded"(7).

WHO has no intention of proposing international standards
except in specific cases, such as those associated with food
safety or the pollution of international waterways; in other
cases WHO and other international organisations have the task
to provide the best available  scientific information and guid-
ance for setting up exposure limits. Such guidance should
include specific reference to  the geographical, climatic,
social, economic and technological considerations as well
as other public health priorities which governments should
take into account in the formulation of regulatory policies
for environmental control and  in setting up of national stan-
dards for the quality of the environment.

More than thirty WHO Member States are collaborating in this
program and an agreement has been reached with the United
Nations Environmental Programme, UNEP, to support these acti-
vities .

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                             1920
                          REFERENCES
1.     The WHO Enyironmenta]^ Health Criteria Programme
       (unpublishe
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                           1921
             DISCUSSION OF THE WRITTEN QUESTIONS
BARTH
What is thejbest way to make available the^results of the
world environmental research to environmental protection
administrators in terms they can understand and use?

BARTH
I believe that the best way to make this kind of information
available is through a process of "agreed-to criteria
documents," very much like Dr. Vouk has just discussed.
In my opinion, an appropriate international body is really
the best kind of organizational entity to put together
such documents. There should be representation from senior
scientists from many different countries so as to have
different points of view.  Hopefully, the documents would ad-
equately reflect an evaluation of the world's pertinent liter-
ature.  Now, I would also hope that, as a result of this and
other meetings and cooperative agreements, that world scientists
and environmental health specialists can get closer together
in terms of the way in which they design and carry out their
experiments and the way in which they report their experimental
data. The better the research, the more comparable the research
and the better the quality assurance techniques, the better
the available data will be for incorporation into criteria
documents which can serve as source documents for administrators
of environmental protection organizations.
TREMOLIERES
Pour que les administratifs ou les decideurs puissent comprendre
veritablement le document scientifique, je crois qu'il faut
qui'il y ait une sorte de culture de base commune et par
consequent les criteres qu'on leur soumet, doivent etre
creuses.  Je prends un exemple:  on a pris des criteres pour
les cyclamates dans la saccharine et on note ces criteres;

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                            1922
11 y a eu un effet cancerigene a des concentrations  extrement

elevees, et par consequent on les a interdit.  Pour  expliquer

veritablement ce que signifiait ce critere il faut aller beau-

coup plus profond.  En prenant 1'alcohol conune exemple  I1 infor-

mation scientifique doit etre comprehensible et la recherche

poussee suffisant pour qu'une dinstinction claire puissent

etre faite entre au processus normal d1oxidation de  1'alcool

et un processus qui est considere a present comme nocif etant

donne qu'il induit an cataboliseme nucleique.  Tout  soit etre

compris par un ensemble de populations, il y un probleme de

culture general qui se pose, il faut sortir de notre hyperspe-

cialisation pour que nous ne soyons plus dans une babel ou

chacun ne peut comprendre le langage du voisin mai ou on a

une culture qui puisse permettre de comprendre de secteur a

secteur effectivement ce que veut dire un travail scientifique
preecis.


If those who take and implement decisions are ready  to under-
stand scientific documents, we must be aware that there has
to be a sort of common basic culture and accordingly the criteria
proposed should be carefully explained.  Criteria were establish-
ed for cyclamates and saccharin.   In the name of these criteria -
very high concentrations were found to be carcinogenic - cycla-
mates were banned.  To give a complete explanation of what
this criteria meant,  one would have had to go much more in
depth.   Taking alcohol as an example,  the information must
be made comprehensible and scientific research must  be taken
far enough for a clear distinction to be made between the normal
process of alcohol oxidation, i.e.  dehydrogenated alcohol,
etc. and processes now considered to be harmful because they
induce nucleic catabolism,  etc.   In other words I think there
is a body of scientific knowledge which we cannot ignore and
this raises a particular problem in our time.  This  must be
understood by the masses and the question of general culture
arises.   We must end our overspecialization and emerge from
a tower of babel where nobody understands his neighbour's lang-
uage into a culture where from sector to sector the  meaning
of a specific scientific study can be comprehended.


EARTH

What forms of pollution are found in the developing  countries

and what environmental research,  with what results have been
conducted in developing countries,  particularly Africa?

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                            1923
 GOLDSMITH
 I  am going to introduce my remarks by further comment on the
 previous  question.   As an occasional reader and occasional
 contributor to criteria documents, I think that the communicat-
 ions need more than the written word.   I think it needs personal
 communication and  it needs especially,  and the question is
 addressed to this,  an appreciation of the fact that conditions
 in different places,  different jurisdictions,  are often very
 different and I especially want to emphasize that a standard
 or even a criterion which is  highly significant and must be
 enforced  and promulgated in a European  country or a North
 American  country or a South American country may be completely
 inappropriate to conditions in other developing areas as in
 Africa.   So the first answer  is that one must not try to copy
 environmental health  regulations uncritically and the best
 protection  is active  interchange with people who know local
 conditions  as well  as the  research.   The specifics of the
 research  in Africa  which I know deal with the effect of housing
 variables on infant broncho-pneumonia mortality which has been
 done by Soffolui, on  naso-pharyngio  cancer by Dr.  Winder and
 in New Guinea by Cleary  which  is also on housing and pollution.
 Then, of course, there are a great many  problems of occupational
 health which  developing  countries  may devote their resources
 to with considerable  benefit and substantial  protection.   Since
 most of the  occupations  are associated with  industries  that
 may also produce community pollution, the  opportunity to combine
 the effort  is  a  very  important  one.  Finally,  in the communic-
 able disease  area,  there is a very interesting  study on cerebro-
 spinal meningitis and its  transmission through  housing.   I
might just  conclude by saying that by stressing the  importance
 of domestic  exposure  to health  in  such research as  has  been
done in Africa,  the scientists  have  called attention to neglected
areas in other parts of the world  and I  think we should recognize
 that this is a very substantial  contribution of  theirs.

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                            1924
BIERSTEKER
I think that basically the problems in the developing countries
and in what we call the developed world don't differ too much/
except that the accent in the developing countries is still
on basic sanitation.  Due to a lack of funds, and maybe also
a lack of interest of the general public, the pressure for
strict control on industries is less than we experience. As
for the studies, I think the amount of money available to conduct
studies in these countries is limited.  I spent about six weeks
in India in December of last year and January of this year
and I think they are willing to do studies; they have planned
to do studies, but so far, have had little financial assistance.
On the other hand, I think if we look at it from a global point
of view, some of the situations there may yield more information
than we can find at home.  I thought India had no smog problems
but when I was in Delhi there was smog for more than a week,
a very bad smog.  The planes couldn't land.  This was really
very impressive.  So our concepts about what's going on in
other parts of the world are sometimes completely wrong.  I
think they are facing the same problems that we are trying
to solve but there is just a time gap.

BEN IN SON
I think that if we go to the primary standards, the standards
relating to tissues, concentration in human tissues, I would
guess the results would come mainly from the more developed
countries where a lot of research is being done.  I think these
values with some qualifications could be used as well and it
would not be very wise to attempt to duplicate research with
less possibilities.  However, when those standards have to
be translated into environmental standards, and to derive working
limits, then the ecological chains are still important.  To
transfer parameters in these chains would be quite different
one place from the other.  Therefore, I think the emphasis
in developing countries would be to study the transfer mechan-
isms in the characteristic environment of that place. For ex-

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                            1925
ample, if studies are done for transfers through  a deposition
mechanism on the grass, passage through milk, and then human
consumption, the results would have  little value  in a place
where milk is not produced.  Emphasis  should be given to the
particular food chain which would be characteristic and critical,
 (critical in the sense of transmitting a larger amount of a
given pollutant).    I think this would be a priority in develop-
ing countries.

VOUK

Since I come also from a developing  country, I would like to
make a brief remark.  There is no clear distinction in the
field of environmental health between  developing  and developed
countries.  Of course, as Dr. Biersteker stressed, the bio-
logical type of pollution resulting  from lack of  sanitation, is
the major problem and will remain so for a long time in the
world as a whole.  But there are regions in highly developed
countries, where lack of basic sanitation is still of major
importance as recent outbreaks of some waterborne diseases
have demonstrated.   Similarly, as Dr.  Biersteker  stressed, there
are soiue areas in developing countries where one  can find
heavy chemical pollution situations  because of very rapid
industrial development and urban development.
EARTH
What specific screening tests or other procedures would you
recommend to delineate individuals or population groups who
are especially vulnerable to certain environmental agents?

GOLDSMITH
Screening tests have not been tried extensively in environmental
health although many of them should and could be. The most
important general yardsticks are that people with inherited
biochemical defects or chronic heart or lung diseases are ex-
ceptionally vulnerable among those on the disease category.

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                           1926
Special attention, I think, has got to be paid to the effects
of pollution on children.  We have had some very interesting
contributions, both experimentally and epidemiologically, to
the importance of early exposure to lead. I think that in the
future, we have to increase the proportion of our attention
that we pay to effects on children on growth and development.

BIERSTEKER
I can think of a number of ways of screening people on the
basis of chemical tests.  So, far, there has not been much
use made of specific screening tests.  The group which deserves
special attention is the group of pregnant women.  Upon further
chemicalization of the environment, pregnant women, not them-
selves, but the unborn fetus may well act as a monitor to pick
up other kinds of risks.  A group which has been used for these
studies is, of course, the group of patients with chronic bron-
chitis.   On what basis they have bronchitis is not as relevant
as their acting more or less as a barometer in the environment.
Some of the most valuable work is being done in England and
other areas.  This has been possible thanks to the cooperation
of these people who act as monitors in these studies.

BARTH
Why has so much time been spent at this Symposium on air
pollution research and^is it possible to apply data from
studies alone for decisions on the protection of human
health?

EARTH
To start with, we had to con ider those papers which were
submitted for this Symposium.  Clearly, there is a great deal
of interest in the various nations conducting environmental
research in conducting research in air pollution.  There just
happened to be a predominantly larger number of air pollution
research papers which were submitted to us for the Symposium.
It was not feasible to go through and completely balance out

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                            1927
 the  papers  that  were  in  the  area  of  air  pollution control,
 water pollution  control,  noise, etc.   If we  were  to maintain
 a  strong scientific integrated  approach  to the  program.   The
 question, "Is  it possible to apply data  from such studies alone
 for  the decisions on  the  protection  of human health" —  certainly
 not. The information  which one  gets  from air pollution studies
 will help insofar as  air  pollution is  concerned.   It has aspects
 not  only with  air pollution  control  for  public  health but also
 for  occupational health.   It certainly is applicable for occupat-
 ional health and to public health, but when  one is concerned
 with protection  of the environment,  one  must be concerned
 with the total environment.   It is never enough to look  just
 at air pollution.  One must  look  at  the  collection of air
 pollution, water  pollution,  soil  pollution,  food  pollution
 and so forth —  the entire aggregate has to  be  looked at in
 order to develop adequate  protection guides.

 TREMOLIERES
 Cette question est tres pertinente car c'est finalement  sur
 un plan metabolique que se juge une  toxicite et laisser  toute
 cette toxicologie dans des secteurs  specialises c'est precise-
ment s'ecarter de cette espece de background commun  qui  permet
 finalement de repondre aux divers secteurs specialises qui
 ne sont interesses qu'aux  applications.

This is a very pertinent  question because tcxicity  is ultimately
assessed on a metabolic level and to divide  the vhole of toxi-
cology into specialized fields is to ignore  this  kind of common
background nhich  in the long run,  benefits the  various specializ-
 ed sectors,  uhich tend to  concentrate  on  applications.
BENINSON
I will agree entirely that air pollution alone cannot be the
basis for protection.  In many cases, levels in air of a given
pollutant may be quite low, but the deposition of that low
concentration in air onto tlie ground and into a significant
food chain may cause a substantial contamination of man and,
therefore, the overall picture has to be studied for each

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                           1928
pollutant — all the chains leading from the source to man.

EARTH
Would it not be desirable to encourage the use of data
accumulated from the provision of health services in the
conduction of environmental health research?

RECHT
Depuis un certain nombre d'annees 1'OMS a docifie les causes
de deces; et classe les maladies; les nomenclatures existent et
1'analyse des donnees epidemiologiques peuvent done etre faites.
Un point souleve dans cette question de referrait a la
possibilite de comparer actuellement des donnees depuis
I960 a des donnees plus anciennes.  II est certain que ces
difficultes existent, mais cela n'empeche pas que cette comparais
peut dormer des information sur 1'evolution du phenomeme;
I1interpretation ne doit pas etre faite en termes absolus.  Il
y a un second point qui etait fort important il concernait le
role du corps medical; le Panel y a ete tres attentif,
1'information, I1education du corps medical est actuellement
insuffisante pour lui permettre de collaborer aux etudes
epidemiologiques necessaires pour I1environment.  Or, conune
nous 1'avons vu, un tres grand nombre d1etudes epidemiologiques
sont basees sur des informations des examens medicaux et re-
quiert, par consequent la collaboration du medecin traitant
et du medecin praticien. Des lors un effort particulier d'infor-
mation et d1education doit etre realise, specialement dans
le domaine de certaines enquetes qui toucheront des regions
caracteristiques et concernant certains domaines que 1'on n'a
pas abordes ici mais qui sont tres importants.  Deux examples:
     - 1'ensemble des facteurs qui interviennent autour de
       naissance; il est impossible actuellement de
       conduire, selon des methodes valables, parce qu'on ne
        1'a jamais fait et on ne c'est jamais mis d1accord
        sur une methodologic, des enquetes relatives a la
       perinatalogie.  Or, la perinatalogie joue un role -

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                            1929
       essential dans 1'apparition des anomalies congenitales
       et 1'environnement est certainement responsable
       pour une partie de ces anomalies;


     - Le second probleme a ete la question de la bronchite;

       la bronchi te chronique qui est consideree conune

       un element important, 1O% de deces au Royaume-Uni, 3 a

       7% en France et des chiffres analogues dans plusieurs

       pays de la Communaute Europeenne, est un probleme

       complexe et que sans la cooperation du corps medical

       on ne peut pas arriver a repondre aux desiderata d'une
       methode scientifique.


For a number of years causes of death have been codified and
a classification of diseases made by WHO; a nomenclature thus
exists and epidemiological data can therefore be analyzed.
Reference was made to the possibility at this stage of comparing
post i960 data with older data.  This problem does exist,
we agree, but such a comparison can still give information
on the trends without having to be interpreted in absolute
terms.   A second very important point concerned the role of
the medical profession; the Panel has devoted much attention
to this and it felt that the present state of information and
training of the medical profession is inadequate for it to
participate effectively in environmental epidemiological studies.
As we have seen,  very many epidemiological studies are based
on information from medical examinations and thus require the
collaboration of the specialist and the family doctor.  A special
information and training effort must thus be madet especially
in connection with certain surveys concerning characteristic
regions and certain fields which have not been mentioned here
but which are nevertheless very important.  I quote two.  The
first concerns all the factors which occur around the time
of birth.  At present it is not possible to conduct surveys
by valid methods because this has never been done and there
has never been any agreement on a methodology for perinatal
studies.   let, perinatal observations play an important part
in the detection of congenital abnormalities, and the environ-
ment is certainly responsible for some of these.   The second
field concerns bronchitis.  Chronic bronchitis is considered
an important factor (it is responsible for lo% of deaths in
the United Kingdom and 3-7% in France; the figures are similar
for several Community countries).  Bronchitis is certainly
a complex problem and without the cooperation of the medical
profession we cannot arrive at a scientifically sound answer.

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                           1930
EARTH
What is the importance of understanding biological mechanisms
of environmental pollutants in recommending required control
measures?

BUTLER
Control measures usually involve placing limits on the releases
of environmental pollutants so that the amounts reaching the
most sensitive receptor are below those causing unacceptable
risks of harm.  Thus one must know whether the effects of
interest are short-term and caused by a concentration exceeding
some critical value for a brief period, or whether they are
long-term and caused by the prolonged accumulation of the
pollutant or its effects in the receptor.  The limits to be
imposed will be quite different for the two modes of action.

EARTH
In estimating environmental hazards how important is an  under-
standing of environmental transformations which the pollutants
undergo?

VOUK
Man's exposure to environmental agents is either direct, such
as by contact with chemical or exposure to certain physical
hazards, or Indirect, involving pathways through one or several
environmental media (air, water, soil and food). In most situations,
there will be various, and often complex, pathways by which
pollutants may reach man.  Experience has shown that certain
exposure pathways are much more important than others; these
pathways are called "critical."  Identification of critical
pathways is of great practical value because it will usually
indicate which population groups are likely to receive higher
exposure than others, depending on their habits, occupation
or age, etc.,  ("critical population groups").  The knowledge
of critical pathways and critical population groups will simplify
and reduce the cost of monitoring and will make effective

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                           1931
control measures possible.

Often a series of chemical transformations occur before a
pollutant reaches its human target.  In many cases these
chemical or biochemical transformations produce substances
which are more toxic than the original pollutant.  A well-known
example of a complicated pathway through the food chain is
that of methyl mercury, but others may also exist, for instance,
tin, platinum, gold and thallium can be methylated in the en-
vironment, but the methyl group cannot be transferred in bio-
logical systems to lead, cadmium or zinc.  Similarly, biochem-
ical transformations involving methylation are expected for
arsenic, selenium and tellurium.  These biosynthetic processes
are influenced by pH, temperature, and by the presence of other
chemical species; for instance, the biological methylation
of metals is inhibited by some chlorinated hydrocarbons.

Complex chemical and photochemical transformations of pollut-
ants can occur also in air.  Some of these have been extensive-
ly investigated as, for instance, the formation of the photo-
chemical complex.  Much less is known about the atmospheric
transformation of sulfur dioxide and the relationships of nitric
oxide to nitrogen dioxide.

EARTH
What is the importance of developing standardized data services
to make rapidly available results of environmental research,
and what quality assurance procedures would be necessary?

VOUK
A major problem today facing both scientists and regulatory
agencies is how to bring relevant scientific and technical
information from the published literature and miscellaneous
sources to those who need it.  In dealing with pollutants,
it is essential to know their chemical structure; secondary
product formation; pattern of usage;  production data
paths of disposal or environmental  "sinks"; animal and human

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                            1932
toxicology; synergistic effects with other pollutants.  There
are already several centres and data banks where attempts are
being made to store such information.  Most of these data banks
are specialized and cover only a part of the information which
scientists or decision-makers in the field of environmental
health may wish to have. Some of them are computerized search
systems, such as TOXLINE or MEDLINE.  The Toxicological Infor-
mation Response Centre  (TIRC), which has been established in
the USA at the Oak Ridge National Laboratory in 1971 and which
is sponsored by the National Library of Medicine  (NLM), aims
at building up a toxicological data base, to collect and
disseminate toxicity information in the form of bibliographies,
reviews and state-of-the-art reports, and to answer specific
research requests from the scientific community.  Another
example is the Environmental Mutagen Information Centre  (EMIC)
which has been collecting data on the mutagenic action of chem-
icals since 1969 to the present and made this information
available in a number of publications.  Most of the existing
systems, however, are designed in different ways and for specific
kinds of uses, and it is therefore difficult to build up from
them a general data bank and information centre to serve differ-
ent types of users.  An integration of such systems is undoubted-
ly needed, but a rigid framework is unlikely to provide the
most serviceable system.  A flexible approach, as conceived
by the International Referral System for Sources of Environmental
Information (IRS) of the United Nations Environment Programme,
would have the best chance of being useful and acceptable.
Maximum flexibility should be incorporated in planning such
a system, so that changes in uses and needs may be taken into
account, if necessary, for data storage and presentation.

Of course, most of these systems have little or no quality
assurance procedures incorporated, and it is difficult to
see how that can be done without a detailed evaluation of
the original information contained in scientific articles.

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                            1933
This evaluation can only be done by experts in a particular
field, and would give the best results if done on an internation-
al basis.  The WHO environmental health criteria programme  (a
short description was presented in my opening statement) is
an example of a system which aims at providing balanced and
unbiased, internationally acceptable, information on the relation-
ship of exposure to environmental pollutants and hazards and
effects on man's health.  One obstacle to the use of available
information is that it has, in most cases, been obtained by
different methods, both for measuring exposure and for assessing
biological response, and that the results obtained in different
laboratories, in various countries, are sometimes not comparable
at all.  A prerequisite for quality assurance of information
on environmental levels of pollutants and their effects is
the harmonization of methods for monitoring pollutants, for
toxicological testing, and for epidemiological studies.  Several
international organizations, including WHO, are making efforts
in that direction.

EARTH
How does one determine the levels of uncertainty of knowledge
that wiJ.l be acceptable to an Environmental Protection
Administrator for his decision?

BIERSTEKER
There is only one way to find this out and that is in actual
practice.  If the pressure from politicians and  (or) the public
opinion is strong enough, environmental protection is possible
in the form of source control or emission reduction without
too much information on adverse health effects.  A fixed level
of uncertainty  (or certainty) to take action without public
pressure is hard to establish, I fear, as this may be a matter
of character and temperament of the administrator.  In general,
administrators have learned that it is impossible to wait for
complete certainty and that action without it is justified.
This is nothing new in public health administration.

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                            1934
EARTH
It is possible^ to  reach agreement  on  the  definition of
"risk" criteria for various environmental pollutants?

BEN IN SON
I believe risk  criteria will be the trend.   Standards  based
on risk are of  normal use at present  in. radiation  protection
and the same approach can be used  for other  pollutants,  part-
icularly taking into account the stochastic  effects for  which
no threshold can be readily demonstrated.  The panel's intro-
ductory statements should be examined closely.

EARTH
How important are  annoyance reactions and irritation effects
in setting protection guides?

BIERSTEKER
They are in my  opinion very important, for the public  in a
welfare society is less and less willing  to  accept nuisances.
The reason is probably that the public feels that  with a bit
more care and some extra engineering  efforts, such effects
on the environment are avoidable. Consultation with sociologists
and psychologists  who are able to measure public feelings
and establish tolerance levels should be  part of the standard
setting process.   A difficulty is that in the course of  time
the tolerance and  attitudes of people are apt to shift.

BARTH
Is there a practical way to integrate qualitative  dose-response
information with quantitative infojrmatic-n?

BENINSON
Qualitative response-dose information can be used  in setting
standards by assignment of some quantitative character.  For
example, by rank ordering procedures  or selection  of suitable
sample groups of people for frequency assessments  in the case
of annoyance or irritation effects.   Even the absence  of

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                            1935
observations may give upper limits of the slope of a non-thres-
hold relationship, in the given confidence levels.

EARTH
How is it possible to consider and judge the relevance of
several subcritical effects occurring at the same time?

BEN IN SON
When several effects of a given pollutant can occur, it is
possible to give a quantitative indicator of the total by the
use of the concept "detriment" as presented in my introductory
remarks. It requires the establishment of severity weighting
factors for each effect.  If several pollutants, each with
a standard based on risk, act at the same time, safety could
                             /"» 4
be assessed by the relation Z~ < 1, where C. is the level
                             bi             1
of pollutant i and S. is its standard.

EARTH
Once an apparent threshold value has been determined for an
effect in relation to an exposure as measured by conventional
means, to what extent,  if at all, should more sophisticated
measurement procedures  be sought to attempt to  find effects
at lower exposure levels?

GOLDSMITH
The demonstration and acceptance of evidence concerning one
threshold for a pollutant is no reason to think that for another
population or another effect other data are not relevant to
control needs.  These effects need not even be  at a lower  level
of exposure.  In my view, threshold concepts should not be
applied to human population reactions.  I prefer  to look at
a set of criteria, which research continually refines.

EARTH
This concludes the written questions  from the participants
as summarized by  the panel.   The panel now  stands  to  receive
questions from the floor.

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                           1936
                 DISCUSSIONS FROM THE FLOOR

NEEDLEMAN  (U.S.A.)
How can we achieve a scientific data base upon which to make
health decisions, if we are not provided with a complete,
unbiased presentation of the data?  For example this morning
we heard a report of neuropsychological performance in children
in Smeltertown USA.  The report found no deficit in children
with blood lead levels greater than 4O pg/lOO ml.  some of us
have had the opportunity to read the Center for Disease Control
Bulletin of May 1, 1974, which showed precisely the opposite,
studying the same sample.  What are the causes for such discrep-
ancy, and why were they not aired in the presentation of the
paper?

GOLDSMITH
This may be aside from the assignment of the panel, but in
a free scientific society, all types of communication by people
of scientific stature, of course, are welcome.  Some of them,
of course, will not agree with others and, in this case, I
think the disagreement is apparent.  I want to refer back to
my opening remarks in which if you are going to use information
for establishing policy decisions, then the peer review process
of publication confirmation and having authoritative scientists
accept the fact that the document is relevant is a very important
part of the process.  I think that this is one of the ways
to make sure that scientific controversy is recognized and
resolved in the most suitable way. We cannot, however, have
a free exchange of scientific information without the hazard
that papers will give conflicting information some of which
will be either biased or will be suspected of bias.  Only the
insistence on open availability of research results, on peer
review and validation, and on calibration and confirmation
of the data base, can assure that scientific results will be
dependable.   Concerning the particular session, no doubt the
lack of time not lack of desire or intent was responsible

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                           1937
for the superficial discussion of the paper in question.

BUTLER
I would just like to add a word to this.  Mr. Chairman, this
example just reinforces what you and Dr. Vouk have said about
what is required for establishing a solid scientific base of
criteria for environmental standards.  That is that you need
a committee of experts, preferably international, where all
points of view and all conflicting bits of information can
be brought together and the differences resolved.  In my view,
this is the best answer that one can give to the question.

HIKE (U.S.A.)
At this time I wish to comment on matters pertinent to the Plen-
ary Discussion Group's deliberations.  I consider it mandatory
to maintain a system of checks and balances so that we can
arrive most precisely at acceptable community levels of pollut-
ants.  Those who work in the gathering of data should have
a limited role in the critical evaluation of their own conclus-
ions.   They should have a still lesser role in the setting
of standards.  Criteria documents should not ignore significant
negative data nor should they cite positive findings if they
are not confirmed by others or are of questionable relevance.
The experimental base should not be located solely in government
laboratories.
There should be an effective mechanism for review of all data
used for establishing air quality criteria.  Not only should
the opinions of qualified scientists in industry and academia
be solicited, but their opinions should be carefully considered,
evaluated and where appropriate, included.  Past experience
leads to the inescapable conclusion that these comments, though
sometimes solicited, are almost always ignored.  Finally I
raise the question of the propriety of any group to function
in all three activities of data gathering, standard setting
and enforcing these standards.

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                            1938
RECHT
Je suis un peu etonne de ce type de reflexion car tout ce que
nous avons appris au cours de ce Symposium a permis de cons-
tater que 1'investigation epidemiologique etait une science
difficile a laquelle ne pouvaient pas participer des amateurs;
a partir du moment ou sur le plan scientifique, sur lequel
nous sommes, on applique des methodes qui sont bien connues
en statistique et en epidemiologie, tout ce qu'on a dit se
revele etre soit de la politique ou de la polemique mais pas
de la science.  Or nous savons que, sauf peut-etre aux Etats-
Unis ou en Grande-Bretagne et dans quelques secteurs isoles
des pays europeens, nous sommes sous-developpes en ce qui
concerne les etudes epidemiclogiques et ce sera une constatat-
ion vraisemblablement de ce Symposium.  Nous le sommes et nous
devons suivre les examples qui nous sont donnes, qui ont ete
donnes par ceux qui ont participe aux reunions mais je suis
tout-a-fait d*accord avec 1'orateur precedent que la question
du contrfile est un probleme qui est tres important, mais
ce contrSle n'est pas un probleme scientifique.  A partir
du moment ou vous avez une equipe qui est constituee,  elle
doit pouvoir repondre aux lignes d'une recherche scientifique
bien etablie.
I am rather surprised at this attitude because everything we
have learned during this Symposium has pointed to the fact
that epidemiologiaal investigation is a difficult science un-
suitable for amateurs*  As long as proven methods in statistics
and epidemiology are applied at the scientific level, every-
thing just said turns out to be either politics or polemics
and not science.   yet we "know that with the possible exceptions
of the United States or Britain and some isolated examples
in Europe we are underdeveloped in the field of epidemiological
studies and this will probably be one of the conclusions of
our Symposium.  Ve are underdeveloped and must follow the ex-
amples given during this Symposium.   I quite agree with the
last speaker that the problem of surveillance is very important
and that this is not a scientific problem.  From the moment
a team is established it should be able to follow the lines
of well-defined scientific research.
MOLLARET (France)
Le richissime catalogue de documents recueillis par le sympo-
sium ne doit-il pas conduire a proclamer les deductions pra-

-------
                           1939
tiques a imposer d'urgence a I1attention des autorites
gouvernementales Internationales et avant tout sur la sauve-
garde du capital en eau potable?
Should not the wide selection of documents gathered by the
Symposium be used as a basis in order to choose the practical
measures which national and international bodies should be
asked to take as a matter of urgency?  One example of an item
requiring immediate safety measures is our most valuable and
most threatened asset, drinking water.

EARTH
The papers presented at this Symposium will most certainly
be used in the development of criteria, standards or means
of choosing practical measures of abatement of pollution.  As I
understand there are plans in the European Communities to have
one or more colloquia regarding drinking water pollution.  In
the United States, our Congress will be preparing legislation
to enable the Environmental Protection Agency to develop regul-
ations and conduct research on drinking water supplies.

KJELLSTROM (Sweden)
I think the panel has presented some interesting views.  I
would however like to take up three points.

First, we had this morning some small discussion about the
carcinogenics and the priorities  - which ones we should
study, etc.  I think it is very important that we as scient-
ists say what the priority should be on whether new possibly
carcinogenic compounds should be introduced at all in the en-
vironment. If we can stop them before they are out in the en-
vironment, so much the better.  It is the ideology of negative
proof as we sometimes call it in Sweden, that has been accepted
by the National Food Administration; e.g., they had in prin-
ciple forbidden all synthetic colors to food.

Second, I think the documentation of any TLV values or any
standards that we set must set forth the basis for the

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                            1940
reasoning behind such standards otherwise people in other
countries cannot make their own evaluation.  I think the Czech-
oslovakians and the Americans have been broadminded when they
published the documentation of their industrial TLVs, whereas
it is extremely hard to get the Soviet Union documentation.
In Sweden there is no documentation at all, but I hope that
some will be available very soon because the TLV list has been
redone.  So I would say that it is necessary that we have a TLV
or a standard for the environment which is completely medical;
that the criteria for this medical decision be defined exactly;
and that the documentation be published so that any country
can determine if the documentation supports the TLV or the
standard. Then if some country wants an administrative TLV
which is higher or lower, it is up to them.

I understand that the developing countries have problems with
their environment and these problems did not have a very high
priority in the panel discussions.  I would say that there
is a great need for some kind of future research where concern-
ed scientists who know the methodology in environmental research
sit down and think out what can happen in these developing
countries when industries are introduced.  I think that this
could prevent a lot of disease caused by pollution of the en-
vironment. Also let me say that methyl mercury incidents should
show us that there is a great need for us as scientists to
push for banning of dangerous substances, which are not really
so necessary to us. We have banned methyl mercury in Sweden.
Why have we not banned it everywhere so that they won't get
a new kind of poisoning say in Central Africa?

EARTH
I would like to first comment myself on our being taken to
task for the amount of time devoted to a single subject.  With-
out question we could discuss any of the questions that have
been asked for the entire two hours.  We had to make a decision
about the amount of material we wanted to cover in the panel
discussion. We concluded that it did not make much sense

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                           1941
to limit our discussions to one single question.  Now on the
other questions that you are raising, I believe Dr. Beninson
would like to make a remark.

BENINSON
I don't think you can directly ban anything which is consider-
ed carcinogenic in the sense that you may do away with some
of the benefits from the use of these materials.  In every
case I think it is a cost/benefit analysis.   Take a very old
and known example; if it were known at the beginning that
X-rays would be carcinogenic, as we know at present, they would
have been banned at the time.  It is possible that we have
cases of induced leukemia from X-rays, but quite surely many
more people would have died from TB and other diseases.  In
the medical sense as a whole, there has been a marked benefit
over the risk.  I feel that in every case a cost/benefit analy-
sis should be done.  Banning for the sake of banning of a potent-
ial carcinogenic agent may be doing something of a disservice
if the benefits outweigh the risks or potential health impact.

MOKEMATKEMGUEMBA  (Republique Centrafricaine)
Nous attirons 1'attention des hommes de sciences car les re-
cherches dans le domaine de 1' environnement doivent e"tre
objectives et universelles d'autant plus les maladies causees
par les effets de 1'environnement peuvent etre  transmissibles,
done il ne faut pas seulement limiter les recherches sur I1en-
vironnement dans les pays developpes.

Il faut egalement aider les chercheurs des pays en voie de
developpement afin que les efforts dans le domaine de 1'envi-
ronnement soient conjugues.
We seek to attract scientists because research on  the environ-
ment must be objective and universal; especially diseases
caused by the effects of the environment can be transmitted,
research must not be confined to the environment in advanced
countries*
Researchers in developing countries must be assisted so that

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                              1942
efforts in this field can be coordinated.

GOLDSMITH
Perhaps this meeting has not emphasized enough the problems
of developing countries.  Possibly the pollution and environ-
mental health problems of developing countries deserve a meeting
which could stress such problems as the tendency to "export"
pollution problems from advanced countries, the unique oppor-
tunities for mutually worthwhile research in developing countries,
and the wax in which the potential conflict between development
and environmental quality can be transformed into strategies
for effective efforts to encourage both.  In this way
researchers of both developing and developed countries
can approach their respective problems together.

LAFONTAINE (Belgique)
Je voudrais ramener le debat a la serenite necessaire et arriver
Si ce que nous voyions chaque chose selon son importance et son
urgence.  Je souhaiterais aussi que 1'on distingue les accidents
des risques lies a un usage normal comme je demanderais qu'on
n'oublie pas que pour agir centre la pollution de I1environnement
il faut disposer de ressources naturelles et ce proble*me est
plus a considerer encore lorsqu'il s'agit de pays en voie de
developpement.

Je souhaite poser trois questions

- d'abord lorsque des mesures ont ete mises en oeuvre/
  comment se rendre compte de leur efficacite a 1'egard de la
  sante et n'est-il pas important de prevoir a cet effet des
  etudes epidemiologiques prospectives ou le generaliste
  doit, si possible, jouer un rSle;

- par ailleurs,  les approches sanitaires ont trop souvent suivi
  les aspects sectoriels des milieux d'o£ vient 1'agression.
  N'a-t-on pas trop oublie que 1'homme est I1integrateur des
  nuisances et n'est-il pas utile d'avoir des approches

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                               1943
  multisectorielles, sauf dans le cas ou le point d'impact est
  unique  (effet irritant de la pollution de I1air sur les
  poumons par exemple)?

- Enfin, en plus des effets directs sur la sante, il ne faut pas
  oublier les effets indirects provenant des repercussions
  ecologiques coirane par exemple la pollution thermique evoquee
  ou 1'atteinte a certaines productions alimentaires.

We must consider each issue in terms of its significance and
urgency*  I would also like a distinction to be made between
accidents and risks inherent in normal use and I hope also that
it will not be forgotten that natural resources are required
to control pollution of the environment; this problem is especial-
ly acute in developing countries.
I would like to ask three questions:
- Firstly, after measures have been applied, hoi: are we to
  assess their efficiency for health protection?  Shouldn't
  provision be made for prospective epidemiological studies
  in which the general practitioner could also participate?
- Secondly, the health protection approach has too often been
  a sector-by-sector consideration of pollution sources.  Ought
  we not to remember more that man is an integrator of pollution
  and that it is sensible to adopt an overall approach, except
  when there is a single point of impact (e,g, the irritant
  effect of air pollution on the lungs)?
- Thirdly, as well as the direct effects upon health, we must
  remember the indirect effects arising from ecological reper-
  cussions e.g. thermal pollution or effects on certain food
  chains.
BIERSTEKER
We should agree fully with Dr. Lafontaine that if the authorities
plan to take action, they should follow up whether there is
any measurable improvement in populations.  The situation in Lon-
don, which improved so much with the reduced smoke concentration,
was used for this type of follow-up and there are similar
situations in many parts of the world.  I think that all engineer-
ing efforts to improve the environment should have a medical
effort as well.  There is also more need to stress the ecological
effects.  Basically we see that the medical profession is

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                               1944
manipulated into a corner when concerned with the environment.
The experience in Holland is that the environment is now handled
by almost anybody except by the medical profession.  The Royal
Society of Medicine in Holland doesn't have a position on en-
vironmental problems.  The doctors, in general, are not really
interested.  It is really hard to get many students interested
in the environment except for a very short, superficial over-
view.

The vacancies which exist for the medical profession cannot
usually be filled because of lack of interest.  I think unless
the medical profession is well aware that they should and could
play a role in the environment, the health responsibility will
slip out of their hands.  The solutions to the problems in the
environment will be found by the public itself and by other
professions.

TREMOLIERES
Votre deuxieme question me paralt tout a fait centrale et resume
beaucoup de roalentendus des questions d'aujourd'hui.  Vous avez
insiste sur 1'epidemiologie.   Mais je pense que si derriere
1'epidemiologie il n'y a pas une bonne clinique, que si derriere
1'epidemiologie il n'y a pas une bonne clinique, que si
derriere la bonne clinique il n'y a pas une bonne toxicologie,
que si derriere la toxicologie il n'y a pas une etude de la
metabolisation tres profondement vue, on ne se comprend.   Je
m*excuse, on par1ait tout a 1'heure de la comprehension au sein
d'organismes internationaux et je me permettrai de faire une
confidence:  je fais partie depuis 1949 des Comites "calories",
puis "proteines" FAO-OMS.  Il faut d'abord se comprendre entre
experts; on a dit sur les avitaminoses tellement de betises;
je le dis tres directement etant donne qu'on a envoye a des
prisonniers des choses qui ne servaient a rien, qui a vu des
amitaxninoses pendant la guerre, elles ont ete rarisimes.  Alors
la comprehension entre les savants demande une formation des
savants eux-memes, c.a.d. une despecialisation qui permit a

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                               1945
specialise de comprendre ce que veut dire le secteur voisin.

II nous faut batir a 1'heure actuelle une nouvelle heterodoxe,

et meme tres heretique.  On parlait des publications tout a

1'heure; publier quelque chose qui est une approche globale

presente les plus grandes difficultes.  11 faut avoir une

conception globale de cette metabolisation des substances

etrangeres dans laquelle il y a tous les secteurs qui sont

representes ici.


J think that your second question, concerning an overall
approach,  is fundamental and explains many of the misunderstand-
ings in today's questions.   you emphasize epidemiology but I
think that epidemiology must be backed up by good clinical know-
ledge, which must be eeconded by good toxicology, which in turn
must be supported by a very thorough study of metabolization.
Otherwise communication breaks down concerning the metabolic
aspect of health protection and debates like these result.

A moment ago we were discussing comprehension within internation-
al organizations,  I may say that since 1949 I have been a mem-
ber of the FAO-UHO Committees on Calories and Proteins.  First
of all the experts must understand each other; so much rubbish
has been said about avitaminoses, such useless things were sent
to prisoners:  who ever encountered avitaminosis during the
war? It was very rare.  Comprehension among scientists requires
training of the scientists themselves; i.e., despecialization
to permit each specialist to understand the language of the
next.

What we need today is a new approach, a heterodox and very
heretical approach.  Publications were mentioned a moment ago.
It is very difficult to get something which adopts an overall
approach published.  What we need is an overall conception of
this metabolization of foreign substances covering all sectors
represented here.

GOLDSMITH
Professor Lafontaine brought up  a very important subject when

he emphasises the need for epidemiologists to be involved in

monitoring in what happens under a variety of circumstances.

This  is especially important when new technical procedures are

being introduced about which the consequences are not fully

known. Unfortunately there are many  proposals by epidemiologists

that  this be done and very few times does the person concerned

with  the  introduction respond adequately.  Just now the issue

is catalytic control devices and we  see a very great difficulty

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                               1946
in getting a recognition that the kind of background data and
epidemiological monitoring that are indicated are being under-
taken.  With respect to the physician often getting boxed in
a corner, I would like to draw a modest analogy.  I have heard
in the corridors, at least, a great deal of discussion about
environmentalists.  You may or may not like them, or you may
or may not feel that what they are saying should be responded
to directly.  But I would like to insist that at least as a
social phenomenon the influence of environmentalists is an im-
portant symptom, a symptom that the scientific and technical
components of society are being insufficiently sensitive to
problems that are disturbing a great many people outside of
the sceintific and technical communities.  The analogy I want
to draw is that a physician in practice who is a good medical
scientist, as well as a good medical practitioner, has a dual
obligation.  When a patient comes to him, as many in the commun-
ity go to their health and environmental authorities with these
problems — some of them not very clearly stated — his first
responsibility is to respond to the symptoms, to make sure
what the complaint is, to listen carefully, to analyse and to
treat symptomatically; his second responsibility is to carry
out a sufficiently good diagnostic survey so that the underlying
features which cause the particular patient  (in some cases the
community) to have maladies is grasped in a reasonably effective
way. I think that this relationship of the scientific and medical
authorities to the community problems which are brought to them
is often either too symptomatic or too scientific, but both
I believe are necessary.

RECHT
The assessment of the efficiency of abatement methods on improv-
ing the public health is of paramount importance.  We are hamper-
ed at present by the lack of sufficiently accurate data and
sometimes even by the total lack of data regarding the health
condition of the population at least with respect to those symp-
toms which may be influenced by environmental pollutants.  By

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                               1947
sensitizing the general practitioner to these problems and with
his active collaboration much knowledge can be gained.

Man is the integrator of all pollutions, but we do not know
yet how to evaluate the overall impact.  This is one area where
much effort should be placed and where the development lead
loss.

DE ZEEUW  (Netherlands)
I am very impressed with the amount of data which has become
available at this meeting, but I wonder whether the question
how priorities should be decided upon with respect to the collect-
ion of scientific data is not a question which should be asked.
Decisions are based on several different inputs, scientific
considerations, public pressure, etc.  Ideally they should be
based on the requirements needed to set protection standards,
namely quantitative relationships between exposure and effects
(or probability of effects).  Scientists should polarize their
research, giving priority to these quantitative relationships;
an additional responsibility of scientists is to advise
decision makers on the real needs for data underlying any
standard.  Who decides upon priorities necessary for obtaining
the scientific data base required for decisions to protect
human health, and how?  What is the responsibility of the
scientist for these decisions?  Why hasn't the panel introduced
this important problem of the responsibility of the scientist
in priority-decisions?

VOUK
The problem of priorities in science is a very complex question,
and I don't think that we can answer it this evening.  It depends
very much on the interest of the scientist, on the sources of
finance, who does the research, for what purpose and so on.
It is quite true that there is also fashion research.  I have
had the chance to visit a number of laboratories about a year
or two ago where I saw such things going on.  There were very
few institutions which I visited which tried to look ahead

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                            1948
and plan their research accordingly.  As  regards the choice of
priorities for studies, it's a very difficult question.  Some
time ago WHO Scientific groups met in Geneva and certain
criteria were agreed on priorities for research and evaluation
 of existing information on health effects of environmental
 hazards.   These were listed in my introductory remarks.  But
 certainly, there is some need for looking ahead for preventive
 action rather than for corrective action, and if the scientists
 themselves are doing or concentrating on the topics which are
 only of current interest, I don't think that there is any
 prevention possible.  A good example of a preventive view was
 really the development of atomic energy research.  When the
 first atomic reactor was put in operation in December 1942,
 there had been already, a fairly advanced program in biomedical
 research under way.
 BUTLER
 I would like to defend the panel against this accusation that
 they don't think about research policy and give any idea about
 priorities.   Many of us earn a good fraction of our salary by
 doing this.   But it's not easy.  If you don't mind, I will tell
 you some of my own experiences in my own country where in my
 division we are concerned with criteria documents for environ-
 mental pollutants.  One of the most difficult questions we have
 is which substances are we going to study and produce documents
 on. The sort of thing that happens is that on one committee
 you can find a man who is highly respected, who has a very strong
 personality who speaks loudly and longly at great length in
 all meetings and most certainly his pet project is going to
 end up one of the first ones studied.  This is one phenomenon.
 Another one, and I have wrestled with this problem not only
 in Canada but in some international organizations, is to design
 some sort of what we thought was a scientific way of evaluating
 the urgency of problems so that we could attack them in a proper
 order.  We designed a system which we thought was perfect and

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                               1949
as objective as possible. We put it through all kinds of commit-
tees and processes and finally it came to a meeting something
like this where there were emotions and where there were people
who had votes and who changed the whole priority order.   Politics
does enter strongly into the priority rating; perhaps this is
just a fact of life.  The problems that are worrying the most
people in the country are, I suppose, the ones you have to attack
first even though on some subjective scientific rating they
don't seem to present much of a hazard.

BEN IN SON
If I may comment on the question that I think is an exceedingly
important one.  The answer of course would be that the research
for this purpose should be polarized.  On what lines?  There
are three main requirements:  one in the biological line, it
would be to assess more relationships between exposure and effect
or the probability of the effect — the quantitative relations —
and to be able to do so at low levels in tissues.   Probably
the research would start with higher levels and be able to extra-
polate to lower levels.  But the main aim should be to obtain
results linking quantitatively exposure and effect for a given
pollutant. On the ecological line, it should be to obtain values
for transfer coefficients between compartments in the environ-
ment, so one would be able to predict what would be the effect
of a given input in one compartment, what would be the resulting
levels in other compartments and what would the exposure of
humans be.  In this respect I think there is perhaps too much
effort on monitoring without having a clearly defined objective
of the monitoring. The third line of research is more a tech-
nological type.  It would be improvement of procedures to reduce
discharge of waste products on one line and secondly to assess
the feasibility of alternative procedures to be able to do the
cost/benefit analysis, taking into account these alternative
procedures.  These would be the lines.   Now which of the pollut-
ants should be studied first?  I think what we have seen in
the Symposium so far shows that the selection of pollutants
for research has been quite reasonable.

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                                1950
 GOLDSMITH
 I think if we have been led into a trap as a panel, it deals
 with the meaning of the word 'required1 in the title of the
 panel.  I think we are — assuming that we are the ones who are
 capable of making what is required—making requirements for
 the scientific data base.  In fact I think reality would indicate
 that it is a social and a political process which decides what
 the data base is that is required and we are only counsellors
 in that process and not those who set the requirements.

 TREMOLIERES
 Just one word to say that my priority is this metabolic aspect
 of toxicology which means that we can understand what we are
 doing,  and  that we  can understand each other.

 BIERSTEKER
 I  would  like  to make a last  comment,  I think that what so far
 has  escaped medical attention  is  mainly the nuisance problem
 or  the  negative side of  the  beauty of life.   I  think if you
 listen  to what  the  public  wants and what the politicians
 begin to translate  into  action, this  is really  where the field
 of action should be.   As you see  the  number of  publications
 in this, there  are  a few doctors  in Sweden  and  one or  two maybe
 in the United States and that's all the interest  there  is for
this subject.   So,  I think if we  want to talk about  fields
which need exploration and medical attention  I  think this is
a new field which should be explored.

EARTH (Conclusion)
I am sorry but we are  going to have to  terminate  the discussion
here, so I declare  this  session closed.

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                                1951
               QUESTIONS SUBMITTED IN WRITING
             BY THE PARTICIPANTS AT THE SYMPOSIUM

 EDITORIAL NOTE:   These questions were submitted in writing by
 the  participants to supplement questions posed to the panel
 by the  Chairman (see Chairman's note).   The panel took the
 prerogative of condensing and paraphrasing the participants'
 questions in order to fully cover the subject areas assigned
 for  discussion in  the limited time.

 GOOTJES (Netherlands)
 What is the best way to make the hesitating results of many
 expensive scientific projects in lots of countries (for instance
 the  time pattern of concentration of a pollutant in the organs
 of man)  more concentrated and integrated so that the administrator
 can  use them better to convince the  people about the justice
 of his  -in  the eyes of the people — income consuming protection
 directives?

 MOKEMATKENGUEMBA (Republique Centrafricaine)
 On dit  souvent que le  probleme  de la pollution est I1affaire
 des  pays  industriellement developpes,  je voudrais  savoir si
 des  recherches pour determiner  le taux de la pollution ou pour
 deceler les effets de  la pollution sur la sante de 1'homme ont
 etfe  deja  faites  dans  les pays en voie de developpement en general
 et en Afrique  en particulier?  Quels ont ete les resultats de
 ces  recherches dans le domaine  de 1'Environnement?  Sous quelles
 formes  se manifests la pollution dans les pays en  voie de
d eve1oppeme n t ?
 It is often  said that  pollution  is a  natter-  for the  industrial
countries concerned.   What  I i^ould like  to  know is:   whether
any  research  has been  carried out  in  developing countries and
particularly  in  Africa  to  determine  the  level  of pollution or
 to detect the  effects  of pollution on  human  health?   h'hat vas
 the  result  of  such  environmental  research?   what forms  doss
pollution take in  developing  countries?

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                               1952
OMENN (U.S.A.)
What specific population groups or specific screening tests
would you recommend to delineate individuals or groups who are
especially vulnerable to certain environmental agents?

Allow me to mention a few possibilities drawn from "eco-genetics"
studies of inherited individual differences.
  o  inducibility of aryl hydrocarbon hydroxylase by
     polycyclic hydrocarbons:  the induced enzyme converts the
     agent to the active epoxide carcinogen; 47% of population
     are minimally induced, 9% maximally induced.  According
     to Shaw, almost all of 50 patients with lung cancer were
     high or moderate inducers.  Presumably, hydrocarbons in
     the general air, as well as in cigarette smoke, induce
     these enzymes.  High inducers in the population should be
     at greatly enhanced risk.
  o  anti-trypsin deficiency/partial deficiency and enhanced
     risk of early obstructive pulmonary disease — from
     smoking and probably from other inhaled agents.
                                        f*
  o  glucose-6-phosphate dehydrogenase  (^6 PD) deficiency
     and risk of hemolysis from oxidizing agents — anti-
     malarial drugs, sulfa drugs, methroquinone in moth
     balls; likely also the nitrogen oxides in the air.
Recognition of potential high risk groups in design of epidemic-
logical investigations is important; otherwise, no statistically
significant risk may be found in studies of overall population
samples.

SULAIMAN (Nigeria)
In the discussions so far, more time has been devoted to air
pollution than to other forms of pollution.  Is it because
this is a major problem in developed countries, or is it because
this is a field where adequate research has been made, or is
it because of the ease with which the parameters can be measured?
Finally I would like to ask therefore whether it is possible
to apply basic scientific data derived from this field alone
for decisions on the protection of human health in general?

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                               1953
DENNIS  (Canada)
Several speakers during this Symposium have stressed the need
for scientific information for the purpose of political decision
making and implied criticism of the scientist for the fact that
cause/effect information is incomplete.  A health effect implies
an impairment of function; if health services are available
people go to physicians to reverse or alleviate their impaired
function; physicians record the functional impairment.  The
harnessing of this information would allow an ongoing evaluation
of health effects in total populations and allow the identification
of problem areas and a description of the populations within
those areas.  The use of this data source requires a commitment
from which the political decision maker cannot be excluded.
Would it be appropriate for this forum to come forward with
a resolution to this effect?

NEWHOUSE  (Canada)
Could the panel consider the question of mechanisms of the bio-
logical effects of pollutants especially air pollutants since
this may make an important contribution to determining control
measure.  Little is known about this as yet but it should, at
least, be identified as a problem which needs to be tackled.
Understanding mechanisms might result in more specific  (and
thus perhaps more specific and cheaper centra-measures) recommend-
ations to industry, etc.

OLOFFS  (Canada)
Recent scientific progress, especially in the life sciences,
has made older data highly questionable — however valid  they
were at the time.  In his paper, The Environment and  the  Protect-
ion of Human Health, Dr. Burger based his arguments  largely
on the comparison of leading causes of death  in  19OO  versus
I960:  How valid is it  to compare diagnosis made in  19OO  with  those
made in I960?   For example, what percentage of what was called
pneumonia and tuberculosis  in  19OO, may have  consisted  of cases
of malignant neoplasms  of the  lung.  Or, what are  "19OO - gastritis"
and 190O  - chronic nephritis"  Versus  "1960  -  Carcinomas"  or
"I960 - Cirrhosis"?

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                               1954
BRAMAN  (U.S.A.)
How do you plan to make any progress in developing  (evaluations)
information on those hazards which undergo environmental trans-
formations?  For example, almost no information  (except that
developed by myself) is available on arsenic transformations
or on the chemical forms of mercury in the environment.

SCHNEIDER (Netherlands)
What is the importance of the "availability" of the scientific
data base required?  Is there a need for calibration possibilities
of the parameters that determine the relationships between human
health and environmental conditions?  Should the users of the
data base be able to refer to uniform harmonized or standardized
data services?

STEELE  (USA)
How does one go about determining the level or levels of uncer-
tainty  (that is lack of knowledge) that will be acceptable to
the administration or politician in setting policy or in making
decisions?  Presumably, economic  (resource) trade-offs and time
effects are both at work here

SHERWOOD (U.K.)
The health of most communities is related more to economic factors
than fundamental knowledge of environmental factors.  Internat-
ional progress toward common pollution standards will therefore
probably await common economic standards in all countries.  Does
the panel consider that agreement might be reached on "risk"
criteria? The advantage of the risk criterion is that it takes
account of varying threshold for groups of varying susceptibility.
The disadvantage is the demand for more information from exposure
situations,  both occupational and environmental.

LINDVALL (Sweden)
Referring to Dr. Biersteker's paper, my first question is:
How important are annoyance reactions and irritation effects
in setting protection guides?  Annoyance reactions may in them-

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                               1955
selves cover a broad field from annoyance to environmental fact-
ors, over subjectively perceived hazards of,e.g., toxic agents,
to annoyance to completely imaginary risks.  Also annoyance
could be looked upon not only as an expression of discomfort
but also as a precursor of disease or an indicator of increased
stress load.  What is the view of the panel on the practical
way to integrate qualitative dose-response information and quanti-
tative information?  In other words, how should we evaluate
the joint effect of magnitude and frequency?  How should we
be able to consider and judge the relevance of several sub-
critical effects at the same time?  Interactions between such
effects may take place in the biological, psychological, and
social domaine as well.

              QUESTIONS SUBMITTED IN WRITING
            BY THE PARTICIPANTS AT THE SYMPOSIUM
                AFTER CLOSING THE DISCUSSION

EDITORIAL NOTE:  These questions were submitted following the
discussion at the request of the Chairman to accommodate the
participants who were unable to ask questions from the floor
during the limited discussion period.   The Chairman agreed
to have the answers included in the proceedings.

WASSERMAN  (Israel)
The definition__pf_the term "exposure" refers to amount of a
particular physical agent, it could be more valuable to con-
sider the  "time" parameter also.

BUTLER
I agree with your idea and the problem is simply one of the
use of words.  To those who wrote those definitions for the
Stockholm Conference "exposure" meant the amount of harmful
agent reaching the receptor, i.e., the critical organ.  From
this and a knowledge of the amount of the agent in the critical
organ the dose could be calculated in terms of exposure X days
per gram of tissue.

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                               1956
JACKSON (USA)
Would the panel comment in more detail on their view of the
role of the scientist in decision-making and policy debate?
Iri many of the questions and replies at this symposium there
is an implicit assumption that the scientific community has
both an obligation and an inherent right to establish research
priorities and determine policy.  This view of the^appropriate
role for the scientific community is, in my view, not only poten-
tially counterproductive, but also clearly not based on the
reality of modern society.

BUTLER
In my view the role of the scientist is to inform the public
and the administrators as honestly and completely as possible
about the facts.  This may be a repetitive process consisting
of questions and answers.   The final decision, however, must
be made by an administrator with due regard for politics and
public views.   The views of the  public are particularly import-
ant since they control the votes and thereby the budget, and,
after all, research and protective measures are undertaken to
protect them and their environment.

DI FERRANTE  (C.E.C.)
In presenting cheir results in meetings and journals should
scientists try to expj-ain^ the meaning of their research with
regard to health and the environment and also clearly indicate
the limits within which their conclusions ought to be used?

GOLDSMITH
It is a view that a scientist who, in describing his research,
attempts to draw conclusions about a specific standard or criter-
ion is at risk of distorting and hence impairing the value of
his scientific work.  On the contrary I strongly urge that scient-
ists in meetings and journals should explain in the most  lucid
language possible the meanings of their research with regard
to health and the environment but such communications should

-------
                               1957
be separate from communications of research findings themselves.

GHETTI  (Italia)
Si deve rilevare 1'assenza di contribute riguardanti 1'uso di
"Indicatori Biologici" animali e vegetali per lo studio del
deterioramento ambientale.
There is a noticeable lack of contributions regarding the use
of animal and vegetable biological indicators for studying en-
vironmental insult.
BIERSTEKER
Dr. Ghetti raises a very important point, stressing the limitat-
ion of most of our present research in estimating long-term
ecological repercussions on public health.

There is, fortunately, a growing concern with environmental
pollution in, and outside, the medical profession, in fact al-
most more outside than inside, I would say.  The primary role
assigned to medical people by society/however, is to assess
direct health effects in people in both qualitative and quantitat-
ive terms.  This explains the fragmentary approach, which will
probably persist for many years to come.   There are possibilities
to model the risk of environmental pollution on the basis of
animal or plant studies.  This is in fact the main cause of
concern in many action groups.  Whether man will be able to
learn more from these models than from his direct medical
risks, time will tell.  The political impact of these action
groups has certainly grown in the past decennia and this is
a hopeful sign.

KUMPF (W.H.O. Copenhagen)
There are methods being developed designed to permit the pre-
market^ hazard evaluation of chemicals to  forecast their	
possible health and environmental effects, based essentially on
analogies with chemicals of similar ^structure and known health
and environmental effects.  Should consideration be given
to derived or estimated environmental and health effects in

-------
                           1958
setting product and productive standards,  i.e., would  the
panel consider such data a scie-ntif ic__data basis  for decisions
on measures to protect human health?  However, as a long-term
goal, does the panel consider it possible  to arrive at positive
lists of environmentally dangerous jsubstances and specifically
those which have a potential health hazard in the sense  that
such lists are used in food legislation?

BIERSTEKER
In the field of communicable disease control there is  complete
public information on the known causative  organisms. There
is no reason why we should not have, in a  comparable way,
lists of dangerous additives published regularly.  However,
this will probably not prevent people from using  food  with
such additives; see the effect of labeling cigarettes  as
dangerous.  Therefore a legal basis to exclude proven  dangerous
chemicals to appear in food over safe concentrations can
and should be part of modern health legislation.   International
uniformity of such standards is highly desirable.  New additive
should not be admitted without screening for possible  risks.
This can be required from the producer demanding  standard
procedures.  Estimated health effects can  help in setting
priorities in the laboratory programs that are necessary
to screen additives.  In connection with the large amount
of work involved, international coordination of these  activities
is essential. As the environmental impact  of chemicals also
depends on their persistence in nature and their  accumulation
in food chains, testing for potential dangers in  this  case
is still more complex than in food additives.  The positive
approach puts a heavy responsibility on the screening  agencies,
but as the only alternative is the trial and error method
followed so far,  there is little hope that the positive
approach can be avoided.  Compared with the cost  of developing
new chemicals, the cost of screening can not be prohibitive.
The need for international coordination of this work again
is evident.

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                           1959
BERLIN (C.E.C.)
In the case when deliberate reduction of one pollutant
in a given environmental media is likely to lead to other
pollution problems, how can the scientist provide advice
for dej:isign_ making_ regard ing the relative cost/benefits
in terms of environmental health of this situation?
BIERSTEKER
It seems highly improbable that there would be no ultimate
possibility to control the secondary, usually unforeseen
effect too.  In the case of lead in petrol, for example,
it is likely that a less harmful additive can be found.
If a side effect of pollution control has to be accepted
by a public health administrator, this can only be  on  a
temporary basis as a necessary step towards complete improvement
of the situation.  Most side effects are unforeseen, however,
and need correction instead of a trade off.

FREDERIKSON  (UNESCO)
Would it be more appropriate from the start to carry out
environmental health research^^nd measurements in direct
cooperation with the "polluters"  (e.g. the industrial  managers
and engineers, etc.) to get a better understanding	
of and sound optimal solutions to the problems?  Care  must
be taken that those who will suffer from decisions  are heard
and have jpossibility to take remedial actions.   (I  would
like to warn about the use of the word  "standards."  It is
polyvalent:  for in meteorology  "primary  standards" is used
in the meaning of the French word  "etalior.,"  refers to ISO
and the_International Association for Legal Meteorology),
EARTH
Carrying out environmental health research  in direct cooperation
with industries which may have to be  controlled  on  the basis
of data collected is not deemed  to  be a  wise  procedure.
Even though both parties carry out  the  research  fairly and
honestly,  there  is always the appearance of  a conflict of

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                           1960
interest to all outside parties.  A better procedure would
be for the governmental agency to carry out its research
independently, propose the indicated standard and then allow
all affected industries and the public to comment on the
appropriateness of the standard in a public forum prior
to the standard's becoming effective.

STEENSBERG  (Denmark)
Even if the panel is concentrating on the scientific  data
base I would be interested^ in^ haying some more management
oriented aspects covered:  e.g., How should we try to define
the role of the various professionals engaged in environmental
protection work especially the administrator-experts?  At
present there is a risk that health aspects are not integrated
in all stages _of_jthe decision process - sometimes because of
administrative borders and a hesitancy to integrate medical
and biological expertise in the new administrations.  How could
we on the administrative and laboratory (institute) side secure
a closer cooperation between occupational health and environmental
health expertise?  Again administrative divisions tend to oppose
the necessary contact.
BARTH
In order to achieve the necessary interaction between adminis-
trators and scientists it is imperative that each appreciate
the problems of the other.  Once this knowledge base is
attained the interaction occurs naturally since both scientists
and administrators will realise that neither can do his
job well without assistance from the other.   It is clearly
desirable that there be close coordination between occupational
and environmental health.  Perhaps the best way to assure
this is to put both groups organizationally under the same
government executive and then have that executive force
the necessary coordination to the required degree.

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                            1961
 STEELE (U.S.A.)
 At the opening session the importance of methods of measurement
 giving reproducible and comparable results was mentioned.
 Three of the 155 Technical Committees of the International
 Organization for Standardization (ISO) are relevant to the
 subject of this Symposium. JESO/TC43 "acoustics", ISO/TC146
 "Air Quality", and ISO/TC 147 "Water Quality".  These
 committees jure preparing ISO International^ Standards for
 methods of measurement.  I should like to ask the members of
 the^ panel, if such international standards for methods of
 measurement will also be of use for collecting data required
 for making decisions and setting regulations in the protection
 of human health?

 RECHT
 1'application et la credibilite de toute decision et de
 tout reglement dans ce domaine sont basees sur 1'existence
 de methodes analytiques sures. La disponsibilite de
 methodes standards internationales pour la surveillance
 de 1'environnement et la surveillance biologique est
 essentielle, afin que ces methodes puissent etre xitilisees
 comme references lors de la calibration ou de 1'harmonisation
 des methodes couremment utilisees.

 The enforcement and credibility of any decision or regulation
 in this area are based on reliable  analytical methods.
 The availability of international standards methods for
 environmental and biological  monitoring is essential so
 that  they may be used as references when calibrating or
 harmonizing the methods which are normally in use.

ZAPHIROPOVLOS  (Greece)

Assuming that the significance of epidemiological, clinical and
toxicological studies mentioned is equal  how could the harmonious
aad constructive collaboration among the referred  disciplines be
obtained. At national and International level?   Should one
substance be introduced into a community, what criteria and
protection guides should prevail especially when opinions on the

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                            1962
substance in question do not fully agree?  I understand the
significance of the uniformity in distribution and comparability
when contracting out research work and other studies in determin-
ing exposure-effect relationships on which will be based protect-
ion guides, but still have some doubts on the methodology to be
followed; i.e. should "absolutely comparable" or should a "cer-
tain freedom" prevail on large scale studies?

BARTH
Referring to the first question, present procedures for insuring
harmonious and constructive collaboration, at the National and
International level, in epidemiological, clinical and toxicolog-
ical studies, are clearly less than adequate.  Steps which could
be taken at national levels which might be helpful would include:
a) Placing all National governmental sponsored work in these
areas under the control of a single National agency; b) if a)
is not possible, develop interagency agreements among all involve
National agencies to allow for joint planning of critical experi-
ments in the three disciplines.  Maximally effective internationa
collaboration is not possible unless National collaboration is
achieved first.  International collaboration may then be brought
about through institution of bi-lateral or multi-lateral agree-
ments among collaborating nations to allow for joint planning of
critical experiments.
 Referring to the second question,  when opinions  on a substance t
 be introduced into a community differ, the establishment of
 criteria and protection guides may be extremely  difficult.   Impc
 tant questions which must be answered in this regard are:  a)  hav
 criteria and protection guides for this substance already been
 established in other communities,  states or nations?  b)  If the
 answer  to a)  above is yes,  how strong and convincing is the
 scientific basis for the established criteria and protection gui
 In the event the answer to a) above is no, then one must conside
 the following factors.  On what legal basis is it possible to
 set criteria and protection guides for the substances in questic
 Whatever legal basis exists must be followed scrupulously.  In

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                            1963
 many cases this will call for the holding of public hearings to
allow the public to express its views.   The responsible govern-
mental agency must then weigh all the evidence it has at its
disposal and render a decision for or against the establishment
of criteria and protection guides for the substance in question.
Referring to the third question, in striving for uniformity and
comparability in studies designed to determine exposure-effect
relationships one must, of course, exercise rules of reason.   It
is not necessary, and probably not desirable, that all studies be
"absolutely comparable".   Such an approach could stifle inno-
vative experimental designs.   To the extent possible, however, I
believe that all studies should be "relatively comparable" in
the sense that it is possible to tie them together through some
systematic procedure.   For example, two different methods of
measuring exposures to air pollutants would be acceptable if both
methods were calibrated against a single reference method.
STUPFEL  (France)

What has been learned through cooperation between air pollution
monitoring networks, meteorological stations and epidemiological
surveys?  Are there experimental investigations in that field?

EARTH
Much has been learned through the cooperative use of air pollution
monitoring networks, meteorological stations and epidemiological
surveys. As an example a common method for selecting study and
control groups for epidemiological studies involves proceeding
from a determination of the important air pollution sources
through a study of the local meteorology to a rough estimation of
the different population exposures to be expected based on pre-
diction transport models of one sort or another.   All of these
input data are of considerable value in designing an adequate air
pollution exposure monitoring network for both study and control
groups.   Of course if there has been an adequate air pollution

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                            1964
monitoring network in place initially, some of the above con-
siderations may not be necessary.   In most cases, however, this
has not been the case.

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                 EXPERIMENTELLE FORSCHUNG



                EXPERIMENTAL INVESTIGATION



                 RECHERCHES EXPERIMENTALES



                   INDAGINE SPERIMENTALE



                  EXPERIMENTEEL ONDERZOEK
Vors-itzender - Chairman - President - Presidents - Voorzitter
                    E.M. MRAK (U.S.A.)

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                             1967
       THE STATE OF THE ART REGARDING THE EXPERIMENTAL
         INVESTIGATIONS OF THE EFFECTS OF POLLUTANTS
                       DAVID V, BATES

Faculty of Medicine, The University of British Columbia,
Vancouver, Canada
ABSTRACT   (prepared by  the editorial board)
     In answering  the question about  "the  state of  the art"  in
 (this field) one should begin by differentiating  between  the
 "state of  the  art" and  the "state  of science".    Th\e scientific
 knowledge  consists in the  thousands of experiments  which  have
 been conducted and on which  our present understandings are based;
 the  "state of  the  art"  consists in putting this  information  to-
 gether allowing for factors  which  are  particularly  difficult to
 measure, but yet which  we  feel  instinctively should be  taken into
 account, and in making  the best judgment we can  on  the  questions
 which  many puzzled lezis'atcrs  in  different countries constantly
 ask  us.

      While admitting  that  there are  still gaps in our knowledge
 and  much  useful work  remains to be done, we can be  reasonably
 satisfied  at  the general state of the individual sciences (animal
 and  human  experimentation in controlled environments - epidemic-
 logical  studies).
      The  statue of the art of decision-making and of  synthesis
 has  still a long Way to go and I believe  it is our responsability
 as scientists   to be participants in this  process and give it
 every assistance.

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                                  1968
 INTRODUCTION

     In his introduction to the conference on the health effects of air
pollutants organized by the National Academy of Sciences in Washington in
October 1973, Senator Muskie said "We must base environmental standards
on the best evidence that the scientific community can develop, and we
must not allow those standards to be compromised because they are
difficult to achieve or because of costs.  The scientific community,
therefore, has a. great responsibility.  It is you who must show what
levels of air quality are needed to protect the health of the people.  It
is you who must show how your experimental conclusions can be the basis
for public policy decisions.  You must show who is in danger from dirty
air and what pollutants in what combinations and concentrations pose the
danger.  Public policy makers need this guidance in order to ensure that
adequate protection is provided for all our people without creating costs
and dislocations that are not justified by the needs of public health.
As scientists, you are in the first line of defence for national
environmental policy, and in air pollution you are the most significant
line of defence.  It is people, not plants or fish, whose health and
welfare may be affected, either positively or adversely, by your findings,
your doubts, and your questions."
     During this conference many thousands of words will have been added
to those already published on the evidence on which reasonable decisions
can be made.  My title, given to me by the organizers of the conference,
is "The State of the Art".  Perhaps we should begin by differentiating
between the "state of the art" and "the state of the science".  The
scientific knowledge consists in the thousands of experiments which have
been conducted on which our present understandings are based; the "state
of the art" consists in putting this information together and in making
the best judgment we can on the questions which Senator Muskie and many
other puzzled legislators in different countries constantly ask us.  I
would  like to begin, therefore, by making some comments on the state
of the science and conclude by trying to summarize the  state of the art.

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                                 1969
CLASSES OF EVIDENCE
     There are three main categories which are customarily used in the
study of air pollution, and in determining 'acceptable1  levels of human
exposure:
1.   Animal exposures to pollutants.
2.   Human exposures to pollutants in a controlled environment.
3.   Studies of the epidemiology of human disease.
In the process of listening to much discussion during the past 20 years
OQ this issue, I have come to feel that one of our difficulties is that
we are not sufficiently aware of the advantages and disadvantages of these
different methods of study, and we do not analyze often enough the ways in
which the information derived from them can be used.  I think it is
therefore useful to look at all three categories of evidence and observe
the ways in which any particular experimental approach is strong, and the
ways in which it is weak.
1.   Animal Exposures
     Animal exposure experiments have formed the basis of the definition
     of toxic levels of a wide variety of different substances.  In the
     case of exposures to gases, they indicate approximate toxic levels,
     provide very valuable information on the target organs being
     affected by different pollutants, and give us an opportunity to
     study long-term effects.  The latter experiments, particularly if
     conducted on primates, are very expensive but nevertheless do give
     some indication of long-term exposure effects on body tissues.
     Sometimes people speak as if this kind of evidence is the only class
     to which we should pay attention.  But there are formidable
     difficulties of interpretation.  Thus, in experiments in which
     monkeys, for example, are kept at low concentrations of sulphur
     dioxide, all other aspects of their environment are carefully
     controlled.  But what if sulphur dioxide predisposed to respiratory
     virus infections?  Does the fact that there  is apparently no
     histological evidence of damage in monkeys after one year of
     exposure to a given level of S0_ have anything much  to do with
     whether such a level of S0? is or is not an  important factor in
     determining a higher rate of chronic respiratory disease  incidence?

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                                 1970
     Quite  apart  from different effects on species, the average human
     working environment  is a very much more complicated one than can
     be replicated  in the laboratory, and there will always be doubts
     as to  how safely we  can transpose evidence of this kind into the
     human  environnent.   Such experiments are extremely valuable in
     pinpointing  the  target organ, in indicating such things as
     cellular changes in  the lung following gas exposure, and even
     indicating the effects of low concentrations in general on
     longevity.   They will therefore inevitably remain an extremely
     important part of all such work.
2.   Exposures of Human Subjects
     It is  relatively easy to study the acute effects of air
     pollutants on pulmonary function.  Although the terminology and
     methodology  used  to measure pulmonary function are baffling to
     anyone who does  not work in this field, it is fair to say that we
     have now reached  a point of development when we are confident that
     the use of our presently available technology allows us to say with
     confidence whether or not a given exposure to a pollutant has or
     has not had any effect on the function of the lung.   Such studies
     have, of course,  the great advantage that we are dealing with the
     kind of population with which we are most concerned, namely
     ourselves; but there are difficulties in interpretation of such
     work. Such studies tell us nothing about the pathological changes
     that follow such exposures,  and we have to reconcile the
     physiological change measured, with the histological  changes
     observed in animal experiments.   The general concurrence of this
     data is not too bad, but the cause of a reversible effect on
     function may be far from evident.   Such studies also confront the
     major difficulty  that we cannot  use them to study long-term effects,
     and we also may be hesitant  to expose individuals with lung disease,
     in the controlled laboratory setting,  to levels of pollutants which
     they might well encounter if they  were  to fly into Chicago or
     Los Angeles on certain selected  days  of the year.
3.   Epidemiological Studies
     It would seem natural to suppose that  the careful study of  the
     incidence of  human disease would be the final arbiter  of  the
     decision making in respect of all  air pollutants.  Certainly

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                                 1971
     the tremendous and distinguished body of work which has been
     done in this field over the last 20 years has put us in a very
     much better position to try and decide what levels of pollutants
     we ought to agree are probably harmless.  However, these studies
     always confront difficulties, particularly since there are so many
     interacting factors.  Our sophisticated epidemiologists can
     usually handle these, but we are still left with difficulties.
     For example, it is all too easy to obtain negative results in
     such studies because the tools were inappropriate.  There is a
     hazard that we may be content to approach new problems of
     pollution armed only with the weapons that seemed useful in other
     circumstances.  It seems clear that the photochemical smog of
     Southern California does not produce the chronic bronchitis in
     large airways which was certainly a feature of the heavy
     particulate high SO  coal-burning type of pollution.  So to
     approach the population of Los Angeles with questionnaires of
     respiratory symptoms that have been useful in a different
     environment may be asking the wrong questions.  If a very long-
     term effect of air pollutant exposure had to do with
     accelerating the changes in the lung that are associated with
     age, what hope would there be that an epidemiclogical study based
     on incidence of hospital admissions would reveal this to us?  The
     use of the computer, or as some would say, the misuse of it, is
     revealing possible associations between such things as oxide of
     nitrogen levels and hypertensive heart disease.  It is easy to
     dismiss such associations whenever we cannot explain the possible
     intervening pathophysiology, but we have to bear in mind that the
     association between cigarette smoking and lung cancer was
     established nearly twenty years ago, but we still cannot name the
     carcinogen with confidence.  Nevertheless, studies of human
     environmental effects are crucial and will, obviously remain
     exceedingly important for many years to come.
THE STATE OF THE SCIENCE
     Looking back over 20 years we can be reasonably gratified with the
state of the science as it now exists.  Admitting that there are many gaps
to be filled, particularly in studies of combined pollutant exposure,
nevertheless, we have a considerable body of animal experimental

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                                1972
information which has given us a very good idea of early histological
changes, mechanisms of tolerance, and principal histological targets
for a wide variety of pollutants.  These experiments have even
indicated the possibility of long-term effects which are more subtle
and more difficult to attribute to a specific change in any given
organ.  Our knowledge in this field is vastly superior to what it was
20 years ago.  Similarly, we have developed methods of measuring
pulmonary function that are very sensitive, and we have a better idea
today of the functional consequences, or absence of them, of exposures
to low levels of sulphur dioxide, oxides of nitrogen, and ozone, than
we had ten years ago.  There is much more to be learned in this field and,
as I shall be stressing in the paper I am presenting to this conference,
we have to replicate conditions of light exercise in making these
observations.  There are hundreds of epideraiological studies and we have
come a long way since the first of these was published.  We do have
reasonably accurate knowledge of the comparative effects of cigarettes and
air pollution on the incidence of chronic respiratory disease, and we do
have an idea of levels of S0~ and particulate pollution which are likely
to be associated with increased morbidity.  Recently, the demonstration of
respiratory disease in men working in New York  tunnels exposed to high
levels of oxides of nitrogen has put some kind of ceiling on human
exposure levels for this pollutant for the first time.  Admitting there
are still many gaps in our knowledge and much useful work to be done, we
can, I think, be reasonably satisfied at the general state of the
individual sciences which make up this body of knowledge.
THE STATE OF THE ART
     As I indicated when 1 began, the "art" is to put together the data
from all these studies as a basis for decision making.  When people begin
to do this, we encounter a number of common confusions.  Perhaps you will
identify them from your own experience if I comment on them in detail.
Firstly, there is the scientist who says that consideration of these
broader questions is no part of his mandate.  It seems clear to me,
however, that scientists who have produced the data and know the
constraints upon it better than anyone else, should take responsibility
for trying to distil out the importance of the conclusions.  Secondly,
you will find those who say that the process of trying to make decisions
on such disparate data as monkey exposure experiments and human field

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                                1973
studies is a waste of time.  It would be very much better,  these people
argue, if we reduced all air pollution to a graphical series of
mathematical formulae and then established a firm threshold effect line.
Such a mathematical approach would be very desirable, but breaks down
when the requirement is to bring together evidence from three different
fields in order to devise the best current opinion.  Thirdly, one is
confronted by the difficulty that some scientists do not recognize that
science is, after all, "the best available hypothesis".  There are some
scientists who are so rigorous in their methods and methodology, and of
course distinguished thereby, they do not recognize that most of science
ultimately consists in devising a best available hypothesis and
proceeding to attack, it by new experimentation.  The construction,
therefore, of the best available hypothesis as to what levels of
pollutant wight be permitted in an urban environment if certain things
are to be avoided, is in this sense a "scientific" endeavour.  However,
the decision as to what is "acceptable" or "unacceptable" in an urban
environment is not a scientific matter, but a matter of opinion.  I have
listed in Table I some of the possible consequences of photochemical air
pollution most of which have been observed.  The question here is, "where
do you put your personal line to indicate that an unacceptable condition
has been reached?" Scientific experimentation has established these
phenomena and given approximate concentrations for them, but where you
say you are going to draw the line is a matter of opinion.  I may say
that it is my opinion that it is intolerable and unacceptable for a
city to have an environment in which children cannot exercise without
suffering a measurable impairment of pulmonary function.  Having stated
this opinion, I will have to be required to state whether this will be
true on every one of 365 days each year, or whether I might permit such
a circumstance for a few days.  I will then arbitrarily draw some line
in trying to define what I feel is an acceptable or an unacceptable
state of affairs.  In doing  this, I will be influenced by my feeling that
it is not safe to assume that there is no relationship between repeated
insults to  the lung affecting function, and the later development of
irreversible changes of one  kind or another.  Since we do not have proof
that  this is so, others are  free to argue that transient changes in
pulmonary function can be ignored.
     Wherein, then, lies the "art"?  I think  the answer to  this question
has to be that the art consists in making allowance for factors which are

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                                 1974
particularly difficult to measure, but yet which we feel instinctively
we should take into account.   How shall we protect people who may be
particularly sensitive to certain pollutants?   How can we be sure that
we have taken account of combinations of pollutants, particularly gases
and particles, which have not been exhaustively studied?   How can we
be sure that our safety factor has taken account of the fact that any
system of air monitoring cannot be depended upon to detect the regions
of highest concentrations?   Can we make a statistical guess as to the
highest concentration a person might be exposed to, if we know the
measurements made by a given number of monitoring units over an area?
Have we built in enough protection for the person who lives very close
to an identifiable point source of oxides of nitrogen or of S0_?   Have
we taken sufficient account of possible very long-term effects which
existing human epidemiological studies could not have revealed?   The art,
it seems to me, is making a guess at these interrelated factors in order
to provide the politicians with the guidance they need.   The state of the
art is constantly shifting, but is generally facilitated by virtue of the
much larger numbers of people in all countries who are giving these
questions serious attention.   The understanding of the basis of this
kind of decision making is not yet well advanced.   In many countries the
process of input to the politician is obscure or non-existent.   In very
few places has serious study yet been given as to how such dialogue may
become effective in being one of the inputs into policy making.   The
refinement of this process is one of the greatest challenges which we
confront today and one which we have hardly yet begun to solve.
     I would conclude, therefore, that the state of the science is
moderately healthy and certainly much stronger than 20 years ago, and
still growing;  but the state of the art of decision-making and of
synthesis has still a long way to go and I believe it is our responsibility
as scientists to be participants in this process and give it every
assistance.  Maimonides, writing in Cairo in the year 1193, gave some
advice on the "preservation of youth", as follows:
   "Sunshine and Fresh Air
    The quality of urban air compared to the air in the deserts and
    forests is like thick and turbulent water compared to pure and
    light water.   And this is because in the cities with their tall
    buildings and narrow roads, the pollution that comes from their

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                                1975
     residents, their wastes, cadavers, and offal from their cattle,
     and the stench of their adulterated food, makes their entire air
     malodorous, turbulent, reeking and thick and the winds become
     accordingly so, although no one is aware of it.  And since there
     is no way out, because we grew up in cities and became used to
     them, we can at least choose a city with an open horizon,
     especially on the northeast and toward high mountains with
     limited forestation and outside water.  And if you have no
     choice and you cannot move out of the city, try at least to
     live in a suburb situated to the northeast.  Let the house be
     tall and the court wide enough to permit the northern wind and
     the sun to come through, because the sun thins out the pollution
     of the air and makes it light and pure."
With automobile exhaust, the effect of sunlight is not to purify the air,
but to provide energy for a most complex series of chemical reactions,
and thus we have a new problem which might modify Maimonides1 advice.
However, after 780 years, our problems are still urban problems, they
still involve garbage disposal, and we still prefer to live in suburbs
if possible.

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                                1976


                                 TABLE I
                    CONSEQUENCES _OF  AIR  POLLUTION
                  WHAT   IS   ACCEPTABLE?

 1.  OCCASIONAL LOSS OF VISIBILITY SOME DAYS EACH YEAR.
 2.  5Z CROP LOSSES IN ADJACENT AREAS SOME YEARS.
 3.  INTERFERENCE WITH VISIBILITY ON MORE THAN 10% OF DAYS.
 4.  MAJOR CROP LOSSES OR ECOLOGICAL EFFECTS ON NATURAL VEGETATION.
 5-  IRRITANT EFFECTS ON EYES AND SORE THROATS A FEW DAYS EACH YEAR.
 6.  DEMONSTRABLE IMPAIRMENT OF PULMONARY FUNCTION IN SCHOOLCHILDREN
     AFTER OUTDOOR EXERCISE ON AT LEAST 30 DAYS EACH YEAR.
 7.  DEMONSTRABLE MORBIDITY IN SOME SECTIONS OF POPULATION ON HIGH
     POLLUTION DAYS ONLY.
 8.  DEMONSTRABLE EFFECTS ON PULMONARY FUNCTION IN CHILDREN AFTER
     OUTDOOR EXERCISE ON AT LEAST 90 DAYS EACH YEAR.
 9.  DEMONSTRABLE AGGRAVATION OF SYMPTOMS AND MORBIDITY IN SOME
     SECTIONS OF POPULATION ON CONTINUING BASIS.
10.  DEMONSTRABLE ADVERSE MAJOR HEALTH EFFECTS ON POPULATION,
     i.e. INCREASED HOSPITALIZATION FOR RESPIRATORY DISEASE,
     INCREASED INCIDENCE OF LUNG CANCER, ETC.
LEGEND;  These are listed as approximate possible consequences of
         increasing levels of photochemical or mixed air pollution.
         They are given in this order to focus attention on tht
         question of where the line of "acceptability" should be
         drawn.   Science provides the evidence for the presence
         or absence of these phenomena, but where the line should
         be drawn is a matter of opinion.

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                            1977
           THE EFFECTS OF LOW LEVELS OF S0? AND OZONE
       IN THE SAME ATMOSPHERE ON HUMAN PULMONARY FUNCTION

              DAVID V. BATES+ AND MI UN HAZUCHA"*"1"

+  Faculty of Medicine, The University of British Columbia,
   Vancouver, Canada
++ Department of Physiology, McGill University, Montreal, Canada
ABSTRACT

     There have, so far, been few studies of the effects of com-
mon ai-T pollutants on man studied under conditions of light ex-
ercise, and practically none in which the effects of combinations
of pollutants hai)e been investigated.   We have studied the effect
of Q.Z7 ppm of 0  and 0.37 ppm of SO , alone and together, on
                &                   &
young normal subjects.   In an exposure chamber, each subject
undertook light exercise on a bicycle ergometer at a work rate
sufficient to double his ventilation for a period of 15 minutes,
followed by IS minutes of rest.

     When breathed alone, 0.37 ppm of SO„ has no effect on the
maximal mid-expiratory flow rate (MMFR) nor on any other measure-
ment of lung function when breathed during intermittent exercise
for two hours.   When breathed alone, Q.3? ppm of ozone produces
a just significant decline of ventilatovy function at the end' of
a two-hour- exposure.   However, when these two gases are present
together, in eight normal young subjects who were non-smokers,
at the end of a two-hour period the MMFR had dropped to 67% of
its initial value;  the -FEV    was 78% of its initial value;
and the mid-expiratory flow rate (50% VC) was only 54% of the
initial value.   As in all previous studies, there was consider-
able individual variation.   A two-hour exposure to 0.75 ppm of

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                            1978
SO  alone dropped the MMFR to 90% of its control value.   We
  2j
conclude that ozone and sulphur dioxide* when present together,
are exceedingly irritative;  that "standards" cannot be set for
these pollutants unless the presence or absence of the other is
specified;  and that a combination of circumstances appears to
be leading to the presence of these two pollutants together in
a number of urban environments in what may well be hazardous
concentrations.

-------
                                 1979
1.   Introduction
     During the past year, it has become increasingly evident that,  in the
future, we are likely to be confronted with new combinations of air
pollutants.  In some cities, such as London, the major reduction in
particulate pollution and the increasing automobile density have led to
levels of ozone of 0.15 ppm, as recently reported by Derwent and Stewart
(1).  This concentration presumably occurred in the presence of
significant coexisting sulphur dioxide.  In California, the classical
photochemical air pollution, of which ozone is one end product, may  be
complicated by increasing levels of S0_ as a consequence of the use  of
high sulphur fuel to meet problems of energy shortage.  These two gases,
in combination, are known to be more toxic to plants than either alone at
equivalent concentration (2).  The purpose of this communication is  to
report on the effects of ozone and sulphur dioxide, when present
together, on pulmonary function of normal subjects under carefully
controlled conditions.
2.   Methods
     The exposure chamber we have used has been described elsewhere  (3).
Ozone is generated from electrical discharge in oxygen, and S02 is added
to the chamber from a tank.  The sulphur dioxide is removed from the line
to the Mast ozone meter by a scrubber of glass fibre paper impregnated
with chromium trioxide in concentrated sulphuric acid.  Ozone is removed
from the line to the SO  meter (model 67, Scientific Industries,
Springfield) by a scrubber of crystals of ferrous sulphate heptahydrate,
The SO,, is added to the inlet to the exposure chamber, and sampling  of
both gases is from a point about one foot away from the nose of the
subject.  As the exposure chamber is within a laboratory in an air
conditioned building without external opening windows, the temperature
and humidity vary very little from day to day, and the particulate load is
negligible.  The possibility of unwanted contamination from ambient  air
pollutants in the air of Montreal is remote.
     All experiments have followed the same protocol.  Observations  are
made at time zero, and the subject then exercises on a bicycle ergometer
at a work rate sufficient to double his ventilation for fifteen-minute
periods, alternating with fifteen-minute rest periods for the next two
hours.  Pulmonary function tests are performed at 30 minutes, one hour,
one and a half, and two hours after the start of exposure.  Our subjects

-------
                                  1980
 have mostly been university students and, in the experiments of exposure
 to both ozone and sulphur dioxide, all have been non-smokers.
      Forced expiratory flow volume curves were recorded by a Fleisch No. 3
 pneumotachograph connected to a Hewlett-Packard (Model 270) differential
 pressure transducer.   This was coupled to a carrier preamplifier and the
 resulting signal displayed on a storage oscilloscope equipped with dual
 trace amplifiers.  This flow signal was simultaneously recorded on a four
 channel FM tape recorder for later playback and analysis.  Four flow
 curves were recorded  on each occasion, and the definitive  value was taken
 as the mean of the two highest values of the four.   The complete technical
 methodology and necessary precautions and calibration procedures have been
 fully described elsewhere (4).  We have also been recording the "closing
 volume" and "closing  capacity" during these experiments, using Xe133 as a
 tracer gas.   The results of  these  experiments have  been noted elsewhere
 (5).
      Frank,  McJilton,  and Charlson (5)  have computed  the probable
 delivery rates of sulphur dioxide  and ozone to  different parts of the
 human airway.   Exercise,  by  increasing flow velocity,  and  by virtue of
 increasing tidal volume  and  hence,  the  diameter of  small airways,  is
 likely to lead to a higher concentration of these gases deeper in the
 lung  than would  be the case  under  resting conditions.   It  has been a
 deficiency of  many previous  studies of  exposure to  air  pollutants that
 observations have only been  recorded  on resting subjects,  although the
 proportion of  people who  sit on  the street compared to  those who  walk
 along  it  is  low,  and children  are  customarily very active  out-of-doors  -  a
 point  to which we will return.
 3.   Results
     The physical characteristics of  the  subjects who have been studied
with exposure  to  (>3 and S<>2  together  are  shown  in Table I.   None  had  any
past history of  significant chest illness,  none had a family  history  of
asthma or atopic  illness, and all were  non-smokers.

-------
                                  1981
                                 TABLE I
                PHYSICAL CHARACTERISTICS OF NORMAL SUBJECTS
                            NON-SMOKING MALES
          Initials         Age         Ht.(cm.)          Wt.(kg.)
             WB             23           182               82
             MG             21           182               80
             FS             22           188               79
             WW             20           185               72
             JS             25           178               67
             JR             21           178               82
             LD             23           178               79
             TE             19           188               84
      In Table  II are shown the results of the exposures to 0.37 ppm of SO
and 0.37 ppm of 0^y present together in the chamber, using measurements of
maximal mid-expiratory flow rate.  This test of function is the mean
velocity of air flow during forced expiration of the middle half of a
maximal expiration  (i.e. between 25% and 75% of the recorded maximal
expiration).  As we have previously noted, this test together with the
velocity at the 50% expiration point, are the two most sensitive
indicators of change amongst those that we have studied (5).
It will be noticed  that a significant fall in maximal mid-expiratory flow
rate has occurred within 30 minutes of exposure.  The table also shows the
observations expressed as percentages of the initial value.  We have
recorded elsewhere  (7) the variance of these ventilatory tests in normal
subjects after exercise in our exposure chamber without any ozone or SO
being present.
     As in all previous experiments of this kind, there is considerable
variation in the magnitude of the effect observed.   Thus,  at the end of
two hours of exposure, subject JS has an MMFR which is still 91% of the
control value, whereas in TE it has fallen to only 35% of  the initial

-------
                            1982
                           TABLE II
           EXPOSURE TO 0.37 ppm 03 AND 0.37 ppm SO
                 INTERMITTENT LIGHT EXERCISE
               MAXIMAL MID-EXPIRATORY FLOW RATE
                          LITRES/SEC.
Control
Subject Time Zero
WB
MG
FS
WW
JS
JR
LD
TE
MEAN
SD ±
SE
"C"
P
(from initial
value)
WB
MG
FS
WU
JS
JR
LD
TE
7.39
4.37
6.08
6.03
10.92
8.62
4.96
9.91
7.28
2.36
0.83
-

100
11
11
11
II
ri
ti
11
                                     Hours of Exposure
0.5
7.32
3.96
5.06
5.16
10.42
7.27
4.22
7.15
6.32
2.15
0.76
3.30
0.02
As
99.17
90.81
83.15
85.47
95.43
84.29
85.04
72.15
1.0
6.30
4.24
4.17
4.47
9.83
7.38
4.21
5.13
5.72
2.03
0.72
3.17
0.02
Percentage of
85.33
97.13
68.63
74.17
90.04
85.65
84.79
51.82
1.5
3.40
4.18
4.30
3.60
9.30
6.40
3.99
4.64
4.97
1.98
0.70
4.00
0.01
Control
45.99
95.79
70.63
59.61
85.23
74.23
80.36
46.83
2.0
3.39
3.90
4.03
3.78
9.97
5.75
3.92
3.47
4.78
2.22
0.79
3.62
0.01
Value
45.91
89.36
66.26
62.59
91.37
66.71
78.95
35.04
MEAN
86.94
79.70
69.83
67.02

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                                  1983
value. The mean fall in MMFR for the group of eight subjects at the end
of two hours was to 67% of its initial value.  The fall in forced vital
capacity over the same period was to 82% of its initial value; of FEV   ,
to 78% of the initial value; of peak expiratory flow rate, to 79%; of
MEFR at 50% of vital capacity was to 54% of its initial value; and the
static (non-forced) vital capacity was 92% of its initial value at the
two-hour point for the group as a whole. Thus, the fall in the flow
measurements was proportionately much greater than the fall in static
volume.
     In Table III are shown a comparison between the two-hour exposure
values with ozone and SO-, separately and together, by comparing the
effect on a single test of pulmonary function, the MMFR.
                                TABLE III
                        CHANGES FROM INITIAL VALUE
                   AFTER 1 HOUR AND 2 HOURS OF EXPOSURE
                    INTERMITTENT EXERCISE - NON-SMOKERS
                                       PERCENTAGE OF INITIAL VALUE

GAS Composition
0.37 ppm S02
0.37 ppm 0«
0.75 ppm S02
0.75 ppm 0
0.37 ppm 0-
and
Number
of
Subjects
4
3
4
9

8
ricixjuuicia. ri-Lu— C.AJJ.
Flow Rate I/ sec .
1 hour
98
96
91
82

80
2 hours
100
88
89
65

67
MEFR (50% of VC)
1 hour
102
98
82
78

71
2 hours
97
86
76
62

54
   0.37 ppm  SO

 It is to  be noted that the subjects exposed to the two gases together,
 whose characteristics are shown in Table I, are different subjects from
 those from whom the other exposure data have been derived who took part
 in experiments we have reported previously (6, 7).  It is apparent from
 this table that the effect of 0.37 ppm of 0, and 0.37 ppm of SO^ together

-------
                                 1984
is about comparable or slightly more severe than the effect of 0.75 ppm
of 0_ alone.  It is much greater than for each gas separately at a
concentration of 0.37 ppm.  This result is shown diagrammatically in
Figure 1.

              Max. Midexpiratory  Flow Rate
              (Nonsmokers ; Intermittent Exercise'
         O
         O
         VJB
            70
            60
            50
                 • 0-37 ppm SO 2 (N-- 4)
               _ 90-37 ppm O3  (N=3)
                 »0*37 ppm SOg+O-j (N=l

                             EXPOSURE
                                                     RECOVER*
                       0-5       10       15
                                TIME  (hrs)
                                                   2-0
2-5
FIGURE 1
            This figure shows the  effect  on maximal mid-expiratory
            flow rate of S(>2  and ozone, separately and  together.
            Physical characteristics  of the eight subjects exposed
            to 0.37 ppm of  SO2 and 0.37 ppm of ozone, existing
            together, are given in Table  I.  The fall in maximal
            mid-expiratory  flow rate  after two hours exposure to
            0.37 ppm of ozone is just statistically significant
            (see Reference  7).  The much  greater effect of the
            two gases, when present together, is illustrated in
            this figure, the  data  from which is derived from
            Table II.

     Since the exact mechanisms of these  effects on  the human  respiratory
tree are not fully understood, it  is  of  interest and  importance  to  study
the rate of recovery after  exposure to these  gases as  a guide  to the  kind
of pathological process which may  be  occurring.  The  time  course of
function test change during two hours of  exposure  (with the  same exercise
protocol) and two hours of  post-exposure  recovery in non-smokers exposed
to 0,75 ppm of 0_ is shown  in Figure  2.  The observation that none of
these tests of function have  returned to  their  initial  control value  two
hours after the exposure has  ended, requires  emphasis.  Some additional

-------
                                   1985
                                 Non-Smokers
                  EXPOSURE
                                                     RECOVERY
100
                     1-0
                            1-5
   2-0
TIME (hrs)
2-5
3-0
3-5
4-0
  FIGURE  2
            Figure 2 shows  the time course of  changes in five
            pulmonary function tests in a  group  of  non-smoking
            normal subjects during a two-hour  exposure to 0.75  ppm
            of  ozone.   Intermittent light  exercise  continued
            throughout the  exposure period involving light
            bicycling for 15 minutes alternating with 15-minute
            rest periods.   None of these tests of pulmonary function
            has returned completely to  its control  value within two
            hours of removal from the ozone, and a  further period  of
            between two and four hours  was required for complete
            restitution.
experiments which we have  carried out indicate that the control value is
attained between four and  six hours after a two-hour exposure period and
all pulmonary function tests are back at their control values at that
time (4).
     We have noted constitutional symptoms, particularly chest pain,
after exposure to 0.75 ppm of ozone and, on one occasion, one of the
subjects had what might have been a small hemoptysis 12 hours after
exposure.   Clinical and radiological examination of him was negative, but
a similar event has been recorded by others who have exposed normal
subjects to levels of up to one part per million.  In view of the
cellular changes which occur in the lungs of animals exposed to levels
of above 0.5 ppm of ozone,  we would not recommend that the exposure of
normal subjects to  these concentrations under conditions of light
exercise be lightly undertaken - indeed in our view the  scientific purpose
of such measurements would have to be very clearly specified for such

-------
                                  1986
 exposures to be justifiable on an ethical basis.   In our initial series of
 experiments at 0.75 ppm of ozone we used mainly physicians as subjects
 (6), and we have done no further experiments at this concentration.
 4.   Discussion
      It is hardly surprising that these two highly reactive gases,  which
 can coexist for relatively long periods of time at these low
 concentrations, should produce a greater effect on the human lung than
 either alone.  The enhancement of toxicity of SO   and 0 ,  when
 simultaneously present,  has been known to exist in plants  for some  years.
 As a first approximation,  one can imagine the two gases, humidified  in
 the airway and presented with a large surface area,  rapidly combining  to
 "paint" the smaller airways with sulphuric acid.   This probably  explains
 the relatively rapid effect which can be noted from the data in  Table  II.
 The MMFR had fallen to 87% of its control value within 30  minutes of
 exposure.
      We find it hard to  believe that  all the effects  are to be explained
 by edema of airways.   The  slow recovery  after ozone  exposure (see
 Figure  2)  might be due to  slow absorption of fluid,  but in the absence
 of more specific understanding of the dynamics  of  fluid absorption under
 this circumstance,  it  is not  easy to  know whether  resorption would
 require a  four-hour  period.   A purely irritant  reflex effect would have
 been expected to resolve even faster,  however;  the most that can  be said
 is that  the  recovery time  in  excess of two hours should warn us that much
 more  research is required  (probably in animal experiments)  on  the
 mechanism of  the effect  of  these  gases.
      Are these  experiments  reliable as indicators of what may  be
 happening in  the world outside  the laboratory?  The recent  study by
 Lebowitz and  his colleagues (8) has demonstrated a decline  in  FEV    in
 children and  adolescents after outdoor exercise on days of  relatively
 high  pollution  in Tucson, Arizona, as compared to days of relatively low
 pollution.  From the data they present, it looks as if  the maximal
 pollution to which these children might have been exposed would have been
 up to 0.28 ppm of ozone,  0.10 ppm of nitrogen dioxide, 70 microgram/cubic
meter of particles, 4.0 microgram/cubic meter of sulphate,  and perhaps
 C.3 ppm of SO2-   It seems likely that it is the interaction of these
pollutants that has caused  the measurable effect,  in line with the
laboratory experiments we have reported here.  Similarly, it seems likely

-------
                                  1987
to us that  the high morbidity amongst children exercising out-of-doors ir
Japan, to which we have drawn attention  (5), is to be explained by the
simultaneous presence of 03 and S0r  These observations suggest that in
many environments, measurable effects on pulmonary function might be
observable  if they were specifically looked for.
     Are these experiments useful as indicators of "acceptable" and
"unacceptable" environments?  At the very least, they warn us of the
dangers of  setting acceptable levels of  individual pollutants when
combinations are likely to coexist.  The question of whether we should
accept as "acceptable" an environment in which one cannot exercise
without a measurable decrement of pulmonary function occurring, is a
matter of opinion.  However, we cannot safely assume that long-term
consequences can be dissociated from short-term repetitive insults of
the kind that are probably now commonly occurring in a number of
environments in different parts of the world.
     It has been the purpose of this communication to bring this to your
attention.

    (Supported by grants from the Medical Research Council of Canada
                     and the Canadian Thoracic  Society)
                               REFERENCES
     DERWENT, R. G., and STEWART, H. N. M., Elevated Ozone Levels in
     the Air of Central London. NATURE, 241, 342, (1973).
     MACDOWALL, F.  D. H., and COLE, R.  F.  W.,  Threshold and
     Synergistic Damage to Tobacco by Ozone and Sulphur Dioxide,  Atmosph
     Env.  .5, 553-559, (1971).                        	
     BATES,  D.  V.,  BELL,  G.,  BURNHAM, C.,  HAZUCHA,  M.,  MANTHA,  J.,
     PENGELLY,  L. D., AND SILVERMAN, F., Problems in Studies  of Human
     Exposure to Air Pollutants.   Can.  Med.  Assoc.  J. 103,  833-837
     (1970).                                         	
     HAZUCHA, M., Effects of Ozone and  Sulphur  Dioxide  on Pulmonary
     Function in Man. Ph.D. Thesis (Physiology),  McGill University
     (1974).
     Proceedings of the Conference on Health Effects of Air Pollutants.
     National Academy of  Sciences,  (November 1973),  Serial No.  93-15,
     (U.S. Government Printing Office, Washington, D.C., Stock  No.
     5270 -  02105).

-------
                               1988
      BATES, D. V., BELL, G. M., BURNHAM, C. D., HAZUCHA, M., MANTHA, J.,
      PENGELLY, L. D. , and SILVERMAN,  F., Short Term Effects of Ozone on
      the Lung. J. Appl. Physiol. 32,  176-181,  (1972).

      HAZUCHA, M., SILVERMAN, F., PARENT, C., FIELD, S., and BATES, D. V.,
      Pulmonary Function in Man after  Short Term Exposure to Ozone. Arch.
      Environ. Health^, 183-188, (1973).                     ~

      LEBOWITZ, M. D.,  BENDHEIM, P., CRISTEA, G., MARKOVITZ, D.,
      MISIASZEK, J., STANIEC, M., and  VAN WYCK, D., The Effect of Air
      Pollution and Weather on Lung Function in Exercising Children and
      Adolescents. Amer. Rev. Resp. Dis. 109. 262-273, (1974).
                           DISCUSSION
BUTLER (Canada)
      Is the enhanced response  to the combination of SO  and  O
due  to an addition  of two physiological responses or to the
production of a more irritant  chemical by reaction between the
two  gases?


BATES  (Canada)

      I believe that the most likely explanation is the formation
of H2SO4  within the airways of  the  lung.  The  chemists whom  I
have  consulted have pointed out that the drawing of the two
gases  into the lung would have  three consequences favouring  H0SO
formation,  namely humidification, heating and  exposure to a      4
considerable surface area.   So I think the most likely hypothesis
is that the airways are in effect "painted" with sulfuric acid.

     I  have no direct evidence  that this is the case,  however.

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                             1989
      EFFECTS OF SULFUR DIOXIDE ON HEALTHY AND PERIPHERAL
                   AIRWAY IMPAIRED SUBJECTS

           FRANCIS W, WEIR  AND PHILIP A, BROMBERG**

+  Research Laboratories, General Motors Corporation, Warren,
   Michigan, USA
++ Ohio State University, Columbus, Ohio, USA
ABSTRACT

     This report describes an investigation of healthy subjects
and subjects with early signs of chronic obstructive lung disease
exposed to atmospheres containing sulfur dioxide (SO.).    Experi-
                             3
ments were conducted in a 28M  dynamic flow environmental chamber
maintained at 22 + 1°C and SO f 5% relative humidity.   Exposures
to 0, 0.3> 1.0 or 3.0 ppm SO  were ordered in randomized sequences
                            z
for 120-hour periods (Series 1-healthy men) or 96-hour periods
(Series 2-unhealthy men).

     In Series lt no significant dose-related changes were ob-
served in subjective complaints, clinical evaluations, or most
pulmonary function measurements.   Significant, but minimal,
reversible decreases were noted in compliance at high breathing
frequencies at the 3ppm exposure level, but these changes dis-
appeared within 48 hours after cessation of exposures.

     Clinical evaluations of Series 2 revealed a variety of sub-
jective and objective findings.   However, complaints could not
be related to the concentration of S0g.   Inter- and intra-sub-
ject variability in the results from each pulmonary function
test far exceeded the variance due to exposure to 50.

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                             1990
     The evidence suggests that persons with minimal preexisting
peripheral airway disease are probably not more susceptible to
the effects of SO„ than normal persons.   However, the existanae
of prior disease confounded any attempt to define threshold con-
centrations of SO-.

     These studies have demonstrated that most conventional meth-
ods for assessing pulmonary function are insufficiently sensitive
to detect early changes resulting from exposure.to SO„ even in
                                                     &
carefully selected healthy subjects and, further, that assessment
of air pollution effects in the large portion of the population
with pulmonary dysfunction may require development of entirely
new methodology.

     (Research supported by the American Petroleum Institute).

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                                   1991

 1.    Introduction
      Recent  legislative  action,  designed  to  control  the  levels of  sulfur
 dioxide  (S02)  in  the  environment,  has  emphasized  the need  for more complete
 knowledge  of the  biological  effects  of  this  compound.  Reports based on
 faulty or  inappropriate  observations have been widely employed as  bases
 for  legal  standards.  Adequate measurement of cause  and  effect has been
 a major  problem.  Retrospective  epidemiological surveys  of pollution have
 failed to  demonstrate that environmental  levels of SOa are causally related
 to deleterious health effects, since so many alternate,  often confounding
 variables  must be considered.  Further, disease or death seldom, if ever,
 result from  pollution alone.
     Animal  experiments  have likewise yielded little appropriate informa-
 tion.  The acute, reflex bronchoconstriction, studied  in a variety of
 animals, is  a convenient bioassay  but is  in  no way useful in assessing
 the  effects  of chronic exposure  (Nadel, et al.[l], Frank and Speizer [2],
 Amdur and Mead [3]).  Regarding  chronic effects, Alarie  and co-workers
 [4,5] exposed guinea  pigs and monkeys continuously for more than a year
 to levels  of sulfur dioxide of about 0.1  and 1.0 ppm.  These investigators
 found no changes  in a variety of pulmonary and cardiac function measure-
 ments, blood chemistry studies,  gas  uptake,  or histology of the respira-
 tory tract which  they could relate to the sulfur dioxide exposures.  Evi-
 dently, the reflex changes observed  in the acute experiments are not re-
 flected as chronic effects.
     Laboratory investigations employing  healthy human subjects exposed
 to moderate concentrations of SC>2  (<5 ppm) indicate  that the irritation
 effects are not debilitating and are rapidly reversible.  Furthermore,
when sequential exposures were performed, the reexposure to 862 was charac-
 terized by a much lower  response than after initial  administration  (Frank
 [6],  Frank, et al.[7]).
     The present  investigation was designed to quantify  the concentration-
 time exposure levels of  S02 in the atmosphere which  does not produce sub-
jective or objective deleterious effects in either healthy subjects (exper-
 imental Series 1)  or subjects exhibiting early signs of  chronic obstructive
 lung disease (experimental Series 2).
     Experiments of Series 1 provide  data useful for assessing the effects
of S02 on healthy subjects.   Subjects of Series 2 are representative of
a large fraction of the population thought to be more susceptible

-------
                                  1992
than normal to the effects of S(>2, thereby making the results of the
study more applicable to the general population.
2.   Methods
     2.1  tojMsure_Facility_
     The exposure chamber, a 28M  room, utilized in this study was a
design modified by Hinners, et al. [8],  The chamber was a truncated
pyramid (3.5M/side) with straight side walls (1.8M high) supporting the
pyramidal funnel structure.  Sanitary facilities were installed in the
chamber to allow prolonged occupancy.
     Influent gas mixtures entered through a transition duct at the apex
of the pyramid; effluent gas exhausted near the floor of each side wall.
     Ventilation through the chamber was maintained at 0.5 changes per
minute.  All influent air was filtered to remove particulate matter.
Temperature and humidity were controlled at 22 ± 1°C and 50 ± 5% relative
humidity by an air refrigeration system and accessory electric heat sup-
plemented with filtered live steam when necessary.
     Access to the chamber was through a double door air lock system
designed to allow researchers to enter the chamber without disturbing
the gas composition.
     The S(>2, anhydrous research grade, was metered into the influent
duct down stream from the air conditioner.  The flow rate of the S02
into the influent air stream was controlled by a capillary flow meter
with a two-stage regulator system and needle valve.
     Samples of chamber air were periodically collected in midget impingers
and subsequently analyzed with the spectrophotometric technique of West
and Gaeke, as modified by Scaringelli, Saltzman, and Frey [9].
     Series 1 Subjects:  Twelve healthy adult males between the ages
of 21 and 28 were selected for the Series 1 study.  In this series, all
subjects were nonsmokers.  Candidates were screened on the basis of motiva-
tion and extensive physical and psychological examinations.
     Series 2 Subjects:  Criteria for selection of subjects for the second
series of experiments were based on the need for otherwise healthy per-
sons exhibiting preclinlcal chronic peripheral airway obstruction.  Males
between age 25 and 49 years with normal chest X-rays, electrocardiograms,
no history of asthma or significant allergy, no clinically apparent
diseases, and satisfactory results on psychological examination, but

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                         1993
                         TABLE I
PLANNED EXPOSURE SEQUENCE FOR SERIES 1 AND 2 EXPERIMENTS
subject
Group
I
II
III
IV

1
1 . 0 ppm
0 . 3 ppm
3.0 ppm
0.0 ppm

2
0 . 0 ppm
1.0 ppm
0.3 ppm
3.0 ppm
Exposure

3.0
0.0
1.0
0.3
Week
3
ppm
ppm
ppm
ppm


0
3
0
1


.3
.0
.0
.0

4
ppm
ppm
ppm
ppm

-------
                                  1994
with evidence of small airway impairment were accepted.  Airway impair-
ment was defined on functional bases.
     Subjects were accepted with abnormal Closing Volumes  (CV), measured
by methods described by McCarthy, et al.[10], depressed Maximum Mid-
expiratory Flow Rates (MMF), and one-second Vital Capacity (VC) between
70Z and 50% of total VC.
     Only eight candidates from over 400 applicants qualified for the
second series of experiments.  Of these, one subject elected not to return
after one week of exposure.  The remaining seven completed the test cycle.
     2.2  Exposure Protocol
     The planned sequence of exposures to S02 for both series of experi-
ments is presented in Table I.
     During each 16-week series, all experiments were conducted using
the double-blind technique.  None of the operating personnel or the sub-
jects were permitted access to the exposure sequence.  The physicians
responsible for daily physical examinations and overall subject health
were also unaware of the exposure schedule until the end of the exposure
series.
     The design for Series 1 experiments was based on the  assumption
that four groups of three subjects would complete the series.  Each group
was exposed for 5 days  (120 hours),  followed by a rest period of nine
days.  For Series 2 experiments, each group consisted of two subjects,
exposed continuously for four days  (96 hours), followed by a rest period
of ten days.  This pattern was repeated for each of four exposure periods.
A final evaluation of health status  was completed within one week following
the final exposure.
     2.3 Pulmonary Function Testis
     Daily pulmonary function measurements included Airway Resistance,
Functional Residual Capacity, Static and Dynamic Lung Compliance, timed
VC, Maximal Expiratory  Flow-Volume  Curves, CV, and Nitrogen Washout.
     Airway resistance  and  functional residual capacity were measured
by methods described by DuBois,  et  al.[ll].   Static and Dynamic Lung
Compliance measurement  techniques were  essentially  those of Woolcock,
et al.[12).   Vital Capacity  and  Flow-Volume measurements were  determined
with a dry-rolling seal spirometer  coupled  to  an X-Y plotter.   Closing
volumes were  determined by  the  spirometer  and  plotter  coupled  to  a  linear

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                                  1995
nitrogen gas analyzer.  The method was essentially that described by
McCarthy, et al.[10].
     The nitrogen washout procedure was similar to the multiple-breath,
open circuit technique described by Comroe, et al.[13].
     2.4  Statistical Analyses
     The experimental design of these exposures utilized a Latin Square
for the selection change-over sequence but allowed for a sample size
that is not rigid relative to a Latin Square.  The order of administration
of S02 was dictated by the use of a 4 x 4 Latin Square to determine the
change-over sequence in each of the four weeks of treatment exposure.
Treatment sequence was then allocated to four groups of subjects with
three (Series 1) or two (Series 2) subjects in each of the four groups.
It was recognized that this particular design is sensitive to a treatment
carry-over effect, but it was further calculated that a 9 or 10-day resting
period was appropriate based on past experience with healthy subjects.
     Statistical analysis was performed by grouping the appropriate results
to conform to a three factor randomized block design with each of the
subjects acting as a separate block.  Three factors considered were:
subject, concentration, and week of study, with "N" levels of subject,
four levels of concentration, and four levels of week (time).  Each of
the three main factors was tested for its level of significance by comput-
ing the appropriate F-value using the proper two factor interaction mean
square as the error term.  The analysis was repeated using a logio trans-
formation of the data where it was thought that subject response was
not expected to be normally distributed based upon biological considera-
tions.
     Data reduction and analysis of all results from both series of expo-
sures, were aided by use of an IBM 370/165 computer.  All variables in
these experiments were analyzed by the use of the Biomedical Computer
Program BMD08V [14].  This program was designed to compute and analyze
variance of two or more factors.  Mean separation of significant data
was accomplished with the Duncan New Multiple Range Test [15].
3.   Results
     3.1  General Observations (Series 1 and 2)
     Daily examination of all subjects by a physician during both series
revealed a variety of subjective and objective findings.  Most of the
presented complaints were randomly distributed throughout each series

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                                  1996
and could not be related to the concentration of SC>2 in the chamber.
Furthermore, the number and degree of complaints or symptoms appeared
to be randomly distributed throughout any exposure week, with the excep-
tion of the last day of exposure.  Fewer complaints of ill health were
reported during the last 24 hours of exposure than during any other period.
The range of complaints included:  headache, nasal congestion, sore throat,
coughing, mild to moderate gastrointestinal disturbance, nose bleed,
and skin rash.
     On several occasions, a subject began an exposure week with a mild
upper respiratory infection, including cough and sore throat.  There
is some suggestion, from the physicians' notes in the daily log, that
recovery from the infection was slower if this situation occurred during
the 3 ppm SC>2 week than if it occurred in any of the other test cycles.
However, there were insufficient cases to verify this statistically.
     No other dose-related patterns of complaints of symptoms could be
extracted from the daily log or physicians' notes.
     In most cases, subjects could not determine by sense the relative
dose of S02 they were receiving during any week.  Several subjects stated
that they sensed something in the chamber atmosphere by taste during
the 1 ppm exposures.  These same subjects sometimes indicated that they
were receiving the "highest" exposure during the 3 ppm week.  All members
of the investigation teau were instructed to minimize thought and dis-
cussion regarding the probable concentration in order to maintain objecti-
vity.  Therefore, no consistent records were compiled regarding subject
concentration-awareness.
     3.2  Pulmonary Function Tests for Series 1
     No dose-related changes were observed in most pulmonary function
measurements of Series 1 subjects that could be attributed to S(>2 exposure.
Significant, but minimal, reversible decreases were noted in the frequency
dependence of compliance tests as a result of the 1 and 3 ppm exposures.
The greatest decreases in compliances at high respiratory frequencies
occurred after 24 hours exposure.  Both 1.0 and 3.0 ppm exposure produced
significant depression compared to controls at 24 and 48 hours after
start of exposure.  These effects became less pronounced as the exposures
progressed through 72 hours.  After 120 hours of exposure, however, the
3.0 ppm values were again significantly depressed.  The daily pattern

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                                1997
                               TABLE  II
                   SUMMARY OF PULMONARY COMPLIANCE
             MEASURED AT 120 BREATHS/MINUTE FOR SERIES 1
Cone.
ppm
0
0.3
1.0
3.0
Hours of Exposure
24
(0.039)(b)
0.217
(0.052)
0.183(c)
(0.047)
0.064
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                                   1998
 of effects  of  exposure on  the highest breathing  frequency  tested  (120
 breaths/min.)  are  summarized in Table II.
     3.3  Pulmonary Function Tests  for  Series  2
     None of the results from the pulmonary  function  tests  illustrated
 clear evidence of  differential SOa  effect.   In both the normal and logic
 transformed data,  inter- and intra-subject variability in the results
 from each test far exceeded the variance resulting from the exposure
 to S02.
     There were scattered points within the  test data which were  signifi-
 cantly (p <0.05) different from control; however, none of these repre-
 sented a dose  response of the subjects to S02-
     The results of the frequency dependence of compliance  test clearly
 demonstrated that  Series 2 subjects had peripheral airway disease.  In
 these subjects, even at the lower frequencies, the dynamic  compliance
 was depressed  compared to the static compliance.  Healthy subjects (Series
 1) performing  the  same test showed dynamic compliances at low breathing
 frequencies (20-30 breaths/min) not significantly different from  results
 of the static  compliance.
 4.   Discussion
     These studies have shown that conventional methods of  detecting
 lung obstruction, such as measurement of total airway resistance, are
 insensitive and fail to detect peripheral airway obstruction.  The data
 presented indicate that there is a significant but reversible effect
 on the pulmonary function of healthy adult males by 3 ppm S02 exposures
 for 120 hours.   The daily pattern of effect shown in Table  II suggests
 that there are probably two mechanisms operating to produce the increased
small airway resistance observed.   Early after the start of exposure,
both 3 ppm and 1 ppm S02 produced changes in compliance that are signifi-
cantly different from control.   These changes became less apparent as
exposure proceeded.  However,  after 120 hours of exposure,  the 3 ppm
SC>2 data again became significantly different from control.
     We reason that the early changes are the result of reflexive constric-
tion of the airways, similar to those reported by Amdur and Mead  [3].
These changes disappeared with continued S02 exposure.  The later effects
represent a breakdown in the adaptation processes at the 3 ppm exposure
level.

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                                  1999
     We think the early changes observed as a result of 1 ppm S02 repre-
sent a threshold of effects for S0£ in healthy subjects under the condi-
tions tested.  Continued exposure to 3 ppm SC>2 beyond 120 hours could
possibly produce more persistant changes.
     In regard to the Series 2 experiments, there is every reason to
expect that pulmonary responses such as the frequency dependence of compli-
ance response are only linear to the effects of S02 over a limited range.
If the effects produced by SC>2 exposure are due to reflex constriction
of the airways, then it is probable that persons with preexisting, non-
reversible disease could not show further significant constriction in
response to S02,
     The evidence from the Series 2 experiments suggests that persons
with preexisting peripheral airway disease are not significantly more
susceptible to the effects of S02 than normal persons.  However, the
existance of prior disease confounds any attempt to quantify threshold
changes from SO2 that are minimal compared to the preexisting condition.
     It should be noted that the changes reported in these experiments
are minimal compared with the effects of smoking on frequency dependence
of compliance.  We reported as significant changes that were within 10%
of control values.  Smoking, on the other hand, depresses the compliance
at high frequencies to as low as 40% of  control values  (Woolcock,  et al.
[12].
     These studies have demonstrated that most conventional methods for
assessing pulmonary function are insufficiently sensitive to detect early
changes resulting  from exposure to S02 even  in carefully selected  healthy
subjects, and  further, that assessment of  air pollution  effects  in the
large portion  of the population with pulmonary dysfunction may  require
development  of entirely new methodology.

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                                 2000
                                 REFERENCES

      NADEL, J. A., SALEM, H. ,  TAMLIN, B., Y. YOKIWA, "Mechanism of
      bronchoconstriction," Arch. Environ. Health, 10, 175 (1965).
 2    FRANK, N. R. , SPEIZER, F. E., "SOa effects on the respiratory system
      in dogs," Arch. Environ. Health, 11, 624 (1965).

 3    AMDUR, M.E., MEAD,  J., "Mechanics of respiration in unanesthetized
      guinea pigs," Am.  J.  Physiol. , 192, 364 (1958).

 4    ALARIE, Y., ULRICH, C.E., BUSEY, W. M. , SWANN, H.E., MACFARLAND, H.
      N., "Long-term continuous exposure of guinea pigs to sulfur dioxide,"
      Arch. Environ. Health, 21, 769 (1970).

 5    ALARIE, Y., URICH,  C.E., BUSEY, W.M. , KRUMM, A. A. , MACFARLAND, H.N.,
      "Long-term continuous exposure to sulfur dioxide in cynomalogus
      monkeys," Arch. Environ. Health, 24, 115 (1972).

 6    FRANK, N.R., "Studies on the effects of acute exposure to sulfur
      dioxide in human subject," Proc. Ray. Soc. Med., 57, 1029 (1964).

 7    FRANK, N.R., AMDUR, M.O., WHITTENBERGER, J.L., "A comparison of the
      acute effects of S02 administered alone or in combination with NaCl
      particles on the respiratory mechanism of healthy adults," Int. J.
      Air and Water Pollution, 8, 125 (1964).

 8    HINNERS, R.G. , BURKART, J.K. , PDNTE, C.L., "Animal inhalation expo-
      sure chambers," Arch. Environ. Health, 16, 194 (1968).

 9    SCARINGELLI, F.P.,  SALTZMAN, B.E. , FREY, S.A. , "Spectrophotometric
      determination of atmospheric sulfur dioxide," An«ily_tical Chemistry,
      3~9, 1709 (1967).

10    MCCARTHY, D.S., SPENCER, R. , GREENE, R. , MILIC-EMILI, J., "Measure-
      ment of 'Closing Volume1 as a simple and sensitive test for early
      detection of small airway disease," Am. J^ Med., 52, 747 (1972).

11    DUBOIS, A.B., BOTELHO, S., BEDELL, G. , COMROE, J., "A rapid method
      for measuring thoracic gas volume.  A comparison with a nitrogen
      washout method for measuring functional residual capacity in normal
      subjects," J. Clin. Invest., 35, 327 (1956).

12    WOOLCOCK, A.J., VINCENT, N.J., MACKLEM, P.T., "Frequency dependence
      of compliance as a test for obstruction in the small airways," J.
      Clin. Invest., 48,  1097 (1969).

13    COMROE, J.H., FORSTER, R.E., DUBOIS, A.B., BRISCOE, W.A. , CARLSEN,
      E., Tj>e_lunfi_:_ Clinical and pulmonary function tests, Year Book
      Medical Publishers, Chicago (1962).

14    DIXON, W.J. (ed.),  BMP Biomedical Computer Programs, University of
      California Press,  Berkeley (1967).

15    DUNCAN, D.P., "Multiple range and multiple F tests," Biometrics, 11,
      1 (1956).

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                             2001
                       DISCUSSION
BATES (Canada)
     What was the level of physical activity during the exposure?
Since most people walk in those atmospheres rather than sit at
rest in them, it seems to me important that if such studies are
to be used for standard setting/ the protocol should include
exercise.  Using an exercise protocol, Dr. Hazucha at McGill has
been able to demonstrate a change of closing capacity in normal
subjects after two hours of exercise in O.75 ppm of S0_.


WEIR  (U.S.A.)

1.   During the described investigations there was no formal
program of exercise.  However, each subject was encouraged to
continue his personal daily exercise including periods of a)
running inplace, b) push-ups and c) deep-knee bends.  These ex-
ercises were not quantitated.

2.   Regarding Dr. Hazucha's studies, I agree that in normal,
healthy individuals, it should be possible to show effects from
short exposures to about 1 ppm SO2.  However, I contend that
closing volume response to SO_ or other contaminants is probably
only linear over a narrow range.

     Our data suggest that these responses will not be dose
related when super-imposed on the pre-existing disease process
in the more normal population^ many of whom have early pre-
clinical changes caused by other factors.


HAZUCHA  (Canada)

     I would like to ask Dr. Weir several short questions.

1.   The interindividual variability of lung function tests is
expected, however, I am surprised by your large intraindividual
variability since some of the tests you used e.g. flow-volume
curves are highly reproducible.  Can you comment please?

2.   Both dynamic compliance and closing volume measurement are
presently considered to be the most sensitive tests for detection
of the small airways impairment.  However, while your dynamic
compliance was altered significantly, the closing volume was not.
Can you explain this discrepancy?

3.   I certainly cannot agree with your conclusions that subjects
with small airways  impairment "are probably not more susceptible
than normal population"(towards SO2)•  Your data clearly show
that they are more susceptible to SO2' although you were not
able to show statistical significance  (yet?).

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                             2002
WEIR   (U.S.A.)

1.   Tests such as flow-volume curves are indeed highly repro-
ducible in healthy individuals.  There is a vast literature to
support this.  My contention remains that most of these "sensi-
tive* indicators will not give reliable dose-response relation-
ships when applied to the general population, many having pre-
clinical small airways disease.

2.   As indicated during my presentation, the closing volume
test was not applied to the Series I, healthy subjects, but only
to the Series II, unhealthy subjects.  My comments to your first
question apply here.

3.   I am afraid that you misinterpreted my results.  All quan-
tifiable results indicated that the Series II subjects were not
differentially sensitive to SO2 compared to Series I subjects.
Although the pulmonary function tests results were variable, the
very careful clinical examinations conducted daily on these sub-
jects as well as answers to subject inquiry of well being in-
dicated no difference in susceptibility between these two groups
of subjects  in response to SO2-


ZEILHUIS  (Netherlands)

     What should in your opinion be the consequences  for the
threshold limit value for occupational exposure.  Your study
suggests a "breakdown in the adaptation processes at  the 3 ppm
exposure levels" in highly selected healthy subjects, without
more than minimal physical activity.  Gould you discuss the
results of your study, trying  to extrapolate these to occupational
exposure, for say, 4-6 ppm SO,,  40h/week + moderate physical
exercise for workers with less healthy lungs than even your
group  II.


WEIR (U.S,AJ

     Our results indicated that subjects representing at least
some of the  population generally considered to be "more sensitive"
did not appear to respond to SO- to a greater degree  than normal
healthy individuals.

     The  5 ppm 502 used for TLVs previously in the USA should
adequately protect all workers likely to be exposed.  Those
individuals  who are  "sensitive" will not be further protected
by decreasing the TLV to 2 ppra as  has been suggested  recently
by NIOSH.  Those persons would need an almost SO,-  free environ-
ment  (essentially the environmental levels proposed by the EPA)
to be  adequately protected.  I submit that those persons probably
"self-select" themselves from  exposure in our work  environment.

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                             2003
GRUENSPAN (Federal »spublic of Germany)

     What effects do bronchodilators, antihistamines or atropine
have on the lung function described?

     Is the reduction in lung function due to the action of
acetyl choline or histamine?

     Are the phenomena observed attributable to bronchocon-
striction caused by another factor?


WEIR  (U.S.A.)

     Unfortunately we had neither the opportunity nor permission
from the University Human Research Committee to conduct what
would essentially be pharmologic experimentation.  It is probable
that the observed effects could have been countered with drugs.
I am of the opinion that at least some part of the response
observed is reflexive and central nervous system controled.

     The experiments your suggest should certainly be conducted
to properly assess the importance of findings such as were pre-
sented in ray paper.


BERLIN (C.E.C.)

     Dr. Weir has just shown that exposure to SO- levels below
1 ppm do not lead to any apparent effects on pulmonary functions',
it was further indicated that this may be due to the insufficient
sensitivity of the conventional methods used.

     In the previous presentation Dr. Bates had demonstrated that
for SO,, levels of O.37 ppm combined with 0- at the same concen-
trations lead to significant effects.

     Both these conditions are not representative of the real
concentration and spectrum of pollutants in an urban atmosphere.

     To use the differenciation made by Prof. Bates between the
"state of the science" and the "state of the art" how should this
double information be used in the "state of the art" context for
the protection of human health from environmental pollution?


BATES  (Canada)

     To answer Dr. Berlin's question,  there are at  least three
conclusions:
1.   Sampling and measurement of O-  and SO- must be synchronous.

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                              2004
2.   Environments which already have  considerable  concentrations
of either SO  or O., must be very  careful that  they do not acquire
the other.

3.   Epidemiological studies must include measurements of pul-
monary function of children before and after exercising in these
environments, as measured decrements  are likely  to be the basis
of standard setting.

     There are at least another three conclusions, of which one
is the necessity of further studies of lower concentrations of
the two gases together.  I do not know the lowest  concentrations
of the two gases together-which might have a significant effect.


WEIR  (U.S.A.)

     The response by Dr. Bates as well as my response to earlier
questions satisfactorly answers these comments.

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                            2005
       EMERGENCY POPULATION EXPOSURE:  A METHODOLOGICAL
                           APPROACH
      (WITH A REPORT ON A HUMAN EXPERIMENT WITH CHLORINE)

              P, E, JOOSTING+ AND M, M, VERBERK^

+  TNO Research Institute for Environmental Hygiene, Delft,
   The Netherlands
++ Coronel Laboratory, Faculty of Medicine, University of
   Amsterdam, The Netherlands
ABSTRACT

     A philosophy on public emergency exposures and the need for
public emergency limits  (PEL) is presented.   PELs can be used
for the purpose of planning settlements, storages, transport,
organizational measures, etc.   PELs apply  to tolerable short
term emergency exposures of the general population which is nei-
ther homogenous in its composition, nor in  its responses upon
foreign agents.   The criteria, used in setting PELs are dis-
cussed, with special attention  to  the more  susceptible part of
the population, as well  as the  question whether there exist ob-
jective physiological criteria  that could be  indicative for the
population group at highest risk.

     A report  is given on an  experiment with  human volunteers
that aimed at  checking up a postulated  tentative  PEL for chlorine.
Eight  subjects, aged 28  - 52, were exposed  for  two hours to 0.5,
1,  2 and  4 ppm.   VC, FEV and FIV  measurements  before and  after
showed no significant differences.  Subjective phenomena, re-
corded with  15 min. interval, showed individual,  time and  concen-
tration dependent characteristics, as  well  as frequency distribu-
tions  to  which arbitrary tolerance criteria could be applied.

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                             2006
The problem of extrapolating the find-ings to susceptible subjects
and the use of a "safety" factor is discussed.   The elaborated
PEL value of 1.5, 1 and 1 ppm chlorine for 0.5, 1 and 2 hours
respectively, complies with comparable PELs from abroad, but is
not consistent with the very small number of literature data
that pertain specifically to subjective responses during short
term exposures to moderate concentrations of chlorine.

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                                  2007
1.   PHILOSOPHY
     1,1  Introduction
     In areas with high population density, industry, storage and trans-
port of dangerous materials, a growing need exists for knowledge about the
medical consequences of unforeseen sudden exposures to toxic substances
during accidental release of gases and volatile material from tank bursts,
traffic collisions, fires etc.
     So it is not surprising that in The Netherlands, with an average po-
pulation density of 396 per square kilometer, a Committee for Prevention of
Disasters by Hazardous Substances was appointed in 1964. The recommendat-
ions to the government are based a.o. on the work of a Subcommittee Toxic-
ity that submits proposals for maximum concentrations of toxic substances
that can be tolerated by the general population at short durations of expo-
sure .
     The present report dwells on the basic philosophy of public emergency
limits and gives an example of an experimental check-up of a postulated
limit  for chlorine that had to be tested because of  lack of uniform and
adequate literature data.
      1.2  Public Emergency Limits
     Emergency  exposures have already been a  recognized problem  in the
philosophy of hygiene since AlHA made issues  on that topic  j_l J-  Fundament-
ally  emergency  exposure  limits  (EEL) only  refer to  abnormal and  accidental
working  conditions,  and  single  exposures of  short  duration  to  one specified
 substance; the  worker's  capacity of  judging  an emergency  situation and
 taking co-ordinated  action  should not be empeded.  Possible  unexpected or
 unknown  hypersensitivity is  not taken into account.  Generally  industrial
 workers  are  supervised  as  to  their  state of  health with regard to  the
 work  situation.
      An  analogous  philosophy has recently  been developed in the USA  with
 regard to  protection of the general  public in case of unpredictable  emergen-
 cies  in  which toxic substances  are  released, in an uncontrolled manner at
 unpredictable times  and places, as  the  result of  accidents  such as  damage
 to transportation equipment or fire in a chemical-storage facility L2J.
      The NAS-NRC document outlines  a philosophy that practically parallels
 the ideas  of the Dutch Subcommittee Toxicity. In the following, some
 aspects  will be touched upon that have rendered difficulties in the
 practice of the elaboration and the implementation of public emergency
 limits (PEL).

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                                 20O8
     The main purpose of recommended PEL'S is serving the planning of.  in-
dustrial settlements, storage, processing and transport of toxic substan-
ces; planning of residential areas; revaluation of existing situations and
development of technical and organizational measures in potential emergen-
cy cases.
     It is reasonable t.o assume that for the greater part of the accidents,
referred to in the above, the atmospheric concentrations of the hazardous
substance will have reached their maximum and have dropped again within
two hours, e.g. as a sequel of measures that have been taken in the mean-
time. For this reason PEL values, as they have been worked out, apply to
exposures of maximally two hours. Additionally, limits for half an hour
and one hour are given, too. For seven substances of the list, dealt with
by the Dutch Subcommittee, PEL*s have been published [3J.
     1.3  Choice of Criteria
     The prime dilemma in the issuance of PEL is the question to what
extent a given effect may be regarded as  tolerable under emergency con-
ditions. Without doubt there will be a common consent that the following
three criteria must be appreciated as unacceptable:
     a -  increased mortality
     b -  symptoms and signs of illness,  either caused or aggravated by
          the exposure, which are irreversible or only reversible by inter-
          vention, or slowly reversible  
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                                  2009
general population in a given set up essentially represents a composition
of sex, age, endogenous factors, intrinsic liabilities, explicit ill con-
ditions and other variations of human health parameters. There are small
children, weak people, pregnant women, subjects suffering from respiratory
diseases, cardiac impairment, aged people, etc. If one would try to protect
at least the greatest part of those who on the above grounds are expected
to be intrinsically more prone to fall a victim under definite emergency
conditions, one should recognize the fact that each population comprises a-
bout ten to twenty percent weak people for whom a special estimate should
be made as to what can be afforded from them. It is logic that PEL values
for the general public will be substantially lower than EEL values for work-
ers in industry under medical supervision.
     In every population there will be a number of hypersusceptible, except>
ionally vulnerable individuals, like people with a serious stage of chronic
bronchitis, pneumonia, cor pulmonale, myocardial infarction, asthma and
comparable conditions, of whom it is hardly possible to predict in which
way and to what extent they will react upon sudden and outspoken changes in
atmospheric quality. Despite the voluminous epidemiological literature
available, it is very difficult to give an appropriate estimate of this part
of the population that cannot be taken into account in a concrete evaluat-
ional procedure. Van der Lende [4J states that in the age group 40 to 64
about 8 percent are affected by chronic nonspecific lung disease (CNSLD) to
such a degree that, in his opinion, regular treatment is required. He
estimates about 1 to 2 percent to be badly handicapped. Personal communic*-
ions by other lung physicians indicate a smaller part of the population to
be in such a stage of illness or hypersusceptibility that their reactions
upon inhalation of a foreign gas can never be predicted. Their estimate
would be between one and five per thousand of a population at risk. Addit-
ionally Van der Lende remarks that exogenous factors (urbanization, air
pollution) seem to have less effect on airway obstruction than on product-
ive cough.
     There is one feature that could receive special attention. It is the
hyperreactivity of the airways as reflected by the results of a histamine
threshold test (VC or FEV decrement more than 10 percent upon inhalation of
a histamine solution of 8 or 4 mg/ml or less). For some population groups
the distribution of histamine thresholds are known [4j.
     If one would know any existing relationships between the response of

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                                  2010
volunteers to defined exposures and their histamine sensitivity, It could
be possible to make an estimate of reactions among the general public on
the basis of the histamine threshold distribution. It would be desirable to
have at hand for these purposes a standardized group of volunteers who re-
flect to a large extent the endogenous factors that may be responsible for
the reaction upon an agent. According to our opinion experimentation with
people with, an explicit clinical condition (bronchitis etc.) does hardly
comply with current thoughts in medical ethics.
     1.5  Other aspects
     Whereas the subject of PEL is still in a more or less developmental
stage, a number of topics are rather relevant but cannot be discussed here.
Some of these have already been touched upon in another publication [3J, as
there are incompleteness of adequate literature data; safety factor in
extrapolations from "healthy subjects" to "general public" and from animals
to human beings; and qualification and quantification of the risks if a PEL
still would be exceeded.

2,   CHLORINE EXPERIMENT
     2.1  Introduction
     When the Subcommittee Toxicity tried to elaborate a PEL for chlorine,
the adequate available literature data were so scarce and displayed on the
other hand such a diversity of dose-effect relationships that it was felt a
need to check the tentatively postulated PEL by human experiments.
     Whereas the Subcommittee is of the opinion that its responsibility
implies that PEL values, postulated with regard to the general public, at
least must be acceptable for its own committee members, it was decided to
set up a small experiment to check whether a tentatively elaborated PEL of
0.5, 0.3, and 0.3 ppm for 0.5, 1, and 2 hours exposure respectively would
stand. These values were based upon experimental work by Beck |_5J and Rupp
and Henschler [&J. Their young and healthy volunteers were exposed for 30
minutes to 0.02 to 0.9 ppm chlorine. Although their results manifested a
broad variance in perceptions, it seemed prudent to assume that 0.5 ppm
during 30 minutes would be a limit to subjects with sensitive mucous
membranes.
     Further the Subcommittee liked to be convinced that at the postulated
PEL exposures no trivial respiratory function disturbances would occur.
Therefore the study at issue was not limited to subjective feelings only.

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                                  2011
     2.2  Materials and Methods
     Eight subjects, aged 28 to 52 and all members of the Subcommittee,  were
available. Some were not able to attend all sessions. With one week inter-
val, twice a day a group of 4 subjects was exposed during two hours to 0.5,
1, 2 and 4 ppm chlorine respectively. Because of a technical failure one run
with 4 ppm had to skipped so that information on only three persons is
available. One of these did not sit out the two hours.
     During the exposure the persons were seated in a chamber of 4 x 2.5 x
2.5 cubic meters (a small lock-space included). The test gas perfused the
chamber at a rate of 100 cubic meters per hour. Mixing was ideal.
     Chlorine gas was generated electrolytically from a. saturated and sour-
ed NaCl solution. Concentrations inside the chamber were monitored by means
of an instantanuously reading coulometric detector. The concentrations re-
mained constant within a range of + 5%, although the test persons entered
and left the chamber at 15 minutes interval to facilitate physiologic tests
before and after the exposure.
     The latter included: a) EGG and b) breathing frequency in rest over
two minutes, and c) spirometry (Pulrootest-Godard): VC, FEV, and FIV. During
the exposure a) and b) were also recorded with intervals of 30 minutes.
     Each person recorded his subjective feelings qualitatively and
quantitatively according to a checklist of symptoms, making use of an
absolute and a relative scale of qualifications: 0= no sensation; 1= just
perceptible; 2= distinctly perceptible; 3= a nuisance; 4= offensive; 5=
unbearable; and + is increasing; - is decreasing; = is constant.
     2.3  Results
     2.3.1  Heart rate
     Although the EGG readings displayed a wide range of individual heart
rates, two interesting observations can be made.
     Before the first experiment the average heart rate was 87 as people
were not yet accustomed to the situation. At the three other runs (with
higher concentrations) the before-exposure rates were 72, 72, and 77; i.e.
significantly lower (sign test, p <0.05). After 15 minutes of exposure the
heart frequency in the first experiment had dropped to 77 already.
     During the exposures the heart frequencies stayed rather constant with
non-significant differences between the four runs.
     The second remarkable phenomenon was observed after cessation of the
tests. The mean heart rates, recorded within the first minutes after leav-
ing the chamber, dropped to 66, 64, 64 and 64 respectively. In each case

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                                   2012
the individual heart rates after exposure were lower than the mean rate
during the session  (p <0.05).
     Although real  basic heart rates for each individual were not known, it
is probable  that  the group showed an "off" effect at cessation of the trial,
independent  of the  concentrations during exposure. Question remains whether
this may be  explained only on account of the mental stress exerted by the
test situation, or might reflect a vegetative response upon cessation of a
physical stimulus that triggers autonomic regulatory mechanisms.
     2.3.2   Rate of respiration
     The respiration rate, as recorded before, during, and after the trial
did not show anything of special interest: no changes over time and no in-
fluence of concentration, neither with regard to the individual values, nor
the group means, could be observed.
     2.3.3   Spirometry
     Comparison of measured VC, FEV and FIV before exposure with the read-
ings after the trial, did not yield remarkable changes. At neither concent-
ration an important decrease could be detected. Only FEV gave a small de-
crease of 90 ml if all readings were pooled (S.D. 180 mil) at a signific-
ance level of p ^.Ol (sign test). But there was no demonstrable associat-
ion between decrease of FEV and magnitude of the concentration. Neither did
individual readings reveal a consistent personal reaction pattern.
     It can  therefore be postulated that exposure to 0.5, 1 or 2 ppm chlor-
ine, during 2 hours does not result in significant changes of ventilatory
capacity among healthy subjects. The 4 ppm experiment did not allow a con-
clusion (only 2-3 subjects).
     2.3.A  Subjective phenomena
     The 6-8 subjects recorded every quarter their responses for ten sens-
ations, resulting in a maximum of 64 fifteen-minutes readings per item.
These could be arranged as mean scores per quarter of an hour to demonstrate
the dynamics of responses during the course of the exposure. Graphical pre-
sentation and group mean scores over two hours would allow to compare re-
sponse patterns with regard to concentration. This has been done in figures
1-6 for the exposures with 0.5, 1, and 2 ppm chlorine (the data of the 4
ppra experiment were too incomplete to allow comparison).
     2.3.4.1  Smell (Fig. l)
     Initial perception of smell is about the same for the three concent-
rations. There is a tendency towards decrease of sensation, particularly
during the first hour. Individual differences of scoring between the

-------
o
o
in
       o-


       X-


       A-
-o s 0,5 ppm (n*6 )

-x * 1  ppm(n = 7)

-a « 2  ppm(n*8)
                              SMELL
             15     30    45    60     75    90    105

                               Exposure time (min)
o
V
z:
                                              120
    Figure 1   Smell perception - Course of mean scores during the


               experiments; means over the 2-hr exposure at the right.
 *>
 fc.
 o
 u
in
    _  1
              ' 0,5 ppm(n =

              > 1  ppm(n =

              • 2  ppmfn:
                                                     C
                                                     O
                                                     u

                                                     Z
                                                               X


                                                               o
             15    30     45     60     75    90    105    120

                                Exposure time (min)

    Figure  2



     Taste perception - Course of mean scores during  the

     experiments;  means over the 2-hr exposure  at the right,

-------
 o
 u
             *0,5ppm(n»6)

             »  1 ppm(n»7)

             «  2 ppm(n«8)
  EYES
                  30    45
                        120
                          60    75    90    105

                          Exposure time (mm)

Figure  3


Eye  irritation - Course of mean  scores  during the

experiments; means over the  2-hr exposure at the right,
o
u
in
             Q,5ppm(n«6)

              1 ppm(n*7}

              2 ppm(n*8)
NOSE
                               o
                               V

                               £
      0     15     M    45    60    75    90     105   120

                             Exposure  time (min)


    Figure 4


    Nose irritation - Course of  mean scores during the

    experimentsf means over the  2-hr exposure at the right.

-------
                                  2015
subjects remained consistent over the concentrations.
     2.3.4.2  Taste (Fig. 2)
     Scores were low, more than half of the test subjects did not perceive
anything. There was a consistent individual response pattern.
     2.3.4.3  Eye irritation (Fig. 3)
     Concentration has a clear-cut influence on eye irritation.  While  at
0.5 ppm, individual scores remained between 0 and 1, they figured between 0
and 2 at 1 ppm. At 2 ppm three subjects recorded 3 with hardly any moment-
ary 0 code. The gradient of the line at 1 ppm was due to a few subjects who
turned out more "responsive" than the others, with an interindividual
consistency over the concentrations.
     2.3.4.4  Nose irritation (Fig. 4)
     No differences can be observed between 0.5 and 1 ppm. At 2  ppm there
is a sharp rise during the first hour and to a less degree in the second
hour, scores then reading 1-3. At 4 ppn the gradient of the first hour was
still steeper.
     2.3.4.5  Throat irritation (Fig. 5)
     The scores demonstrate a definite association with the concentration
level, a.o. reflected by an increasing number of subjects scoring ^0,
Curiously enough, the person with the highest scores, recorded 0 during  the
first hour at all three concentrations. The scores at 4 ppm were still high-
er than at 2 ppm. Individual consistency could not well be observed.
     2.3.4.6  Cough (Fig. 6)
     Irritation of the airways, resulting in coughing, was experienced by
only a few subjects. At 2 ppm three of eight responded. At 4 ppm two of  the
three subjects scored nuisance. These two were the same who already had  re-
acted at 1 and 2 ppm. There seems to exist a consistency of response.
     2.3.4.7  Other phenomena
     Headache was experienced by some subjects, generally at the end of  the
exposure. There were indications for individual consistency.
     Perception of chest irritation was observed by one subject at three
concentrations. At 4 ppm one out of the three responded with code 1.
     Other signs of unfavourable influences upon the well-being of the
subjects were not be observed by themselves.
     2.3.4.8  Intercomparison of phenomena
     In Fig. 1 mean group scores, ranges of individual means and individual
maximum readings are presented.
     At 0.5 and 1 ppm all the group means were below the level of just per-

-------
u
U*
e »0,5ppm(n-6)
» - 1 ppm(n»7)
                            THROAT
            15
                                                          o
                                                          1)
                                          120
              30    45     60     75    90    105
                          Exposure time(min)
Figure 5

Throat  irritation  -  Course of mean scores during  the
experiments; means over  the 2-hr exposure at the  right.
            «0,5ppm(n»6)
            * 1 ppm(n»7)
            • 2 ppm(n«8)
                COUGH
           15
       30
                    45
                                                c
                                                o
                          SO     75    90    105
                          Exposure time  (min)
Figure 6

Cough - Course of mean scores during the
experiments;  means over the 2-hr exposure at the right,

-------
       »^X/J
o
VI
 0.5  ppm (n = 6 )
 1   ppm (n = 7 )
 2   ppm (n = 8 )
 It   ppm (n = 2 )
                                )>
                                  I
                                                    - /:
       Smell
Taste
Eyes
Note
                                                             II
Throat    Cough
Headache  Chest
                 Figure 7   Mean scores (height of  columns), lowest and highest
                           individual means (}	»_———))» and incidental
                           maximum scores (• ) in exposures of two hours.

-------
                       EYE  IRRITATION
   100
 M
 Vf
   so
 o
 u
 c
 4*
 U
0.   0
 0 » 0.5 ppm  Cl2 ( n * 6 )
 « « 1  ppm  Cl2 (n = 7 )
 * * 2  ppm  Cl2 (n« 8)
	otter  30 mm. exposure
	after  60 min. exposure
	after  120 min exposure
                                   X
                                   A
                                                                50
                                   o
                                   u
                                   VI
   C
   tj
   u
   &.
00 a
                                                Score
     Figure  8   Eye  irritation  - Cumulative  frequency  distribution
                of individual scores  at  three  times of exposure  to
                three concentrations.

-------
                                   2019
 ceptible (code 1), and the individual means figured below distinctly per-
 ceptible (code 2). At 2 ppm the group means for smell, eye-, nose- and
 throat-irritation increase above the level of minimum perceptibility. The
 highest individual means figure about code 2, while incidentally nuisance
 (code 3) is  recorded.
      At 4 ppm particularly irritation of nose and throat, and coughing in-
 crease in intensity.  Some other symptoms tend to diminish. The latter tend-
 ency  is probably associated with an increased secretion by mucous membranes
 of  nose and  eyes as was observed by some subject: rubbing one's eyes in-
 creased the  irritation. To prevent extra irritation of the throat and cough-
 ing,  one did not speak or laugh during the second half of the experiment.
 All three, of whom one performed only 75 minutes, experienced the exposure
 at  4  ppra as  a limit,  mainly because of irritation of the throat.
      2.3.4.9  Presentation in  frequency distributions
      To  get  an impression about the variation of individual scores,
 cumulative frequency  distributions  were made  of  the readings  at definite
 intervals, which  reflect  the momentary condition of the  group after  0.5   1
 and 2 hours  of  exposure.  Figures  8-10 display  such distributions for
 eye-, nose-  and throat  irritation.
     With increasing  concentration  or exposure time, the lines  shift  to
 the right, reflecting an  increase of  response by level or by  number of
 reacting subjects.
     The graphs display the phenomenon that in several instances where the
 -,oup mean score  (50 percentile)  levels  at code  X,  there may  exist a  rather
 l*rge percentage  (35 -  40%) with readings up to  code (X  + 1), whilst  10-15%
 even reach at a level of  code  (X +  2). Although  the number of test subjects
was  too small  to be representative in any respect, this distributional
phenomer m should be taken into account  seriously when discussing the
acceptabi) ty of PEL for a population  as a whole.
     2.4  Criteria and, elaboration  of a PEL for  chlorine
     The present investigation has  not produced  any evidence that a 2-hour
exposure to 2 ppm chlorine would have  deleterious effects on heart rate
and breathing frequency. Apparently these objective parameters do not limit
the acceptability for the eight persons under investigation.
     From the point of view of checking the PEL values, postulated by the
Subcommittee, this finding is reassuring and suggests the possibility of a
somewhat higher PEL value for chlorine.

-------
                                  2O20
     If one would elaborate criteria for acceptability from subjective find-
ings, the frequency distributions of paragraph 2.3.4.9 could serve as a tool
to estimate and fix tolerance levels based on the percentage of a population
that one would allow to suffer from a definite inconvenience.
     To exemplify this by means of a theoretical model, it is imaginable to
postulate that under emergency conditions it could be tolerable if 50 per
cent of the subjects would perceive the agent in terms of just perceptible
(code 1), 35 per cent would record more distinct sensations (code 2), 12
per cent would be bothered by a real nuisance (code 3), whilst 3 per cent
would be allowed to suffer from any still more troubling experiences.
     Such a procedure is displayed in Fig. 11. The guide is drawn accord-
ing to the above mentioned percentages of tolerance. Some of the frequency
distributions, as presented in Fig. 10, have been brought up in the same
diagram: Throat irritation showed to have the best discriminating power
between the various exposures and was one of the main arguments of the
tested subjects to estimate A pptn a limit.
     According to the guide-line model, an exposure to 2 ppm chlorine during
two hours would not be acceptable because not 50 per cent, but 75 per cent
of the people experience more than code 1, and 25 per cent instead of only
15 per cent are bothered by a nuisance (code 2). The frequency distributions
of the two other dose-response relationships are acceptable.
     Although this principle might be correct, a number of incertainties re-
mains. The number of subjects was small. A true upper level of unbearable
(!) exposures has not been set. The subjects were motivated and were
familiar with the situation. They were under medical supervision and were
aware of the finiteness of the experiment. They could leave the chamber
when they wanted. Accordingly their tolerance will have been greater than
might be expected for the general public during an emergency. In addition
health of the test subjects was good and certainly not representative  for
the general population.
     If one assumes that about 20 per cent of a population is in a condit-
ion of reduced resistance  (asthma, chronic bronchitis, emphysema, cardiac
distress, pregnancy), one might expect that  these would represent the  tail
ot a frequency distribution. Even more complicating is the  fact  that  the
healthy part of  the population (cf. the group of  test  subjects) may  dis-
play a broad range of   responses, upon which  the  proportion  of the above
mentioned 20 per cent of sensitive people should  be superposed.
     As  long as  it  is unknown  in which way and  to what extent  the more sens-

-------
              NOSE IRRITATION
 100.
X
\f
  50
o
o
w
+*
M
Cl
Cl
L»

t>
Q.  0
                                                         %
                                 0.5 ppm Ct2 ( n * 6)

                                 1  ppm Cl2 (r» = 7)

                                    ppm Cl2 (n«8)

                                         30 min. exposure

                                         60 min. exposure
                               = 2
                                  otter

                                  otter
                                  otter  120 min. exposure
                                                         50
                                                             x
                                                             A
                                                             o
                                                             o
                                                             vt
o»
o
Arf
c
V
u
                                                         100 tL
                                           Score
       Figure 9


       Nose irritation - Cumulative frequency distribution

       of individual scores  at three times  of exposure

       to three concentrations.

-------
                 THROAT IRRITATION
 100
X

\f
o
u
*

t*
•*
  SO
o
*»
c

u
                               » O.S ppm  Cl2 (n »6)


                             * * 1 ppm  Cl2 (n«7)


                             A » 2 ppm  Cl2 (n*8)

                            	otter  30 min. exposure

                            	 after  60 min. exposure


                            	 after  120 min. exposure
SO
   x

   A
   o
   u
                                                            O



                                                            JC
   o

   V
   o»
   o
                                                        100
                                           Score
      Figure 1O


     Throat irritation -  Cumulative frequency distribution


     of individual scores at three times  of exposure


     to three concentrations.

-------
                                  2023
itive groups of the general public will react vpon short term exposures  to
the agents for which PEL are required by the authorities,  application of a
"safety"factor should be used, although this term, admittedly,  is  a  little
bit queer in case of emergencies.  It seems realistic to divide  the con-
centrations that are associated with a tolerated frequency distribution of
symptoms among healthy test subjects, by a factor two, in order to expect
that the true frequency distribution of the general population be  more  or
less in line with the experiment.
     According to our findings, obviously an emergency exposure of 2 hours
to 4 ppm chlorine would not easily be tolerated by non-informed healthy
subjects. An exposure of 2 hours to 2 ppm appeares reasonably bearable  for
healthy subjects.
     If this exposure would be appreciated as marginal and a "safety"
factor of 2 would be applied, a PEL for the general population would read
1 ppm chlorine for 2 hours. According to  the frequency distributions of
Fig. 8 -  10 it may be assumed that an exposure of  1 hour to 1 ppm and 0.5
hour to 1.5 ppm would not result in outspoken deleterious effects either.
     It should be stressed that extreme susceptibility cannot and has not
taken into account.
     2.5  Comparison with existing PEL
     The NAS/NRC Committee on Toxicology  [?] has  given PEL values for
comparable exposures. Although  the philosophy is  about the  same,  the
reasoning is not quite  clear  in the NAS/NRC paper.  The more therefore  is
it  interesting that both  issues are  sufficiently  matching.  The Pennsylvania
Department on Occupational Health  [s]  has also  issued a limit, viz.  3  ppm
during 5  minutes, which also  fits  the  overall pattern;
                           PEL for  chlorine (ppm)

exposure  time  (min.)    5      10        30       60       120
Pennsylvania
NAS/NRC
Netherlands
3 - -
3 2
1.5
_
2
1
_
-
1
      Both American PEL values are in agreement with our findings  in the
 human experiment with healthy subjects. The latter, on the other  hand,

-------
                                              Score
  100
   97
   85
   SO
o
V
01
o
t*
c
v
u
u.
•r
0.
        *'
1 ppm.  60 min.
2 ppm,  30min.
2 ppm, 120min.
GUIDE
                          °/o
                          0
                          3
                         *•  s
                             u
                                                                   O

                                                                   -C
                                                      3    3.3
                                                               100
                                              Score
     Figure  11  Example of  an arbitrary  guide  for  throat  irritation.

-------
                                   2025
 differ widely from the response patterns found by Beck [s] and Rupp  and
 Henschler 1.6J, who probably have worked with very sensitive and differently
 motivated volunteers. In our opinion our first tentatively postulated  PEL
 values of 0,5, 0.3 and 0.3 ppm, which were based on their work, must be
 judged as unrealistically low.  On the basis of the underlying study  the
 members of the Subcommittee Toxicology had no difficulties in changing
 their mind into 1.5,  1,  and 1 ppm respectively.
      2.6  Final remarks
      To develop further  methodology for the purpose of elaborating real-
 istic PEL, some more  information is needed about:
      -  qualitative composition of the general public  with regard to its
         sensitivity for  irritating gases in general.
      -  effects among sensitive groups by certain gases.
      -  relationship  between parameters of hyperreactivity (e.g. histamine
         sensitivity)  and response  mechanisms on irritating substances  in
         general.
      -  influence  of  physical stress (= increased  ventilation)  on the
         influence  of  irritant gases.
         influence  of  emotional  factors, fear and ignorance of the toxicity
         of the  agent.
      Research  in these fields should be stimulated and co-ordinated.
      2.7  Post scriptum observation
      Some weeks before the presentation of this paper observations were
made that should not remain unrevealed, because they might throw a light
on the above mentioned differences between the German findings and our
own.
      In order to check a PEL for ammonia the subcommittee members ex-
posed themselves in the same manner as to cholrine. Because it was
felt a need to check the possible hypertrophy of motivation arrL the
outsposen familiarity with the subject, a group of eight healthy students
was selected to participate in the experiment, in separate sessions.
      The students turned out to score systematically higher than the
committee members had done, for a great number of sensations.  An example
of extreme differences between the frequency distributions of both
groups is revealed for throat irritation in figure 12: the maximum
score among the committee members (code 2+ ) is lower than the mini-

-------
* x
v/ AMMONIA, UO ppm, 30 MINUTES A
u
LU 0-
g 100 -i
\mf
O
en
X
t
CO
a BO-
CD

U
O
1

' s
COMMITTEE ' St
MEMBERS^-' 	 	 •*" /
-' r /
A / x^
/ / ^-"^STUDENTS
J
1 Jfc^^
/ / COMBINED
* ^
V /
^

I 1 1 i I i
r° 8
o
m
• \f 9
i^
l
cn
•
-50 a
3
to
LU
O
g
•100 S
£ 0 1 2 3 A 5 £
£ SCORE £
  Figure 12
Exposure to ammonia :  difference between cumulative
frequency distribution of committee members and students

-------
                                   2027
 mum score (code 3)  of the students. Two of the latter  (2556) scored
 unbearable (code 5)  after half an hour, already.
       If one pools  both parties,  the result  can be illustrated by the
 combined frequency  distribution  in the centre of the graph. Effectively
 this procedure means a big shift  to either of the two  original distri-
 butions.  This phenomenon  induces  questions about the students data anl
 throws doubt on the question whether our  group of committee members
 still is an appropriate panel  to  test tentatively postulated PEL's.
       According to  our opinion this observation does not undermine
 essentially the principle of a phenomenological and experimental ap-
 proach in the setting of  standards, particularly the elaboration of
 public emergency limits in case  of simple irritating substances.
Acknowle dgement
     The Directorate General of Labour, under the auspices of which the
Subcoramittee Toxicity fulfils its task, gave a grant for the experiment
that was performed in the test chamber of the TNO Research Institute for
Environmental Health, medical supervision and physiological tests being
performed by Dr. B. Bink, Netherlands Institute for Preventive Medicine TNO,
Leyden.
     The authors wish to express their special gratitude towards the other
members of the Subcommittee who participated as test subjects in the ex-
periment and who were so kind as to allow publication part of the basic
philosophy that was developed by the joint effort of present and past mem-
bers, whose names are: K. Biersteker, A.W.M. Balemans, H. Eilers, M.J. van
Logten, J.H. Koeman, T. Rooyakkers-Beemster, E.H. Siccama and R.L. Zielhuis
(Chairman).

-------
                                  2028
                                REFERENCES

1   AMERICAN INDUSTRIAL HYGIENE ASSOCIATION (AIHA),  TOXICOLOGY COMMITTEE.
    "Emergency Exposure Limits", Amer. Industr. Hyg. Ass. J., 25,  578
    (1964).

2   NATIONAL ACADEMY OF SCIENCES - NATIONAL RESEARCH COUNCIL (NAS  - NRC),
    Basis for establishing guides for short-term exposure of the public
    to air pollutants, U.S. Dept. Coranerce, Nat. Techn.  Inform. Service,
    Washington DC, (May 1971).

3   SUBCOMMITTEE TOXICITY OF THE DUTCH COMMITTEE FOR PREVENTION OF
    DISASTERS BY HAZARDOUS SUBSTANCES, "Public Emergency Limits",
    Staub-Reinhaltung der Luft, 34, 85 (1974).

4   LENDE, R. VAN DER, Epidemiology of chronic non-specific lung disease
    (chronic bronchitis), Thesis Groningen University, Van Gorkura  Comp.,
    Assen, The Netherlands (1969).

5   BECK, H., Experimentelle Ermittlung von Geruchsschwellen einiger
    wlchtigen Reizgase und Erscheinungen bei Einwirkung  geringer
    Konzentrationen auf den Menschen, Inaugural-Dissertation, Wurzburg,
    West-Germany (1959).

6   RUPP, H., HENSCHLER, D., "Wirfcungen geringer Chlor-  und Brom-
    konzentrationen auf den Menschen", Arch. Gewerbepath. Gewerbehyg.,  23,
    79 (1967).

7   COMMITTEE ON TOXICOLOGY OF NAS/NRC, Guides for short term exposures
    of the public to air pollutants. III.Guide for chlorine, Washington
    DC, (1973).

8   PENNSYLVANIA DEPT. OF HEALTH, DIVISION OCCUPATIONAL  HEALTH, Short
    term exposure limits, Harrisburg, Pa., (1966).

-------
                             2029
                       DISCUSSION
HINE (U.S.A.)
     In using human sensory response panels two basic principles
are followed by our group.  First the panel must be carefully
selected by trials to eliminate persons giving false positive
response in the absence of stimuli and those failing to give
a response where 9O% of subjects rate the exposure as + 3 on a
O-5 scale.  gecondly the panel must be trained by repeated ex-
posures to the same concentration so that they will have a con-
sistent grading of the degree of irritancy.  Of course all ex-
posures after the training sessions at known concentrations are
done in a double blind fashion.
JOOSTING  (Netherlands)

     Thank you for your constructive comment.  We were fully
conscious about the fact that our method would not stand your
criteria.  It was not our aim to set PEL's on a  scientific basis,
but we would check what the impact could be of the postulated
PEL for chlorine which we did not yet  feel sure  enough about.
Therefore we started with the low concentration.  In the recent
experiment with ammonia we kept the procedure of increasing the
exposure in the next session on the basis of the findings in the
preceding one, but without telling the subjects  to what actual
concentrations they would be exposed.  The students were told
that they would be exposed to an irritating agent, without any
hazard.


LEFEVRE  (Belgium)

     Did the patients exposed to chlorine vapours by Dr. Joosting
subsequently show dyspnoea or a tendency to pulmonary oedema?


JOOSTING  (Netherlands)

     From the literature that we had studied before starting the
experiments with ourselves, we had not learned that an exposure
during 2 hours to e.g.  4 ppm chlorine  would result in clinical
symptoms like dyspnoea  or signs of a tendency  to pulmonary oedema.
One of us experienced a sensation of irritation  within the chest
at some moments during  the exposure to 2 ppm', but this phenomenon
disappeared at cessation of the exposure.  No  one of us complained
about the symptoms you mentioned.  If  we would have expected
such serious effects to happen as a sequel of  the well controlled
exposure, we would never have started  the trial.

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                              2031
     L'INFLUENCE DU BRUIT SUR LE SOfWEIL DES TRAVAILLEURS
     DE  NUIT ET DES TRAVAILLEURS PAR EQUIPES ALTERNANTES

                  P, KNAUTH ET J, RUTENFRANZ
Institut fur Arbeitsphysiologie an der Universitat Dortmund, BRD
RESUME

     Dans 1e cadre de travail par roulement realise en labora-
toire, nous avons etudie lfinfluence du bruit et du travail par
roulement sur le sommeil.   Le travail par roulement experimen-
tal nous a servi de mefhode nous permettant d'&tudier I'influence
de la combinaison de ces deux facteurs sur le sommeil.

     Nous basant sur le resultat d'enquetes effectuees aupres de
travailleurs en equipes alternantes, nous avons perturbs le som-
meil des sujets par des bruits de circulation (45~70dB(A)) et
par des bruits d''enfants (20~60dB(A)).   Pendant le sommeil, nous
avons enregistre les EEGf EMGf EOG et ECG ainsi que les mouvements
du lit afin de pouvoir deceler les influences du bruit sur le
sommeiI.

     Etant donne que les travailleurs de nuit ne dorment apres
le travail de nuit en moyenne que 6 heures, nous avons limite le
travail de jour de deux des sujets a 6 heures pendant une periode
de travail de nuit de trois semaines, tandis que deux autres su-
jets jouissaient d'un sommeil illimite.

     La perturbation du sommeil par le bruit ne d&pendait pas
seulement de I'intensite de ce bruit, mais surtout  de I'informa-
tion qui en ressort.   La duree totale du sommeil ainsi que cells
du sommeil PMO etait en moyenne plus courte quand le  sommeil

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                              2032

etait perturbs par des  bruits d'enfants  que  tors  d'une  pertur-
bation par des bruits de  circulation.

     Les parametres EEC,  EOG, EMG  et mouvements du  lit  ne fai-
saient pas apparattre une  habituation au  bruit  au  cours  de I'ex-
perience*

     La limitation a 6  heures du sommeil  de  jour, comme on  la
rencontre habituellement  chez les  travailleurs de nuit,  avait
pour consequence une reduction des phases PMO  et  IV par rapport
au sommeil de jour illimite.   En  ce qui  concerne le pourcentage
des stades PMO et IV par  rapport £ la totalitS du sommeil,  nous
n'avons pas constate de difference statistiquement  significative
entre le sommeil limits et le sommeil illimite.

     L'evaluation subjective de la qualitS du  eommeil presentait
au coura des trois semaines de travail de nuit un trac£ different
de aelui des stades de  sommeil PMO et IV.

     Etant donn& qu'en general le  sommeil de jour des travailleurs
de nuit est plus court  et plus perturbe par  des bruits  que  le som-
meil normal de nuitr il a ete recommandS de  ne pas  utiliser de
plans de roulement comportant beaucoup de travail de nuit conse-
cutif.
ABSTRACT

     Shift work uas organized in the laboratory to study the ef-
fects of noise and shift work on sleep.   Experimental shift
work served as a means of studying the combined effects of these
two factors on sleep.

     Using as a basis the results of surveys carried out on shift
workers, we disturbed the subjects' sleep by traffic noise (45-

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                             2033
70 dB(A}) and noise made by children (20-60 dB(A)).   We recor-
ded the EEG, EMG, EOG and ECG and bed movements during sleep in
order to determine the influence of noise on sleep.

     As night workers only sleep an average six hours after night
work, we limited the day sleep of two of the subjects to six hours
during three weeks of night work, while two other subjects enjoyed
unlimited sleep.

     Sleep disturbance owing to noise did not depend solely on
the intensity of the noise, but above all on its information con-
tent.   The overall duration of sleep and EEM sleep was on the
whole shorter when it was disturbed by children's noise than when
it was disturbed by traffic noise.

     The EEG, EOG, EMG parameters and bed movements showed no
evidence of inurement to noise during the experiment.

     The fact of limiting day sleep to six hours, as is usually
the case for night workers, caused a shortening of the REM stage
and stage IV compared to unlimited day sleep.   As regards the
percentage of the REM stage and stage IV in relation to the over-
all  period of sleep, we did not discover any statistically sig-
nificant difference between limited and unlimited sleep.

     Subjective assessments of the quality of sleep during the
three weeks of night work showed a different pattern to that of
the REM stage and stage IV.

     In view of the fact that the day-time sleep of night workers
is shorter and disturbed by more noise than normal night-time
sleep, it has been recommended that work schedules involving a
lot of successive night work should not be used.

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                              2034
1. Int roduct i on
On des problemes essentials du travailleur en equipes alter-
nantes est le sonaneil. Dans une etude precedente  (fig. 1) nous
{KNAUTH et al. [l]) avions, dans le cadre de travail par equi-
pes en laboratoire, compare premierement le soiraneil de jour au
sommeil de nuit, deuxiemement le sommeil perturbe au soiraneil
non perturbe. La fig. 1 represente quatre exemples de sommeil
d'un merae sujet. Nous avons constate ces memes tendances chez
les autres sujets. Le dessin du haut represente le sommeil de
nuit sans aucune perturbation d'un sujet dormant dans une
chambre insonorisee. Pendant la nuit perturbee par des bruits
(second dessin), la duree du sommeil est plus breve, 1'image
generale est plus raorcelee et le stade IV en particulier est
plus souvent interrompu. Un decalage de phase du sommeil vers
la phase de jour donne lieu a un racourcissement supplementaire
du sommeil. Les conditions de sommeil de jour avec perturba-
tions sonores sont particulierement defavorables, car il en
resulte un deficit important de sommeil par rapport au sommeil
de nuit non perturbe.
Nous nous somes done demandes quels fitaient les ef fets sur le som-
meil de jour d'un travail de nuit de plusieurs semaines  : est-
ce qu'il en resulte une accumulation du manque de sommeil, ou
bien est-ce que le corps humain peut compenser ce manque de
sommeil ? Cette question ne pouvait etre etudiee que dans le
cadre d'un cycle d'experience de travail par roulement d'une
longue duree. Etant donne que de telles experiences de plu-
sieurs semaines necessitent beaucoup de personnel et de  temps,
il n'existe - a notre connaissance - pas encore d'etude  compa-
rable.

2. Methods
Quatre sujets de  sexe masculin  ages de  22 a  25 ans ont vecu  5
semaines chacun a 1'Institut. A une semaine  de travail de  jour
succedaient  trois semaines de travail de nuit et  une  semaine
de repos sans travail.  Deux des sujets  pouvaient  dormir  autant
qu'ils  le  voulaient apres  le travail  de nuit. Nous  avons limi-
 te le sommeil apres le  travail  de  nuit  des  deux  autres  sujets
 a 6  heures,  puisque des enquetes aupres de  travailleurs  de
 nuit ont revele une duree  moyenne  de  sommeil de  6 heures.

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                             2035
          sftxfes Or sonwnert
                                         cfeiul sons trufe
                            ur   un
                                                  to
                                         Of nu* one trafe
            R£M fPMOi
      Fig. 1   Stades de sommeil d'un sujet

Les sujets dormaient dans une chambre insonorisee.  La partie
gauche de la fig.  2 represente un sujet couche dans un lit
dans la chambre insonorisee.  La frequence du pouls et la tem-
perature du corps etaient enregistrees pendant le somweil et
continuellernent pendant la totalite de 1'experience. De plus,
les enregistrements suivants etaient effectues pendant le som-
meil :
          2  EEC  (potentiels electroencephalographiques)
          1  EOG  (poteritiels oculaires)
          1  EMG  (tonus musculaire)
          raouvements du lit

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                              2036
               Fig.  2   Disposition experimentale

Nos enquetes  (KNAUTH et al.  [2] , RUTENFRANZ et al.  [3] ) ayant
revele que le sommeil de jour des travailleurs de nuit est per-
turbe la plupart du tenps par le bruit cause par la circulation
et par les enfants, nous passions alternativement des enregis-
trements de ces deux genres de bruit pendant le sonuoeil dans
la chambre insonorisee- Tandis que les bruits d'enfants vari-
aient fortement dans leur intensite entre 2O dB(A) et 6O dB(A)
environ  (fig. 3), 1'intensite des bruits de circulation restait
relativement constante entre 45 dB(A) et 7O dB(A) {fig. 4). Le
bruit standardise etait passe a 1'aide d'un magnetophone et par
1'intermediaire d'un haut-parleur environ toutes les 8 minutes
pour une duree de 3 secondes dans la chambre. Les effets de ce
bruit sur 1'EEG, 1'EOG, 1'EMG et sur les mouvements du lit
etaient enregistres. A la fin du sonuneil, les sujets remplis-
saient un questionnaire par lequel on leur demandait d'evaluer
leur dernier sommeil.

-------
                              2037
            dB(A)    bruits  d'enfants
            60
            50
            30-
            20-
            10
                     0        1       2       3 Sec

            Fig.  3   Intensite des bruits  d'enfants

Pendant les heures de travail, les sujets devaient effectuer a
1'Institut un travail de montage a une place de travail indus-
trielle.  Le travail etait interrompu a intervalles reguliers
(toutes les 2 heures 1/2) pour 25 minutes environ. Pendant
cette interruption, on mesurait la vitesse de reaction des
sujets a 1'aide d'un appareil a choix multiple. En outre, les
sujets devaient temoigner de leur forme ressentie subjective-
ment en remplissant un questionnaire scalaire. Nous recueil-
lions enfin a intervalles reguliers des echantillons de 1'urine
des sujets pour en definir ensuite le taux en catecholamines
et en electrolytes.
Les stades de sommeil ont ete determines de facon visuelle d'-
apres RECHTSCHAFFEN et KALES. Us ont de plus ete compares aux
resultats d'une analyse de sommeil automatique  (developpee par
GAILLARD [4] ) .

-------
                              2038
        dB(A)     bruits  de  circulation
        70-

        60-

        50-

        40-
        20
         10
                   0       1        2        3 Sec.

        Fig.  4   Intensite  des  bruits de  circulation

Je voudrais ne presenter ici que quelques-uns des premiers
resultats se basant sur le depouillement des enregistreraents
objectifs du sommeil et sur 1'evaluation subjective du sommeil
de jour aucours des trois semaines de travail de nuit.

3. Resultats
I/es differents parametres du sornraeil temoignent d'une forte
instabilite interindividuelle et intraindividuelle. II est
toutefois possible d'y reconnaltre certaines tendances.
Corane nous avions perturbe le soraweil alternativeroent un jour
par des bruits de circulation et 1'autre jour par des bruits
d'enfants, nous etions en mesure de comparer les differents
effets de ces deux genres de bruit. La fig. 5 represente la

-------
                              2039
duree moyenne du sommeil de jour des deux sujets auxnuels nous
permettions de dormir sans limitation. Le sommeil perturbe par
des bruits de circulation durait en moyenne 49O,9 minutes. D'-
une facon statistiquement significative (p
-------
                              2040
La fig. 6 represente des examples de sommeil d'un  sujet qui
pouvait dormir sans limitation. Les trois dessins  du haut roon-
trent tin raccourcissement progressif du sorameil, en partant  du
sommeil de nuit sans bruit, en passant par le  sommeil  de nuit
avec bruit et en allant jusqu'au premier sonuneil de jour avec
bruit. Pendant la periode de travail de nuit  Her,  9eme et
19eme jour, fig. 6), la duree du sommeil augmentait de nouveau.
Toutefois deux des sujets n'avaient pas la possibilite de de-
terminer la duree du sommeil a leur gre durant la  periode de
travail de nuit.
                                     j    nn	rrmi
                         Itr sonvnt* df iaur one bnrto
                                             AMT«> cfe sommtH
                                                    {La 11
          Fig. 6   Stades de sonraeil de deux  nuits
          et de trois jours d'un sujet

-------
                             2041
La comparaison du sommeil de jour illimite avec le sommeil
limite a 6 heures revelait des differences significatives
(p < O,O1) dans la duree totale du sommeil profond (fig. 7) et du
soauneil PMO.  En cas de sommeil illimite,  le soi^eil PMO durait en
moyenne plus longtemps (1O1,2  min)  que  dans le  sommeil limite
(76,2 min). Alors que les sujets passaient en moyenne 43,6 min
en sommeil profond  en cas de  sommeil illimite,  le stade IV
ne durait en moyenne que 34,5  min dans  le cas de sommeil limi-
te.  Le calcul du pourcentage des stades PMO et  IV de la tota-
lite du sommeil n'a toutefois  pas revele  de differences signi-
ficatives entre le sommeil limite et le sommeil  illimite.
                                          PMO   (REM)
                      St. IV
            50
sommeti limite (360 min)
                                  tOO                [min]
                                    sommeil sons limitation
            Fig.  7   Duree des stades PMO et IV
            pendant le sommeil limite et illimite
                    (x ± e- ? P < O,01)

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                              2042

 II  n'existe  pas jusqu'ici  d'opinion unitaire  sur  les  criteres
 de  la qualite  du soromeil.  On  accorde toutefois dans le  sommeil
 un  role  important aux stades  IV et PMO. Nous  avons done compa-
 re  sur une duree de  trois  semaines de travail de  nuit le  pour-
 centage  des  stades PMO et  IV  de la totalite du sommeil  a  l1-
 evaluation subjective de la qualite du sommeil exprimee par
 chacun des sujets. Ce n'est que dans quelques cas isoles  qu'il
 etait possible de reconnaitre une  tendance unitaire des trois
 parametres.  Alors que les  deux  parametres objectifs variaient
 le  plus  souvent dans la meme  direction, 1'evaluation  subjective
 de  la qualite  du sommeil temoignait d'une evolution divergeante.

 4.  Discussion
 II  est pratiquement  impossible  de  separer les effets  des  deux
 facteurs : bruit et  travail de  nuit.  Le changement du rhythme
 de  vie consecutif au travail  de nuit a dans une certaine  mesure
 une influence  sur les nombreuses fonctions du corps humain
 sound.s a un  rhythme  circadien.  Le  sonroeil se  trouve egalement
 change a la  suite du decalage des  phases des  activates, du fait
 que le cours de chacun des stades  est lie a 1'heure de  la jour-
 nee.
 Le  bruit etant en general  beaucoup plus fort  le jour  que  la
 nuit, il faut  s'attendre   en  cas de  sommeil de jour des
 travailleurs de nuit a une reduction de la duree  et a une
 degradation  de la qualite  du  sommeil.
 Dans notre experience, 1'intensite des bruits d'enfants n'-
 atteignait pas la valeur maximale  des bruits  de circulation.
 Cependant les  bruits d'enfants  - sans doute parce qu'ils  sont
 plus varies  et plus  riches en informations -  perturbaient plus
 fortement  le sommeil  que les  bruits de circulation.
 La  reduction de  sommeil a  6 heures, habituelle chez les tra-
 vailleurs de nuit, diminuait  la duree du sommeil PMO  de meme
que la duree du  sommeil profond. Ces deficits n'ont pas ete
compenses au cours de  la jseriode de trois semaines de travail
de nuit.  C'est pourquoi nous estimons que les plans de roule-
ment ou se succedent plusieurs nuits de travail  ne sont pas
recommandables.

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                             2 043
 Bibliographic

 1.  Knauth,  P. und Rutenfranz,  J.,  "Untersuchungen zum Pro-
    blem des Schlafverhaltens bei  experimenteller Schicht-
    arbeit", Internationales Archiv fur Arbeitsmedizin, 3O,
    1  - 22 (1972)

 2.  Knauth,  P. und Rutenfranz,  J.,  "Untersuchungen iiber die
    Beziehungen zwischen Schichtform  und Tagesaufteilung",
    Internationales Archiv fur  Arbeitsroedizin, 3O, 173 - 191
    (1972)

 3.  Rutenfranz, J., Knauth, P., Hildebrandt, G. und Rohmert,W.,
    "Nacht- und Schichtarbeit von Triebfahrzeugftihrern,
    1. Mitteilung. Untersuchungen tiber die tSgliche Arbeits-
    zeit und die Ubrige Tagesaufteilung", Internationales
    Archiv fur Arbeitsroedizin,  32, 243 - 259  (1974)

 4.  Gaillard, J.-M., Simmen, A.E. et Tissot, R.,  "Analyse
    automatique des enregistrements polygraphiques de
    sommeil", Electroencephalography and Clinical Neuro-
    physiology 3O, 557 - 561 (1971)
                         DISCUSSION
STUPFEL (France)
     Pourquoi avez-vous utilise des bruits d'enfants et non
des bruits de circulation ou des bruits d1ateliers?

KNAUTH (Republique federale d'Allemagne)
     Nos enquetes par questionnaire effectuees aupres d'ouvriers
effectuant un travail par roulemment ayant revelS que le som-
meil de jour des travailleurs de nuit est perturbe" la plupart
du temps par les bruits de la circulation et les bruits causes
par les enfants, nous avons lors des essais perturbe le sommeil
des personnes par ces deux genres de bruit.

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                              2044
JANSEN (REpublique fEdErale d'Allemagne)

1.   Quels Etaient les niveaux acoustiques continus Equivalents
enregistrEs lors de vos essais?
(Les valeurs guides mentionnent toujours des niveaux acoustiques
continus equivalents)

2.   Est-ce que ce sont les niveaux maximaux ou les niveaux
sonores constants Equivalents qui sont plus importants?


KNAUT (RSpublique fEdErale d'Allemagne)

1.   Nous n'avons pas encore calculE des "niveaux acoustiques
conti nus Equ ivalents".

2.   D'apres les premiers dEpouillements, ce ne sont pas les
pointes d1intensity sonore qui semblent avoir jouE un r61e
plus important mais les variations du niveau de pression sonore
et le contenu d'information de bruit.  Les bruits d'enfants
avec une intensity maximale de 6O dB(A) (figure 3) perturbaient
beaucoup plus fortement le sommeil que les bruits de la circula-
tion avec une intensity maximale de 7O dB(A) (figure 4).  La
durEe totale du sommeil, la durEe du sommeil REM (PMO) ainsi que
du sommeil profond Etaient nettement plus courtes quand le
sommeil Etait perturbE par des bruits causes par des enfants
que lors d'une perturbation par le bruit de la circulation.


VAN MEIRHAEGHE (Belgique)

     Je voudrais avoir quelques precisions sur 1'Evaluation
subjective du sommeil par les sujets et, notamment, si ceux-ci
dEclaraient par exemple que leur sommeil avait EtE troublE par
des reves en rapport avec la nature de bruit.


KNAUT (REpublique fEdErale d'Allemagne)

     A la fin du sommeil, les sujets devaient remplir un question-
naire dans lequel on leur demandait d'apprEcier la qualitE de
leur sommeil, d'indiquer comment ils s'Etaient endormis, les
perturbations ressenties pendant le sommeil et leur condition
personnelle.  Les rEsultats de ces Evaluations subjectives du
sommeil correspondaient rarement avec les enregistrements ob-
jectifs.

     Alors que les sujets dEclaraient par exemple s'etre habituEs
au bruit au bout de quelques jours, aucun indice d* aocoutumance
Eventuelle n'Etait dEcelable dans les enregistrements des EEC,
EOG et EMG ainsi que dans les mouvements du lit.

     Un sujet a dEclarE apres une pEriode de sommeil perturbE
par des bruits de la circulation avoir revE plusieurs fois de
bruits d"avion.

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                              2045
CHAMBERS (Irlande)

     II m"interesserait de savoir comment le stimulus acoustique
a €t£ applique; est-ce que le bruit a 6te emis au hasard ou S
intervalles specifiques?  Je pose cette question parce que si
la frequence du stimulus correspondait a des p&riodes de sommeil
REM  (mouvements oculaires rapides), il se peut que se produise
un surcrolt de sommeil REM similaire a celui que d^crit Dement
dans son ouvrage sur la privation de sommeil REM.  Meme si vous
n'avez pas conside're' cela comme un phenomene, il devrait vous
etre possible, en examinant vos notes, de verifier s'il y a eu
coincidence entre le stimulus et le sommeil REM afin de confirmer
ou d'infinner cette hypoth&se et, le cas §cheant, d'eliminer cet
effet possible.


KNAOT  {Rgpublique fedSrale d'Allemagne)

     Lors de cette €tude nous avons passe1 un bruit toutes les
8 minutes dans la chambre insonoris§e de fa^on a ce que tous les
stades du sommeil soient perturbes.  Nous avons choisi ce bruit
standardise et rfigulier pour placer les sujets dans les memes
conditions d'essai.  Toutefois, nous envisageons pour nos essais
futurs de perturber Sgalement le sommeil par des bruits presented
au hasard.

     Jusqu' a present, dans nos essais, la dure"e totale du sonuneil
REM  a  toujours €t6 nettement plus courte dans le cas d'un sommeil
perturb^ que dans celui d'un sommeil non perturbe".   Si toutefois,
on compare sommeil REM au sommeil total, on ne constate pas de
difference significative entre la proportion du sommeil REM dans
le cas du sonuneil perturb^ et cette poportion dans le cas du
sommeil non perturb^.


 KREUZER (Rgpublique f£d§rale d'Allemagne}

      Se trouvait-il des pSres de famille parmi les sujets exposes
 aux bruits  d 'enfants?

      Ces sujets-la devraient au moins reconnaitre qu'ils sont
 plus sensibles aux bruits d1enfants qu'aux autres bruits de
 meme intensitS.   Le cot€ psychique doit probablement jouer ici
 un grand role.

      Cela explique peut-etre aussi, dans le cas oQ des p^res de
 famille se trouvaient parmi les ouvriers travaillant par roule-
 ment, que I1on ait observe une amplification in^gale de la sen-
 sibilit§ aux bruits d'enfants.


 KNAUT (Rfipublique f^d^rale d'Allemagne)

      Sachant que les parents r^agissent aux bruits de leurs
 propres enfants autrement qu'aux bruits d'autres enfants, nous
 n1avons choisi que des sujets qui n'etaient pas peres de famille.

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                              2047
      ALTERATION IN URINARY D-GLUCARIC ACID EXCRETION AS
          AN INDICATION OF EXPOSITION TO XENOBIOTICS

             W, R, F, NOTTEN AND P,  TH,  HENDERSON
Institute of Pharmacology, University of Nijmegen, Netherlands
ABSTRACT

     Many compounds foreign to the organism, particularly those
which due to their high lipophilicity tend to remain for a long
time in the body, are capable of stimulating a number of enzymes
in the liver.   On the one hand this affects the  enzymes involved
in the conversion of lipophilic toxic substances  into more hydro-
philic, non-toxic products.   On the other hand the metabolism
of "normal" body constituents can also be influenced.   This
stimulation, as a rule, becomes manifest in an enhanced excretion
of metabolites by the urine.   A well-known example is the stim-
ulation of the D-glucuronic acid pathway in the liver, which
leads to an enhanced excretion of L-ascorbic acid (in mammals
excepting primates and the guinea-pig) and D- glucaric acid.

     In view of  this, it  can  be questioned whether the measure-
ment of an enhanced urinary D-glucaric acid - as  a reflection  of
an altered  liver metabolism - can serve  as a reliable  test for
the  effect of  exposure to body-foreign compounds.  Such  a test
might  be especially  important when  exposure  takes place  chronic-
ally to known  agents at  subtoxic  concentrations,  for  which spe-
cific  test methods  are absent or  hard  to perform, and in  those
cases  where  exposition occurs to  a  varied  spectrum of unknown
xenobiotics.

-------
                              2048
     During the present study a number of xenobiotic compounds
(some drugs, organic solvents, pesticides and other environmental
pollutants) were screened for their capacity to enhance urinary
D-glucaric acid excretion in the guinea-pig and rat.   The ex-
perimental results support the usefulness of the D-glucaric acid
test as a measure of general exposition.

     As far as the mechanism of the stimulation is concerned, it
appears that the increased synthesis of D-glucaric acid is dir-
ectly related to an acceleration of the conversion of UDPglucose
to UDPglucuronic acid.

     The applicability  of the D-glucaric acid test for man in
the field of industrial and environmental hygiene, is discussed.

-------
                                  2049
I-                         INTRODUCTION
    In industrial  and  environmental  hygiene  it  is  of  great  importance
to  estimate exposition  to  toxic substances  as  early  as  possible and
at  concentrations  which do  not yet  lead  to  intoxication phenomena.
Periodical diagnosing of persons or animals  for exposure to xenobio-
tics at subtoxic  concentrations may serve a  double purpose. On the
one hand one can  guard  against unacceptable  toxic damage by excluding
from further exposition, on the other hand  indications  can be found
in  order to take  adequate  measures  with  respect to the  sources of pol-
lution.
    As amatter of  fact,  when exposition to certain known agents is the
case, as often occurs in industry,  specific  tests are relevant. Houi-
ever, if suitable  specific  test methods  are  lacking  or  hard to per-
form, and in those cases where exposition occurs  to  a broad spectrum
of  unknown chemicals, it mould be of great  value  to  have the disposal
of  one or more non—specific test methods.
1-1.                  The D-glucaric acid test
    Evidence has accumulated regarding the ability of many compounds,
particularly those which due to their high lipophilicity tend to
remain for a long  time  in the body, to induce  the activities of the
                                 1—3
varzous drug-metabolizing enzymes   . It appeared also  that exposure
to  chemicals foreign  to the organism often leads to  a stimulation of
the glucuronic acid pathway, in which by sequences of biochemical
processes hexose is transformed via D—glucuronic acid to L—ascorbic
acid, 0-xylulose and D-glucaric acid (see figure l). Already in 1940
Longenecker et al.  reported that various drugs administered to rats
enhance the urinary L-ascorbic acid excretion. Further  investigations
extended this phenomenon to other compounds and up to now a stimulat-
ive influence is known  of a large variety of xenobiotic  agents '  '
From this the suggestion was made that an enhanced urinary excretion
of L—ascorbic acid may be used as an indicator of drug—induced alter-
                                                                1 B
ations in the activities of drug-metabolizing enzymes in animals '  .
It is of particular importance for man,  who is unable like other
primates and the guinea-pig to synthesize L—ascorbic acid,  that the
stimulative effect of drugs on the glucuronic acid pathway also re-
                                                         11-14
suits in an enhanced urinary excretion of D-glucaric acid
   It was the purpose of the present study to find out if an altered

-------
                                  2050
                       D-galactooe
                                  *
                              UDPG
                                   NAD
                                   UDP6 dehydrogenctse
                              UDPGA
                                  \
                              se  VOPglumronyl tmnsfc
                         *             »
                     D-«flucurOr»e         gfucurpnidea
                   add-1- phosphite
                                       L-«acorbic fcid
             Figure 1. The D-glucuronic acid pathway.
urinary D—glucaric acid excretion, in its  turn,  can  be considered as
an indication  of  previous intake of certain  chemicals. In that case,
the so—called  0—glucaric acid test might be  of  value as a non-select-
ive exposition test.
2.                          METHODS
2.1.    Aninals
   •ale Wistar rats (175-230 g) and male guinea-pigs (195-205 g) mere
utilized. All  compounds were administered  i.p.  or  p.o. (see tables).
Control animals received saline or sesame  oil in equivalent quanti-
ties.
   For the Measurements of the in vitro activities of the liver en-
zynes, the animals were killed by decapitation.  The  livers uiare rapid-
ly removed and cooled in ice. Portions of  liver  mere weighed, finely
minced, and transferred into 4 vol. of ice-cold  0.25 (ft sucrose sol-
                        ^Q
ution containing  5 x 10~ M tris(hydroxymethyl)aminomethane-HCl (pH
7.4). Homogenates were prepared using a Teflon-glass homogenizer.

-------
                                 2051
TABLE I. EFFECT OF CHRONIC TREATNENT WITH XENQBIOTICS ON IN VITRO

ACTIVITIES OF HEPATIC N-DEHflETHYLASE AND UDP-GLUCURONYLTRANSFERASE
compound dose*
N-demethylation
of aminopyrine
control
hexachloro—
benzene
heptachlor
disulf iram
Perthane
toluene
tatraethyl-
lead
Aroclor 1260
ethanol
n-hexane
Dodin
Atrazine
dimethoate
nitrobenzene
aniline
benzene
10
4
300
100
60
1.5
1
100
3
14
10
7
0.5
1
1
phenylmercur— 1 .5
ic acetate
77
77
95
90
85
90
108
90
108
77
83
77
108
85
108
90
+
+
+_
±
^
j+_
+
+
+
+
+
+
±
+
±
+
9
9
6
5
8
5
9
5
9
9
4
9
9
8
9
5
treated
77
81
143
123
108
105
94
111
91
37
86
55
107
114
123
95
± 1
i 2
?xxxx
xxxx
+. 8
±3X
± 5
± ?X
± 4
?xxx
± 4
±lx
± 5
±4X
± 7
± 8
UDPglucuronidation
of p-nitrophenol
control
52
52
58
45
46
45
49
45
49
52
46
52
49
46
49
45
+
i
+_
+
+_
+_
+
+
+_
+
±
+_
+_
+
+
3
3
2
4
2
6
2
6
3
5
3
6
4
6
2
treated
64
46
73
46
44
46
67
54
70
44
41
47
77
60
69
26
± 1
xxxx
± 1
± 4
± 1
±3X
+ 2XX
±4"
± 4
± 5
_+ 2
XX
+ o
±5X
±5X
.XXX
+ 4
 in mg/kg/day; all compounds were administered i.p. in sesame oil,
 except for disulfiram, which was suspended in saline and given p.a.
_Ths duration of treatment is given in tablell.
 Expressed as 10~8 moles formaldehyde produced per hour per mg of
 microsomal protein.
'Expressed as 10"^ moles p-nitrophenol conjugated per hour per mg of
 microsomal protein.
 Values are means (_+_ S.E.RI.) of six animals.
xxxx: indicates significantly different from control at P < 0.001;
 xxx: idam at 0.001 < P < 0.01; xx: 0.01 < P < 0.02; x: 0.02 < P <
 0.05; for all further values P > 0.05 (Student's t-test).

-------
                                 2052
Crude microsomal fractions were prepared from the homogenates by
centrifugation at 9,000 g for 20 min at 2 °C.
2.2.    Determination of D—glucaric acid in the urine
                                                                 19
   0—glucaric acid was estimated according to the method of Marsh
from the inhibitory effect of D-glucaro-l,4-lactone, to uihich it is
converted by heating at pH 2, on p-glucuronidase.
   Creatinine concentrations in the urine were estimated according
                        20
to Gorther and de Graaff
2.3.    Enzyme assays
   •icrosomal N-demethylation of aminopyrine was measured in 9,000 g
                                                  21
supernatants as described by Henderson and Karsten
   UDPglucuronyltransferase activities were determined in 9,000 g
                                   22
supernatants according to Henderson  , except that for activation of
the enzyme the eupernatants were treated with the detergent Triton X-
100 (0.25JC, final cone.) instead of sonication. p-Nitrophenol was
used as acceptor substrate.
2.4.    Protein assay
   Protein concentrations were determined following the method of
            *}*l
Lowry et al.  . Bovine serum albumin (Sigma) was employed as refer-
ence standard.
3.                  RESULTS AND DISCUSSION
3.1.    Comparison of the effects of chronic treatment with xeno-
biotics on drug metabolism and on urinary excretion of D-glucarate
   It has been postulated by Hunter  *  , Latham   and others that
the urinary 0—glucaric acid excretion might act as an indirect estim-
ate of hepatic enzyme activity. In the present study, therefore, we
tried to find out if alterations in enzyme activities involved in
drug metabolism appear after treatment with various compounds, and
•nether such alterations are paralleled by similar changes in urin-
ary 0—glucarate levels.
   Guinea-pigs were treated with relatively low doses of different
xenobiotics, including some drugs, pesticides, hydrocarbons, for a
fa* weeks. The effect on the in vitro activities of the oxidative
N-demethylation of aminopyrine and the UDPglucuronidation of p-nitro-

-------
                                 2053
TABLE II. EFFECT OF CHRONIC TREATfflENT WITH XENOBIOTICS ON THE URINARY

           EXCRETION OF 0-GLUCARIC ACID IN THE GUINEA-PIG
compound dose
D-glucaric acid
(microgram/24h)
control
haxachloro—
benzene
heptachlor
disulfiram
Perthane
toluene
tetraethyl-
lead
Aroclor 1260
ethanol
n-hexane
Dodin
Atrazine
dimethoate
nitrobenzene
aniline
benzene
phenylmercur-
ic acetate
10
4
300
100
60
1.5
1
100
3
14
10
7
0.5
1
1
1.5
80
80
86
76
72
76
74
76
74
80
79
80
74
74
74
76
± 6
± 6
± 3
± 7
± 7
± 7
± 5
± 7
± 5
± 6
± 6
± 6
± 5
i. 5
± 5
± 7
treated
372
301
166
122
96
103
98
101
95
103
94
94
86
82
78
46
XXX
± 25XXX
± axxx
±12XX
±*X
± ?X
± 6X
±6X
±*X
±?X
± 2X
± l:L
± B
± 8
± 9
±iox
per cent
of
control
465
376
193
157
133
132
132
129
128
128
119
118
116
111
105
51
treatment
period
(days)
A B
21
21
e
21
21
21
35
21
35
21
21
21
35
35
35
21
7
7
3
14
21
21
35
14
35
21
21
-
-
-
-
14
 +in  mg/kg/day;  all  compounds  mere  administered  i.p.  in sesame  oil,
  except  for  disulfiram,  which vuas  suspended in  saline  and  given  p.o.
 •Expressed as  microgram/24 h/100  gram body weight;  values  are  means
  (^ S.E.ffl.)  of  six  animals.
 B:  indicates the day at  which the  first significant  (P < 0.05) alter-
  ation in  the  urinary excretion of D-glucaric acid  mas observed.
 A:  indicates the total duration of the treatment. The  D-glucaric acid
  levels  mentioned were measured during the last 24  h of this period.
 xxx: means significantly different from control at  P < 0.001;
  xx: idem, at  0.01  < P < 0.02; x:  0.02 < P < 0.05.  For all further
  values  P  >  0.05 (Student's t-test).

-------
                                  2054
TABLE III. EFFECT OF  SHORT-TERM TREATItlENT UIITH RELATIVELY  HIGH DOSES
OF SOBIE XENOBIOTIC AGENTS ON THE URINARY EXCRETION OF D-GLUCARIC  ACID
compound
barbital sodium
phenobarbital
nikethamide
chlorbutol
DDT
Perthane
D-glucaric acid++ .
control
104 ±
104 +.
104 ±
95 ±
95 _+
151 ±
8
6
B
B
8
14
treated i
330
224
272
135
188
423
™CXXX
+ oo
+ 24XXX
± 36XX
±14X
+ 20XX
± 49XXX
ser cent
of
control
317
215
261
142
198
324
treatment
period
(h)
8
8
8
B
8
24
*barbital sodium (150 mg/kg, i.p.), phenobarbital (130 mg/kg, i.p.),
 nikethamide (130 mg/kg, i.p.), chlorbutol (150 mg/kg, P.O.), and
 DDT (60 mg/kg, p.o.) dissolved in saline tuare administered to male
 Wistar rats (175-230 g) in turn doses, at zero time and at 4 h. Con-
 trols received an equivalent volume of saline. Perthane dissolved in
 sesame oil utas administered i.p. (500 mg/kg) to male guinea-pigs
 (195-205 g); control animals received an equivalent volume of sesame
 oil.
  Amount of D-glucaric acid present in the urine collected during the
 whole period. Values represent means (*_ S.E.RI.) of six animals.
xxx: significantly different from the control at P < 0.001; xx: at
 0.001 < P < 0.01; x: at 0.02 < P < 0.05 (Student's t-test).
phenol (two typical examples of drug-metabolizing reactions)  is
demonstrated in table I. Only disulfiram, ethanol and aniline  treat-
ment led to an enhanced rate of N-demethylation and glucuronidation.
Perthane, tetraethyllead, Oodin and dimethoate increased only  the
N-deraethyJating activity, whereas hexachlorobenzene, Aroclor  1260,
n-hexane, nitrobenzene, and benzene enhanced only the activity of
UDPglucuronyltransferase. In contrast, the glucuronidating activity
was found to be reduced after treatment with phenylmercuric acetate.
Heptachlor, toluene and Atrazine exhibited no influence on the in
vitro activities of these drug-metabolizing enzymes. From these re-
sults it can be concluded that with regard to drug metabolism  sever-
al of the compounds can act as specific inducers.

-------
                                 2055

   As is seen in table II, treatment of the guinea-pigs mith most of
the chemicals under investigation led to a significantly enhanced
excretion of D-glucaric acid. Only dimethoate, nitrobenzene, aniline,
and benzene did not exhibit a measurable influence, whereas phenyl-
mercuric acetate markedly inhibited the urinary D-glucaric acid
excretion.
   Obviously, induction of the drug-metabolizing system is not always
accompanied by an enhancement of the urinary D-glucarate excretion,
as is demonstrated by the action of dimethoata, nitrobenzene, aniline
and benzene. On the other hand, heptachlor, toluene and Atrazine
stimulated the glucuronic acid pathway, but left the drug-metabol-
izing enzymes unaffected.
3.2.  Effect of short-term treatment on the D-glucaric acid excretion
    The conclusion that stimulation of the D-glucuronata pathway,
 leading to an enhanced D-glucaric acid excretion,  is not necessarily
 related to an induction of the drug-metabolizing enzymes is further
 supported by the results from some short-term experiments.
 Rats ware treated for 8 hours with relatively high doses of barbital,
 phenobarbital, nikethamide,  chlorbutol, or DDT. Guinea-pigs ware
 treated with Perthane for 24 hours, as indicated in table III.
    No alteration in N-demethylase and UDPglucuronyltransferase act-
 ivities could be observed at the end of the treatment period. However
 as appears from the data given in table III, all compounds caused a
 significant enhancement of the D-glucaric acid excretion within 8,
 or 24 hours, respectively.
    Further experiments pointed out that stimulation of the D-glucur-
 onate pathway presumably is based on an accelerated synthesis of
 UDPGA. It appeared also that, at least in cases of short-term treat-
 ment, this stimulation is independent of a drug-induced de novo
 synthesis of LJDPGdehydrogenase, the enzyme which catalyzes the  con-
 version of UDPG into UDPGA
 3.3.  Normal values of the urinary D-qlucaric  acid excretion in man
    On account of the results obtained from the  animal  experiments
 as mentioned above, the D-glucaric acid excretion  can  be  used as  an
 index of  hepatic enzyme induction  only in some  particular  cases.
 But,  as it appeared that, irrespective of an  enzyme  induction,  in  the
 majority  of  cases  treatment u/ith  xsnobiotics  caused  a  clear  alterat-

-------
                                 2056
TABLE IV.  NORMAL  VALUES  OF URINARY  D-GLUCARIC  ACID  EXCRETION  IN  R1AN
 urine
 urine
                          D-glucaric acid/mg  creatinine
                 mean
                          inner limits  for
                                                outer limits for
                            2.5
                                     97.5
 2.5
                                97.5
collected
for 24 h
(A)
collected
overnight
(B)
A minus 8
9.9 5.4 - 15.5

9.8 5.2 - 15.3
10.3 4.3 - 17.6
3.7 - 17.2

3.4 - 16.0
2.5 - 20.7
jig D-glucaric acid/hour
                 mean
                          inner limits for
                                                outer limits for
                            2.5
                                     97.5
                       2.5
          97.5
 collected
 for 24 h
   (A')
 collected
 overnight
   (B')
 A'minus B'
                 635
                 603
                 663
349  -  999
260  -  1150
307  -  1083
                                                 219  -  1373
177  -  1253
                                                 191  -  1437
The confidence intervals for the percentiles P    and P__
                                              2*3      y § •
constituted u/ith f = 90%. The number of samples was 110.
                                                            are
ion in the D— glucaric acid level, the usefulness of the D— glucaric
acid excretion as a non-selective exposition test has to be serious-
ly considered. This prompted us to extend our studies to the D-gluc-
aric acid excretion by the urine in man.
   Normal values were determined according to the procedure described
                    1 fl
by RUmke and Bezemer  . In this method for statistical analysis of
normal values two types of tolerance limits are distinguished. The
inner limits for percentiles are recommended as warning limits, the
outer limits as limits for more descriptive purposes,
   With respect to the general applicability of the D-glucaric acid
test collecting of urine for 24 hours may encounter some practical

-------
                               2057
                  jjg D glucarate /mg creatinine
                  3O
                  20
                  IO
                            • •

                           t
                         CONTROL
                                     ENDFMN
Fig. 2.  Comparison of the D-glucaric acid  excretions of some
         industrial workers exposed to endrin  with those of
         some  office workers.   The broken  horizontal line
         indicates the upper limit of the normal  values
         (Table  111).    Note:  nearly all values  of the Endrin
         group are beyond this limit.

-------
                                  2058
difficulties. Therefore, normal values of D-glucaric acid related to
the urinary creatinine concentration, have been compared with the
D-glucaric acid excretion per hour, both for 24 hours urine collect-
ions and urine samples collected overnight. 110 Young healthy human
subjects mere tested in this study. The results are presented in
table IV. No substantial fluctuations in the daily D-glucaric acid
occur. Further, it can be derived that reliable values of the D-gluc-
aric acid excretion can be obtained if expressed per mg creatinine.
Further investigations, in u/hich persons working in different bran-
ches of chemical industry are tested for their urinary excretion of
D—glucaric acid, in order to assess the validity of the test in
industrial and environmental hygiene, are in progress now.  (Fig. 2)
                    Acknowledgement
   This work mas supported by a grant from the Prevention Fund of the
Dutch Ministry of Health and Environmental Hygiene.
                      REFERENCES
1   CONNEY, A .H.,"Pharmacological implications of microsomal enzyme
    induction", Pharmacol. Rev.. 19, 317 (1967).
2   ZEIDENBERG, P., ORRENIUS, S, ERNSTER, L., 3. Cell Biol., 32,
    528 (1967).
3   BIULDER, G.3., Biochem. 3.. 117, 319 (1970).
4   BURNS, 3.3., EVANS, C., 3.Biol. Chem., 223, 897 (1956).
5   BURNS, 3.3., EVANS, C., TROUSOF, N., 3. Biol. Chem., 227, 785
    (1957).
6   LONGENECKER, H.E., FRICKE, H.H., KING, C.G., 3. Biol. Chem..
    135, 497  (1940).
7   CONNEY, A.H., BURNS, 3.3., Nature. 184, 363 (1959).
8   AARTS, E.M., Biochem. Pharmacol., 17, 327 (1968).
9   CONNEY, A.H., BRAY, G.A., EVANS, C., BURNS, 3.3., Ann.  N.Y.  Acad.
    Sci.. 92,  115 (1961).
10  CONNEY, A.H., GILLETTE, 3.R.,  INSCOE, 3.K., TRANS, E.R., POSNER,
    H.S., Science. 130, 1478  (1959).
11  BARSH, C.A., REID, L.B.,  Biochim. Biophys. Acta.  78, 726 (1963).
12  HUNTER, 3., CARELLA, IB.,  BIAXIKELL, 3.D., STEWART,  D.A.,  WILLIAMS,
    R-» Lancet  I, 572  (1971).
13  OKADA, «.,  MATSUI, IB., KAISU,  T., ABE, F., Chem.  Pharmac. Bull..
    17, 2625  (1969).
14  HUNTER, 3., WAXltfELL, 3.D.,  STEU/ART, D.A., UJlLLIAfflS,  R.,  Nature.
    237, 399  (1972).

-------
                               2059
 15  HUNTER,  :).,  IflAXUJELL, D.D.,  STEWART, D.A.,  WILLIAMS, R.,
    Biochem. Pharmacal., 22, 743  (1973).

 16  AARTS, E.flf!.,  Biochem. Pharmacol.. 14, 359  (1965).

 17  NOTTEN,  U/.R.F., HENDERSON,  P.Th., submitted  for publication.

 18  RUIT1KE, Chr.L.,  BEZEfflER, P.O.,  Ned. T. Genessk., 116, nr. 35,
    1559 (1972).

 19  NARSH, C.A.,  Biochein. 3.. 87,  82 (1963).

 20  GORTHER, E.,  de GRAAFF, U/.C.,  Klinische DiagnostiBk. 7e druk,
    p. 440,  Stenfert Kroese N.V.,  Leiden (1955).

 21  HENDERSON,  P.Th., KERSTEN,  K.J., Biochein.  Pharmacol.. 19,  2343
    (1970).

 22  HENDERSON,  P.Th., Life Sciences. Part II.  9, 511 (1970).

 23  LOli/RY, O.H.,  ROSEBROUGH, N.3., FARR, A.L., RANDALL, R.J.,
    3. Biol. Chem.. 193, 265 (1951).

 24  LATHAm,  A .N.,  3. Pharm. Pharmac.. 26, 285  (1974).
                        DISCUSSION


de BRUIN  (Netherlands)


1.   In connection with the observation  that several non-enzyme
inducing  agents  also cause anoumalous  excretions, which mechanisms
could be  responsible for the augmented elimination of L-glucaric
acid, apart  from microsomal enzyme  induction?

2.   Is there  any evidence that compounds  which inhibit drug
metabolizing enzyme systems, either directly or by way of hepa-
totoxic damage,  might have a lowering  effect upon L-glucaric acid
output?

3.   Can  the sensitivity of the glucaric acid test be indicated,
especially as  compared to similar enzyme-inductive tests, such
as alteration  of drug disposition or change in endogenous steroid
hydroxylation?


NOTTEN  (Netherlands)

1.   As mentioned already the stimulation of the glucuronate
pathway presumably is based upon an enhanced synthesis of UDPGA
from UDPG.   In case of chronic treatment this may be due to an
increased synthesis of the non-microsomal enzyme UDPG-dehydro-
genase.   From  our short-term experiments it appeared that urinary
glucaric  acid  has been elevated even within 8 hours.  This indi-
cates that another mechanism is involved.   One can imagine, that
in this case,  the conversion of UDPG into UDPGA can be accelerated
as a consequence of altered enzyme  biometrics.  On the other hand,
since it  was found in some preliminary studies that an enhanced

-------
                                2060



 synthesis of D-glucaric acid is accompanied by an inhibition of
 the glycogen synthesis, the possibility exists that more UDPG
 becomes available as substrate for the glucuronate pathway.

      So, it is likely that stimulation of the D-glucuronate
 pathway and induction of microsomal drug metabolism are regulated
 by different mechanisms.

 2.   Yes, phenylmercurie acetate lowers the glucuroxidation rate
 (table I) and at the same time already inhibits the D-glucaric
 acid synthesis (table II).  The same phenomenon has been observed
 with disulfriam after a 24-hour treatment.

 3.   As far as the sensitivity in comparison with other tests is
 concerned, no studies have been done.

      The D-glucaric acid test, however, is preferable to the
 tests mentioned by you, because analysis of D-glucaric acid is
 rapid and easy to perform.  This in contrast to a determination
 for example 6 -/J - hydroxy steroids  (enzyme induction test) .
 On the other hand, in the D-glucaric acid test no drugs have
 to be administered to the persons being tested, as is the case
 of a drug disposition test (antipyrine).  Hence, the D-glucaric
 acid test is more suitable as a general test.


 ZIELHUIS (Netherlands)

 1.   How long does increased  glucaric acid excretion persist
 after exposure?  In the Netherlands Jager found increased ex-
 cretion in ex-endrin workers.   Endrin has a very short biological
 half life.   Could you explain that?

 2.   In our experience the intra-individual coefficient of var-
 iation of ALA-excretion if measured per gram of creatinine is
 about similar as if measured  per hour in case of overnight urine.
 What is your opinion on the comparative coefficients of variation
 of both methods?   In my opinion it is certainly possible in
 occupational health studies to apply the time-correction method-,
 it only demands some extra organisation and it does not load
 the laboratory with creatine measurement.


 NOTTEN (Netherlands)

 1.   We have  not measured how  long  the  increase of  D-glucaric
acid excretion  lasts  after stopping  the treatment.   Therefore,
we cannot explain this.

2.   We also  found comparable variation coefficients of D-glucaric
acid excretion both if related to the urinary creatinine concen-
tration and if expressed per hour.   The only reason  for expressing
 urinary D-glucaric acid per mg creatinine, is a practical one.
We prefer the very simple routine-assay of creatinine to the extra
risks of making mistakes in a more complex sampling  scheme of
urines.

-------
                               2061
          THE ASSESSMENT OF ENVIRONMENTAL CARCINOGEN
               RISKS IN TERMS OF LIFE SHORTENING

              ROY E, ALBERT AND BERNARD ALTSHULER

Institute of Environmental Medicine, New York University Medical
Center, NY, USA
ABSTRACT

     This paper presents an approach to the assessment of carci-
nogen risks in which the dominant effect of carcinogen exposure
is life shortening and the impact falls both on those individuals
who would have got cancer without the carcinogen exposure as well
as the new cancer cases.   This analysis is based on the inter-
action of age-specific tumor incidence rates and population sur-
vival in terms of age-specific mortality rates without the in-
duced risk from carcinogen exposure;  the analysis yields estimates
for life-time probability of developing cancer, average life-span
lost by the entire population, the average age of cancer occur-
rence and the average life-span loss of cancer cases.   The
approach utilizes the animal response data to assign, to the
existing human cancer occurrence, an equivalent dose of the same
carcinogen which is under consideration in terms of risk evalua-
tion.   The approach has the advantages of (a) keying the estimates
of carcinogen risks to those which already exist in the environ-
ment, (b) avoiding large extrapolations from animal data, and
(c) encompassing the variability in susceptibility and carcinogen
exposure in humans.

-------
                              2062
      Carcinogens can pollute the environment from techno-
 logical processes that are too important to abandon, as for
 example, the combustion of fossil and nuclear fuels for
 production of electricity.  Hence, it is necessary to
 estimate the magnitude of cancer risks from environmental
 contamination as an essential part of the process of weighing
 the  costs  of controlling the release of carcinogens against
 the consequences of deleterious health effects and thus to
 make a rational choice between alternative technological
 processes that achieve the same ends.

      The assessment of cancer risks from exposure to known
 environmental carcinogens is an exceedingly difficult
 problem.  Carcinogen exposures will never be tolerated unless
 they are expected to cause negligible increases in the
 existing burden of cancer; unless there is a grave miscalcu-
 lation in the formulation of exposure limits,  the actual
 risks could never be feasibly measured in human and certainly
 not in experimental animals.   There are additional uncer-
 tainties associated with differential sensitivity amongst
 humans and  between humans and test animals.

      In principle,  the  only feasible basis for  risk extrap-
 olations to very low levels of carcinogen exposure is  to
 develop a sound understanding in  animals  of  the component
 processes that  determine the  dynamics of  tumor  formation and
 thereby establish the general principles  for making risk
 extrapolations.   It is  also necessary to  use epidemiologic
 data  on human cancer in response  to  define levels of exposure
 in  order to equate  the  relative sensitivities of  human  and
 animal  for  particular target  organs  and carcinogens.  The
 conventional  method of  assessing  carcinogen  hazards  is  done
 by  relating the  level of  dose to  cancer incidence.  This
 implies that  the extra  cancer cases bear  the full effect of
 the carcinogen  exposure and the rest  of the  population  suffers
 no  ill  effects.   No  attention is  paid to  the age  at which  new
 cancer  cases  occur or to  the  possibility  that additional
 carcinogen  exposure  could  affect  individuals who  were going
 to  get  cancer from other  causes.  The  shape  of the dose-
 incidence curve  used for extrapolations is arbitrary as  for
 example  in  the case  of  the Mantel-Bryan approach  which uses
 a log-normal  dose-response  curve with  a slope of  one probit
 per log dose  (1).  The  thrust of  this  paper  is that the
 temporal patterns of mortality-corrected  tumor incidence
 should  be used as the primary basis for characterizing tumor
 responses from chronic  carcinogen exposure since  they are
 more directly related to the time-dependent processes of
 neoplastic cell transformation and growth of transformed
 cells into tumors; the  incidence and age of at which tumors
 form depend on the interaction of the temporal tumor response
 patterns with population survival.  This approach provides  a
more complete characterization of carcinogen risks since the
effects are defined not only in terms of the excess cancer
 incidence but also in terms of life shortening.   Furthermore,
the approach provides a way to link the responses observed  in

-------
                             2063
test animals to  that already occurring in the  same target
organ in humans  and to circumvent the need for very large
extrapolations.

     Strong evidence exists for a simple and systematic
relationship between the magnitude of chronic  life-time
carcinogen exposure and the temporal behavior  of tumor
incidence when corrected for intercurrent mortality by
conventional life-table techniques.  The early work by Blum
on the induction of skin cancer in the mouse by ultraviolet
radiation  (2} and the later studies of Druckrey with various
chemical carcinogens on a variety of target organs (3) gave
a mathematical formulation to the common experience that the
higher the level of carcinogen exposure the earlier the
appearance of tumors.  These investigators showed that at a
given dose level, the cumulative incidence of  tumors can be
represented by a log-normal distribution of time to tumor
occurrence.  Thus, the overall response can be expressed in
terms of the median time, t, for tumor formation.  The
geometric standard deviation, O-, of the cumulative
incidence provides a measure of the temporal dispersion of
the individual response times, i.e., the larger the
geometric standard deviation the more heterogenous the
response.

     The relationship between the daily dose of carcinogen,
d, and the median time of tumor induction t was dtn = c
where n and c are constants for a particular carcinogen and
test system and  the standard deviation oq was  insensitive to
dose rate.  As shown elsewhere, the interaction of the log-
normal tumor incidence curve with the population survival
curve yields the cancer incidence, the average age at which
cancer develops, and the average amount of life lost by the
individuals developing cancer and the amount of life lost
averaged over the whole population; the values of n and Og
have important effects on the dose-response relationships
(4).

     The above formulation implies that a given dose level
of carcinogen,  every exposed individual would develop cancer
if he lived long enough.  The individuals that actually
develop tumors are the more susceptible members of the
exposed population; those that do not develop cancer are
simply less susceptible and die from extraneous causes
before they have a chance to develop cancer.  The contrast
between the effects of higher and lower carcinogen exposure
levels is that the time of tumor occurrence is shortened by
the higher dose  level in all exposed individuals by the same
proportion.  Thus, the susceptibles develop cancers earlier
than at the lower exposure and the additional cancer cases
occur because they now die before other causes instead of
afterward {in principle).

     The applicability of dtn = c formulation to human cancer
is illustrated by the comparison of stomach cancer in USA,

-------
                             2064
Germany and Japan  (5, 6).  In all three areas, the cumulative
mortality-corrected incidence is log-normal with the same Oq
of 1.5.  It is a reasonable supposition that dietary factors
are responsible for the differences in cancer experiences in
the three countries.  The parallel log-normal cumulative
incidence curves supports the notion that incremental
carcinogen exposure shortens cancer development by a constant
factor in the entire population.  On the assumption that
n = 2, the equivalent carcinogen exposure is thirty percent
higher in Germany and twice as high in Japan relative to
New York State.  The large value of the geometric standard
deviation for stomach cancer in all three countries (oq =
1.5) suggests the existence of a considerable heterogeneity
in the combined effects of susceptibility and carcinogen
dose.

     The epidemiological data on stomach cancer illustrates
the generally accepted notion that much of the current
cancer experience in humans is due to exposure to
environmental carcinogens.  It follows that the impact of
additional carcinogen exposure should be described in terms
of its interaction with the existing cancer experience and
presumed carcinogen exposure.  The biggest impact that a
small additional carcinogen exposure could have on a
population (that is already substantially exposed to
carcinogens)  would occur when the additional and existing
carcinogens were the same agents.  This would shift the
entire log-normal response curve to an earlier age.  By
contrast, if the actions were entirely independent, the
small additional carcinogen exposure would have its own
log-normal incidence curve occurring at a later time than
that of the existing carcinogen exposure.  The summated
effects of the two log-normal curves would result in a
deformed log-normal curve with tumor development fore-
shortened only at the more advanced ages.

     A simple and conservative approach for estimating
cancer risks could involve the use of the tumor response
data in test animals (for the particular carcinogen whose
risk is under consideration) to assign an equivalent
carcinogen dose, do, to the existing cancer experience in
the human population to be exposed.  The d,-, dose is thus one
that produces in animals a comparable temporal response to
that currently experienced by humans when the two species
are normalized for differences in life-span and under the
assumption of equal average susceptibility in humans and the
test animals.  The impact of an additional carcinogen dose,
d, is evaluated in humans in terms of the do + dtn = c
formulation.

     To illustrate the application of animal data to the
assignment of an equivalent carcinogen dose do to human
cancer and the subsequent estimation of risks, let us
suppose that we are concerned about the risks from the
carcinogen diethylnitrosamine (DENA)  which is assumed to

-------
                              2065
   Table  I   Effect  on  various  response  parameters of  the
             indicated  percentage  increments  in  equivalent
             carcinogen dose  for liver cancer occurrence
             in Connecticut males,  1962-64.
Percent
Increment in
Equivalent Carcinogen
Dose for Current
Liver Cancer
0
2
5
10
Experience t
350
347
342
334
P x
18
20
21
24
10-4-
.9
.0
.7
.6

71
71
71
70
X
.27
.20
.07
.87
A x
2.
2.
2.
2.
in-2
12
25
51
92

11
11
11
11
6
.21
.24
.32
.43
t - median age of cancer occurrence  (years)

p = probability of cancer

x = average age of cancer occurrence  (years)

A = average life-span loss in entire population  (years)

6 = average life-span loss of cancer cases  (years)
    Table II  The effect of 0.2 pg/kg/day of DENA on the
              incidence, average age of tumor occurrence
              and life-span loss of spontaneous and new
              hepatic cancer cases.

"Spontaneous" Cases (no DENA)
"Spontaneous" Cases (with DENA)
Extra Cases
Total Cases
Average Life-Span
Incidence Age Loss
per 10^ (years) (years)
18.9 71.27 	
18.9 67.85 3.42
5.7 80.75 1.99
24.6 70.87 	

-------
                             2066
produce only primary liver cancer in humans.  A log-normal
temporal response for liver tumors over a wide range of dose
rates of DENA with an n equal to 2.2 has been reported by
Druckrey for an inbred strain of rats  (3).  The cumulative
incidence of primary liver cancer reported by the
Connecticut State Tumor Registry for males in 1962-1964 (6)
is log-normal with a o~ of 1.7 and an extrapolated median
time, t, of 350 years.  For purposes of illustration, it is
assumed that one year of human life-span is equal to 1.52
weeks of life-span in rats.  The median time t of 350 years
corresponds to 535 weeks in rats.  The plot of log d vs
log t for the DENA response of rats is linear and extrap-
olates to a dose of about 2 yg/kg/day at 535 weeks which is
taken to be the equivalent dose do for the background
primary liver cancer experience in men.  Computer calcula-
tions have been done for normalized tumor responses covering
a range of values of n and aq to obtain values for
incidence, average age at cancer occurrence and the average
amount of life-span lost by cancer cases (4).  In these
calculations, d was normalized to unity for t equal to the
62 year mean life-span of humans.  It is assumed that
humans exposed to DENA will show the same n value as that
observed in rats, i.e., n = 2.2, but Og which is a measure
of heterogeneity of response, is taken to be that for the
observed primary liver cancer occurrence in humans, namely,
Og = 1.7.  Table I presents, for various percent increments
in the DENA equivalent dose do for background liver cancer
occurrence, the calculated values of t (median age), p
(life-time probability of developing cancer), A (average
life-span lost by the entire population), x (the average .age
of cancer occurrence) and 6 (the average life-span loss of
cancer cases).

     To understand the significance of these figures let
us examine, for example, the effects of the 10% increase in
the equivalent carcinogen dose, i.e., 0.2 yg/kg/day.  This
carcinogen exposure reduces the time of tumor occurrence
by 4.8%.  The effects in the average age of cancer occurrence
and the corresponding losses in life-span are tabulated in
Table II.  The average age of the 18.9/10* cases which occur
at background exposure will be reduced by 4.8% from 71.27
years to 67.85 years.  These spontaneous cancer cases would
therefore lose 3.42 years of life due to the carcinogen
exposure.  Furthermore, the incidence rises from 18.9/10*
to 24.6/10* with the carcinogen expcsure so that there are
an extra 5.7/10* cancer cases.  However,  the average age of
cancer development for all the cancer cases which occur in
association with the carcinogen exposure (24.6/10*)
decreases only slightly from 71.23 years at background
exposure to 70.87 years with the carcinogen exposure.  Since
the carcinogen exposure reduces the average age of the
original 18.9/10* cancer cases to 67.89 years, the 5.7/10*
extra cancer cases must have developed at an average age of
80.75 years since the overall average age of cancer develop-
ment in association with the carcinogen exposure is

-------
                            2067
70.87 years.

     As a result of the carcinogen exposure and the
consequent foreshortening of the cancer time, the extra
cancer cases die somewhat before other causes of death
instead of somewhat later {in principle) so that the actual
life shortening is roughly one-half the full 4.8% life-span
loss of the original cancer cases.  Hence, the average age
of death at 80.75 years represents a 2.4% reduction from
what it would have been without the carcinogen exposure,
i.e., 82.74 years and, consequently, the extra cancer cases
suffer a 1.99 year loss of life due to the carcinogen
exposure.  This is substantially less than the 3.42 years
lost by the original cancer cases and it also occurs later
in life so that the major brunt of the life shortening is
born by those cancer cases who would have gotten their
disease without the additional carcinogen exposure.
References

1.  MANTEL,  N.,  BRYAN, W.R.,  "Safety"  testing  of  carcinogenic
    agents,  J. Nat.  Cancer  Inst.,  27,  455-470  (1961).

2.  BLUM, H.  F.,  Carcinogenesis  by ultraviolet light,
    Princeton University  Press,  Princeton,  New Jersey,
    1959.

3.  DRUCKREY, H.,  "Quantitative  aspects  of  chemical
    carcinogenesis",  Potential carcinogenic hazards from
    drugs, evaluation of  risks,  UICC Monograph Series,  Vol.
    7, pp. 60-78,  (Rene Truhaut,  Ed.), Springer-Verlag,
    New York, 1967.

4.  ALBERT,  R. E., ALTSHULER, B.,  "Considerations relating
    to the formulation of limits for unavoidable  population
    exposures to environmental carcinogens", Radionuc1ide
    carcinogenesis,  (C. L.  Sanders, R. H. Busch,  J. E.
    Ballou and D.  D.  Mahlum,  Eds.), AEC  Symposium Series,
    CONF-720505,  NTIS, Springfield, Virginia,  June, 1973,
    pp. 233-253.

5.  Cancer incidence in five  continents, Vol.  II, (R.
    Doll, C.  Muir and J.  Waterhouse, Eds.), (Geneve,
    International Union Against  Cancer,  1970), Distributor:
    Springer-Verlag,  Berlin,  Heidelberg, New York.

6.  Cancer incidence in five  continents. A technical
    report.   (R.  Doll, P. Payne  and J. Waterhouse, Eds.),
     (Geneve,  International  Union Against Cancer,  1966) ,
    Distributor:   Springer-Verlag, Berlin,  Heidelberg,
    New York.

-------
                               2068
                       DISCUSSION
CROCKER  (U.S.A.)
     Do you have data  for  life  shortening  in  animals  that  do  not
develop tumors?  The reason  for this  question is  that life shor-
tening as such may be  an important  concomitant of exposure to
carcinogens.  Premature aging and life  shortening are associated
with chronic, low-grade X-ray exposure  and in turn with  increased
lesions  (renal glomerular  sclerosis,  for example)  commonly re-
cognized as age-related.   X-irradiation is also "carcinogenic"
though  doses used for demonstrating  this  property are usually
high.  It would therefore  seem  particularly useful to include
calculation of non-specific  life-shortening in animals experi-
ments in which the exposure  regimen has produced  tumors  in some
animals but not in all animals  under  exposure.


ALBERT (U.S.A.)

     The point is well taken, but so  far we have  not  looked at
the temporal aspects of the  dose-response  relationships  for non-
specific life-span shortening to examine the  similarities  and
differences with respect to  tumor formation.


MAGE (Denmark)

     Any data can be plotted on log probability paper, and
scatter will exist.  If one  makes the assumption  that these data
are random samples from a log-normal distribution  then the  line
must pass through the  geometric mean  of the data  at 5O%
            n

           I,    log X.
            n                                  with Xgm = e/u.

     The 5O% occurrence must be plotted at a value Xg.m = e/u.
The slope of the line is determined from the standard geometric
deviation of the data.

     In your equation was your "median" value determined in this
manner?  If not, on what statistical basis can you justify, or
better, with what confidence level can you state that the dis-
tribution is lognormal?

-------
                               2069
ALBERT  (U.S.A.)

     Whether the cumulative incidence of tunriors  at  a given dose
level is truly log-normal is an important question.  considering
the uncertainties in the temporal response data  from most ex-
periments and the general lack of data below  the 5% response
level, we have examined the matter  only to the extent  of satis-
fying ourselves by an eye fit that  such data  as  we  have looked
at, is consistent with the log-normal pattern.

-------
                             2071
           ERFORDERNISSE IM HINBLICK AUF MESSUNGEN



                      MONITORING NEEDS



        BESOINS EN MATIERE DE MESURE DE L'EXPOSITION



      NECESSITA'  RELATIVE ALLA MISURA DELL'ESPOSIZIONE



  EISEN VOOR HET TOT STAND BRENGEN VAN TOEZICHT OP EXPOSIE
                            Panel








Vorsitzender - Cnairman - President - Presidente - Voorzitter






                  T.  SCHNEIDER (Nederland)

-------
                              2073
           POLLUTANT-ORIENTED INTEGRATED MONITORING
             SYSTEMS AND LEAD EXPOSURE ASSESSMENT

            E, A, SCHUCK, 6, B,  MORGAN/ D, S, EARTH

Environmental Protection Agency, National Environmental Research
Center, Las Vegas, Nevada, USA
ABSTRACT

     A recent USEPA sponsored workshop panel of medical and mon-
itoring experts concluded that the quantities of lead emitted
into the environment have significantly degraded the quality of
man's food, water and air supplies - as well as contaminating
all exposed surfaces - and that motor vehicles using leaded
gasolines contribute about 90% of these airborne and settled
lead particles.

     Evidence indicates that blood-lead concentrations increase
with proximity to central urban areas and heavy traffic.   Young
children, particularly in central urban areas, are more suscep-
tible to lead poisoning, absorb more lead, and may have a higher
body burden (5 to 10 times more) on a body weight basis than do
adults.

     Little precise information is known concerning the subclini-
cal pathological effects of lead poisoning.   The destruction or
partial incapacitation of the brain and other nerve tissues are
of particular concern.   Furthermore, permanent neural and organ
damage may occur prior to birth, for lead has long been associ-
ated with aborted fetuses and stillborns.   On the basis of pub-
lic health surveys, it has been estimated that between 5,000 and
25,000 children per year suffer adverse effects from lead in the
United States with known deaths at a rate of about 200 per year.

-------
                              2074
     Jn conclusion, the panel strongly recommended that an inten-
sive research monitoring system be designed and implemented to
quantitate the exposure and effects of chronic environmental lead
exposures to man.

-------
                              2075
1.  Introduction.

     Medical  and  environmental  monitoring  experts   were
convened  by  the National Environmental Research Center-Las
Vegas (NERC-LV) in March 1974  to  discuss  and  assess  the
present state-of-knowledge about lead--its sources, exposure
pathways,  and  physiological and environmental effects--for
the purpose of developing concepts necessary for determining
the feasibility of establishing an integrated  environmental
research monitoring system and its requirements.
     A   practical   and   effective   integrated   research
monitoring system is needed to complement basic research  in
determining  the  physiological and environmental effects of
numerous pollutants, to identify and map areas  of  concern,
to  provide  reliable trend data, and to establish realistic
control measures.
     The need for environmental research monitoring  results
from   recognition  that  man  and  his  activities  have  a
substantial modifying effect on the  environment.   Assuming.
the  existence of a well-planned research monitoring program
suggests an ability to prevent the occurrence of detrimental
or undesirable environmental  effects.   To  date,  however,
most research monitoring programs have developed for reasons
other  than  prevention.  Thus, research monitoring programs
are  instigated  only after  environmental   pollution   has
resulted in substantial detrimental and undesirable effects.
     In such a situation, the demand for information becomes
intense  concerning  the  nature of the problem--the causes,
the consequences, and the solutions.  The resulting research
monitoring program cannot  meet  these  demands  within  the
short  time  frame  required.   Consequently,   such programs
contain very little planning and  tend  to  focus  on  those
aspects assumed to be most important.  Solutions, therefore,
are  frequently  based  on  a grossly inadequate data base--
inadequate to the point of leading to control actions  which
have  dubious value and which are executed at great costs in
resources.  This situation cannot be permitted  to  continue

-------
                               2076

and  leads  to  the  necessity  for  adequate  planning  and
execution of environmental research monitoring activities.
     Recognition of this need has resulted in the assignment
of resources at NERC-LV  to  investigate  the  design  of  a
pollutant-oriented  integrated  monitoring  system.   Such a
system  would  take   into   account   sources,   transport,
interactions,  exposure,  and  dose in all applicable media.
Also involved are  sampling  site  selection,  methods,  and
analysis,  data  collection  and  presentation,  and quality
assurance for the system and its individual components.
     In recent decades, there has been a tremendous increase
in  both  the  number  and  concentrations  of  man-produced
substances  entering  the environment.  The effects of these
substances at various exposure and dose levels and rates  is
often  unknown  or is only poorly defined.  While any one of
these substances may be harmless in itself,  it  may  become
harmful in combination with other materials or under certain
environmental conditions.  For instance, one effect which is
inadequately known that could lead to widespread detrimental
consequences  to  man  is  the potentiating effect which one
substance might induce in  others.   The  present  state-of-
knowledge  about  these  substances  and  their  effects and
interrelationships  is  inadequate.    The   known   harmful
effects, and the large number of potentially harmful effects
not  presently  known, necessitate the establishment of some
form of integrated monitoring system to map, study, and  aid
in  controlling  these substances.  Many scientific research
projects are also needed.
     In  order  to  develop  a   practical   and   effective
monitoring  approach,  this  workshop  was  held  to develop
concepts and to determine the feasibility of such  a  system
using lead as a test case.   Lead was selected because of the
copious  amount of data available on it, because it presents
a worldwide problem, and  because  it  enters  man  and  his
environment   in   many   forms   and   by   various  media.
Consequently, research  monitoring  concepts  developed  for

-------
                               2077
lead   should   also   apply  to  many  other  environmental
pollutants.

2.  Discussion.

     Lead, which is found in the earth's crust, also  occurs
naturally  in  the atmosphere and hydrosphere as a result of
both physical and chemical processes.  Man's activities,  of
course,  introduced the greatest quantities of lead and lead
compounds into the atmosphere and hydrosphere.  For example,
lead or its compounds can enter the environment at any stage
during the mining, smelting, processing,  and  use  of  this
metal and its derivatives.  Additionally, lead is stable and
incrementally accumulates as a waste or contaminant.
     Increases  in lead use have been on the order of 3% per
year over the last decade, and the total annual  consumption
of  lead  in  the United States is approximately 1.5 million
short tons.  Fortunately, most of this lead is not in a form
or use likely to pose an environmental or health threat.
     Major sources  of  lead  causing  or  likely  to  cause
adverse environmental or health effects include exhaust from
the use of leaded gasoline  (this contribution alone accounts
for  about  90%  of the airborne and settled lead particles)
and some other common fuels such as coal; the use of  paint,
ink,  coloring,  and other pigments containing lead; the use
of lead in treating, processing, and packaging food; the use
of lead in water pipes and pesticides; the use of lead as  a
stabilizer  in  the manufacture of certain plastics; the use
of  lead  in  candle  wicks;  and  the   mining,   smelting,
processing,   and   terminal  disposal  of  lead  containing
materials.  These and other sources bring  large  quantities
of lead into man and his environment via the air, water, and
food  cycles.   Figure 1  portrays  the  major pathways to a
critical receptor.
     During the past few years, environmental lead has  been
the  subject  of several publications and reviews.  Based on
these efforts, the population at risk,  i.e.,  the  critical

-------
                                2078
            MAJOR PATHWAYS AFFECTING  LEAD BURDEN
                      IN CRITICAL RECEPTOR
,4 —
1
FOOD
WATER
MOTHER


T
DUST
SUSPENDED
PARTICLES










INGESTION
INTERMITfENT &
VARIABLE ABSORPTION

PLACENTAL TRANSFER
INITIAL BURDEN


INHALATION
CONTINUOUS &
VARIABLE ABSORPTION

*
FOOD WATER
PA (NT DUST
I URBAN
CHILD EXCRETION
(CRITICAL
•"' RECEPTOR)
T
DUST
SUSPENDED
. PARTICLES
k.
	 f
                       FIGURE I.
receptor,  in  the  United States has been identified as the
urban  child  of  one  to  three  years  of  age.   However,
neurological damage may have actually occurred at an earlier
period  with the effects first appearing at a later stage of
development.  In addition, these effects may be  too  subtle
to  be  easily  recognized since they may gradually progress
with growth and maturation.  An example of the acute effects
of high blood-lead concentrations in a pregnant  mother  and
her  newborn  infant are well illustrated in a case reported
by Palmisano et al. [1] in which these  authors  also  state
that  lead  has  a  "devastating  effect on reproduction and
pregnancy since it most commonly causes sterility  or  early
spontaneous  abortion.   It has been clearly shown that lead
crosses the human placenta and may cause untoward effects in
the fetus (Karlog and Holler [2]).  Around the beginning  of
this  century  it  was recognized that women employed in the
lead trades often produced live-born infants who were small,
weak, and neurologically damaged (Cantarow and  Trumper [3];
Angle and Mclntire [4])."

-------
                             2079
     The  detailed  exposure  pathways  contributing to lead
burdens in a receptor are shown in  Figure 2.   It  was  the
objective  of  this  study to quantitate these pathways and,
thus, to determine total exposure and its variability  among
individuals  and  populations. •  Additionally, the relations
between exposure, dose, uptake, and effects  were  assessed.
The  diagram  in  Figure 3  illustrates the pathways of lead
within a critical receptor.  These pathways were assessed to
determine the best indices of body lead burdens.
     A hypothetical example of comparative lead body burdens
based on calculated absorption is  shown  in  Figure 4.   It
should  be  noted  that this comparison cannot be correlated
with blood-lead levels, and only represents an unknown total
body  distribution  after  absorption,  but  prior  to   any
excretion.   From  this  illustration,  however, it is clear
that young children must be given top priority in  designing
a viable monitoring system for lead.
     In   spite   of   the  availability  of  an  impressive
information base, the pathways leading  to  and  within  the
critical  receptor  (Figures 2  and 3) cannot be quantitated
without additional substantial investments in  research'  and
monitoring.   One  result  of this lack of quantification is
that current  review  documents  tend  to  overestimate  the
contributions of leaded paint chips and to underestimate the
importance of airborne lead which continually settles out on
all   surfaces   and   is  available  for  resuspension  and
subsequent inhalation and ingestion.
     Once  sound  integrated  research   monitoring   system
concepts  are  developed  and  applied to environmental lead
surveillance, as well as for other pollutants,  it  will  be
possible  to  further delineate areas of concern, to provide
reliable  trend  and  research  data,   and   to   establish
efficacious and economical control measures.

-------
                                 2080
        LEAD  EXPOSURE PATHWAYS TO CRITICAL RECEPTOR
                    OTHER SOURCES
                    PAINT. INK, ETC.
   Pb &
   Pb COMPOUNDS
MAJOR
    ROUTES
MINOR
   ROUTES
HIGHLY
  VARIABLE
     ROUTES
   AUTO
   EXHAUST

AIR


-»
1


OUST




*"


            WATER
                    WATER
                     PIPES
                  FORAGE
                  CROPS
          CONSUMER
            CROPS
                       FOOD
                    PROCESSING
I ANIMALS L
ANIMALS L.............—-'
                                     tNGESTION
                                        AND
                                    INHALATION
                                         BY
                                      CRITICAL
                                      RECEPTOR
                       FIGURE 2.
                PATHWAYS OF LEAD IN  CRITICAL RECEPTOR
                INHALATION
                   UPPER
                RESPIRATORY
                  SYSTEM
                  LOWER
               RESPIRATORY
                  SYSTEM
                 Gl TRACT
    • RATES
     VARIABLE,
    DEPENDING
    ON MANY
     FACTORS
INGESTION
                       B
                       L
                      O
                      O
                      D
                                        SOFT
                                        TISSUE
                                      BONE
                                      HAIR
                                      NAILS
                                     NERVES
                                      BRAIN
                                      URINE
                                     SWEAT
                                      FECES
                       FIGURE

-------
                                2081
                    HYPOTHETICAL
    LEAD  BODY  BURDEN  COMPARISON
         BETWEEN  ADULT   AND  CHILD
                                               ONE YEAR OLD
                                                 MAY  HAVE
                                             TEN  TIMES GREATER
                                                BODY BURDEN
                                                THAN  ADULT!
          '*•   * • 0.7Mg Pb/fcg
                 dose  per day
                                       7.5Hg Pb/kg
     ADULT (70kg)
Daily Intake

Food - 300 pg
Water - 20 pg
Air - 2.5 ug (23

TOTAL
                                      dose per day <
                    CALCULATED ABSORPTION
                                      CHILD  (10kg)
       Lead Absorbed

        30 ug (101)
         2 pg (101)
        21 pg (37*)
Daily Intake

Food - 130 vg
Water - 10 yg
Air - 2.5 pg (6 m3)

TOTAL
Lead Absorbed
      Vg (50%) (a)
      ug (50%) (a)
      yg (37S)
                                                    ±75.5 pgPb
   S3 pgPb

   70 kg man
=  0.7 wgPb/kg dose
   per day to man
   75 wgPb
   	  =  7.5 ugPb/kg dose
   10 kg child  per day to child
     (a)  Based on study entitled "The Uptake and Fjccretion by Children
of Lead and Other Contaminants" by Alexander et al. (1973).  Also refer
to "Intestinal Lead Absorption" by Karliausen (1973) for further infor-
mation.
                            Figure A

-------
                             2082
                         REFERENCES
1   Palmisano,  P.  A.,  Sneed,  R.  C.,  Cassady,  "Untaxed
    Whiskey and Fetal Lead Exposure," J. Pediatrics, Vol. 75
    No. 5., 1969.

2   Karlog, D. , Holler, K. 0., "Three Cases  of  Acute  Lead
    Poisoning," Acta Pharmacol., Vol. 15 No. 8., 1968.

3   Cantarow, A., Trumper,  M.,  Lead Poisoning,  Baltimore,
    The Williams and Wilkins Company, p. 143, 1944.

4   Angle, C. R., Mclntire, M. S.,  "Lead  Poisoning  During
    Pregnancy," Am. J. Pis. Child, Vol. 108:436, 1964.

-------
                              2083
    INTEGRATED  ASSESSMENT  OF  HEALTH  EFFECTS  OF  AIR POLLUTION

      L.  R,  BABCOCK JR,+,  N,  L,  NAGDA+  AND A, T,  ROSSANO++

 +   University  of  Illinois, Chicago  and Argonne National Labora
    tory, USA
 ++  University  of  Washington, USA
ABSTRACT

     This presentation is directed  to  individuals who seek a
comprehensive methodology for assigning air quality management
priorities.   A comprehensive system of indices is being devel-
oped which relates population exposure and health-related damage
back to individual emission sources.   The system Dill be used
to rank source severities and at the same time will enable im-
provement of the experimental design of health effect studies.
This presentation addresses one aspect of our continuing research:
a simulation names "popex" is described which integrates earlier
air-quality-index concepts with population exposure considerations.

     The prime purpose of most air  quality management strategies
is to reduce the health risk to exposed human populations.   The
situation is complex with many kinds of sources, pollutants, and
human receptors, all ties together  by uncertain source-receptor
pathways (1).    Much needed is a quantitative framework which
considers both costs of improving air quality as well as health
and other damages associated with exposure of people to various
air pollutants.

     A limited number of studies analyzing direct and indirect
control costs  are available,  but much less is known about damages
caused by air  pollution exposure.    Many more health-effect studies

-------
                              2084
are needed before all facets of health damages due  to air pollu-
tion can be defined.   Generally, the individual studies that
are presently available are restricted to single pollutants and
are quite limited in terms of time-concentration variables.
Seeds are veil recognized* and health effects studies are inc-
reasing, both in number and in level of sophistication.

     However, health effect data are not in themselves sufficient
for relating emissions to health damages.   One of several mis-
sing links is knowledge of exact pollutant concentrations received
by human receptors as a function of time.

-------
                                    2085
1.  Introduction
    Despite complexities and uncertainties, the need for an overall frame-
work for assessment of integrated air pollution health effects is quite
evident.  Such a framework should be able to relate air pollution damages
back to sources and should enable the ranking of source severity.  The
system would be expected to have some gaps wherein the relationships be-
tween some parameters are not known rigorously.  In such instances, our
strategy would be to use the available not-so-rigorous relationships until
better information is available.
    It may seem unusual to describe uncertainty so early and prominently
in our paper, but it seems appropriate that the complexities of the under-
taking be defined before presenting our model:
    Pollutant transport mechanisms.  Dispersion models with various levels
of sophistication can be employed for the estimation of pollutant concen-
trations at receptor sites.  Such models all require some form of calibra-
tion and have never been verified for complex urban situations.  Compound-
ing the difficulty, the large amounts of prerequisite meaningful meteoro-
logical data are virtually non-existent.
    Interaction of pollutants.  Pollutants react with other pollutants as
well as with other materials.  These reactions could produce more- or less-
potent intermediates, leading to particulate matter which is eventually re-
moved from the atmosphere, or to innocuous decay end products, such as car-
bon dioxide and water.  Some systems are understood better than others.
Oxidant formation has been subjected to intensive study, but similar work
is just beginning to shed light on the mechanisms involved in the atmo-
spheric conversion of sulfur dioxide and nitrogen oxides into possibly-
roore-toxic particulate sulfates and nitrates.  None of the systems are
understood sufficiently to permit the reliable prediction of actual atmo-
spheric reaction rates.  Recent research has tended to identify increasing
complexity rather than develop useful models with general applicability.
    Source-receptor geometry.  In order to arrive at an estimate of pollu-
tant dosage, source-receptor geometry must be considered.  Residences are
not distributed evenly throughout a region, and inhabitants do not spend
all their time near home.  Indoor-outdoor pollution differentials add
further complexity.
    Health effects.  There are numerous health-related uncertainties.

-------
                                      2086
 Questions remain for even the more-direct adverse health effects such as
 emphysema and bronchitis, and thus, with the present state of our knowl-
 edge, quantification of probable indirect psychological or genetic effects
 seems almost impossible.
 2.  Rationale and Approach
     Our approach is stepwise, starting with simplistic relationships,
 adding complexity as it seems justified.
     An emission inventory serves as the starting point for ranking sources
 of air pollution; early evaluations indicated that the automobile was re-
 sponsible for over 50 percent of the gross mass of emissions within many
 urban regions.   Based upon this evidence, control programs were initiated
 within the USA  and elsewhere.  However, an emission inventory ignores both
 the relative toxicity of the pollutants as well as the source-receptor
 geometry.
     Our next step was to develop an air quality index which considered sul-
 fur dioxide, nitrogen oxides, carbon monoxide,  particulate matter,  hydro-
 carbons,  and oxidant.   The index,  called pindex,  enabled  the direct com-
 parison of multi-pollutant emissions.   When toxicities are considered,
 automotive pollution is reduced significantly in  relative importance.   De-
 tails as to the development and application of  pindex are described else-
 where (2).   Pindex  concepts have been  employed  in a 2u-category cost-
 benefit analysis  (3)  and have also  been extended  to address ambient air
 quality (4).
     Pindex adjustment multiplies the meaningfulness of  an emission  in-
 ventory,  but the  emission-directed  pindex ignors  factors  involving  source-
 receptor  relationships.   General land-use-people-pollution  relationships
 have been addressed  elsewhere (5,6); our  first  generation attempt at
 specifically integrating population exposure with pindex  is described below:
 3.   Population-Adjusted-Pollution Index:  Popex
     The pindex methodology  is being expanded to incorporate important con-
 siderations  such  as meteorological  dispersion modeling and  variables re-
 lated to  the  source-receptor  distances and population distribution.  We
 call  this population-adjusted-pollution index,  "popex", for  short.
    Utilization of a complex  and time-consuming meteorological model in
popex could have increased the sophistication of  the methodology.  But we

-------
                                      2087
have chosen  to first construct and evaluate a simple "pseudo-meteorologi-
cal" model.   The concentric ring, gaussian plume model has a  level  of so-
phistication beyond that of a box model,  and we feel it well  balances with
the present  levels of complexity and  accuracy of other aspects  of the
methodology.
    The geographical area considered  in popex is the Air Quality Control
Region  (AQCR), and in our present version, counties are the smallest areal
elements considered.  Our initial application will evaluate sources within
the Chicago  AQCR which consists of 8  million people distributed among
eleven counties.
    First  calculated in popex are total  dosages of the six pollutants
reaching each of the counties from all the sources within the AQCR.  Then
the levels of each pollutant, including oxidant formed by simulated photo-
chemical reactions, are combined into single effect numbers by  pindex.
Finally, the percent contributions of individual sources  (to  the total
pollution-population exposure) are used to assign a priority-ranking to
each source.   The diagram summarizes  the  popex model (7).
                             POPEX  FLOW  DIAGRAM
                                          CONCENTRIC RING
                                          DISPERSION MODEL
                                               ftlLLUTANT
                                               CONCENTRATION
                                               AT RECEPTORS
                                          MODIFIED PINDEX  )
                                               PINDEXED SECEPTWI
                                               CONCENTRATION
                                          POPULATION
                                          EXPOSURE MODEL
                                               TOTAL ROLLUTION-
                                               POPULATIOK [FFECT
                                        PRO-RATING EFFECT TO
                                        INDIVIDUAL SOURCES
                                          RANKING SYSTEM

-------
                                      2088
 4.  Discussion and Conclusions
     The popex model yields the expected results when applied to contrived
 situations.  However, it will be necessary to carefully evaluate the model
 with actual geographically-specific data; we look forward to presenting in
 the near future, our assessment of the 1600 sources within the Chicago
 AQCR.
     Our preliminary results indicate that short-distance source-to-recep-
 tor exposures account for the dominant fraction of total air pollution
 damages.  (The population affected may be low, but the exposure-per-person
 is very high.)  Thus, small errors in this part of the simulation may
 yield large errors in the overall results.  For example, our first-genera-
 tion source-receptor configuration locates all the receptors in a county at
 a single distance or radius from the point sources,  at a "ring of average
 exposure".   The correct location of this ring is a complex function of
 stack height,  population distribution,  and meteorological variables.  The
 assumptions made must receive careful appraisal.
     With popex,  we are attempting to integrate population exposure param-
 eters and toxicity of pollutants to arrive at a system for hazard-based
 ranking  of  sources.   Our present version must be  considered a  "first gen-
 eration"  model.   Most of the  complex variables mentioned in this paper
 have not  yet been included  in popex.  Considerable revision, extension,
 and  verification  will  be required before  popex can become a meaningful
 policymaking tool.  Toward  this  goal, we  seek communication and inter-
 action with  others  involved in similar  endeavors  (7).
 5.  Acknowledgements
    Gratefully acknowledged are  the  computer  services provided  by the
Computer Center at UI, Chicago Circle,  and the continuing  financial  sup-
port provided by  the U.  S. Environmental Protection Agency  (Grant R-802111)
and by the National Science Foundation  (RANN program AG-352 through
Argonne National  Laboratory and Grant GK-27772 through the University of
Illinois).

-------
                                     2089
References
1.  A. T. Rossano, Jr., ed.. Air Pollution Control; Guidebook for Manage-
    ment, E.R.A., Inc., Stamford, Connecticut, 1969.
2.  L. R. Babcock and N. L. Nagda, "Indices of Air Quality," Indicators
    of Environmental Quality (W. A. Thomas, Ed.), Plenum Press, New York,
    1972, pp. 183-197.
3.  L. R. Babcock and N. L. Nagda, "Cost Effectiveness of Emission Con-
    trol •" Journal of the Air Pollution Control Association, 23, March
    1973, pp. 173-179.
4.  W. A. Thomas, L. R. Babcock and W. D. Shults, Oak Ridge Air Quality
    Index, (ORNL-NSF-EP-8), Oak Ridge National Laboratory, Tennessee,
    September 1971.
5.  N. L. Nagda and L. R. Babcock, "Integration of Land Use Considerations
    into Air Quality Management Strategies," Papers and Proceedings Third
    Pacific Regional Science Conference, August 1973, Honolulu, Tokyo
    University Press (to be published, 1974).
6.  D. V. Lamb, F. I. Badgley and A.  T. Rossano, "A Review of Diffusional
    Modeling Techniques for Predicting Air Quality with Relation to Trans-
    portation," study prepared for Washington State Highway Commission
    {Contract 63-1062), Seattle, Washington, 1973.
7.  Additional information may be obtained from the authors.  Write Prof,
    L. R. Babcock, Jr., School of Public Health, University of Illinois,
    P. O. Box 6998, Chicago, Illinois  60680, USA.

-------
                             2091
                  MEXICO CITY'S PINDEX LEVEL

    H, BRAVO A,, M, E, ESPINOSA, A, C, LOWE AND S, SALAZAR

Institute de Geoflsica, Universidad Nacional Aut6noma de Mexico
Mexico, D.F.


ABSTRACT

     Up to now, an attempt to evaluate Mexico City's air pollution
as a total has not been made.   A more comprehensive way of eva-
luation,  where concentration as well as tolerance limits are
interrelated is needed.   Also, the effect of solar radiation
has never been considered as a direct parameter in the dynamics
of the air pollution problem.

     Due to the special location of Mexico City, in an altitude
of 2,268 m above sea level, and a latitude of 19°20' North, we
strongly believe that solar radiation has to be considered if a
good understanding of air pollution is desired.

     The use of the Pindex approach will be useful in the study
of the expected epidemiological and health effects as a result
of air pollution and it will, as well, be a tool for the selec-
tion of control strategies.

-------
                               2092
      In order to evaluate the existing air pollution moni-
 toring data, the tendency in Mexico is by unit mass.  It is
 well known that different air pollutants have different
 toxicity levels, based on mass-time relationships.  Babcock1'4
 introduced these tolerance levels and other parameters to
 develop an index.  if we follow this new approach, we will
 be able to calculate a similar pollution index, although
 not 100% correct for Mexico City and other Mexican cities,
 it would, at least, be the beginning to point out the need
 for a new way to evaluate the existing data and obtain more
 realistic regulations and control policies.

      Mexico City is located in the southwestern corner of an
 elevated basin (2,268 m above sea level),  at a latitude of
 19°20'  North, with a very high incidence of calms and verti.
 cal gradient inversions of temperature during the year2.   ~
 The above parameters make of Mexico City a unique case of
 study regarding air pollution and its  effects.

      The City is part of the metropolitan  area,  with a popu
 iation  close to nine million inhabitants,  about 700,000 ve-
 hicles,  and containing about 20.5% of  the  total  industrial
 activity of the country.  As Mexican petroleum  crudes are
 very high in sulphur,  the oil fuel used contains between  3
 to 4% in weight of sulphur.3

      The reliable  air  quality data in Mexico City is,  at  this
 moment,  quite limited,  but most of the  information used in
 this paper,  although  short in number, has  a  high degree of
 confidence.   Some  of  the  data used is the  only existing one;
 its  source  is quoted  in the paper.
                used here is the one suggested by Babcock
et al.  •  to estimate the total air pollution pindex, al-
though  we feel that for Mexico City a more complex index,
with specific tolerance levels and the introduction of sev-
eral new parameters is needed.

     Table No. I shows the air quality data used for the
development of Mexico City's Pindex and their corresponding
pindex  levels.

     Table No. II shows the method carried out,  in order to
obtain  the individual pindex levels c.nd the total index
from the given information.

     Mexico City's data was reduced when it was  necessary,
using Larsen approximation.7

-------
                              2093
       The tolerance factor used for particulate matter was
 derived from the primary 24 hour standard?.

       For SO2,  the tolerance factor was derived from the
 secondary 3  hour standard7.

       The N02  factor was the one used by Babcock, but extrapo
 lated  to 1 hour.4                                           —

       Carbon monoxide's factor was derived from the 1 hour
 standard.

       The hydrocarbon's factor used was the one employed by
 Babcock1'^.

       The  oxidant's factor was derived from the standard used
 by Larsen  .

       Table  III  lists  the pindex obtained for Mexico City and
 some U.S.A.  cities.  The pindex for the U.S.A. cities was re^
 calculated using the concentrations reported by L.R. Bab-
 cock , and the same tolerance factors used for Mexico City's
 Pindex.

       It  is  clear from Table III  that Mexico City ranks first,
with a typical photochemical problem in addition to a high
 concentration of particulate matter.

      Once more,  this  index for Mexico City is just a prelim-
 inary approach,  which  shows the need to obtain specific
tolerance  factors  for  pollutants  at Mexico City's conditions.

      Besides, regulations  and control strategies have to be
reviewed and designed  based on a  specific index for Mexico
City.
                     TABLE NO. I
                  Mexico City's Air  Pollutants

          Concentrations and Corresponding Pindex levels
P.M.* SO_ NOV
2 x
Wg/m3) (ppm) (ppm)
Concentrations 227 0.021 0.113
Pindex levels 1.060 0.030 0.049
CO HC Oxidant
(ppm) (ppm) (ppro)
13.4 10.52 0,048
0.290 0.27 0.750
    *P.M.  = particulate matter

-------
                            2094
                       TABLE NO. II
                   Pindex Calculations
        Available information for Pindex Calculation
        Particulate matter5  (PM)   = 277^g/m3
        Sulfur Dioxide*      (SO2)  =  42.26
        Nitrogen oxides*     (NOX)  = 132.5
        Carbon monoxide^     (CO)   =  11.655
        Hydrocarbons6        (HC)   =5231
        Oxidant*             (O3)   =  71.1
        Solar radiation**    (SR)   = 591  cal/cm2day

Convert reactants to-X/mol/m
        NOX    =    132.5/46    =     2.88/(Tnol/m3
        HC     =    5231/16     =   326.9
        03     =    71.1/48     =     1.48
Determine limiting reactant for oxidant synthesis
(NOX or HC):  NOX is limiting
Create oxidant:
        O3     =   0.0006 x SR x  (limiting reactant)
        03     =   0.0006 x 591 x 2.88 = 1.02/fmol/m3
Determine total oxidant and excess HC and NOX
        03     =    1.48 + 1.02  =    2.50 JU mol/m3
        HC     =  326.9  - 1.02  =  325.88
        NOX    =    2.88-1.02  =    1.86
Convert reactants back to weight basis
        03     =    2.50 x 48    =  120.00 M g/m3
        HC     =  325.9  x 16    = 5214.00
        NOX    =    1.86 x 46    =   85.56
Apply tolerance factors
      •  PM     «  277/260    =   1.060
        S02    =  42.26/1330 =   0.030
        NOX    =  85.56/1720 =   0.049
        CO     =  11655/40000=   0.290
        HC     =  5214/19300 =   0.027
        03     =  120/160    =   0.750
Determine Synergism term (SYN)
        SYN    =  SO2  or  PM (whichever is smaller)
        SYN    =  SO2   =   0.03
Sum terms to determine pindex
    Pindex = PM + SO2 + NOX + CO + HC + O3 + SYN
    Pindex =2.48
     *  Departamento de Contaminaci6n Ambiental. institute
        de Geoflsica, U.N.A.M., Mexico
    **  Departamento de Radiaci6n Solar, Institute de Geo-
        flsica, U.N.A.M., Mexico.

-------
                    TABLE NO. Ill

          Mexico City* and U.S.A. cities**
                    Pindex levels

Mexico City
Chicago
Los Angeles
Philadelphia
Denver
San Francisco
San Diego
P.M.
1.060
0.477
0.450
0.590
0.480
0.260
0.260
S0x
0.030
0.307
0.044
0.170
0.020
0.020
0.020
NC-
J^
0.050
0.115
0.144
0.066
0.057
0.110
0.040
CO
0.290
0.345
0.316
0.190
0.227
0.090
0.080
HC
0.270
0.101
0.134
0.067
0.080
0.100
0.200
Oxidant
0.750
0.531
0.987
0.350
0.570
0.650
0.510

2.48
2.18
2.12
2.04
1.45
1.25
1.13
  *  Calculated from existing data
 **
* **
                                Annual average,
Annual average
This term includes the synergistic effect
                                                                               K5
                                                                               o

-------
                          2096
                     REFERENCES

 Lyndon R. Babcock,  Jr.,  "A combined pollution index
     for measurement of total air pollution",  J.  Air
     Poll. Control Assoc..  20 (10):653 (October 1970).

 H.  Bravo, A.  et al.,  "Contribution of stationary
     combustion sources to  the horizontal  sulphur.
     Dioxide concentration  in the Mexican  Valley.
     Paper No.  68-35.   Presented at the 68th Annual
     Meeting APCA, Saint Paul Minnesota, June  1968.

 Serie Estudios/1.   Medio Ambiente Humano  Problemas  Eco-
     16gicos Nacionales.  Secretaria de la Presidencia.
     Cuadernos  de Documentaci6n.   Serie Estudios/Num.1.
     Segunda Edici6n Ampliada.  Pa"g.  42,  1972.

 Babcock,  L.R.  and Nagda, N.L.,  Letter  to  the Editor,
    "Rating  of  Pollutants by  Effect", J. Air Poll. Con-
     trol  Assoc.  (22)  9:727,  (September 1972).

 Bravo, H.,  Corona,  L., "La Contaminaci6n atmosferica y
     su relaci6n  con el flujo  de vehfculos en la Ciudad
     de Mexico".  Inst. de Ingenierfa,  227. Mayo 1969.
    Mexico.

Panivino, N.."Determinaci6n de Hidrocarburos Totales en
    el aire por el M£todo de Ionizaci6n de Flama", Te-
    sis, Facultad de Ciencias Quimicas, U.N.A.M., (de-
    sarrollada en el Institute Mexicano del Petr6leo),
    Mexico, D. F.,  1973.

Larsen, R.I.,  "A mathematical model for relating air
    quality Measurements to Air Quality Standard, U.S.
    Environmental Protection Agency. Research Triangle
    Park, North Carolina, November 1971.

-------
                              2097
      A METHOD FOR SIMULATING THE TRUE HUMAN EXPOSURE OF
         CRITICAL POPULATION GROUPS TO AIR POLLUTANTS

               WAYNE R, OTT+ AND DAVID T, MAGE"*"*"

   Environmental Protection Agency, Washington D.C., USA
   San Jose State University, California, USA
ABSTRACT

     There are serious problems in obtaining an accurate assess-
ment of the true population exposure to air pollutants.   In the
United States, the primary means for monitoring air quality is
by measurements at fixed locations in urban areas, or air rnoni-
tor ing stations.   It is becoming increasingly apparent that data
from such stations provide a relatively poor measure of the true
exposure of members of the general public to air pollutants, be-
cause these stations are not necessarily located where the public
is exposed to the highest concentrations.

     To demonstrate a method to obtain a more representative
measure of the human exposure of a critical population group -
pedestrians and shoppers - a technique was developed for simul-
ating the true exposure of pedestrians to carbon monoxide (CO)
by collecting integrated CO samples over the routes that they
valk.    A total of 425 of these simulated exposure samples (SES)
Here collected on congested downtown streets in a major urban
area.    The results were compared statistically with conventional
measurements from a nearby fixed monitoring station.   Signifi-
cant differences were observed between these two sampling metho-
dologies - fixed and SES - and the significance of these findings
for setting air quality standards is discussed.

-------
                              2096
1.   Introduction
     Serious problems arise in accurately assessing the true
human exposure to air pollutants.  In the United States, the
usual practice for monitoring air quality is to measure
selected air pollutants  (carbon monoxide, hydrocarbons, sulfur
dioxide, particulates, etc.) at fixed locations in each urban
area.  Unfortunately, concentrations at these air monitoring
stations are not necessarily representative of concentrations
to which a significant proportion of the population actually
may be exposed. ' '   This occurs because the stations are not
always located where concentrations are highest and where
large numbers of people  also congregate.  The most serious
problems arise when monitoring stations are located high—for
example, on the 5th or 7th floor of a downtown building—or at
the outskirts of a city's central business district.  Such
monitoring locations are common in the United States.
2.   U.S. Air Quality Standards
     In any environmental monitoring effort, the  first  step is
to decide whose exposure the data  are  intended to measure.
In the United States, national ambient air quality standards
have been adopted  for the major  air pollutants.   The  standards
for carbon monoxide  (CO),  for example, specify the  following
concentrations  and averaging times:*
                      35  ppm CO for 1 hour
                      9 ppm CO  for  8 hours
The U.S.  standards "are  not to be  exceeded more  than once  a
year"  in  the  "ambient air," which  is defined  as  "that portion
of the  atmosphere,  external to buildings, to  which the general
public  has  access."
     The current U.S.  definition  excludes special occupational
groups  (traffic policemen,  taxi  drivers, etc.),  but it
apparently includes shoppers,  housewives, businessmen, school
children,  or any others  who are  outdoors and are, thereby,
exposed to ambient air pollutants.

 *One ppm (part per million by volume)  is equal to 1.14 mg/m
  at  24 C and 1 atmosphere pressure.)

-------
                              2099
3.   Methodology of Study
     In our study, we define a "critical population" to be any
subset of the general population who share common characteris-
tics and, as a result, are exposed to similar atmospheric
concentrations.  To demonstrate how the exposure of one par-
ticular critical population may be more adequately monitored,
we focused on one critical population—downtown pedestrians.
     To determine their true exposure, we developed a
Simulated Exposure Sampling (SES) technique.  In this
approach, the investigator walks along the sidewalk while
collecting an ambient air sample in a specially prepared,
plastic bag.  The intake tube is held at "breathing height,"
and the bag is filled by a small, portable electric pump.*
The result is a "linear average" of CO concentration over
several city blocks or more.  Enough samples are collected to
provide a statistically valid measure of the human exposure
along a given route and over a designated averaging time.
     We collected 425 SES bags on 21 days in midwinter in the
downtown area (South First Street) of San Jose, California.
Each bag was filled continuously during walks over each of
four different routes.  Generally, each sample required about
5 minutes.  While walking, the investigator generally main-
tained the same pattern of movement as pedestrians—for
example, halting for traffic signals.
4.   Results of Study
     The SES values were significantly higher than values
simultaneously recorded at an official air monitoring station
approximately 2.6 kilometers from the downtown area  (Table 1) .
The 425 SES values averaged 9 ppm—1.6 times the average value
of 5.5 ppm recorded at the air monitoring station.  Individual
SES values ranged as high as 10 times the monitoring station
•Breathing height was defined as 5 ft. +1/2 ft. above the
 ground.   Bags were constructed of Mylar and required about
 5 minutes to fill.  All bags were analyzed in a central
 laboratory by nondispersive infrared absorption.  The pump
 was powered by nickel-cadmium batteries and was constructed
 with inert Teflon parts.

-------
                                             TAiLI 1




      MSULTS OP IIMULATID IXPOWRI •AMU.INQ (MS) IN DOWNTOWN IAN JOtf, GROUPED BY DAT!


DM
11/10/70
11/11/70
11/17/70
12/11/70
12/22/70
12/24/70
12/30/70
1/03/71
1/06/71
1/07/71
1/18/71
1/19/71
1/21/71
1/26/71
1/27/71
1/28/71
1/29/71
2/03/71
2/04/71
2/24/71
3/19/71
Owrall
Rtwito;

No. of
tmftutl*m
••n^fW
27
10
IB
36
40
40
41
a
4
11
12
8
10
18
6
30
9
10
12
34
41

429

••••I*—*
•ffnpung
jbjlnjj I*..
rwHJUf nra«
.0
.3
.3
.0
.0
.5
.0
2,0
0,3
2.3
3.3
1,3
1.3
4.0
1,0
6,3
1.3
2,0
2.0
7.3
8.3

97,0
CO Conowwitton, ppm*
Hmutettd lnpoMM tamp*
fcA^^M
WMVI
7.4
4.3
10.0
a. 6
13.0
14.2
9.1
6.4
3.2
8.8
10.3
7.3
3.3
9.9
11.4
s.e
8.3
5.3
7.0
5.2
7.6

9.0
ltd. Dm..
2.2
3.8
3,3
3.1
3.9
3.9
2.7
1.6
1.3
3.2
3.3
2.3
1.6
3.0
2.8
2.9
2.3
1.8
4.0
2.1
3.1

4.4
iinnttinr
MQnnQr
Mm
3.4
2.1
7.9
5.3
4.4
6.2
5.6
5.8
3.2
4.3
7.3
7.0
3.2
7.6
8.5
6.8
5.9
6.2
3.8
2.4
5.0

5.5
idViltNt
ltd. Dm.
1.0
0,1
2.4
1.3
2.2
1.9
1.5
3.5
0.6
0.3
2.1
0.6
0.5
1.8
2.3
1.7
2.0
1.5
1.3
0.6
1.1

2.1

Ratio
(818/Mon.)
1.4
2.1
1.3
1.6
3.0
2.3
1.6
1.1
1.0
2.1
1.4
1.1
1.0
1.3
1.3
1.3
1.4
0.8
1.2
2.1
1.5

1.6
•ppm • putt par million by volum.

-------
                              2101
values.  The correlation coefficient between SES values and
official monitoring station values was positive, but low
(r = +0.20).
     The seven dates on which SES values were collected over
at least 8 full hours are of particular interest because the
U.S. ambient air quality standard specifies an 8-hour period.*
On these dates, the 8-hour average of SES values ranged
between 1.4 and 3 times the values observed at the air moni-
toring station (Table 2}.  Application of the sign test found
the difference in means to be statistically significant in all
seven cases (p < 0.01).
     On three of these dates, downtown pedestrians or shoppers
spending 8 hours in the area were exposed to CO concentrations
above the U.S. standard of 9 ppm, while the monitoring station
reported values below the standard.  On the two dates with
highest concentrations, there was considerable variance in the
SES values (Figures 1 and 2).  These were dates of heavy
downtown traffic, and the SES values, which are indicative of
concentrations to which downtown pedestrians are actually
exposed, were strikingly greater than values reported at the
same time at the official air monitoring station.
     SES data also illustrate the feasibility of obtain-
ing true measures of the exposure of critical population
groups with relative ease and economy by the SES approach.
5.   Implications for Setting Air Quality Standards
     In any given city, at any instant of time, actual pol-
lutant exposures occur both above and below the values
observed at a fixed monitoring station.  Figure 3 shows a
hypothetical distribution of 8-hour CO exposures for the pop-
ulation of a particular city during any given 8-hour period.
The shaded area under the curve to the right of x* is the
probability that a given individual will be exposed to an
8-hour CO concentration greater than x*.
*The average value of 30 or 40 discrete, 5-minute bag samples
 collected over an 8-hour period can be used as a valid
 estimate of the overall average during the period.

-------
                             TABLE 2
           RESULTS OF SIMULATED EXPOSURE SAMPLING (SES)
          OVER EIGHT-HOUR PERIODS IN DOWNTOWN SAN JOSE*
                      SIMULATED      MONITORED
DATE
11/10/70
12/11/70
12/22/70
12/24/70
12/30/70
2/24/71
3/19/71
NO. OF
SAMPLES
27
36
40
40
41
34
41
EXPOSURE SAMPLE
(MEAN CO, ppm)
7.4 (± 0.9)
8.6 (±1.1)
13.0 (± 1.9)
14.2 (± 1.9)
9.1 (i 0.9)
5.2 (± 0.7)
7.6 (± 1.0)
VALUE
(MEAN CO, ppm)
5.4 (± 0.4)
5.5 (t 0.4)
4.4 (± 0.7)
6.2 (± 0.6)
5.6 (i 0.5)
2.4 (± 0.2)
5.0 (t 0.3)
RATIO
(SES/Mon.)
1.4
1.6
3.0
2.3
1.6
2.1
1.5
•VALUES IN PARENTHESES ARE 95% CONFIDENCE INTERVALS.

-------








,li






-
*j







-„
*~*
>:oo i :oo



H1






Air
M





*•-••
io;oo
,••
S
CO,

>ni»ormg Stotion -
yi
O
i
9:00

^Cl

V-

~\
\
icr
e:oo 7:
ppm
40
30
20
10
0
00
                        -«— Time, am
     Air Monitoring
   5:00
   4:00
3:00
2:00
1:00
                                                    CO, ppm
                                                         40
                                                         30
                                                         20
                                                         10
12:00
FIGURE 1.
            ••— Time, pm

          DECEMBER 22, 1970

CONCENTRATION MEASURED BY SIMULATED EXPOSURE SAMPLING ON
DOWNTOWN FIRST STREET IN SAN JOSE, CALIF., ALONG WITH CONCEN-
TRATION MEASURED AT THE AIR MONITORING STATION. (NOTE: TIME
IS PLOTTED FROM RIGHT TO LEFT.)

-------
                                                          CO.ppm
                                                              40
           Air Monitoring Stotion
                                                              90
                                                              20
      12:00       11:00      10:00       9:00
                               •*- Time, am
8IOO
7:00

5.



Air Monitori


•v^
x>

"v


-^—
rig Station — j
T
j

-

If
,-X




ta/w
4:00 3:00
-A
•5
J.





^ ^^^v
2:00


-


• • •
CO,




^_ i
:

:1P
1:00




T
I.
12
ppm
40
30
20
10
0
:oo
•• — Time, pm
DECEMBER 24, 1970
FIGURE 2.   CONCENTRATION MEASURED BY SIMULATED EXPOSURE SAMPLING ON
           DOWNTOWN FIRST STREET IN SAN JOSE. CALIF.. ALONG WITH CONCEN-
           TRATION MEASURED AT THE AIR MONITORING STATION. (NOTE: TIME
           IS PLOTTED FROM RIGHT TO LEFT.)

-------
   .40
   -30
o
QC
a.

w  .20
oc

CO
X
LLJ
   .10
   0.0
PROPORTION OF EXPOSURES

ABOVE THE STANDARD
                                                                    12
                  14
                                X, CONCENTRATION (ppm CO)
        FIGURE 3.   HYPOTHETICAL DISTRIBUTION OF CO 8-HOUR EXPOSURES IN A GIVEN CITY.

-------
                              2106
     The area also may be interpreted as the proportion of the
general population exposed to pollutant concentrations above
x* during a particular 8-hour period.  If, for example, x*
is the U.S. CO standard of 9 ppm, the goal of air pollution
control efforts would be to shift the curve as much as
possible to the left, minimizing the area under the curve to
the right of x*.  With such a model, of course, some propor-
tion of the population, however small, will always be exposed
to concentrations greater than x*.  This is why air quality
standards should state more precisely the proportion of the
population to which they are intended to apply.  An ideal CO
standard, for example, might state, "the value applies to the
highest 5 percent of 8-hour exposures of the general popula-
tion, which consists of ..."  Or the standard might apply to
the highest 20 percent of the exposures of a given critical
population group, such as downtown pedestrians.
     In our view, a more precise, probabilistic definition
would permit designing monitoring networks that more
faithfully reflect true human exposures.

-------
                              2107
                         REFERENCES
    OTT, WAYNE R., "An Urban Survey Technique for Measuring
    the Spatial Variation of Carbon Monoxide Concentrations
    in Cities," Ph.D. Thesis, Department of Civil
    Engineering, Stanford University, Stanford, California,
    October 1971.
2.  OTT, WAYNE R. and ELIASSEN, ROLF, "A Survey Technique
    for Determining the Representativeness of Urban Air
    Monitoring Stations with Respect to Carbon Monoxide,"
    Journal of the Air Pollution Control Association,
    Vol. 23, No. 8, August l9?3, pp. 685-696.


3.  OTT, WAYNE R. and MAGE, DAVID, "The Representativeness
    of Urban Air Monitoring Stations with Respect to
    Carbon Monoxide," Proceedings of the Second Annual
    Environmental Engineering and Science Conference,
    Speed Scientific School of the University of Louisville,
    Louisville, Kentucky, April 20-21, 1972, pp. 379-394.


4.  "National Primary and Secondary Ambient Air Quality
    Standards," Federal Register, Vol. 36, No. 84,
    April 30, 1971, Washington, D.C., pp. 8186-7.

-------
                              2109
              PERSONAL AND INDOOR EXPOSURE METERS

            A, P, ALTSHULLER AND ANDREW E, 0*KEEFFE

Chemistry and Physics Laboratory, National Environmental Research
Center, Environmental Protection Agency, Research Triangle Park,
N.C., USA
ABSTRACT

     Current community health studies generally have to utilize
results of pollutant monitors in fixed or mobile sites to deter-
mine pollutant exposures of individual subjects.   Pollutant ex-
posures over 24-hour periods involve exposure not only to community
pollutant concentrations, but also to commuter and indoor pollu-
tant concentration levels.   The exposures of a discrete and
mobile group of subjects to community pollutant concentration
levels are not readily derived from monitoring results at a
limited number of fixed sites.

     The limitations in the present situation can be resolved by
the development of small portable "exposure meters" for measuring
an individual's exposure to air pollutants.   The individual in
the study could carry the instrument and operate it at home, in
a vehicle, office, laboratory, etc.   Projects are being developed
by the Chemistry and Physics Laboratory, National Environmental
Research Center, RTP, for exposure meters and as an alternative
simple portable samplers.   Emphasis will be on measuring nitro-
gen oxides, sulfur oxides and oxidant.   Because of the available
capability to do carbon monoxide in blood samples, there is no
current need for a carbon monoxide device.   Particulate species
such as sulfate and nitrate probably could be collected on appro-
priately designed mini-samplers.

-------
                              2110
     A state of the aft survey will be conducted, especially
concerned with fine partioulate species, noise and other environ-
mental pollutants.   This document will be used to develop the
long-range program on exposure meters in the U.S. Environmental
Protection Agency.   The status of available personal monitors
for physiological parameters will be evaluated by the Human
Studies Laboratory, National Environmental Research Center,
Research Triangle Park, N.C.

-------
                                     2111
1.  Introduction
Our present knowledge of the effects of pollutants on the human population
is very largely based on estimates of the dosage received.  These estimates
are derived from measurements of pollutant concentrations at fixed sites,
about and among which sites the individuals making up the population con-
tinually swirl in patterns so complex as to defy analysis.  The Environ-
mental Protection Agency is conducting a program to develop a capability
for recording the exposure of individuals to selected pollutants, without
placing constraints upon the subjects' normal activities. This paper will
examine the several strategies available for approaching that ideal, and
will consider the compromises demanded by each in the interest of practi-
cality.
2.  Physical Methods
2.1  Principle;  A physical property of a gas is measured and related to
concentration.
2.2  Scope;  Spectral absorbance is the only physical property which cur-
rent technology can measure with sufficient accuracy for the present appli-
cation.  The requirement of a very long optical path to attain parts-per-
billion sensitivity appears to be incompatible with size constraints
inherent in the term "personal exposure meter" and (because of the extreme-
ly precise collimation needed) with the inevitable exposure of such an
instrument to inertial disturbances when carried on the person.  In light
of these limitations, absorbance methods have been given low priority in
the present program.
3.  Energy-Transfer Methods
3.1  Principle;  Reaction of the pollutant of interest (usually with a
second reactant) results in a transfer of energy quantitatively related to
concentration.
3.2  Scope;  Three major forms of such devices are presently known to exist:
     3.21  Chemiluminescence:  Gas molecules interact with production of a
           new molecule in an excited state.  Excited molecule relaxes to
           ground-state with release of light energy:
                     A + B  	t C*
                         C*	> C + hv

-------
                                    2112
     3.22  Fluorescence:  Gas molecule absorbs light energy of a specific
           wavelength and is raised to an excited state.   This relaxes to
           ground-state with release of light energy of (ordinarily) greater
           wavelength:
                     A + hv  	> A*
                           Y
                         A*	>hv ,
     3.23  Electrochemical:  Gas molecule is oxidized (or reduced) at an
           electrode of a galvanic cell, transferring electrical energy to
           (or from) the opposing electrode:
                     A° __, A* + e
4.  Sorptton Methods
     4.1  Principle;  A sorbent collects the gas of interest at a rate
dependent upon concentration.  Subsequent desorption and measurement of the
gas (or of a stoichiowetrically related product) is usually performed in an
off-site laboratory.
4.2  Scope;  The great majority of pollutant gas measurement methods in use
a few years ago were of this type.  Classic examples would be the lead
candle method for S0_ and iodometric methods for ozone.
5.  Research Strategy
5.1  General Considerations:  Ideally, the device that is sought will be
the size of a pocket radio  receiver, and will store within itself a con-
tinuous record of the instantaneous concentrations of a given pollutant
experienced over an extended sampling period - say 24 hours.  It should
have sufficient sensitivity to detect the pollutant at or near background
levels, and should be free  from interference from other substances  likely
to be encountered in an urban atmosphere.  Unit cost must be kept at a level
compatible with eventual use on a  scale sufficient to support meaningful
health-effect studies.
     It is apparent that the above ideal will not be reached in a single
step.  Thus, we must choose those  parameters that can best be compromised
with reasonable assurance  that either  (a) later developments will eliminate
the compromise, or  (b)  health-effect studies will not be rendered impracti-
cal.  On such considerations, the  ideal requirements pertaining to  size and
to time-resolution  of educed data  have been  considerably liberalized  so
that a start can be made on assembling and evaluating prototype devices
representative of the several types of methods previously discussed.

-------
                                     2113
 Initial prototypes are  expected to be no  larger than a briefcase in size,
 and to be capable of measuring the integrated average concentration of a
 pollutant for a period  of  time in the range  0.01-1.0 day.
 5.2  Specific Plans for Initial Phase
     5.21  Ozone:  The  potential of continuous instrumentation will be eval-
           uated through the medium of a  miniaturized ethylene-ozone chemi-
           luminescence device.  It is expected that this will include
           substitution of a less volatile substance for the ethylene, thus
           obviating the need for a pressurized cylinder.  A parallel effort
           will examine the feasibility of reacting ozone with a solid
           organic substrate to form a metastable adduct that subsequently
           undergoes chemiluminescent decay  upon heating.
     5.22  Sulfur Dioxide:  A commercially available electrochemical cell
           designed for measurement of ambient SO  will be miniaturized and
           provided with integrating electronics, as a candidate method.
           Also, an efficient sorbent will be identified, whose SO. adduct
           can later be  thermally decomposed, the product being measured
           with a flame photometric sulfur detector.
     5.23  Nitrogen Dioxide:  Miniaturization of chemiluminescence instru-
           mentation for NO  is possible, but has the drawback that power
           is required  to convert N02 to NO.  Hence, the choice will prob-
           ably go to an electrochemical device.  The alternate method will
           involve sorption, subsequent thermal desorption, and eventual
           measurement through chemiluminescence.
 6.  General
     Wherever possible,  especially in the sorption methods outlined above,
 the need for accurately metering the air sample will be eliminated by inter-
posing between sample and sensor a membrane permeable by the gas to be
measured.   This renders  the sensor responsive to partial pressure of the
analyte rather than to the absolute quantity contained in a sample of
known volume.  Relief from the requirement of a precision metering pump will
greatly simplify the  construction of a miniature instrument.  We are also
assessing,  by means of aerodynamic modelling calculations,  the magnitude of
the error  that will be introduced by eliminating the sample pump altogether
and depending entirely on convection-diffusion for sample acquisition.
Should  the  error prove to be acceptably small, this offers  still another
major  simplification.

-------
                                    2114
     Development of personal monitors for particulate pollutants is a more
difficult task.  It Is obvious that power must be supplied to move, separate
and collect such materials; this fact alone poses major problems.  Active
work on this segment of the program will not start for another year or so.
7.  Summary
     EPA has an active program under way to provide personal monitoring
devices for Important pollutants.  The instrument development program is
being thoroughly coordinated with the efforts of the Human Studies Labora-
tory In order to assure optimum response to their needs.  Our target is
the narrow zone that lies between the inadequate and the over-sophisticated.

-------
                              2115
     DIE AUTOMATISCHE UEBERWACHUNG DER KRAFTFAHRZEUGABGAS-
             IMHISSION IM ARBEITS- UND WOHNBEREICH

                     D, JOST,  W,  RUDOLF

 Bundesstelle fur Umweltangelegenheiten,  PilotPtetion Frankfurt/M
 BRD
KURZ FAS SUNG

     Ein vollautomatisches Messnetz zur Ueberwachung der Luft-
verunreinigung im Arbeits- und Wohnbereioh der Stadt Frankfurt
wird,  ausgehend von den Erfahrungen bei der Sahwefeldioxidiiber-
uachung, sur Ueberwachung der kraftfahrzeugbedingten Abgase
erweitert.   Aufbauend auf langjdhrigen Messreihen versehiedener
meteorotogisaher Grossen und von LuftT)erunreinigungen werden
mit Bilfe von Absehatzungen atmoaphdrischer Ausbreitungsvor-
g'dnge die Luftverunreinigungsmeasdaten bsw. Daten der luft"
verunreinigungsdosis fur die Bewohner des betraahteten Raumes
ermittelt.
     Folgende Komponenten werden dabei erfasst:  SO f CO, SO ,
                                                   2j        30
Oxidantien,  Gesamtstaub, Blei und weitere Metalle im Staub.
     Mit Hilfe der Untersuahung der Kon>elation verschiedener
Luftverunreinigungen ist ea moglich, die Zahl der Mesetellen
und die Zahl der zu untersuahenden Komponenten zu optimieren*
wobei die Anforderungen folgender Untereuahungsziele zu beruak-
siahtigen sind:
Fldohen- und zeitbezogene Belastung mit Luftverunreinigungen,
Trend verschiedener luftverunreinigender Komponenten, Smog-
Varnung  Optimierung von Messnetzen, Daten fur epidemiologische
Untersuahungen,
         diesen Aufgabenbereichen werden Untersuahungsergebniese
mitgeteilt,  die aueh eine Bilanzierung von Belastung durah
Luftverunreinigungen und Erholung in Ballungsgebieten gestatten.

-------
                              2116
ABSTRACT
     A fully automatic network for monitoring atmospheric pol-
lution in the industrial and residential areas of Frankfurt is
being extended to include vehicle exhaust gases, after experience
with sulphur dioxide monitoring.   On the basis of meteorogical
and pollution measurements made over several years*  pollution
data and data on the exposure of inhabitants of the area under
survey to pollution will be determined by using estimated
atmospheric spread patterns.
     The substances measured are:  SO-,  CO,  NO * oxidising
                                     b          X
agents, total dust, lead and other metals in the dust.
     By investigating the correlation between different pollu-
tant* it is possible to optimise the number of measuring points
and also the number of elements to be examined; the conditions
governing the following items to be examined, must be considered:
Area - and time - related pollution levels, patterns of different
pollutants, smog warning, the optimising of monitoring networks,
and data for epidemiological investigation.
     This type of data will produce results which also make it
possible to draw up a balance sheet of pollution and anti-
pollution factors in densely populated areas.

-------
                                2117
1.  Lage der MeBstellen und  Datenverarbeitung

Die bereits  1966 errichtete, seit dem 1.1.1974 in die Bundesstelle
fur Umweltangelegenheiten ("Umweltbundesamt") integrierte "Pro-
bemefistation Frankfurt/M  - Pilotstation fur Luftreinhaltung" be-
steht z.Z. aus einer Zentralstation und sieben "Unterstationen
(Abb. 1).

Die Zentralstation liegt ca. 1,5 km von der Stadtmitte entfernt in
einer Wohn- und Biirogegend. In ca. 6 km Entfernung von der Zen-
tralstation befinden sich an der Peripherie des eigentlichen Frank-
furter Stadtkemes die beiden grofieren Unterstationen (Ost- und
Weststation) mit drei (SO  , CO, CO ) bzw. zwei  (SO  , CO ) Mefi-
komponenten. Die vier kleineren Unterstationen mit nur einer Mefi-
komponente  (SO ) sind am  Rande des Frankfurter Verdichtungsge-
bietesund meist auBerhalb besiedelter Gebiete errichtet. Zur Uber-
wachung der kfz-spezifischen Komponenten CO, NO,  NO9, O^ und
                                                      Z.    o
Blei vurde eine Station in  der Frankfurter City eingerichtet.  Alle
MeBstellen sind iiber eine Fernwirkanlage und standig durchgeschal-
tete Datenleitungen der Deutschen Bundespost mit dem Prozefirechner
der Zentralstation verbunden.  Die Fernwirkanlage arbeitet nach dem
Frequenznmltiplexverfahren, hierbei konnen bis zu 18 MeBgeber an
eine Leitung angeschlossen werden. Dieses Verfahren ermoglicht
eine Mehrfachausnutzung der einzelnen Leitungen.

An der Zentralstation werden die meteorologischen Parameter
Windrichtung, Windgeschwindigkeit, relative Feuchte,  Luftdruck,
Temperatur  und Strahlungsbilanz,  sowie die Spurenstoffkom-
ponenten  Kohlendioxid,  Schwefeldioxid, nitrose Case,   Kohlen-
monoxid,  Gesamt-Kohlenwasserstoffe und Schwebstaubgehalt gemes-
sen. Zur Vermeidung grofierer Totzeiten und um Absorptionsver-
luste in den Leitungen auszuschlieBen, sind samtliche Cerate zur

-------
                               2118
Messung der Spurenstoffe an eine Ringleitung angeschlossen, die mit
     3
1,5m /min beliiftet wird.  Jedes Gerat wird darm nochmals durch eine
eigene Pumpe mit der zur Analyse notigen Luftmenge versorgt. Die an-
fallenden MeBwerte werden  iiber Zwischenverstarker generell in einge-
pragten Strom von 0-20 mA umgesetzt, auf einem Kontrollschreiber
kontinuierlich. registriert  und iiber einen Bemessungswiderstand an den
Eingang des Analog-Digitalwandlers (0 - 1 V) angepaflt. Die von  den
AuBenstellen zur Zentralstation iibermittelten MeBwerte werden ebenfalls
von der gepulsten Frequenz (2 -  12 Hertz) in eingepragten Strom umge-
setzt und parallel zur Rechnereingabe auf einem Analogschrieb regi-
striert. Auf diese Weise ist eine optimale Kontrolle der Aufienstellen
von der Zentralstation aus moglich. Analog-Digital-Vergleiche wer-
den im Stichprobenverfahren fiir alle Komponenten durchgefuhrt.

Alle 23 Stunden  werden  die Meftgerate  durch Rechneransteuerung dy-
namised geeicht. Durch  einen rechnerseitig gesetzten SchlieBer werden
iiber Relais und  Magnetventile zunachst die unteren Eichwerte (Eich-
gase fur CO und COO, Nullpatrone fur SO,,) aufgegeben: nach 6 Minu-
                   
-------
                            2119
ners kommen zwei Schreibmaschinen (Teleprinter), zwei Lochstrei-
fenstanzer, ein Lochstreifenleser und eine Grofirechnerkompatible
Magnetband-Einheit hinzu. Auf der ersten Schreibmas chine erfolgt
die Ausgabe des Routineprogrammes mit Zeilenkennung, Datum (Jah-
restag), Uhrzeit, Halbstundenmittelwerte, Varianzen und 3 Minuten-
Maxima. Die zweite Schreibmaschine dient der Korrespondenz mit
dem Rechner (Gestortsetzen einer Meftstelle,  Eintragen neuer Eich-
werte usw.), der Ausgabe von Stormeldungen durch den Rechner (z.B.
Leitungsausfall, Uberschreitung eines unteren oder oberen Grenzwer-
tes usw.) und der Protokollierung von Sonderprogrammen. Nach der
Ausgabe der Protokolle auf den  Schreibmaschinen erfolgt die gleiche
Ausgabe unter  Wegfall der Blanks auf Stanzer und Magnetband. Die
Lochstreifen und Magnetbander  werden auf dem Groflrechner des Deut-
schen Wetterdienstes (CDC 3 400) weiter  verarbeitet.
1m einzelnen nimmt  der Prozeflrechner folgende Aufgaben  wahr:
    1. Umsetzung der Analog- in Digitalwerte •
    2. Priifung dieser Digitalwerte auf Richtigkeit (es kommen hier
       5 Kriterien  zur Anwendung, wie z.B.  Priifung ob die Anzahl
       der Einzelwerte fiir einen statistisch gesicherten Mittelwert
       ausreicht).
    3. Verarbeitung der gepriiften Einzelwerte zu statistischen Kenn-
       groBen.
    4-. Automatische Eichung der MeBgerate.
Durch die Punkte 1-4 1st eine optimale Sicherstellung der Richtig-
keit der auf den Lochstreifen ausgegebenen Meftwerte  gegeben.
Nachtragliche und zeitraubende  Korrekturen werden auf ein Mindest-
maft beschrankt.
In der Routineverarbeitung werden z.Zt.  die Haufigkeitsverteilungen
und mittleren taglichen Gange  aller Komponenten fiir jeden Monat oder
beliebig wahlbare Zeitraume errechnet. Fiir Sonderauswertrungen und
den Nachtrag von MeBkollektiven,  der z.T. durch Leitungs- oder Rech-

-------
                               2120
   nerausfall bei Wartungsarbeiten notwendig wird,  steht ein Tischrech-
   ner der Firma Hewlett Packard Typ 9810 A mit 2036 Programmschritten
   und 111 Datenregistern zur Verfugung. Diesem Tischrechner sind  als
   periphere Einheit ein Lochstreifenleser, ein Plotter und ein Teleprin-
   ter mit Stanzer angeschlossen. Eine umfangreiche Anzahl von festver-
   drahteten Funktionen fiir mathematische und statistische Zwecke,  sowie
   fur die Plotter-Programmierung ermoglichen eine leichte Weiterverar-
   beitung der angefallenen Mefiwerte. 1m Sinne einer Zeitersparnis wird
   da mit auch die Eingabe eines nach speziellen Gesichtspunkten vom
   Grofirechner erstellten Lochstreifens in den Tischrechner sinnvoll.

2. Lage und Instrumentierung der Mehrkomponenten-Meflstation zur Uber-
   wachung und Analyse der Immissionen von Kraftfahrzeugabgasen in der
   Frankfurter City.
   Zur Messung der durch Kraftfahrzeugverkehr verursachtenImmissio-
   nen in der City Frankfurts wurde im Juli 1972 in der Frankfurter In-
   nenstadt zwischen Hauptwache und dem Eschenheimer Turm eine Mehr-
   komponenten-MfcBstelle eingerichtet (Abb. 1). Die im weiteren als City-
   Station Rundschauhaus bezeichnete Me6stelle liegt an einer dreispurig
   befahrenen  HauptstraBe mit hoher Verkehrsfrequenz. Die Analysenge-
   rate sind in 5 und in 20 m Hohe in im Freien stehenden MeBschranken
   untergebracht.
   Die Ansaugstelle ist 40 m von der nachsten Strafienkreuzung entfernt
   und befindet sich in der Mitte des Biirgersteiges 3,5 m iiber Strafien-
   grund  bzv.  3,5 Meter Dachhohe (ca. 23 m iiber  Strartengrund).
   An der MeBstelle wird der Verkehr durdx Einbahnregelung in siid-
   nordlicher Richtung gefuhrt. Die Verkehrsfrequenz ist in den Tages-
   stunden standig sehr stark ( ca. 1300 Kfz/Stunde ) und erreicht in den
   Nachmittags- und Abendstunden ihr absolutes Maximum (ca. 2000 Kfz/h).
   Zur Zeit der abendiichen "rush-hour" wird die Strafie, bedingt durch
   Ampelsteuerung und Verkehrsaufkommen,  bis an die Grenze ihrer Auf-
   nahmefahigkeit belastet. Die Bebauung ist in der Nahe der Mefistelle

-------
                                   2121
 V"-- '-•• ~* •
 n •          -t  ' • i K ''  •
3.  •    • ^ -.- A -  -. •» - -^ ...
                       •    &:^':-rl       -«
                  j;.     H^_^-   ::\je      •  (
                    -T >              I*r.  -s  Uf
                     i-
          ':x?tfb-^,;- ...%•,

   i|       p
   =yr -,r
&  ^
                                                     ^C3
    :"":-ss:
                s£
^•%S£/3i---          Tk
H'b: ^fJt*••-'  --  -       -r-4«
1*1 •>••:.-      vr^
 It     |-«:H-jrbiL:
                             Q ZENTRAL.ST»TIO«
                               -"i SJi IO>
                               *»irkanbge
in

til
^errrwirkoTiage
Aunenstatiorw



Analog-
Digital-
Umwand
Zentraleinheit d
Prozessrechners
AEG 60-10
IBK)
I 	






	 * 	 ,
BecJienurvjsfetd
des Rechners

Lochslreifcnleser

1 Sc^reibmascMne
(Kso!e)

1 Lochstreifensfanzar

2_ Sctveibmcs^iine
I Koosofe )

2. Lochstreifenstanzer
                            Abbildung 2

-------
                                 2122
 geschlossen und einbeitlich ca. 20 m hoch.

 Im Moment erfolgt die Messung von Kohlenmonoxid (CO), Stickstoff-
 monoxid (NO), Stickstoffdioxid (NO?) und Ozon in beiden Ebenen mit-
 tels kontinuierlicher Infrarot- und Chemoluminiszenz-Verfahren. In
 der ersten Etage warden zusatzlich Filterproben (low volume) zur
 Analyse der Schwermetallkonzentrationen (Blei, Eisen, Kupfer,
 Cadmium) mittels Atomabsorptionsspektrometrie gezogen. Analysen-
 methoden, Geratetyp und Herstellerfimra sind der folgenden Tabelle
 zu entnehmen:

1. Etage:
Ansaughohe
3.5m iiber
Straften-
grund


Dach
Ansaug-
hohe
3.5 m iiber
Bebauungs-
hohe und
ca. 23 m
iiber
StraBen-
grund
CO
Infra -Rot
Uras II
Hartmann
+ Braun



wie
1 . Etage




NO
Chemolu-
miniszenz
NOx-Moni-
tor
Bendix


wie
1. Etage




N02
wie
NO



wie
1.
Eta-
ge




°3
Chemolu-
miniszenz
Ozon-
Monitor
Bendix


wie
1. Etage




Schwermetalle
Filterproben
mit low volume
bei 24 Stunden
Sammeldauer
Atomabsorp-
tionsspektro-
metrie
Perkin+Elmer
entfallt




Die Unterbringung der MeBgerate im Freien ermoglicht relativ kur-
ze Ansaugleitungen und gute Wartungsbedingungen, die insbesondere
die Ozon- und NO -Messung wesentlich erleichtern. Der EinfluB der
schwankenden Auftentemperaturen und der Aufheizung der MeUschran-
ke durch Sonneneinstrahlung konnte durch eingebaute Liifter inner-
halb der werksseitig angegebenen Toleranzgrenzen iiir die einzelnen
MeBgerate gehalten werden.

-------
                                     2123
IE PPM
IB PPK
 C PPM
                                                             \
TRSCSM1TTEI-   OEfJ  CD-KDN2EKITRRTIDNEN  ZENTRRUSTr^-INNENSTR&TSTBTIDN 	

TRSCSMITTCl-   I>ER  W!N»SE5CMWINOISKEIT  ZEKITRRl-STHTlDM  » I

    Abb.  3.  Pilotstation des BMI  Frankfurt/Main  - Juni  1973.
                  SEPTEMBER 1973 ALLE TAGE  RUNDSCHAU-HAUS
                  • 1. STOCK
                  + DACH
      Abb. 4.   Pilotstation  fiir Luftreinhaltung.

-------
                                2124
3. Das horizontale und vertikale CO - Konzentrationsfeld.

Wahrend bei der Uberwachung der Schwefeldioxidkonzentrationen in
Gebieten, die vorwiegend durch. Hausbrandemission beeinfluftt sind,
ein relativ homogenes  Konzentrationsfeld angenommen werden kann,
treten bei den durch. Kraftfahrzeugemissionen verursachten Immissio-
nen in der Straflenluft starke horizontale und vertikale Konzentra-
tionsgradienten auf.
So betragt der Unterschied zwischen den an der Zentralstation und
den in der City gemessenen mittleren SO2-Konientrationen ^n8efshr
25%. Die CO-Konzentrationen an der Zentralstation liegen jedoch oft
um den Faktor 10 niedriger als die in der Innenstadt im Straflen-
niveau ermittelten CO-Konzentrationen. In Abb. 3 ist beispielhaft
der Verlauf der Tagesmittelwerte an diesen  beiden Stationen fiir den
Juni 1973 dargestellt. Es zeigen sich deutlich die oben beschriebenen
Konzentrationsunterschiede, wobei auBerdem nur eine schwache Kor-
relation zwischen den Einzelwerten besteht.
In Abb. 4 sind fur den Monat September 1973 die mittleren tagli-
chen CO-Konzentrarionsvariationen dargestellt, fvir die MeBhohen
3-50 m und 23 m iiber StraBenniveau, die bestatigen,  dafi auch in
den City-Bereichen der GroBstadte die von Kraftfahrzeugen emittier-
ten Priroa remission en sehr schnell mit der Hohe abnehmen. Nach Un-
tersucb-ungen in Berlin (Lahmann,  1972) und Frankfurt (Busch,
Georgii, Weber,  1967)  kann   die CO-Abnahme mit der Hohe in Stras-
senschluchten expontiell angesetzt werden, wobei wegen des "Street-
canyon effekts", d.h. der Ausbildung einer rotorartigen Stromung bei
Windrichtungen quer zur Bebauungsflucht, noch eine starke Abhan-
gigkeit der CO-Konzentrationen von der Windrichtung zu beobachten
ist. Abb. 5 zeigt, dafi eine Winddrehung um 180  die gleiche Konzen-
trationserhohung bzw.  -erniedrigung bewirken kann,  wie eine Steige-
rung der Windgeschwindigkeit von 0.5 auf 4.5 m/sec.

-------
                                       2125
     SO
:
     zc
      :

        CO
        ppm
            Abb.  5-  CO Konzentration  als
            Funktion der Wlndgeschwindig-
            keit  aufgeschliisselt  nach  Wind-
            rich tungssektoren.
                                                           WIXDRICHTUKG:
                                                           J-J-LXORD (315- n)
                                                           	 OST  ( 4S-U4)
                                                           	st'D  (135-224)
                                                           	WEST (22S-3H)
                  2       3      i       5       6
                   CO RUXDSCHAU-HAUS VIXIER 1972
                                                                     8 m/s
 . M. ? (n«

. TToP   ff . ?.l «/=ee

  Iiollnian der CD - Konjvitrmtlan

-------
                               2126
 Die starken vertikalen und horizontalen CO-Konzentrationsunter-
 schiede im Stadtgebiet, denen durch die Variation der Emission und
 der Ausbreitungsparameter im Tages- und Jahresverlauf noch ein
 zeitlicher Gang iiberlagert ist, machen eine ortlich und zeitlich re-
 prasentative Immissionsbestimmung aufierst schwierig. In Frankfurt
 wird zur Losung dieser Frage versucht, die im Stadtgebiet vorkom-
 menden oder auch augenblicklich herrschenden CO-Konzentrationen
 nach unten und oben abzugrenzen. In der Umgebung der City Station
 konnen, bedingt durch Verkehrsaufkommen  und Bebauung die maxi-
 malen CO-Belastungen sehr gut erfaBt werden.
 An der Zentralstation werden fur Biiro-und Wohngegenden reprasen-
 tative Konzentrationen erfaflt. Damit ist sichergestellt,  daft die Ein-
 wohner Frankfurts  meistens Konzentrationen ausgesetzt sind die
 zwischen den an diesen beiden Stationen gemessenen Werten liegen.

 4. Modellierung des CO - Konzentrationsfeldes.
 Zur Unterstutzung der meBtechnisch.en Erfassung der CO-Konzen-
 trationen wird z.Z. in Frankfurt versucht, die durch Kraftfahrzeu-
 ge verursachten Luftverunreinigungen durch Modellrechnungen abzu-
 schatzen. Ausgehend von Verkehrsmengenzahlen und mittleren Ge-
 schwindigkeiten in Rastern von 500  x 500 m konnte mit Hilfe von
 Emissionsfaktoren die mittlere tagliche CO-Emission pro Einheits-
flache errechnet werden. Diesen Emissionsfaktoren liegen umfang-
reiche Untersuchungen in Koln zugrunde (Emissionskataster Koln
1972), bei denen durch Mefifahrten unterschiedliche  Fahrzyklen und
deren Haufigkeitsverteilung ermittelt  werden konnten.  Korrela-
tionsrechnungen zeigten, dafi jeder  Zyklus  in guter Naherung durch die
mittlere Zyklusgeschwindigkeit definiert ist. Die einzelnen Zyklen wur-
den durch Priifstandversuche simuliert und die dabei emittierten Men-
gen der Kfz-Abgase gemessen.

-------
                                                      2127
                                               Ylo.
Stab. Hi. ?
EM - ?70°   ff - 3.1 «/
—— Isollilnn oer *CK - K^n^cn

-------
                                2128
 Aus den Verkehrsdaten und den Emissionsfaktoren konnten fur das
 Stadtgebiet von Frankfurt folgende Emissionen ermittelt warden:
               CO     :    154   Tonnen / Tag
               NO     :      7.1
               CnHm   :      4.6
 Nach Berechnung der Emissionen erfolgte in einem weiteren Schritt
 mittels eines Diffusionsmodells (Baltrusch 1972^ die Berechnung der
 Immissionskonzentrationen.  Dabei wurden die in den Flachen  (500 x
 500) emittierten Mengen als Punktquelle in der Rastermitte zusam-
 metigefafit und fur ein gleichliegendes I mmiss ions raster durch Su-
 perposition der einzelnen "Rauchfahnen" die Schadgaskonzentra-
 tionen errechnet.
 tJberpriifungen der errechneten Ergebnisse zeigten,  daB die zuerst
 gewahlte Dimensionierung von Emissions- und Immiss ions raster
 (jeweils 500 x 500 m) nicht die Bedingungen einer Flachenquelle er-
 fullte,und daS  deshalb zu hohe Konzentrationen errechnet wurden.
 Erst eine Unterteilung der pro 500 x 500 m Einheitsfiachen errech-
 neten Emission auf 25   im jeweiligen Raster  gleichverteilte Ein-
 zelquellen ermoglichte  die Errechnung korrekter Konzentrationen.
 Die Berechnungen wurden fur die vier Hauptwindrichtungen (Nord,
 Ost, Siid, West) und fur unterschiedliche atmospharische Stabilitat
 durchgefuhrt.
 Das hier beschriebene Modell kann als "stadtisches Backgroundmo-
 dell" bezeichnet werden, bei dem Emissionen aus den StraBenschluch-
 ten herausgehoben und in einer Hohe von 20 Metern als Punktquellen
 angenommen werden. Fiir diese Hohe sind auch die spater angegebenen
 Konzentrationswerte berechnet. Die Konzentrationen im Stralienniveau
 werden auf diese Weise nicht erfaBt und miissen durch ein besonderes
 Strafiermodellr das in Vorbereitung ist, berechnet werden.
In Abb. 6,  7 und 8 sind  fiir die Stabilitatsklasse 2 (nach Klug) West-
wind und fur eine mittlere Geschwindigkeit von 2.1 m/sec die Konzen-
trationsfelder fur Kohlenmonoxid (CO), Stickoxide (NO ) und Kohlen-

-------
                                  2129
«  04  06  08   10
Abb. 9»  Prozentuale Verteilung
des Tagesverkehrs in Frankfurt/
Main.
Abb. 10.  Hittlerer Tagesgang
Kohlenmonoxid (gemessen am Inst,
fiir Meteorologie u. Geoph.).

Abb. 11.  Korrelation der CO-
Werte an der Zentral-Station
mit den CO-Werten der Stadt-
rundfahrten zum 1700 Terrain.
                                                    	a* vtni
                                                    	c,«,
Abb. 12.  Summenhaufigkeit  der
     CO Mittelwerte.

-------
                                2130
 wasserstoffe (HC) dargestellt.  Die Linien gleicher Konzentration zei-
 gen deutlich den EinfluB der City und des Autobahnkreuzes im SiLd-
 westen Frankfurts (Maximalwert 4 ppm CO). Zu bemerken ist, daB die
 hier errechnetaiTagesmittelkonzentrationen gut mit den im Dachni-
 veau der City-Station gemessenen Konzentrationswerten iibereinstim-
 men.
 Zur Simulation des Tagesganges der CO-Konzentrationen  wurden, ge-
 maB dem Verlauf des Verkehrsaufkommens (Abb. 9), die errechneten
 Konzentrationen gewichtet und der Tagesgang der atmospharischen
 Stabilitat iiberlagert. Durch diese Methode konnte  der an der Zen-
 tralstation gemessene mittlere tagliche Verlauf der CO-Konzentra-
 tionen (Abb. 10) sowohl in der Hohe der Konzentrationswerte als auch
 in seiner Form gut angenahert werden. Insbesondere die in den Som-
 mennonaten beobachtete Verschiebung des abendlichen CO-Maximums
 in die  spa ten Abendstunden ergab sich aus dem Gang der Stabilitat.  Wie
 Abb. 9 zeigt  sinkt das Verkehrsaufkommen von seinem Spitzenwert
 urn 17 Uhr bis 20 Uhr auf die Halfte ab, sodaC bei konstanten Stabili-
 tatsbedingungen auch die Konzentration auf die Halfte  absinken mufke.
 Unter der Annahme,  daB im Mittel die atmospharische Stabilitat zwi-
 sch.en diesen Zeitpunkten um eine Klasse stabiler wird, ergibt sich.
 eine Erh.6h.ung der Werte um Faktor 3. Dadurch wird das sinkende Ver-
 kehrsaufkommen kompensiert und das abendliche CO-Maximum auch
 rechnerisch um 20 Uhr ermittelt.

 5- Mobile Messung der Kohlenmonoxid-Konzentration
 Als Erganzung zur stationsren Kohlenmonoxid-Messung, und zur Un-
 tersuchung der Belastung des Autofahrers wahrend seines Weges von
und zur Arbeitsstelle,  wurden 1969und im Winterhalbjahr  1970/71
 CO-Messungen mit einer mobilen Mefistation durchgefiihrt.
 Die Streckenfuhrung wahrend der Mefiperiode  1969 erfafite den west-
 lichen  Teil Frankfurts, das Gebiet um den Hauptbahnhof und die City.

-------
                               2131
Die Messungen wurden von 17.oo bis 17-3o Uhr durchgefUhrt.  In der
zweiten MeBperiode (1970/71) wurde jeweils morgens zwischen 7-3o
und 9.3o Uhr und abends zwischen 17.oo und 19-oo Uhr ein west-ost-
liches Profil gefahren. Verwendet wurde das Gasspurgerat der Firma
Drager  mit Priifrohrchen "CO - 5 b".
Die Ansaugstelle befand sich jeweils in Hohe des rechten, vorderen
Ausstellfensters des verwendeten Kraf£ahrzeuges.
Eine Korrelationsrechnttng der durch mobile Messungen ermittelten
Kohlenmonoxid-Konzentrationen mit den an der Zentralstation gemes-
senen CO-Werten wahrend der ersten MeBperiode (1969) ergab einen
Korrelationskoeffizienten von 0.72 (Abb. 11), fiir die Winterhalbjah-
re 1970-71 lag der Korrelationskoeffizient bei 0.6. Daraus ergibt sich
fur beide Untersuchungen ein. signifikanter,  statistischer Zusammen-
hang der Mefiwertkollektive, der wesentlich durch den taglichen Gang
von Verkehrsaufkommen und CO-Konzentration bestimmt wird. Die
Abweichungen der Einzelwerte von den Regressionsgeraden sind
aber  so groft, daB fiir den Einzelfall die CO-Konzentrationen in der
Innenstadt nicht aus den an der Zentralstation gemessenen CO-Kon-
zentrationen ableitbar sind. So treten z.B.  1969 bei CO-Konzentra-
tionen zwischen 2 und 3 ppm an der Zentralstation in der Innenstadt
im fliefienden Verkehr CO-Konzentrationen zwischen 17 und 70 ppm auf.
Fiir die CO-Messungen in der Winterperiode 1970/71 wurden die er-
mittelten Werte nach Tageszeit (morgens bzw. abends) und Stadtbe-
zirk (Ausfallstraflen, AuBenbezirke, Stadtzentrum) getrennt unter-
sucht.
Die Darstellung der Meflwerte in Form von Summenhaufigkeitsver-
teilungen (Abb. 12) ergibt folgende 50%-Werte:
fiir das  gesamte Wertekollektiv              38 ppm
Konzentration auf den Ausfallstrafien        14- ppm
Konzentration im Citybereich               44- ppm

-------
                         2132
     CO-HOtlZBITMTIOII UNO VEBCEHRSAST.III .BHAN6IOKETIT VON
             OURCHFAHRBBI STAOTGEB1ETEW
Ifl
                     Abb.  1?
PUDTSWTIX
                                      **>   wn  mi
                 Abb.

-------
                               2133
Die Trennung der MeBwerte nach den durchfahrenen Stadtbereiclien in
AusfallstraBe, AuBenbezirk und Stadtzentrum sowie nach morgens und
abends ergibt die in Abb.  13 dargestellten CO-Konzentrationswerte.
Abb. 13 zeigt, daft  die 3 Bereiche (AusfallstraBe, AuBenbezirk, Stadt-
zentrum) im Mittel deutliche Unterschiede der CO-Konzentration und
der Fahrtanteile mit Verkehrsstau aufweisen (rechte Seite der Abbil-
dung).  Die hochsten Konzentrationen wurdeu abends im Stadtzentrum
gemessen. Der arithmetische Mittelwert aus 20 Einzelwerten ergab
52 ppm. Die Konzentrationen der  anderen Stadtgebiete sind der Abb. 13
zu entnehmen.

6. Trendanalyse der CO - Konzentrationen.
Neben den bereits beschriebenen  mittleren  taglichen Konzentrations-
variationen und den Summenhaufigkeitsverteilungen  ist die Trendana-
lyse eine  weitere wichtige HilfsgroCe bei der Beurteilung der Ent-
wicklung der Luftverunreinigung. Auch hier ergeben sich, gerade bei
der Anwendung dieser Methoden auf die durch Kraftfahrzeuge emit-
tierten Komponenten, grofte Interpretationsschwierigkeiten,  well der
Trendverlauf sowohl durch eine groBraumige Entwicklung (z.B. Er-
hohung des allgemeinen Verkehrsaufkommens, Erniedrigung der Emis-
sionen durch technologische MaBnahmen) als auch durch Anderungen
der Verkehrsfuhrung in umnittelbarer Nahe der MeBstelle verur-
sacht werden kann. Die in Abb. 14- dargestellte Trendanalyse fiir die
Schwefeldioxid- und Kohlenmonoxid-Konzentrationen,  gemessen an
der Zentralstation, zeigt ein deutliches Absinken der  SO9-Konzentra-
                                                     £,
tionen ira  Verlauf der MeBperiode, wahrend die Kohlenmonoxid-Konzen-
trationen  zwischen 1969 und 1970 einen deutlichen Trend zu hoheren
Werten zeigen. Es  kann zwar angenommen werden,  daB diese Erhohung
eine Folge der steigenden Zulassungszahlen der Kraftfahrzeuge ist,
doch ist der Absolut-Wert der Anderung so gering,  daB wegen der  MeB-
ungenauigkeit eine gesicherte Aussage nicht moglich ist.

-------
      21:34
Abb. 15
Abb. 16
Abb. 1?

-------
  2135

Abb. 18
**"


•f 	

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__fi
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•-TICK F RANKFUlT/hJ
C1TT-STAT1OW lUWDSCKAy-KA
e JUKI
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B AUGUST




















Bf
UKC STICISTDFFBKHtTD
US 1. STC5C*


2DQ 300 pi*
Abb.  19
 Abb. 20

-------
                                  2136


'• -
If
/ ,
i s
' ///
. Ill
• ///
•ft—
:f
//
f



J 6
/
7






















































( NO . NO )
6 JUNl
7 JUtl
8 AUGUST

TTON





20 M 60 ft} 100 120
	 1 — . 	 L— 	 1 	 1 	 ^_ 	 1 	 , 	 i . i . ,
                           Abb. 21



7- Kohlenmonoxid als Leitsubstanz  fur die Uberwachung von  Kraftfahr-


zeug-Emissionen.



Die in Kap. 3  in der Form der mittleren taglichen Konzentrationsvaria-


tionen beschriebene Abnahme der kdurch Kraftfahrzeuge verursachten


CO-lmmissionen mit der Hohe und dem Abstand zur City gilt in gleicher


Weise fiir die Immissionen nitroser Case. Zur Verdeutlichung dieses


Sachverhaltes sind aus den Darstellungen der Summenhaufigkeitsver-


teilungen fiir die MeBorte "City-Station  1. Stock", "City-Station Dach"


und "Zentralstation" fur die Komponenten CO, NO und NO  bzw. NO
                                                      £,         A

(Abb. 15 bis 21) die 50 und 97-5 Perzentile in Tab. 1 zusammenge-


stellt.



Bei der Beurteilung dieser Werte ist zu  beachten, daB diesen Auswer-


tungen die Gesamtkollektive, also auch die niedrigen Nachtwerte, zu-


grunde liegen. Dadurch werden insbesondere die 50%-Werte stark


herabgesetzt.



Somit bestatigt sich die eingangs gemachte Feststellung, daB sich die


fur die CO-Konzentrationen ermittelten Unterschiede  zwischen den


einzelnen Stationen in  den Konzentrationen der Stickoxide wiederho-


len.
Dies bestatigt zunachst,  dafi das Kohlenmonoxid als Leitsubstanz fur

-------
TAB    1
Monat


C O



N O




N O
2



N O
X



Juni
Juli

August
September
]uni
Juli
August
September
Juni
Juli

August
September
Juni
Juli

August
September


City Station
50%
4.9
4.5

7
5
23
28
40
60
42
30

51
34
1. Stock
97.5%
19
19.5

21
23
145
190
200
320
100
90

130
135

City
50%
Station Dach
97.5%

6
7
7
13
17
15

16
12







28
40
53
83
43
43

54
33





Zentralstation
50% 97
1 4
1.2 2

1.4 7
1.7 7

.5%
.4
.9

.0
.0


10 38
6 30

7
__
I
I







                                                                                                                                                                                                                                                                   rsj
                                                                                                                                                                                                                                                                   M
                                                                                                                                                                                                                                                                   Ul

-------
                                 2138
die ebenfalls durch die Kraftfahrzeuge emittierten Nitrosen Case gel-
ten kann. Betrachtet man die in Abb. 22 dargestellten mittleren tag-
lichen Konzentrationsvariationen der Komponentai CO, NO, NO^, ge-
messen an der "City-Station 1.  Stock" im Juli 1973 so zeigt sich al-
lerdings eine deutliche Abweichung im Verlauf der NO-r und CO-Kon-
zentrationen. So ist der der morgendlichen "rush-hour" entsprechende
Konzentrationspeak beim NO deutlich beim CO dagegen gar nicht aus-
gepragt und erst gegen 11.oo Uhr ist zwischen CO und NO ein weit-
gehend paralleler Verlauf zu beobachten. Umgekehrt dazu ist der Zu-
sammenhang zwischen CO und NO_  in den Morgenstunden relativ gut,
in den Nachmittags stun den dagegen  schlechter ausgepriigt.
QualitatLv konnen diese Unterschiede zwischen NO und CO durch die
wnterschiedlichen Emissionsverhaltnisse im Abgas erklart werden,
die eine Funktion des Fahrzyklus bzw. der mittleren Fahrgeschwin-
digkeit sind. Fiir das Konzentrationsverhalten der Komponenten CO
and NO  sind in den Sommennonaten photochemische Prozesse maB-
gebend, die je nach Einstrahlung unterschiedliche Verhaltniswerte
verursachen.
Aus diesen Untersuchungen folgt, daB  das Kohlenmonoxid zwar fur
viele Problemstelbangen als Leitsubstanz bei der tjberwachung der
Kfz-Abgase dienen kann, fur spezieile Untersuchungen, z.B. fur
photochemische Prozesse, miissen  allerdings die relevanten Kom-
ponenten mitanalysiert werden.

8. Abschatzung des CO - Dosisangebots
Wie to den vorhergehenden  Kapitehi beschrieben,  liegen fur Frank-
furt fur den City-Bereich und den Wohnbereich kontinuierliche Meli-
reihen der Kohlenmonoxidkonzentrationen vor, die durch mobile
Messungen  und Modellrechnungen vervollstandigt werden. Mit diesem
Datenmaterial lafit sich das Dosisangebot abschatzen,  das als zeitli-

-------
                                                 TABELLE  II
•ittl.Ionz.
Exposlt.
Dauer
Dosls-
angebot
Anteil an
d.Gamt-
dosis

Mittl.Konz.
Cxposit.
Dauer
Dosis-
angabot
An tell
an der
Gesamt-
dosls
Fahrt zur Arbeit jjt.

-------
                                Pilotstation fiir Luftreinhaltung
CO    A-A
NO    	
N0    o-o
Abb. 22
                                    t-IHf*
                                                    I X.  UMf*
                                                                       IB. t-IHH
Mittlerer  Tagesgang CO, NO, NO,,,  0

Rundschau-Haus  1.  Stock  Juli  1973
                                                                                              20 ppm
                                                                                               16 ppm
                                                                                              12 ppm
                                                                                               8 ppm
                                                                                               4- ppm
                                                                                       ro
                                                                                       I—I

                                                                                       C:

-------
                                  2141
ches Integral iiber die Konzentration definiert 1st. Bei dieser Ab-
schatzung tritt zu dem Problem der zeitlichen und ortlichen Varianz
der Kohlenmonoxid-Konzentrationen die Schwierigkeit hinzu, die
Aufenthaltsdauer von Personengruppen in den unter schiedlichen Kon-
zentrationsniveaus  zu bestimmen.
Im augenblicklichen Stand ist es nur moglich, Grenzfalle fur maxima -
le und minimale Expositionen im Stadtgebiet anzugeben.  Folgende
drei Falle werden betrachtet:
1. Berufstatiger  Pendler mit Arbeitsplatz im City-Bereich.  Der Ar-
   beitsplatz ist
   a) im StraRenniveau
   b) in ca. 20 Metern Hohe
   Pie  Fahrzeit von und zur Arbeitsstelle betragt jeweils eine halbe,
   die Arbeitszeit  selbst 9  Stunden.
2. Berufstatiger  Pendler v/ie unter 1. mit Arbeitsplatz in einer Biiro-
   und Wohngegend.
3. Berufstatiger  der auf seinem \Veg  zur Arbeit nur Wohngegenden
   passiert und auch in einer Biiro- und Wohngegend arbeitet.

Fiir jeweils zehn Stunden Aufenthalt im Stadtgebiet von  7-17 Uhr er-
gibt sich fur die  Falle 1-3 die in Tab. 2 dargestellte Belastung.

Bei diesen Schatzungen wurde von mittleren Sommerkonzentrationen aus-
gegangen, weil in dieser Jahreszeit die CO-lmmission in Frankfurt/M
fast ausschliefilich durch den Kraftfahrzeugverkehr verursacht \vird.
Im Winter liegen die CO-Konzentrationen durch Feuerungsemissionen
und grofiere atmospharische Stabilitat hoher. In einzelnen Fallen, d.h,
an Tagen mit austauscharmen \Vetterlagen konnen die Mittelkonzentra-
tionen iiber 8  Stunden bei 30 ppm liegen, auch dieser Fall ist hier nicht
beriicksichtigt.

Unter den in 7 • genannten Einschrankungen lassen sich mittels der be-
kannten Emissionsverhaltnisse im Kraftfahrzeugabgas (Emissionskata-
ster Koln) diese  Abschatzungen auf die Nitrosen Case und Kohlenwas-
serstoffe iibertragen.

-------
Literatu rverzeichnis
                              2142
LAHMANN, E., 1972
GEORGII, H.W., 1968
BUSCH, E.
WEBER, E.
EMISSIONSKATASTER 1972
Koln

BALTRUSCH, M., 1972
CO Concentrations in the Urban Area of
Berlin, VD1 Berichte, Band 180, S. 40 - 45

Untersuchungen iiber die zeitliche und raum-
liche Verteilung der Immissionskonzentra-
tion des Kohlenmonoxid in  Frankfurt a.M .,
Ber. Nr.  11 Inst.  f. Meteorologie und Geo-
physik der Universitat Frankfurt a.M.

Minister fur Arbeit, Gesundheit und Sozia-
les des Landes Nordrhein-Westfalen

Dreidimensionale Analyse  des- CO-Konzen-
trationsfeldes iiber einer Flachenquelle.
Diplomarbei^lnstitut fur Meteorologie und
Geophysik der Universitat Frankfurt a.M.

-------
                              2143
       INCORPORATION OF POPULATION EXPOSURE CONCEPTS IN
                AIR QUALITY CRITERIA DOCUMENTS

                ARTHUR C, STERN AND YUJI HORIE

Department of Environmental Sciences and Engineering, School of
Public Health, University of North Carolina at Chapel Hill, USA
ABSTRACT

     Short, intermediate and long term responses of human, vege-
tation, animal and material populations are categorized with
respect to the pollution concentrations and durations associated
with them and are located as zones on arrowhead charts.   The
thrust of the paper is that these mayor categories and response
terms need separate documentation in air qualtiy criteria docu-
ments ,

-------
                                     2144
    1.  Introduction
       There  are  a number of different populations exposed to air  pollu-
 tion,  populations of  people, trees, crops, animals and materials.   Each
 such population  responds to air pollution in a number of different  ways.
 Each such response correlates best with a specific range of concentration-
 duration of  exposure  combinations.  Although this is not a new concept, it
 has not heretofore been fully elaborated.  The purpose of this paper is to
 show how to  differentiate these several concentration-duration combinations
 en  a concentration-averaging time -frequency of  occurance (arrowhead) chart
 (1).
       It has been noted before (2,3) that to be meaningful air quality
 standards stated in terms of different averaging times should be on the
 same percent!le line on the arrowhead chart.  While standards tend  to be
 points on or near such lines, criteria documents deal in ranges of  con-
 centration-response data rather than in points, and,in addition .reflect
 such factors as the fatigue and acclimitization responses of human  organs.
   2. Short. Intermediate and Long Term Responses
      Population responses  tend to separate themselves into short,  inter-
mediate and long term responses,  the short term ones being characterized
by response times in seconds  or minutes, the intermediate term ones by
response times  in hours or  days,  and the long term ones by response times
in months  or years.   Each of  the  principal  population categories, human,
animal, vegetation and material have such tripartite response groupings
(Table  I).
                                   TABLE I
               Examples of Population  Category  Characteristic
                              Response  Times

Population
Category
Human
Vegetation
(Animals)
Materials
Charact
Short Tern
(Seconds-Minutes )
Odor; Visibility;
Nasopharyngeal
and eye irritation
Field crop ai
plant damage
Acid droplet pitting
Nylon hose destruc-
tion
eristic Response Tinu
Intermediate Term
(Hours-Days)
Acute respiratory
disease
id ornamental
Rubber cracking;
Silver tarnishing;
Paint blackening
es
Long Term
(Months-Years)
Chronic respirato-
ry disease; Lung-
cancer
Fluorosis of
livestock;
Decreased fruit
and forest yield
Corrosion; Soiling;
Materials deteriora-
tiou

-------
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-------
                                    2146

   3. Short Term Responses (Fig.l)
      The short term responses of the several populations are quite diverse.
Odor is characterized by the ability of the nose to detect extremely low
concentrations but, because of odor fatigue, to rapidly lose the ability
to respond to ambient concentrations.   Point, a, is the odor threshold,
which is known.  Since a single inhalation, requiring approximately a
second will establish the presence of odor, point, a, is located at 1
second.  The ability to recognize the odor is lost due to odor fatigue at
point, b, which is on the maximum ambient concentration line at a duration
that has not been well established by experiment but can be at about 10
minutes.  The shape of line a-b, which depends upon the odor fatigue time-
concentration relationship is also not well established experimentally.
        Nasopharyngeal and eye irritation have higher thresholds than odor
  and exhibit the  fatigue phenomenon to a lesser, or even negligible,
  extent.   Nasopharyngeal Irritation and odor thresholds for substances
  with  irritant properties have not been well differentiated.  The instan-
  taneous exposure  threshold  point, c, is arbitrarily placed at 1 second.
  If we assune some acclimatization of the eye to the instantaneous thres-
  hold  after extended  exposure, the effective threshold after a number of
  minutes will be point, d, which  is at a concentration somewhat higher
  than  the  instantaneous threshold value, c.  Neither the exact location
  of, d, nor the shape of line c-d are known from experimental data.
        Visibility  is  a human response since it  is what the eye can see.
  It correlates with suspended particulate matter concentrations in the air
  and has a threshold  value of suspended particulate matter for each value
  of visibility, expressed in kilometers or miles.  Thus line e-f, for
  a hundred mile visibility is at  a lower pollutant concentration than line
  e'-f  for a ten mile visibility, or en-fM for  a one mile visibility.
  If there  is a functional relationship between  the concentration and size
  distribution of suspended particulate matter,  it would affect the locus
  of these several  lines.  As shown on Figure 1, the possibility of such a
  functional relationship is  ignored.
        Typical short  term responses of materials are pitting of metal and
  painted surfaces  and destruction of nylon hosiery by acid droplets, and
  acid  bearing solid particles, such as acid smuts.  Since damage rapidly
  follows attack,  it can be depicted as starting at one second, line g-h,
  and terminating  in a matter of minutes of exposure, line i-j.  The thres-
  hold  concentrations  of droplets  or particles,  line g-i, is arbitrary and

-------
                                  2147
 is at a low level,  below which damage from pollution cannot be dif-
 ferentiated from that  from other  sources.  Line g-i, is not known
 from experimental data.

    *+• Intermediate  Term Responses (Fig. 2)
       The short and intermediate term response of vegetation to higher
 levels of certain pollutants can be measured in two ways, biologically
 and socio-economically.  Air pollution damage to field crops and orno-
 mental annual plants is better measured in socio-economical terms
 than in terms of more subtle biological response.  In these terms, up
 to a certain level of biological response,k, there is no socio-econom-
 ic loss in the sale value of the crop or the ornamental value of the
 plants.  At a higher level of biological response, 1, there is 10056
 loss of sales value of the crop and of ornamental value of  the plants.
 These responses are related to dose, ie., concentration times duration.
 When these response (loss) curves k-k'  and 1-1', are transferred to an
 arrowhead chart,  they depict the short and intermediate term responses
 of field crops and annual ornamental plants to ambient pollution levels,
 It should be recognized that the arrowhead chart must be for the
 location where the crop or plants grow,  and that there is a paucity
 of such data for  rural agricultural  areas.
       The principal intermediate term human health response is acute
respiratory disease  such as has occurred during air pollution episodes.
The biological response involved is a dose-response curve, which for
constant concentration  becomes a duration-response curve.  The shape
of a duration of constant concentration-response curve for acute
respiratory disease  reflects the ability of the human body to cope
with short term ambient concentration respiratory exposures and the
overwhelming of the  body's defenses by continued exposure.  When
transferred to an arrowhead chart the curve has an induction point,
m,  at the maximum concentration line at about a day and continues to
the concentration level, nf where continued exposure can no longer
induce acute disease.
       A number of manifestations of material response, e.g., rubber
cracking by ozone, require an exposure duration long enough for the
adverse effects to be significant economically, ie., attack for just
a few seconds of minutes that does not affect the utility of the
material for its intended uses is not significant, concentrations

-------
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-------
                                   2149
lees than point, o.   This would be  the case for  rubber exposure  to
ambient  concentrations of ozone.  Conversely utility can be destroyed
by a number of days  of exposure (point p).

   5,  Long  Term  Responses  (Fig.  3}
       Fluorosis  of  livestock from ingestion of forage with high fluoride
 content  has a  point of induction, q, which does not occur until there has
 been a long enough  period  of deposition of a high enough ambient
 concentration  of fluoride to build up the level  of fluoride in  the  forage.
 Since  the forage is either eaten by livestock or cut for hay at least
 once a growing season, the duration of desposition ends after the growing
 season (point  r).  The longer the duration of the season, the greater the
 time for deposition, hence the shape of line q-r.
       The long term human responses to air pollution-chronic respiratory
 disease  and lung cancer; the long term vegetation responses-decreased
 yield  of fruit and  forest; and the long term material responses-corrosion,
 soiling, and materials deterioration, all occupy the same zone, s-t,  on
 the arrowhead  chart.
       It is not possible to closely differentiate intermediate and  long
 terra respiratory response so that there is overlapping of these cate-
 gories.

   6.  Conclusion
       A  characteristic of  all the charts presented in this paper is that
 the response curves stay put on  the concentration-duration axes, but
 the arrowhead  chart moves  up and down depending upon whether a com-
 munity's air is  polluted or not  polluted.  This is most apparent in the
 case of  long term responses where in a non-polluted community the long
 term response  zone  can be  completely above the maximum line of the  arrow-
 head chart, or in a highly polluted community the response zone can com-
 pletely  engulf the  arrowhead chart down  to the minimum concentration line.
       There are  important  aspects associated with the geographic dis-
 tribution  of these  several populations with respect  to the geographic
 occurrence  of  short,  intermediate and long term exposure which could not
 be covered  in  this  paper due to  space limitation.  The general thrust of
 this paper is  that  there are three major population  categories and three
 major  response terms,  short, intermediate and long,  that each need sep-
 arate  documentation in air quality criteria documents.

-------
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-------
                                   2151
References
1.  R. I. Larsen, "A mathematical model for relating air quality measure-
    ments to air quality standards", U. S. Environmental Protection
    Agency, Research Triangle Park, North Carolina, Publication No. AP-89
    (Nov. 1971).

 2.  R.  I.  Larsen,  "Important  Factors  for  the  Sulfur Oxide  Concentration
    in  Central  Stockholm", Atmospheric Environment, Vol. 6, pp.  423-426
    (1972).

 3.  H.  Shoji  and T. Trukatani,  "Statistical Model  of Air Pollutant Con-
    centration  and  its Application  to  the Air Quality Standards", Atmo-
    spheric Environment, Vol. 7,  pp.  485-501  (1973).

-------
PANEL DISCUSSION

-------
                                2155
                   SUMMARY OF DISCUSSION
SCHNEIDER  (Netherlands)
     Following the short statements made by the  panel members,
I feel that the discussion should now center on  three questions;
     - do we need monitoring at all;
     - the way of monitoring;
     - what kind of monitoring we need.

     I hope we can also have a short discussion  on  the way
we use the data that we acquire and receive from the monitoring
systems.
                         DISCUSSION
     Before we talk about monitoring needs, we actually have
to ask ourselves the question do we need monitoring?  I know
the feelings of the panel on this so I do not  have to ask
them if they think it is worth while to monitor but I know
there is a difference of opinion within the panel about the
way the monitoring system should function or the way monitoring
should be done; and I think it is worth while bringing it up
later on. I would like at this stage to ask the audience whether
some of the participants have a very strong feeling about the
need for monitoring, if there is someone who says I do not
need your monitoring at all, let him stand up and say so because
we would like to solve his problem first.

-------
                                2156
LAUER (U.S.A.)

  Is it better to have source monitoring with an appropriate
model or monitoring of the population exposure burden?

SCHNEIDER (Netherlands)
    I understand your question, and I am glad you asked it
because now I can use my prerogative as a Chairman. I did ask
the question, "do we need monitoring?" And you answered, "yes."
You only questioned the kind of monitoring, this is the second
part of the question.  I really want to know if someone questions
the monitoring at all.  If not, I am willing to answer the
second part.

MAGE (Denmark)

     Like all questions I can argue equally well on both sides,
so let me just say that the answer to Lauer's question is yes
and no.  I believe that it is better not to have any data at
all about air pollution levels than to have bad data which
is non-representative, or which had been taken by instruments
which have not been calibrated properly, or which are taken
at a location which is non-representative and you have no idea
of the distribution of air pollution.  You don't know whether
its a high location or a low location.  When one has bad data,
it confuses the issue considerable.  We need good data which
is designed properly, and not bad data.
GOLDSMITH (U.S.A.)

  Monitoring should not be done under certain circumstances:
     A.  If one has several year's data from an unvarying matrix
         of emission sources, the only source of variation will

-------
                                2157
         be meteorological and measurement of pollutants  is
         much more costly than meteorological data.
     B.  If the purpose of monitoring is to obtain information
         for protection against some hazard and there is  no,
         or poor linkage between measurement and protective
         services, monitoring may give a false sense of security
         and should not be done.
     C.  If the purpose is to provide data for health purposes,
         then the health reactions must either be predictable
         or monitored along with environmental exposures  and
         if both are not being done in a balanced fashion, it is
         better not to make measurements.

HOGGER (Switzerland)

      Immissionskontrollen sind zweckmaGig zur Ueberwachung
der Entwicklungstrends der Luftverunreinigung.  Zur Erfassung
der Exposition, resp.  der GefShrdung der Bevolkerung sind sie
jedoch nur sehr beschrankt brauchbar. Das AusmaB der Verunrei-
nigung wechselt sehr stark von Ort zu Ort und im Lauf der Zeit;
vor allem aber ist die BevSlkerung mobil und wechselt den Standort
im Lauf von 24 Stunden immer wieder.  Die Exposition kann deshalb
nicht mit stationaren  MeBstationen erfasst werden.  Kohlenmonoxid-
und Bleigefahrdung werden sehr viel besser durch Blutuntersuch-
ungen erfasst  als  durch Immissionsmessungen.

     Monitoring stations  are useful for assessing trends in
air pollution;  for the determination of exposure or hazards for
the population  they have  very limited value.   The extent of
the pollution  varies greatly from place to place and from
hour to hour;   above all3  the population is mobile and moves
from one  locality  to another many times during a period of
24 hours.   The  exposure cannot therefore be established by
stationary  measuring devices.   Carbon monoxide and lead pol-
lution are  much better determined by blood examinations than
by monitoring  stations.

-------
                                2158
SCHNEIDER  (Netherlands)
     It seems that we have drifted already  into the  second
phase, nobody actually wants to say that he does not want moni-
toring.  All questions are comments about the way of monitoring,
and what kind of monitoring we should have. When talking about
the way of monitoring we can start discussing whether  it is
worth while doing ambient air measurements  using stations as
it has been done so far, or using exposure  meters, or  going
into a more biological monitoring of man himself. First of
all I would like to ask, if there is anyone from the panel
who would like to say anything about the integrated  monitoring,
because this is a new approach.

MAGE (Denmark)

     Again one has to define what one is monitoring  for.  For
instance as Dr. Goldsmith recently mentioned, if we  are monitoring
for data, for an epidemiological study we have to design the
experiment very carefully to simulate man's exposure.  A quick
example:  The dose one gets of any pollutant, such as  sulphur
dioxide.  The number of grammes per day that one inhales is
an integral over 24 hours of the respiration rate  (so  many
litres per unit time) times an efficiency factor  (we do not
absorb everything) integrated over time. When sleeping, the
respiration rate is only four to six litres per minute; sitting
at a table, not working too hard the respiration rate  is only
1O litres per minute; when walking on the street the respiration
rate is between 1O and 2O litres per minute.  However, most
of the samples are taken by sampling at a constant flow rate.
Therefore one has to treat the data and look and say well,
is it worse at night, or is it worse during the day.   Air pol-
lution is not a random variable, air pollution is very highly
correlated.  Maximum values for air pollution occur, for carbon
monoxide usually during high traffic periods; in the winter
in San Jose, we have a radiation inversion  at the surface and
we get very high carbon monoxide levels, 24,25 parts per million

-------
                                2159
between midnight  and one o'clock in the morning.   But  if  I
am only breathing 6 litres per min. why should  I  rate that
exposure as highly as an exposure during the day  when I am
breathing at 2O litres per min.  So one possible  modification
could be, a variable pumping system, pumping at 1 litre per
min. during the night and 2 litres per min. during the  day,
and then you would get the integrated average which would  cor-
respond to an exposure.  The danger of a monitoring system
which does not take this into account in the beginning,  is
that the epidemiologist has data now, which he  is going to
analyse and look for significant correlation but  the data  have
been biased because the data had not been collected the same
way that the exposed person of a target population is influenced
by it. When we design monitoring systems we have  to take many
things into account and this is an example of a thing which
makes a set of data very bad.   If one is interested in  corrosion
of statues, the weight loss would be independent  of the breathing
rate and the corrosion of the statue would be as  if sampled
at a constant rate,  but if you are interested in  human  exposure
then you have to vary the rate.
SCHUCK (U.S.A.)

     I would agree that there are many factors in determining
the exposure of  an individual or population at risk.  However I
suggest that it  is up to persons responsible for data inter-
pretation to make that simple adjustment or correlation.  This
can be done from a continuous monitor record, without further
complexing the monitoring equipment.  I am wondering if the
term 'integrated1 does not include more than one definition.
My definition of integrated is multimedia.  It is a multi-media
because there are many pollutants that reach us from air, water,
and food.   An integrated approach is to devise a system which
measures  some critical point in these exposure pathways, and
therefore defines exposure to the receptor.   Ideally it v?ould

-------
                                2160
be nice if you had a simple test for the human, that you could
test his blood and find out what his exposure is to a given
pollutant. With something like lead and many other materials
this is not possible because once you study the entire system
of the exposure pathways plus the rate of transfer of this
lead, once it is absorbed into the blood, you have another
entire system that is concerned with the deposition in bone,
the elimination in urine, the deposition in hair, transfer
into the central nervous system. Thus, this measure is very
crude and has not been related to exposures.
BRAVO A (Mexico)
     I wish to comment on the type of monitoring that has been
in operation in Mexico and South America for at least 9 years
operated by the Pan American Health Organization and the countries
involved.  There has been since its conception, a serious doubt
as to the validity of some of the measurements of the PAHO
Network such as suspended particulates in the air.

     The method chosen by PAHO and used in Mexico for this
particular parameter was the Warren Spring Laboratory  (U.K.)
method based on reflectance of the particulate matter collected
on a filter, using inexpensive instrumentation.
     This method was suitable for U.K.  or countries which
use coal as a source of heat; however, as your are well aware,
in Mexico we have very little heating and the fuel we use is
not coal. In a research work undertook at the University that
at the same location in the same time period, the PAHO network
reported an average particulate loss of 80-9O mg./cm  ; however,
the estimation of this parameter with a High Volume sampler
by the University indicated an average of 250 mg/m  .  Consequently,
the data published by PAHO is misleading since Mexico would
not be considered a city with a suspended particulate problem.
Therefore, this data should be used with a great reserve if
used at all for health studies, control engineering studies

-------
                                2161
or in definition of air quality.
     For monitoring in countries with little money and qualified
human resources I believe it is better to collect fewer data
but data that can be used with some sense of validity.
     Source sampling applied to mathematical models in Mexico City
would require a great amount of research due to our unique
meteorological and topographical conditions.
BRAVO-A (Mexico)
     Monitoring is needed, but the validity of the measurements
should be a must.
SCHUCK (U.S.A.)
       I believe that we did not answer the original question
which was, how best to monitor?   Why could we not use source
monitoring and then a model to predict.  Well there are many
models that have been developed and for some point sources
they are reasonable, but in general these models, even knowing
quite a bit about the source and the meteorology of the area
can one predict within 1OO% of the real value 5O% of the time?
That is not good enough for defining exposure pathways for
example.   So there is a need for all types of monitoring.
Actually it depends on the mandate for why, when you ask the
question why are you monitoring?  Usually you will find a
basis in law, and if that law requires source monitoring,
then that is what you will do.  On the other hand, many of
the laws are stated in terms of air quality. Disregarding for
a moment the unreasonableness perhaps of some of the air quality
measurements depending on their location, when this is called
for,  you must monitor and you cannot depend on the model.

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                                2162
 SCHNEIDER (Netherlands)

      Before opening again the discussion. I would like to make
 just  one  distinction.   There are clearly two points brought
 up here.   One by Dr. Mage; when we talk about exposure monitoring
 do we not have to simulate in some way the intake of the pol-
 lutants,  or do we go on using a constant flow.   The other
 point which was brought up by Dr.  Schuck was the integrated
 monitoring in the sense of the multi-media monitoring.  I
 think these are two different questions, the first case is
 more  or less restricted to inhalation of air pollutants,  the
 second case is indeed  a  multi-media problem, when you have
 the same  kind of pollutant in air,  water, food  and maybe even
 other media.   So I would like to divide it up into two so as
 not to confuse the issue.   To start with the first one raised,
 I would like  to hear from the audience what they think about
 the question  raised by Dr.  Mage of  a simulated  flow rate.

 BATES (Canada)
       It seems to me  that the purpose of contrasting 'personal1
monitoring and 'station1  monitoring is to give  us an indication
of  the safety factor that has to be built into  a standard.
This  leads  to two  questions:

      1.   Does the  personal average  level of exposure generally
          correspond with  the  station measured average,  but
          with swings both above and below it?
      2.   Do you find any  general 'constant'  that relates the
          maximal pollutant exposure you  may get,  given a
          normal level  of  activity of the person,  to  the station
         measured  average?
MAGE (Denmark)
     I would like to return to figure 2 of my paper.  The bars
represent the actual collection of the sample.  The periods
between the bars were when the bags were being put away in

-------
                                2163
 the car, so what  you  see  there  is  approximately 3O five minute
 random samples  during that  period,  and  from the number of samples
 from the arithmetic average and the standard deviation about
 the mean of this  distribution,  we  could reject the null hypo~
 thesis that the air quality on  the  street  level,  2.6  km.  away
 from the monitoring station is  the  same as  at the monitoring
 station.  The other question: is there  in general constant
 ratio between maximum exposure  and  average  exposure?  One  must
 sample the distribution.  In order  to define most distributions
 one must know two parameters.   The  arithmetic mean which  you
 could get by bubbling for example  through a sulphur dioxide
 reagent for a year, will be a very  fine measurement but you
 will have no information at all of  the  standard deviation of
 daily averages  about  that mean.  So the answer to your question
 is, that if you tell  me with what confidence level you will
 wish to know the maximum value  that a person can  be exposed
 to, whether you want  to know it at  99%,  95%  or 9O%, then  I
will tell you how well you have to  monitor.   If you want  a
 100% confidence level, as is required in radiation exposure,
 every employee of the reactor must  wear a radiation badge,
but in air pollution  I do not believe we have  to  have  an  in-
 finite number of monitors; we can take  the  data and treat  it
 statistically and make estimates as  to  the  maximum exposures.
LAUER (U.S.A.)
     Prior to modulation of the intake volume it is necessary
to understand the uptake mechanism for the actual pollutant
in question;  the mechanism may be 1st, 2nd or O order with
respect to the  concentration.

-------
                               2164
MAGE  (Denmark)

      Yes, one has  to  know  the  uptake  kinetics  if  the model
is that the  intake is simply a function of  what you have in-
haled, and this  is really  what you  are measuring  with a constant
volume flow-rate like a  24 hour OECD  type measurement,  you
are getting  an average but it  is a  non-weighted average.   If
one is breathing twice as  fast during the day  than at night,
then  one will breath  in  twice  as many carbon monoxide molecules
in the same  time.   The 8 hour  CO standard is based on an up-
take  ventilation rate of sedentary  people.  A  pedestrian who
is walking on the  street is breathing at around 15 to 20 litres
per min. and so  he will  reach  a higher level of carboxyhaemo-
globin than  the  sleeping person who is exposed to the same 'CO1
concentration.
O'KEEFFE  (U.S.A.)
     I suggest that it is appropriate  for  the epidemiologist
to attempt to refine his data according to the variable-intake
concept upon a given concentration exposure; that  it would
be usurpation for the physical scientist to attempt to  force
that calculated modulation on the epidemiologist.
SCHUCK  (U.S.A.)
     Is it correct Dr. Mage that those 5 minute bag  samples
represented the condition for a person walking along a street,
in other words a pedestrian on or near a street and  if so  the
average for the entire area may be much closer to the continuous
measurement than those high peaks shown by these 5 min. bag
samples?

-------
                               2165
MAGE  (Denmark)

     The  five minute  samples were  taken,  as  it  stated in the
paper at  breathing  height which was  defined  at  approximately
1.6m.  The man would  walk and hold his  air sample intake at
nose level. He was  walking  just like the  pedestrians  stopping
for traffic lights, walking across the  street,  passing cars
which were idling and walking on the street  again.  This exercise
was aimed at finding  out really how  well  monitoring station
measurements relate to the  data from such a  random approach.
We have also done another study.   We randomly sampled,  by laying
a grid over a city  (San Jose and Copenhagen), using random
numbers we generated coordinates on  the grid and  we went to
the randomly selected locations and  we  sampled  randomly.   We
do not get the same value at each  place,  but we get a distribution
of values, and of course if one did  this  for a  very large number
of samples you will find that you  can demonstrate at  a  fairly
high level of significance  that there is  not a  constant  average
value.
GOLDSMITH (U.S.A)
     While I agree with the principle that a monitoring station
Is most valuable when it "breathes" the same way a population
sample does, I do not support the proposal to weight the sampling
rate by the diurnal variation of the respiratory or ventilation
volume.  As Prof. O'Keeffe has said epidemiologists are willing
to weight the set of valid data obtained.  In addition with
use of twenty-second breath holding, the expired air analysis
for carbon monoxide permits use of human blood in the body
to be read as a monitor.

     The most obvious problem of unrepresentative monitoring
is in connection with emergency action based on a monitoring
reading. It is of great importance to know whether the high-
reading station, which is proposed to be used, represents

-------
                              2166
with validity the exposure of a real population.

     I do not agree with Dr. Schuck that blood lead  levels
are not useful for monitoring lead in air.  As long  as the
exposure is steady over a period of months, blood  lead levels
may vary with atmospheric exposures  (above a level of about
2 yg/jn .  This makes them useful for monitoring population
levels in population groups which have only slowly changing
exposures. This illustrates the point that long-term average
levels for lead are all that needs to be known in order to
estimate the health impact of exposures.  Hourly or  daily aver-
ages add little or nothing to 3O day average as a source of
information related to health effects.
SCHNEIDER  (Netherlands)
     We should discuss now the use, or the potential use of
exposure meters, the ones described in the paper by Altschuler
and O'Keeffe.  Is there anyone in the panel who would like to
comment on the use of exposure meters versus let us say ambient
air monitoring or any other kind of monitoring?
O'KEEFFE  (U.S.A.)
     We discuss in the paper three general types of  such exposure
meters that are imaginable. One would be a physico-chemical
type, which would include the optical devices.  We ruled the
optical devices out as being mechanically not adaptable to
personal dosimetry.  I cannot imagine an optical absorption
based instrument that could be put together with sufficient
mechanical stability that it could stand the stresses of being
carried around on a person; nor could it be of a size adaptable
to that use.  Therefore these were given quite low priority.
A second class depends on the transfer of energy triggered by
    presence of the pollutant to be measured.  This  would

-------
                               2167
 include such  devices  as  the  ethylene-ozone  device which operates
 on the principle  of luminescent  reaction  between ozone and
 ethylene.  This obviously  can  very  readily  be  miniaturized,
 perhaps-hopefully-to  the point where  it can be readily carried
 around on the person. Other  members of that family of  lumine-
 scence instruments pose  much greater  problems  in miniaturization,
 but it is not beyond  the bounds  of  imagination that they could
 be so miniaturized.   The last  category in the  physico-chemical
 class would be that of the so  called  electro-chemical  devices;
 the Ecolyzer would be an example.   These  measure either ampero-
 metrically or coulometrically  the oxidation or reduction of  the
 given pollutant.  They lack  sufficient sensitivity for  dosimetric
 use at the present stage, because such use  requires  the
 ability of responding to background levels  of  the pollutants
 of interest.   I would safely predict  that this  minor defect
 can be overcome.  They have  not  yet been  fully  miniaturized,
 but I consider that a purely mechanical or  engineering  problem,
 I think there is great hope  for  this  type of instrument.
Now a third category is  that of  the sorption device.   In
 principle it goes back to the  type of air monitoring we did
 5, 6, or 1O years ago.   In our present thinking it tries
 to avoid the difficulty of working with aqueous solutions,
because these would pose obvious difficulties  in  a personal
dosimeter.   Instead it proposes  to use solid sorbents which
would either  ad-or absorb the  pollutant,  with  a modest  degree
of specifity.   After a given period of exposure which can be
 selected,  they will be transported back to  the  laboratory and
 there,  in the simplest case,  they would simply  be  heated to
effect desorption and  the product would be  carried into one
of the present day ambient air monitoring instruments for an
 instantaneous  read-out.

-------
                              2168
SCHNEIDER  (Netherlands)
     I think we are here  at  an  important  point,  a starting point
in the development of not a  new technology  but  let us  say  a
new generation of monitors.  There  has  been  in the past,  and
certainly  in recent times, many comments  on the  use of existing
and maybe  even on the planned monitoring  systems with  respect
to the correlation with health  data, and  I  think quite correctly
so in many cases. You heard  fron Dr. Bravo,  for  instance,  of
problems he met in Mexico.   The original  monitoring systems
were not planned essentially for the purpose of  correlating
their measurements with health  data and explain  them.   It  is
different  I think for the type  of  monitors  that  Dr.  O'Keeffe
has described. These are  especially intended for the combination
of environmental monitoring  and health data.  It would be  very
valuable if we could hear some  comments on  this  new approach.
MAGE  (Denmark)
     Dr. O'Keeffe indicated a concern about  low  levels.   I do
not think that we really have to worry about measuring  such
low levels.  One can in terms of statistical analysis work with
a censored distribution and use that part of the distribution
that one can measure; at low levels there is little  effect.
I really do not want to know when an instrument  is in the noise
level near zero whether it is 0.1 or 0.2 ppm, it is  sufficient
for me to know that it is less than 1 ppm.   This should not
really be much of a bottle-neck in the development of instruments.


O'KEEFFE (U.S.A.)
     In our early planning on this subject we have arbitrarily
assumed that a minimum integration period of one hundreth of
a day or 15 minutes would be satisfactory.

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                                2169
 BENARIE (France)
      Monitoring networks allow four-dimensional mapping of
 the isopleth contours (one dimension being the time).  The usual
 ten or twenty station network gives a very weak spatial resolution,
 Imagine now an isopleth map with a perfect, an almost infinite
 resolution.  This, like the 1 to 1O.OOO surveyor's map can be
 used to give a road, a land use, a political...etc., map.
 In the same way a perfect isopleth map can give information
 about human exposure (the subject moving on the four dimensional
 surface)*  an imperfect one will not.
      Regarding personal monitors, we shall look for the "old"
 methods also:   rubber cracking for ozone,  sulphation plates
 for the S02, etc.
O'KEEFFE  (U.S.A.)
      We are  quite  aware  of these older methods,  and it is ad-
vancement  on these existing methods that I have  in mind when
talking of the  solid  sorbent type of collector for return to
the  laboratory,  for analysis.  You mentioned the  rubber as a
collector  for ozone.   We actually have a very small project
under way  in which rubrene,  which is a polycyclic hydrocarbon,
reacts with  ozone  instead of the rubber.   It is  a simplified
analogue of  the  rubber:  upon return to the laboratory,  when
we get a objective instrumental  read-out,  rather than a human
observation.  A  read-out in  terms of the  rubrene would  be that
its  adduct with  ozone  is theroluminescent,  and upon heating
to 5O°C it gives off  light  in  proportion  to the  integrated
concentration of ozone to which  it has been exposed.   I  cite
that as an example  to say that we are  aware of the  old waysj
we think that they  need  to be developed to  the point  where they
give rapid simple  read-out methods.

-------
                              2170
MAGE  (Denmark)
     Regarding the question whether  it  is  necessary  to  know the
location I think it is yes.   If one  is  looking  at  something
which is not an ambient pollutant but which  is  generated  within
the home such as cigarette smoking where there  might be some-
body lighting a match which will give sulphur dioxide close
to your exposure meter.  Something like this might happen
but you would not want to tell the big  refineries  that  they
are putting out too much sulphur dioxide because somebody was
lighting matches near his exposure meter and so I  think one
should know something of where the person  has been,  so  that
one can estimate whether the  air pollution which one is meas-
uring is ambient or whether it is residential or occupational
exposure.  The other question is to  whether  one has  to  know
something about the movements of the people. I  think you  might
look at it this way, if the air pollution  monitoring station
which is fixed in space operates in  an  "eulerian"  framework
because it is fixed in space  and different gas  puffs come to
it in time a person is rather "lagrangian" because he is  moving,
he has a moving frame of reference,  and so as one  can talk about
lagrangian correlation coeficients and  eulerian correlation
coefficients, it should be possible to,from the  analysis of the
monitoring system.get some kind of information  on  the movement
of the person in the system.  So since  we  certainly  are not
going to do this with a 1OO%  confidence level,  we  have  to be
able to make some assumptions and accept some reductions  in
confidence of our results.
STEELE  (U.S.A.)
     One cannot argue the premise that  "good" data may be bene-
ficial for a designated information use, and that "bad" data
may be detrimental or misleading.  The  difficulty comes in  setting
criteria (specifying purpose or use, pertinent variables  (pol-
lutants) , statistical measures of concern relative to each

-------
                               2171
variable, frequency  and  schedule  of  sampling  to  achieve  those
measures, and  the  latter  leading  to  desired or required  accuracy
of measurement) on what  constitutes  "good" or "bad"  data.   Speci-
fication of  criteria depends  greatly upon use of the obtained
information  (that  is in  this  case impact on policy,  on decisions
involving environmental  health effects  or standards).  Also
economics plays a  critical note.   At some point  in evaluating
data-monitoring programs, one reaches a point where  the  incre-
mental allocation  of resources to improve the accuracy of  some
measure or to  provide additional  information  may not be  "worth"
it  (worth commonly being  a value  judgment based  upon some  de-
signated consensus).

     In the  field  of water resources, for which  I am more
familiar, several  studies here have  been carried out to  help
in determining the "worth" of certain hydrologic data, including
those for water quality variables.   Are there examples of  paral-
lel studies  (completed or ongoing) in the field  of assessment
or monitoring  of air pollutants?
SCHUCK  (U.S.A.)
     I  will attempt to answer the question concerning the
modeling.  There is a great deal of effort within EPA, within
the modeling field both in terms of specific models for point
sources for example and more general atmospheric models that
would apply to an urban area.  There is a very major EPA effort
called the RAPS in St. Louis, in which a data basis is being
generated with an array of stations for the very specific purpose
of testing but these very complex urban models that have been
developed.  St. Louis was chosen because it was the simplest
case in the United States and somewhat representative of many
other cities, it is not near a large body of water, it has re-
latively a flat terrain, it does not have other major sources

-------
                               2172
close by.  The object was to find the  simplest  case  first  because,
simple as it is it is extremely complex.
O'KEEFFE  (U.S.A.)
     I would like to comment from a  somewhat different point
of view on the same question.  Basically we have  two different
types of monitoring, one of these is that done to determine
whether or not we have attained a legally required  level  of
air quality.  This is a legally motivated operation and the
tendency is to demand the extreme of precision in such measure-
ments because after all it has a legal background and eventually
it will get to court and the attorneys will argue over the last
significant figure until they tire themselves out.  On the other
hand we have, and my feeling was that today's discussion  was
confined to this, the use of monitoring as a tool in epidemi-
ology.  There I believe that we can  tolerate rather a larger
uncertainty, plus or minus 25% perhaps would be quite tolerable.
SCHNEIDER  (Netherlands)
     We are supposed to be talking only about monitoring needs
in relation to health.  I know it is difficult, I already warned
both the panel and the audience about the trap that one can  fall
into when talking about monitoring needs.  Monitoring  is a
whole field and we are only supposed to be talking about part
of it.  Although I know that there are some economists within
this audience, I think first comes the question that is being
asked by the epidemiologists or others from the medical field.
One finds any kind of monitor or monitoring system one needs
to answer the question and if this interferes with finance or
economy one starts to design it again.  But I think the answering
the question should come first.  Although I know it is a very
interesting subject, the subject of the exposure meter, looking

-------
                               2173
at the clock, I would  like to close  this  one  and  go  to  the
next.  Moving to  integrated monitoring, in  the  sense of the
multimedia approach mentioned by Dr.  Schuck.    I  know of a few
other institutes  and agencies besides EPA who are working in
this field. Here  also  we need to put in a large effort  to get
some small results out of it, and here also it  is important
to know, or to get a feeling whether  it is  worthwhile.
SCHUCK  (U.S.A.)
     This subject of integrated monitoring design  is  a fairly
all encompassing one, it does take  into account cost  benefit
for example, it does take  into account the number  of  stations.
I think if you reflect on  our environment and our  present con-
cerns they seem to be increasing each year.  If we  do  not take
a careful look at, and plan our monitoring we will be spending
unreasonable resources on  such activities and that really is
probably the basic point behind this charge  we have in Las
Vegas to examine in detail monitoring systems and  this includes
existing monitoring systems.  Are they adequate?   Are they  (they
were planned many years ago) still  fulfilling their mandate?
Are there too many stations involved for example?   There are
indications from some preliminary work that  perhaps in large
geographical areas, because of the  over-riding influence of
gross meteorological factors perhaps one station instead of
25 in several thousand square miles might be sufficient, de-
pending on how accurately  you wish  to define the level (in cases
where there is a very definite proportionality, between the
reading at one station and another  station 5O miles away). This
may not apply to all geographical areas.  The geographical area
that I am speaking about is in southern California where stable
conditions prevail, but within reason I believe that  we will
find many areas where we can reduce the number of  sampling stations.
Certainly we have to take  a hard look at how much  monitoring
we do.  Again there are some requirements (legal)  but do we

-------
                               2174
 have  to  go  beyond  that?   If  we have in fact enough research
 information to  have  defined  the exposure pathways, need we sample
 continuously? If  indeed  there is a need for research perhaps
 that  has long since  been satisfied, and now one is merely
 fulfilling  the  requirements  of the law, in that one is watching
 the levels  to see  whether one's control measures are having
 their effect.   One may then  be able to get by with much less
 expenditure in  funds for monitoring activities, when viewed
 from  that angle.

     We  have this  concern because of the very large cost of
 monitoring  the  environment.
MAGE  (Denmark)
     Talking about costs,  there  are  two different  magnitudes
involved.  If one talks about  costs  to the  American  automobile
industry of the model changes  and  to the  industries  throughout
the European area who have to  modify their  vehicles  to  be  sold
in the American market, it is  a  very large  cost.   Much  of  these
modifications are made in relation to air pollution  air quality
data in terms of what reduction  of emissions  is necessary  to
meet an air quality standard.  If  one had better information
perhaps one might be able to reduce  the emission reduction.
For example, if one spends, lets say one hundred thousand  dol-
lars, and got more precise measurements then  one might  not have
to spend a billion dollars but only  9OO million saving  one hun-
dred million for an expenditure  of only one hundred  thousand.
I think perhaps more monitoring might indeed  be good if its
designed properly and one knows what one is going  to do with
the data once one gets it.
SCHNEIDER (Netherlands)
     While cost-benefit analysis is very interesting and impor-
tant, our concern at present is the multi-media approach.

-------
                              2175
DUNCAN  (U.K.)
     Epidemiology requires accuracy of monitoring to a high
degree and this then has to be for a limited programme. If you
talk of routine monitoring the accuracy may not be so important,
it depends on the realism of the standards to be met.  We must
be careful not to search for knowledge with routine observations,
We must keep our purpose clearly defined in planning monitoring
programmes.
MERIAN  (Switzerland)
     Does the panel agree that we should primarily monitor the
most relevant pollutants?  I believe that in traffic oriented
monitoring hydrocarbons are more relevant than carbon monoxide
and nitrogen oxides. It is however interesting to note that
in this session hydrocarbons are not mentioned.  Perhaps the
reason is that it is more difficult to measure hydrocarbons
than for instance the less relevant carbon monoxide. Should
we not develop a simple instrument to monitor hydrocarbons?
It is a question of responsibility to do research in fields
which are recognized to be relevant.
SCHNEIDER  (Netherlands)
     Regarding the question on accuracy I feel that 25% accuracy
might be enough for epidemiology and maybe we could do with
the same accuracy for other purposes.  What is necessary is
that we answer the question asked, and so far the questions
being asked by our medical colleagues is to give and answer
plus or minus 25% as a rough guess.  If you talk about other
questions you may need a much higher accuracy to answer the
question.
     Regarding hydrocarbons, I think Dr. Merian perfectly right,
at least for certain regions.  I can answer you by naming a

-------
                              2176
 few regions  in my  own  country and  I  know many more in western
 Europe and all over  the world.   The  point is that here again,
 we do not have at  the  moment  the possibility of measuring hydro-
 carbons with exposure  metres.   But if  there is a question,  and
 if there is  a problem, I  think  that  such a monitor should be
 developed.
SCHUCK  (U.S.A.)
     Generally  speaking  the  choice  of  the  pollutants  measured
in routine monitoring at least  in the  United  States,  has  to
do with the selection of pollutants which  have been studied
in detail, and  for which national air  quality standards have
been established.  Beyond that  much of it  is  still  in a research
stage.  I would think that what should be  the highest priority
on the list might depend to  some extent on what region, what
country we are  talking about.
MAGE  (Denmark)
     To respond to the question as  to why do  we  not  measure
hydrocarbons, I would first  like to point out that in  the  tech-
nique which was involved in  the paper which I presented  were
samples collected using aluminized  mylar bags.   Assuming that
it is nothing like ozone or  nitrogen dioxide  which would chemicalli
react in the dark atmosphere, the same technique could be  used
for anything, since you can  go back to your laboratory with
the bag and put it on the gas chromatograph.   When you are
talking about hydrocarbons one has  to be careful because these
are personal type exposures.  If one is interested in  a  break-
down into carcinogens or chemically reactive  species in  the
photo-chemical oxidant network you  are asking for somebody to
walk around with at least a  gas chromatograph, and to  my know-
ledge those instruments are  not quite portable and vibration
free, but I would suggest that one  could do the  type of  analysis

-------
                               2177
which we had authored  as a means  of  sampling.   Once  the  sample
is back in the  laboratory, if  you are  only  interested  in total
hydrocarbon you  can use a flame ionization  unit,  if  you  are
interested in a  breakdown you  can use  a  gas chromatograph.
SCHUCK  (U.S.A)
     I would  like to comment  that  I  so  not believe  that  any
instrumental  technique  is easy.  I would agree  that yes  we have
developed instruments for certain  pollutants  and  those are the
ones we intend to monitor.  I would  further agree that we have
research techniques for  just  about any  other  pollutant you wish
to name or group of pollutants, and  people are  working to make
practical instruments out of  these.  At the moment  they  are
either much too expensive techniques, they are  unreliable  or
require the complete attention of  a  well trained  technician
at all times, if not a professional.  Many methods  have  been
applied but not in a routine  monitoring sense.  The area of
aerosols for  example is  one field  where much  work is being done
and probably more research is required  in that  field than any
place else.  Work is going on on acid aerosols  but  at the moment
our best technique is a  rather gross measure, it  is a total
measure of the amount by weight of particulates in  the air,
and that is about the least amount of information,  the least
useful amount of information  that we can get.   Yes  there are
complex techniques that  can go beyond that, but they cannot
be applied routinely yet.
OREL  (U.S.A.)
     Much has been said at this conference about the problems
of sulphates therefore I address my question, are we monitoring
the right pollutants to assure public safety? when high sulphur
fuel oil was burned early this year, we needed much more data
on particulates in sufficient locations and sufficient frequency
with this in mind?

-------
                               2178
MAGE   (Denmark)
     When you are asking "do we have enough monitoring,"  one
has to know something about the standard deviation  of  particulate
measurements.  If there is a very  low  standard deviation  (within
a city) of lets say 5 or 6 stations, one does not need as many
monitoring stations as when you have,  in a particular  area,
a very high standard deviation which is a measure of the  spread
of the total population about the  arithmetic mean of the  pop-
ulation.  So to answer your question I would have to look at
                                                                 /
the information from existing networks, look at  the standard
deviation of sulphate measurements, if they are  all,   lets  say
3Oyg/m  every day then I think we  have enough; but  if  some  are
one some 1OO , some 20 and some 4o then we perhaps  do  not have
enough stations.
SCHNEIDER  (Netherlands)
     To further answer your question about more measurements,
looking all over the world, I think that in general the  answer
is no, there are enough measurements. Especially  if we talk
about the total amount of particulates, but if we talk about
what kind of particulates,  (with sulphates among  them) then
the answer is not straightforward;  it depends on the location.
In general we could say there are enough measurements on amount
of particulates, not enough on composition, size  distribution
and reactive properties of particulates.
SCHWING  (U.S.A.)
     What is the appropriate rollback model to protect the
public health and what are the monitoring needs to  satisfy
the model. Should one monitor for peaks or averages or statistical
deviations from some norm?

-------
                              2179
SCHUCK  (U.S.A.)
     Perhaps I can answer part of  it.   It does depend  in  a  sense
on what pollutant you are talking  about.  This is usually speci-
fied in the law that covers  the pollutant.
GOLDSMITH  (U.S.A.)
     That depends on the health problem which  has  been defined,
often monitoring is done for what is easy  to measure and not
for what is most important to health. Examples include:
1)  aldehydes, which appear to be important in producing
    effects of photochemical pollution.
2}  Monitoring of acid aerosols.
3)  The present monitoring of suspended particulate matter
    without size distinctions or chemical  characterization
    is so inappropriate to health considerations that the
    California Health Department has been  advised  not to
    consider further the standards for particulate matter
    until particle size data to reflect the sizes  with
    high respiratory retention are available.

4)  One needs portable or personal samplers to estimate
    pollution within residential premises  (so  called domestic
    pollution).  Dr. Bouhuys mentioned an  example  of this.
MAGE  (Denmark)
     I just cannot resist quoting something that I have heard
lately which is that "if something is not worth doing, it is
not worth doing well," and I think that Dr. Schwing's question
is a typical example of this type of problem that one gets into
when one talks about — lets say a linear rollback.  I do not
know how many in the audience are familiar with what linear
rollback is; It is a simple assumption that if you reduce the

-------
                               2180
sources by a factor of  2 from what  they  are  this  year  the  highest
level you will have next year will  be  half of  the highest  level
you have this year.  This  is an  incorrect model from the simple
dimensional analysis approach, because the dimensions  for  any
kind of equation must be homogeneous and consistent.   One  cannot
say that concentration  which is  dimensionless  —  lets  say  ppm
is equal to a constant  which has dimensions  times an emission
factor which is something  like mass tons per unit time per
unit area.  There are more than  just one variable in the equation,
and there is no purpose in devising a  monitoring  system and
analysing data to verify the linear rollback model when the
linear rollback model is theoretically incorrect.
SCHUCK  (U.S.A.)
      I would agree that the concept of linear rollback has
its problems/ but at the moment this is the only tool we have
to approach this subject.  What becomes the biggest area of
concern is when you try to apply this to a secondary type pol-
lutant such as ozone in the atmosphere.  Ozone in the atmosphere
is caused by the photo-chemical interaction of oxides of nitrogen
and hydrocarbons, and unless you have a control programme which
controls each of these variables to the same degree, then you
cannot expect linear rollback to predict with any degree of
accuracy what the effect will be.  It is most important that
this be recognized when attempting to reduce oxidants in the
atmosphere, This gets back to the entire question of taking
a systems approach, you cannot say that hydrocarbons are the
most important and ignore what is happening to your oxides of
nitrogen as has been done in some past effort to control the
hydrocarbons in exhaust gases.  They reduced hydrocarbons by
increasing the flame temperature in the combustion chamber and
thus increasing the oxides of nitrogen.  It had a benefit in
that the oxides of nitrogen, in this case nitric oxide reacts
rapidly with ozone to destroy it, so you have an additional
benefit besides reducing the hydrocarbon.  At the same time,

-------
                              2181
this was somewhat offset by the increase levels  in nitrogen
dioxide levels in the atmosphere which is also health related.
Any application of linear rollback has to take into account
a lot of factors, particularly if we are talking about secondary
pollutants.
MAGE  (Denmark)
      would like to point out that it is very dangerous to use
a linear model, even if it is dimensionally correct for a
non-linear system.  Air pollution and photochemical smog is
a highly non-linear system. For instance there are regimes which
have been found in smog chamber studies, where if you decrease
the amount of nitrogen oxides and you increase the amount of
hydrocarbons in your smog chamber you can increase the amount
of ozone that is produced.  You have a non-linear differential
equation and try to linearize it.  The assumptions that you
make are vital in the confidence of the answer you get from
the results of your study, and so one has to know the system.
SCHUCK  (U.S.A.)
     To say that we are using a linear model in our attempts
to control ozone in the atmosphere is not correct.  We did
not use data from chamber systems, for the simple fact that
they cannot be extrapolated.  There is too much effect of the
chambers themselves on the reacting mixtures.  The model that
is used for predicting the degree of control required is based
on actual observations of values in the atmosphere and their
variation, and it is indeed a non-linear model.

SCHWING (U.S.A.)
     Though the mechanisms of photochemical smog are non-linear,
models currently used are linear in that they incorrectly assume
that reducing emissions by 9/lo will reduce peak values by 9/lo.

-------
                               2182
It has been demonstrated  that  the degree  of  nonlinearity  Is
so great that the abatement requirements  differ  from  the  re-
quirements of linear models.
SCHUCK  (U.S.A.)
     There are several non-linear models,  including  the  one
that EPA is currently using, and they all  come up with approximate!)
the same predicted degree of control based on non-linear concepts
including the model that you have mentioned.
SCHNEIDER  (Netherlands)
     We have not touched upon one subject which  is the form of
the data and the use of the data.  I know this is a difficult
subject.  You heard on the one side the approach used by
Dr. Bravo, an index as a measure of pollution, on the other
side you have the automatic and continuous monitoring giving
measurement results every minute or even less, but in view of
the lack of time and importance of the subject we should  leave
it for a latter discussion.
     It is always dfficult to summarize a discussion.  We have
been talking about the need for monitoring, especially in con-
nection with health.  I think that it is certainly worthwhile
to have second thoughts about the design of the  monitoring systems-
I think this was definitely a point that was made this afternoon
There is a general opinion that the use of personal exposure
meters should be promoted.  Also clearly expressed was the wish
fcom the medical side to monitor not only the "easy" pollutants.

-------
                              2183
                       GEWEBSMESSUNGEN
                     TISSUE MEASUREMENTS
           MESURES RELATIVES AUX TISSUS BIOLOGIQUES
                MISURE NEI TESSUTI BIOLOGICI
               METINGEN VAN B10LOGISCH WEEFSEL
                            Panel

Voraitzender ~ Chairman - PrSsident - Preeidente - Voorzitter

                        D.  SZADKOWSKI
                (Bundesrepublik Deutschland)

-------
                              2185
         INTERCOMPARISON PROGRAMME ON THE ANALYSIS OF
        LEAD, CADMIUM AND MERCURY IN BIOLOGICAL FLUIDS
           A, BERLIN"*", R, LAUWERYS"*^/ J,P, BUCHET"
         H. ROELS"1"*", P, DEL CASTILHO"1"1" AND J, SMEETS+

+  Health Protection Directorate, Commission of the European
   Communities, Luxembourg
++ Unit6 de Toxicologie Industrielle et MSdicale, University
   Catholique de Louvain, Brussels, Belgium
ABSTRACT

     An intercomparison programme was undertaken to evaluate the
accuracy and precision of lead, cadmium and mercury determinations
in blood and urine.   Sixty-six laboratories participated in this
study.   For all the analyses a minority of laboratories only
measure  the metals with a satisfactory precision  (intralabora-
tory coefficient of variation of 3 or 4 measurements £?10%).  How-
ever, the laboratories which demonstrated a sufficient precision
in their measurements did not necessarily determine the metal
concentration with a satisfactory accuracy.   It seems that at
least for blood lead determinations the variability found between
laboratories which measure the metals with precision is due to
systematic errors.

-------
                               2186
     Precise and accurate determination of heavy metals in body
fluids is of paramount importance for the correct evaluation of
environmental exposure to these metals.
     Intercomparison programmes have been run and in some coun-
tries are routinely undertaken for controlling the accuracy of
the methods used by different laboratories for the determination
of lead, cadmium and mercury in blood and/or urine.   In some
programmes the precision of the methods has also been evaluated.
In general, these studies have indicated that the reliability of
these measurements is unsatisfactory (1, 2).
     The study performed in 1970 by Kepler et al in the U.S. in
which 66 laboratories participated showed that about half the
laboratories only reported results of acceptable accuracy.   This
study gave also an indication of the precision of blood lead
measurements since the participating laboratories recieved two
identical samples wearing different code numbers.   About 43% of
the laboratories reported similar results  (+ 10^ug%) for two iden-
tical blood samples containing approximately 65wg/100 ml lead.
However, the range of the reported values was considerable:
from 0 to SeOO^g/lOO ml for all the laboratories and after elim-
ination of two "outlyers" still from 30 to 120^%.
     In 1972, the CEC ran a limited intercomparison programme
(26 participating laboratories) of blood lead determination (1),
which also demonstrated a great scatter of the results reported
by the laboratories.   However, the precision of the measurements
performed by each laboratory could not be evaluated since only
one sample of each blood specimen was sent to the laboratories.
A more extensive intercomparison programme was therefore under-
taken in which accuracy and precision of lead, cadmium and mer-
cury determinations performed by 66 European laboratories could
be evaluated by sending to each laboratory 3 or 4 identical samples
wearing different code numbers.

-------
                               2187
MATERIAL AND METHODS
a.   Experimental Protocol
     Only unspiked samples were used.   Using    Pb as tracer,
Kopito and coworkers  (3) have recently shown that because of
differences in-vivo and in-vitro lead binding, recovery of added
inorganic lead may not accurately reflect the fate of endogenous
lead in some analytical procedures.   The following samples were
sent to the laboratories:
     Blood;             Blood C: 4 identical samples  (cow blood)
                        Blood D: 3 identical samples  (cow blood)
                        Blood E: 1 sample (human blood)
     Urine;             Urine A: 4 identical samples
                        Urine B: 3 identical samples
     Aguequs Solution;  Solution 1: 3 identical samples
                        Solution 2: 1 sample obtained by 10%
                                    dilution of solution 1.
b.   Preparation of blood, urine and aqueous solutions
     A.   Blood
     1.   Cow Blood
     The two different bloods C and D were obtained on different
times by withdrawing blood from a cow that was receiving metal
salt solutions orally.   The blood was taken by means of a needle
introduced into a vein of the neck and connected to a 1 litre
baxter in which the blood was collected under reduced pressure.
The first 10 ml were discarded to avoid contamination.   The
baxters were treated beforehand with 10% nitric acid.   After
rinsing with demineralized water, sodium heparin (Roche)  was
added.   The baxters were evacuated at the water pump*   To pre-
vent bacterial growth, blood was stored at 4°C., furthermore,
50 mg streptomycine and 10 mg chloramphenicol per litre of blood
were added.
     2.   Human blood
     The human blood E was obtained from four donors of blood
group A who were not occupationally exposed to heavy metals.   A
total quantity of 1.5 litres was collected on sodium heparin in
two 1 litre baxters under reduced pressure.

-------
                               2188
     B.   Urine
     Urine was obtained from normal persons and from workers oc-
cupationally exposed to Pb or Hg.   The samples were collected
on concentrated nitric acid to yield a final HNO- concentration
of approximately 3%  (w/v).   During storage in the cold room
(4°C) a precipitate was formed.   The urine was filtrated and
mixed to yield two different urine pools, i.e. A and B.
     C.   Aqueous solution
     pb(NO,) and CdCl-.2  1/2 H,0  (pro analysis, U.C.B., Belgium)
          j 2        *•        ^
were dissolved in 1% HN03  (w/v) to yield a solution containing
186yug Pb and 34.7yug Cd per litre  (solution 1).
     The 10% lower concentration solution was obtained by dilution
of the preceding solution with 1% HNO- (solution 2).
c.   Sampling
     For blood and aqueous solutions 10 ml polystyrene tubes were
used (Distrilabo, Belgium).
     For urine 50, 200 and 250 ml polyethylene bottles were used
with internal polyethylene cap and external rad plastic screw
cap  (Belgolabo, Belgium).   All containers were treated beforehand
with 10% HNO. and demineralized water.   Urine and blood were
continually agitated during distribution.   The homogeneity of
the blood samples was checked by measuring the haematocrit value
of several samples taken at regular intervals during the distri-
bution.

RESULTS AND DISCUSSION
     All the results are  summarized in Table I.   Three different
coefficients of variation  (C.V.) could be calculated:  1) an inter-
laboratory C.V. which is  calculated from the distribution from the
means of the independent measurements which were made by each
laboratory on blood C and D, on urine A and B and on aqueous
solution 1, and from the distribution of the single measurements
performed on blood E and  aqueous solution 2.   2) a "true" intra-
laboratory C.V. which is  calculated from the 3 and 4 independent
measurements made by ea9h  laboratory  (blood C, D;  urine A, B and

-------
                                  TABLE I :  Suirjnary, Table - Lead - Mercury ^ Cadmium
Analysis
^Blood C (4)
Blood D (3)
Blood E (1)
Urine A <4)
Urine B (3)
Aq.. sol. 1(3)
Aq..sol.2(l)
nf? -n.1 _ J1. /4V
^3-i.ood C (4)
Blood D (3)
Blood E (1)
Urine A (4)
Urine B (3)
Cd31ood C (4)
Blood D (3)
Blood E ( l)
Aq.soU (3)
N
52
52
55
33
34
51
50
18
18
18
29
29
14
17
17
22
Aq.sol2 (1) I 22
i
Mean
15.0
24.1
23.3
62.6
84.1
209.0
193.8
4.4
2.8
2.6
7-5
13-0
1.4
1.5
1.9
38.2
36.4
Median
12.8
22.7
18.1
62.3
78.3
196.6
190.0
3.6
2.1
2.4
4.6
11.0
0.65
0.9
0.96
35.0
33.0
Range
2-7 - 49.0
10.3 - 87.3
1 -115
5.3 -159
8.4 -185.2
24.7 -669.8
18 -580
1.9 - 9.4
0.8 - 11.3
0.2 - 9.0
1 - 67.5
4.9 - 63-3
0.1 - 9-2
0.03- 7.3
0 - 11.0
18 -116.7
11 -150
Interlaboratory
52.2
77.5
58.8
49.7
54.9
47-9
49-7
86.6
80.5
206
61.8
(168)*" *
(116)
(143)
(51.2)
(72.0)
"True" i
CV (%
Median
12.2
6.6
12.8
11.1
8.3

11.5
15-8

12.0
6.8
29-9
12.8

3-2

ntralaboratory
)
Range
1.5 - 129.2
0.8 - 99-9
1.7- 38.1
0 - 64.0
0 - 89.5

1.1 - 46.8
0 - 86.8

0 - 168
o - 36.5
8.8 - 61.9
3.6 - 28.4

1.3 - 39-0

"Repc
K
37
38
38
19
21
26
24
10
10
10
14
13
8
11
11
9
9
rted" int
C\
Median
7-8
4.U
4.9
4.8
4.0
4.2
2.2
6.6
8.4
1.1
12.3
6.3
4.6
7.4
7-2
4.3
2.0
r alaborat ory
' (*)
Ranse
0.4 - 17.0
0 - 22.6
a - 20.0
0 - 19.6
0 - 21
0.6 - 30.3
0 - 32.8
2.4 - 28.2
0 - 37
0 -104
2.6 - 40
1.6 - 26.6
0 - 22.9
0 - 28.0
0 - 26.1
1 - 10
o - 13.9
  *  jTunber of identical samples sent to each laboratory.
. * *  CV = coefficient of variation
* f  Distribution of neans deviatesaarkedly from normality,  (values vithin brackets)
     The concentrations are expressed in  ^jg/100 al  (blood) or in  jig/1 (urine).

-------
                                   2190
§
*J
2
A/
c
t
O
o
   30
   20
   10
    N
                        • mean •
                        .median
                                    Ct coloriraetry
                                    P: flame atomic absorption
                                    NP: flameless atomic absorption
                                    Oi polarography and anodic stripping
       Figure 1 t Distribution of results of lead analysis on blood
                sample D (all laboratories).  The techniques used
                by the laboratories are indicated by the abbreviations
   aqueous solution  1).    3) a "reported"  intralaboratory C.V.  could
   also be calculated when laboratories  performed several analyses on
   the same sample.   For all the analyses  the range of the reported
   results is very large.  As expected the interlaboratory C.V.  is
   always greater than the "true" intralaboratory C.V.  It is  inter-
   esting to notice  that in general the  "reported" intralaboratory
   C.V. is smaller than the "true" intralaboratory C.V.  This  finding
   demonstrates the  existence of a laboratory bias in reporting  re-
   sults;  when several measurements are performed on the same  sample,
   obviously outlying results are rejected.

-------
                                 2191
            Laboratories
Laboratories
     Figure  2i Distribution of results  of mercury analysis on blood
              C and cadmium analysis on blood D (all laboratories)

     Figures  1 and 2 Illustrate the distribution  of the  mean
values of lead,  cadmium and mercury in the blood  samples in which
their concentrations were the highest  (D for Pb and Cd,  and C for
Hg) .
     For lead determinations (Fig. 1) the analytical methods are
also indicated.    It is evident that the scatter  of the  results
is rather similar  whichever method used in this programme.    We
have found  that  the  interlaboratory coefficients  of variation for
laboratories  with  high (>30 analyses/month) or low degree of ex-
perience are  not significantly different.
     The interlaboratory coefficient of variation  for  laboratories
which obtained a low "true" intralaboratory coefficient  of  vari-
ation «10%)  was lower than that of the other laboratories.    The

-------
                                2192
 ^  30
 o
  en
 "8
  C
  O
 '13
20
  £  10
  O
  O
  u
  i
 JD
 Q_
      C i colorimetry
      F J flame atomic absorption
      UP t flameless atomic absorption
                     NF
                                            n=19
                                            r=0.74
                                          [D]=0.74[CH268
                   5          10           15          20
                      Pb-concentration blood  C (jug%)
    Figure 3  « Correlation between lead concentrations in blood D
             and C measured by laboratories with small "true"
             coefficient of variation. (< 10$)  for both measurements
influence of both  factors on interlaboratory coefficients  of
variation is shown in Table II.    Although combination of  both
factors  improves the interlaboratory coefficient of  variation,
the accuracy of the  results remains unsatisfactory.    These re-
sults suggest that systematic  errors are responsible for the
high interlaboratory variation observed between laboratories that
measure  the metals with  a satisfactory precision.    To test this
hypothesis we have compared the results of the low blood lead (C)
and the  high blood lead  (D) analysis obtained by the 19  labora-
tories which exhibited a "true"  intralaboratory coefficient of
variation 
-------
                                    2193
       TABLE II :  Effect of combination of small "true" intralaboratory
                  variation (CV)  and high experience on interlaboratory
                  mean and variation.
Analysis
^Blood C
'Blood D
; Urine A
Urine B
HgBlood C
Blood D
Urine A
Urine B
Laboratories vith "true"
CV\C 10 % and exper. ^ 30
anal /month.
n
20
2k
5
8
5
3
7»»
11**
- *
X
13.1*
22.6
52.6
78.7
2.81
1.20
U.36
11.00
cv (JO
3H +
20 +
32 +
11* +
17 +
16 +
33
21 +
Other Laboratories
n
30
27
26
26
13
12
21
n
_ *
X
16.5
25. 1*
61.7
85-9
1».98
3.33
lj.69
11.30
CV (*)
55
5*
60
5^
1*6
81*
1*6
50
  *  The concentrations are expressed  in jag/100 ml  (blood) or ug/1  (urine)
* *  1 outlying value excluded
   + Distributions significantly different  (p^0.05)
   Data for Cd are omitted because to  few laboratories have a low "true"
   C.V. and a high experience.

-------
                               2194
hypothesis that the variability found between laboratories that
measure the metals with precision, is due to systematic errors.

CONCLUSION
     The majority of the laboratories which took part in the
studies have not adequately developed the techniques required
for the precise measurement of Pb, Cd and Hg in blood, urine and
water.
     The variability of the results is not due to the use of
different analytical methods or difference in experience since
precise results were obtained by laboratories using different
techniques and having a different degree of experience.
     Furthermore the laboratories that measure the metals with
precision do not exhibit the same degree of accuracy.   Heavy
metal levels in blood and urine cannot therefore be directly
compared when the measurements are performed by different labora-
tories even if the precision of their analysis is satisfactory.
Therefore interlaboratory comparison of dose-response curves for
different parameters (e.g. urinary 5-aminolevulinic acid, ery-
throcyte 5-aminolevulinic acid dehydratase...)  based on the metal
content of biological materials are questionable.   Laboratories
should exchange samples regularly and standardize further their
methods for reducing their systematic differences.

ACKNOWLEDGEMENTS
     While in the impossibility of listing individually the
nearly seventy laboratories who have participated in this pro-
gramme, the authors wish to thank them and acknowledge the fact
that without their enthusiastic collaboration this programme
would not have been possible.   Mme Langevin deserves a special
note of recognition for the coordination of this programme.
     Thanks are also due to Dr. Recht for his advice and encour-
agement during this whole study.

-------
                               2195
REFERENCES
     Berlin, A., Del Castilho, P. and Smeets, J., European
     Intercomparison Programmes, page 1033 in Proceedings Inter-
     national Symposium Environmental Health Aspects of Lead,
     Amsterdam 1972 published by the Commission of the European
     Communities, Luxembourg 1973.
     Keppler, J. F., Maxfield, M. E., Moss, W. D., Tietjen, G.
     and Lich, A. L., Interlaboratory Evaluation of the Reli-
     ability of blood lead analyses.   Amer. Ind. Hyg. Assoc. J,
     31, 412, 1970.
     Kopito, E.  L., Davis, M. A. and Shwachman, H., "Sources of
     Error in Determining Lead in Blood by Atomic Absorption
     Spectrophotometry".    Clin. Chem.  20/2, 205-211 (1974).

-------
                               2 197
      A COMPARISON OF METHODS FOR ANALYSIS OF  CADMIUM  IN
     FOOD AND BIOLOGICAL MATERIAL,   A  CO-OPERATIVE  STUDY
               BETWEEN SWEDEN, JAPAN AND U.S.A.

            TORD  KJELLSTROM+, KENZABURO TSUCHIYA++
         EDYTHALENA TOMPKINS    .  E.IGO TAKABATAKE
                BIRGER LIND+ AND  LARS LINNMAN+

+    Department of Environmental  Hygiene, Karolinska  Institute
     and National Swedish Environment Protection  Board,  Stockholm
     Sweden
++   Department of Public Health  and Preventive Medicine, Keio
     University, Tokyo, Japan
+-H-  Human Studies Laboratory, National Environmental Research
     Center, Research Triangle Park, NC, USA
•H-++ Institute of Public Health,  Tokyo, Japan
ABSTRACT

     The data presented are part of the  "Japanese-Swedish~United
States Co-operative Studies on the Health Effect of Environmental
Cadmium".   In addition certain data are included from Swedish
(Isotope Technics Inc., Stockholm), American  (Dr. Philip D. La-
fleur, National Bureau of Standards, Washington DC) and Japanese
laboratories (Professor Jun Kobayashi, Institute for Agricultural
and Biological Sciences* Okayama University,  Kurashiki, Japan;
Dr. Kentaro Kubota and Dr. Kaauko Shiroishi,  Toyama Institute of
Hygiene and Medical Microbiology, Toyama, Japan), which Originally
     not participating in the co-operative  studies.
     The study is a part of a larger project, including studies
on the accumulation of cadmium on organs with age, on the average
Cadmium excretion in urine and feoes in "normal" populations, and
on the cadmium concentration in blood and urine of highly exposed
Populations,

-------
                                 2198
Introduction

Dependable analytical methods are of outmost importance in studies
on health effects of environmental pollutants. Cadmium is of spe-
cial significance in this respect, because of the low daily intakes
estimated (250-550 ;ig) to induce the earliest effect, tubular dys-
function (1). The concentrations of cadmium commonly reported (l)
in staple food like wheat and rice are below 0.1 ug/g wet weight.
The concentrations corresponding'to the critical daily intakes
would be around 0.4 ^.g/g. Average urinary concentrations of cadmium
among non-exposed persons are usually around 1 ng/g. In cadmium-
exposed workers the levels may Increase about 100 times. Urinary
cadmium is an indicator of both body burden and cadmium-induced
kidney damage.

The methods most commonly used for analysis of cadmium in staple
food, and in urine, are atomic absorption spectrophotometry,
neutron activation and the dithizone colorimetric method. The
latter method is less sensitive than the two former, and is
therefore nowadays not in common use for samples with low con-
centrations. The sensitivity of neutron activation or atomic
absorption is dependent on the type of samples analyzed. Certain
salts and metals will influence the analysis as discussed by
Friberg et al., (1).

When analyses made in different laboratories are compared a poor
agreement in results may be caused by analytical errors, failure
to procure homogeneous subsamples or e.g. wall-effects in the
storage of liquid samples. In the following study efforts were
made to avoid systematical errors in the selection and storage
of subsamplea. The aim was to find possible differences in
results of chemical analysis and not to explain from
whence such differences emerge.

-------
                                 2199
Materials

Grains

Two rounds of analysis were performed. In the first round five
Swedish wheat samples and ten Japanese rice samples were analyzed.
Two of these rice samples were so-called "standard rice", used for
calibrating analytical methods within Japan. They are supplied "by
the National Institute of Public Health to laboratories upon request.

The wheat samples had been stored in glass bottles or paper bags
until collection and then stored in transparent polyethene bags.
Prom these batches 500 g samples were taken, from which sticks
and straws were discarded. The samples were mixed by shaking
thoroughly and subsamples were taken for the different labora-
tories. The rice samples had been stored in transparent un-
colored plastic bottles.

The final data from each laboratory were sent to Dr. Jaroslav
Vostal, Department of Pharmacology and Toxicology, University
of Rochester, N.Y., U.S.A. No  communication between the labora-
tories was permitted before Dr. Vostal produced a table of the
comparison of data.

The second round of analysis included three Swedish wheat samples,
one Swedish oats sample and one Japanese wheat sample from an air
polluted area. The samples were stored and treated in a similar
way as for the first round.

The coding of the second round was performed in a more elaborate
way than that of the first round. The five samples were prepared
from the original batches by the Karolinska Institute and given

-------
                                 2200
code numbers. Dr. Kazuo Nomiyama, Department of Hygiene, Jichi
Medical School, Toohigi, Japan took six subsamples of each sample
and gave them new codes. The key to the codes and all analytical
results were sent directly by each laboratory to Dr. Vostal.
Urine
A pool of 10 liters of urine from 20 employees of the Karolinska
Institute was thoroughly mixed and poured into six 1.5 liter bottles.
Dr. Nomiyama added a standard solution of cadmium chloride to four
of these bottles. A pool of 3 liters of urine from 6 workers occu-
pationally exposed to cadmium in a Swedish Cd-Fi-battery factory was
thoroughly mixed and poured into two 1.5 liter bottles. Another
pool from 6 other workers, not so much exposed, was prepared in a
similar way. From the total of ten 1.5 liter bottles so produced,
subsamples were taken after shaking.

The original ten bottles were coded by the Karolinska Institute.
Dr. Nomiyama divided the urines into subsamples and receded them.
Karolinska Institute receded the subsamples once more before
sending them to the participating laboratories. All codes and
results were sent directly to Dr. Vostal, This procedure should
assure a completely blind analysis.

Methods

Neutron activation analysis was performed by Isotope Technics
Inc., Stockholm, according to a method originally described by
Ljunggren et al., (2), which has now been modified. The sample
is put in a quartz tube, sealed and irradiated in a nuclear
reactor. The whole tube is crushed and washed in a vessel by a
10:1 mixture of H&O, and H^SCK and then 20 mg Cd++ carrier and
20 mg Hg"*"*" holdback carrier are added. The sample is wet digested
in a Bethge apparatus with reflux until complete digestion.
Several separation and precipitation procedures follow among

-------
                                 2201
which ion exchange is used for the final separation of Cd and Zn
(Sjbstrand, unpublished data). From the eluate cadmium is precipi-
tated on a thin gold plate by electrolysis (Sjostrand, unpublished
data) and the final measurement of the radioactivity is made in a
scintillation detector. The recovery of the 20 mg of carrier - Cd,
measured by weighing the plate (SjSstrand, unpublished data), is
usually about 60 $. The radioactive and stable Cd-ions are supposed
to react chemically in exactly the same way.

In their work on wheat and rice, the Institute of Public Health,
and Keio University Japan, used a simplified version of the
standard method for cadmium analysis of rice, designated by the
Japanese Ministry of Health and Welfare (j). The samples are
ashed in a low temperature oven. The ash is treated with HNO,
and this solution is analyzed by AAS, using a flame without back-
ground compensation. The standards are treated like the samples.
For urine the Institute of Public Health used an atomic absorption
method, employing extraction in dithizone/chloroform, which is
the Japanese standard method (4). The standards are treated the
same way as the samples throughout the whole process.

Karolinska Institute utilized an atomic absorption method for
wheat and rice, in which the samples are prepared by dry ashing
at 450°C, and dissolved in 1 M HNO,. The analysis is performed
with a heated graphite atomizer with deuterium background correc-
tion (5). For urine the Swedish laboratory used two different
methods. Both employ regular atomic absorption after extraction
in APDC/MIBK*. One method (the macromethod) uses sample volumes
of about 100 ml. They are wet ashed, extracted and the final
analysis is made with flame AAS on the organic phase. This is
modified from Lehnert, Schaller and Haas (6). Keio University
used a similar method for urine. The other method (micro-

* APDC = Ammonium pyrrolidinedithiocarbamate
  MIBK = Methyl isbbutyl ketone

-------
                                 2202
method)   at the Swedish laboratory is performed on 25 ml samples,
and includes dry ashing, 1 M HNO, treatment and extraction in APDC/
MIEK. The final analysis is made flameless on the organic phase with
the heated graphite atomizer (Linnman, to be published).

Toyama Institute of Hygiene and Medical Microbiology used regular
AAS after wet ashing and DDTC/MIBK* extraction for both wheat and
urine. The Institute for Agricultural and Biological Sciences,
Okayama University, analyzed the wheat samples with regular flame
AAS after dry ashing at 450 C (Kobayashi, pers. conun.).

The Environmental Research Center, USA, contracted Stewart labora-
tories^Knoxville, Tennesee to make the analysis of wheat samples,
The grains were powdered in a mechanical grinder equipped with
a tungsten carbide blade. Two different analytical methods were
employed for the powder. Method I involved digestion of the
sample in H?SO,, dry ashing, packing the ash in a graphite tube
and multielement determination with optical emission spectro-
scopy. In method II the powder waa ashed over HC1 in an 0-
atmosphere, the vapors condensed into the HG1 in an ice bath, and
the HC1 solution was brought to a standard volume. Cadmium concen-
tration in the HC1 solution was determined with standard AAS.

The National Bureau of Standards chose the following method for
wheat samples. Three to five gram samples were oxidized by wet
                                                         _7
ashing with nitric-perchloric acids after spiking with 10   g
of    Cd and the excess acid was removed by evaporation. The
residues were each taken up with 5 ml of 1.0MHC1 and passed
through an anion exchange column (AG 1x8, 100-200 mesh) con-
taining about 4 ml of resin. After washing the column with
25 ml of 1.0MHC1, the Cd was eluted with 4MHNO,. When the
* DDTO « Dietyldithiocarbamat
  MIBK » Methyl isobutyl ketone

-------
                                  2203
 eluates  were evaporated to dryness, it was apparent that there was
 too  much residue to spark directly so the Cd was electrodeposited
 onto gold wires. The altered isotopic ratios were measured by Spark
 Source Mass  Spectrometry (SSMS). The preliminary results showed  that
 sample C (wheat) was drastically underspiked so a second sample  of
                                             ~7   111
 about 0.8 g  was taken and spiked with 2 x 10 ' g    Cd.

 Table 1. Comparison of cadnium analysis in wheat (1st round).
 unit  - ng Cd/g wet weight, duplicate analysis.
Sample Ho,
(harvest year)
P-1 (1945)
P-2 (1971)
P-3 (1970)
P-4 (1947)
P-5 (1970
Estimated
variation
eoeff-tniont m V
laboratory
Heutron Karolinaka Keio Univ. Institute
activation Institute, AAS AAS of Public
range average range average range average Health, AAS
average
14- 24
71- 76
118-171
28- 33
94-106
?1.4 *
t9
73
145
30
100

24- 33
63- 75
133-155
33- 35
96- 97
19. <; t
29
69
144
34
97

120-140
160-180
220-230
120-150
180-220
11.4 <
130 190
170 160
225 230
135 120
200 170
r76ii-i«Y
v_irM\ ffi
                                                              2 a
Results
MS at the Institute  of Public  Health and Keio University gave
values more than  twice as high  as those given by AAS at Karolinska
Institute (Table  1).  The difference is statistically significant
(p < 0.05). There is, however,  a very good agreement between the
two Japanese laboratories,  as well as between Swedish AAS and
neutron activation. The estimated variation coefficient from
duplicate analysis is 12.5$ and 11.4 $ respectively, for AAS
at Karolinska Institute and Keio University,  and 21*4
neutron activation.
The difference in analytical results  between the Institute of
Public Health and Karolinska Institute  is similar in the first
round of analysis of rice samples  (Table  2),
Only one rice sample was analyzed  with  neutron activation, the
result falling in between the  Swedish and Japanese AAS results.

-------
                                     2204
Table 2. Comparison of cadmium analysis in rice (1st  round).
unit - ;ig Cd/g wet weight.
Sample Ho.
R-1*
R- 3
R- 4
R- 5
R- 6
R- 7
R- 8
R-9
R-10
Laboratory
Karolinska
Individual
IAS
0.079 0.081
0.018 0.026
0.025 0.028
0.006 0.008
0.029 0.031
0.062 0.063
0.109 0.113
0.105 0.126
0.103 0.10?

Institute
values ; average
0.089 0.083
0.022
0.027
0.007
0.030
0.066 0.064
0.111
0.116
0. 105

Neutron Institute of
activation Public Health
IAS
0.130 0.19
0.07
0.0?
0.07
0.11
0.16
0.26
0.23
0.20
x - "standard rice, 0.2 ug/g".
Results of AAS at the National Institute of public Health and
Keio University  in round  2 (Table j) differ more than 100 %  from
results of AAS at Karolinska Institute  as well  as neutron activa-
tion analysis.  There is a good agreement between AAS  at Karolinska
and neutron activation, even though the AAS-values in the second
 T.abl».-3. Comparison of analysis of  cadalua concentration in grains (2nd round).
 Duplicate analysis when ranges are  given, unit » ng Cd/g wet weight.
Laboratory
Beatron r
activation a
Keio
University a
Institute of r
Public Health a
Karolinska r
Institute a
ERS , 0SA a
Toyama Inat. r
of Hygiene a
Okayama
University
National
Bureau of
Standards x
Sanple Bo, (harvest year)
1 (1971)
wheat
41 - 48
44.5

120
133 -144
139
20.7- 40.6
30.7
220
44-51
47.5

44


41
3 (1945)
wheat
23-26
24.5

60
67 -100
84
17.1- 18.3
17.7
53
32 - 36
34.0

33


20
4 (1971)
(XI ts
12 -17
14.5

50

84
6.1- 7.2
6.7
210
20-21
20.5

17


12
5 (1941)
wheat
22 - 22
22.0

100
107 -111
109
14.5- 21.6
18.1
65
23 - 28
25.5

79


35
Estiaated
variation
coefficient (V)
14.1 %



16.5 %

27.6*


9.8*






 r - range   a - average
 x - only one figur* significant.

-------
                                   2205
round  are  relatively lower than  in the first  round. AAS  analysis at
Toyama Inst. of Hygiene, at Okayama University as well as  Spark
source mass spectrometry isotope dilutions  at National Bureau of
Standards  and neutron activation analysis generally showed a good
agreement. There  is  a large variation in the  data from the Environ-
mental Research Center, USA and  all values  lie more than 100 $ above
the neutron activation values  (Figure 1).
          AAS. Sweden,
          Japan, USA
          ng Cd/g wet weight
        200  -
                                   WHEAT
        150 •
         100 •
         50
                             A
                             *
               *
               A

             * *
                                     /
                                       /
                                        /
                               /
                              /
                           /
                                                    neutron activation
            0          50

            •  KaroHmka bnt. lit round
            o  KafoUmka tost. 2 ltd round
            A  Kelo Untvenily, 1 it round
            A  Kelo University, 2nd round
                                100          150  ng Cd/g wet weight
                                    *  Nat, Inst, o! PH, 1st round
                                    ft  Nat. Inst. of PH, 2nd round
                                    '  Tbyomo hist of Hygiene
                                    «  Okayama University
                                    a  Environmental Research Center
                                    •  Notional Bureau of Standards
                                       (Note: SSMS-method)
Figure  1.  Comparison of cadmium,  analysis in wheat (1 st and 2nd
round).  Neutron activation analysis used as a "reference  method"
on the  abscissa and the ordinate is giving  results of atomic
absorption (and sparked source mass spectrometry isotope  dilution)
from all participating laboratories.

-------
                                  2206
 The results reported  so  far concern samples with concentrations
 0.01-0.2 )ig Cd/g, which  is the range found in basic foodstuffs
 in non-polluted areas. Two samples from polluted areas with
 considerably higher concentrations were also «oopared (Table 4)*
 Table 4. Conarlion of eadftlva analyst! of polluted wheat and riea.
 unit - jig Od/g wet weight.
Laboratory
Neutron
activation
Karolinaka
Institute
Inititute of
Public Health
JUio
University
IBS, USA
Toyaaa last.
of Hygiene
Okayua
University
Rational Bureau
of Standards
General average of
laboratoriti
standard dtTiatlon
TYB* uf luplt
wheat froa Annal
indiTldnal
mluas
2.95
2.71
2.5*
2.59
1.89
2.40


2.94
3.20


all

Tariatlon coefficient

ca, Japan Japanese "ftandard rice, 2 ug/g"
indlTldual
average value* average
2.83
t C7 2. 1 1 '}•}•)
2.57 2>32 2.22
2.15 2.21
3.20
2.58
3.07
3.46
1.6
2.68
0.60
22.4 %
The  average of the results of analysis of wheat by 8  laboratories
was  2.68 ug Cd/g (range 1.6 - 3.46 ;ig/g, variation coefficient
22.4 #). The systematic difference between the Institute  of  Public
Health, Keio University and ERS, ITSA on the one hand  and  the
remaining  laboratories on the other hand as was seen  in low-level
samples did not occur for these higher-level samples.
As seen in Table 5 and Figure 2 there is generally a  good agree-
ment between the results of urine analysis apart from the two
samples (A and D)  with "normal" Swedish urine. Neutron activation
gave 2 ug/liter and ^0.8 >ig/liter on duplicate subsamples.  In the
calculations  of linear regression lines depicted in' Figure 2,
instead of <. 0.8 the  number 0.5 was used,  y » AAS ^ug  Cd/literf
x =• NA jig  Cd/liter. The correlation coefficients between neutron

-------
                                         2207
Table  5. Comparison of cadmium analysis  of urine.
unit -g Cd/lHar.
Sample Ho.
(Cd added
^g/liter)
                 laboratory	
                 NeutronKeioInstitute        lHarolinsIta last.
                 activation   University   of Publ. Health   micro   j   nacro
i a flanelees

xx a in
               , other method than  the regalAe oa« used in Tuyata.
          calculations the figure 0.5 is  used.
                    AAS.
               40-
               30-
               20-
               10-
Toyama lust.
of Hygiene
A CO)
D (0) = A
B (12)
f (12) -B
B (20)
E (20) = C
Exposed workers
0 (0)
Ff (0) = C
I (0)
K (0) •> 1
2
iP.a"
13
13
22
18

11
1
26
28
1.0
1,1
13.3
'2.5
21.0
22,4

8.9
8,6
31.2
28,7
1.0
1.9
13.9
'3.2
22.7
20.2

9.6
B.6
38.4
29.7
0.6
0.3
10,3
10.4
18.2
18.1

7.0
7.1
26.2
25.1
0.2
0.2
8.4
B.4
11.8
14.0

4.*
5.8
19.5
18.0
0.43*
0.35*-1.B
9.9
9.95
19.2
19.75

7.2
7.65
27.2
29.75
                            10
                                                  30
                 I * •StocMm.Mocro method.watef standard
                 Jo. -ii-  .Micro melhod.
                   'Insllluic of Public Health.
Figure  2.  Comparison of  cadmitua  analysis  of urine.  Feutron
activation analysis  used as  a "reference  method" on the
abscissa and the ordinate is giving results of MS  from
the five laboratories. Linear regression  lines depicted.

-------
                                   2208
 activation analysis and MS analysis  at each  laboratory vary
between 0.96 and  0.98 (Table  6)  and are all statistically signifi-
cant  (p < 0.001).  Calculations were based on the average results
of duplicate analysis. The  slope of the regression  line of AAS
(macro-method) at Karolinska  Institute  was significantly less
than  1  (at p < 0.05,  b = 0.69+0.14).  The Institute of Public
Health had a relatively steep slope (b  = 1.21) but  it was not
significantly higher than 1  (at  p < 0.05, b = 1.21  +0.24).
Table 6. Linear regression  lines for  comparison of  neutron
activation analysis and atomic absorption analysis  of cadmium
concentration in  urine. Number of samples =10. y = AAS value
for each laboratory,  x = neutron activation value.

  I   Karolinska  (macro)        y = 0.69x - 0.62       r = 0.97
 II   Karolinska  (micro)        y = 0.96x -1.18       r = 0.98
III   Keio University           y = 1.10x - 0.58       r = 0.98
 IV   Inst. of Publ. Health     y = 1.21x - 1.11       r = 0.96
  V   Toyama                     y = 1.05x - 1.6?       r a 0.98
As the subsamples of urine  used consisted of  pairs  of duplicate
 samples and  some  laboratories made duplicate  analysis of each
 subsample, the repeatability of urine analysis  can  be calculated
 in two different  ways  (Table 7)  . In these calculations samples
A and D were excluded.
 Table  7. Estimated variation coefficient of  cadmium  analyst* of urine
 (concentration range 7  -
 Laboratories from which duplicate analysis of each subsaaple was reported:
                                        In tra« ample variation coefficient
                                        (8 duplicate*)
 Karolinska Institute
 micro- «ethod                              4.3 jf
 macro-aethod                              12.0 f (7 duplicate! )
 Toyama Institute of Hygiene                   3.2 %

 Intersample estinated variation coefficient between 4 pairs of similar urine samples :
 Heutron activation                         17.4 $
 Karolinska Institute
 aicro-aethod                              1,6 %
 oacro-method                              11.8 %
 Keio University                            4.5 £
 Institute of Public Health                   10.8 %
 Toyama Institute of Hygiene                   4.0

-------
                                 2209
The recovery of added  cadmium to samples B, C, E, and P varied
between 60 and 100 $ (Table  8).
 Table 6. Relative recovery of added cadaiun to urine tuples, liiueed Cd concentration
 in original urine • 1.25 vg/liter. Average* of two autoaiplea.
Laboratory
Neutron activation
Kelo Dnivertlty
Institute of Public Health
Earolinska Institute
•iero-aethod
nacre-Method
Toyama Institate of Hygleut
fie, cadniun
12
98.0*
97.4 *
102.3 *

78.1 *
O.4*
74.9*
added/liter urine
20
93.8 %
102.1 %
100.9 *

85.4*
60.7*
91.9*
The recoveries  agree with the slope of the linear regression
lines calculated from all the samples.

Discussion

A methodological study on the Japanese standard method performed at
the Institute  of Public Health (5) showed a good agreement between
low temperatur ashing (average of 8 runs = 0.188 jig Cd/g,  variation
coefficient *•  2.7 $) and APEC/MIEK extraction  techniques (average of
8 runs  =  0.189 ug Cd/g, variation coefficient  = 7.4 #) when the same
rice  sample was analyzed. 9 different laboratories  analyzed samples
from  this rice batch employing APDC/MIHK-extraction and the average
concentration  was 0.176^ug Cd/g with a variation coefficient of
10.2 ?&.

No detailed methodological study on the various steps in neutron
activation analysis has been published. If the digestion of the
irradiated sample is incomplete losses may occur, Sjostrand (8)
has described the wet digestion boiling reflux technique in detail
and states that the distillation cycle is repeated  until the
solution is completely clear and colorless when boiling.
                                    t

The AAS-method used at Karolinska Institute has been studied in
detail  (5) by addition of radioactive cadmium. The  recovery was on

-------
                                 2210
an average 95 % with an average of 4.4 % losses in the second dry
ashing step. The method was compared with neutron activation on
59 wheat samples ranging 29 to 257 ng Cd/g (7) giving a correlation
coefficient of 0.946 and equation for linear regression AAS = 1.01 •
NA - 1.65.

Also in the present study there was a good agreement between analy-
sis of wheat by neutron activation and by AAS at Karolinska Insti-
tute as also by AAS at Toyama Institute of Hygiene and Okayama
University and by SSMS at National Bureau of Standards, Keio
University, Institute of Public Health and Environmental Research
Center got systematically higher results, The lack of background
correction after low-temperature ashing may explain the relatively
high results from Keio University and Institute of Public Health.
The great scatter of the results from Environmental Research Center
make them hard to evaluate.

Similar systematic differences between Karolinska Institute, and
Institute of Public Health can be seen for rice (Table 2). Further
data on rice are now under processing with a larger number of
samples analyzed by neutron activation.  Rice and wheat have differ-
ent composition of the ash, which may be of importance for the
recovery of the various ashing procedures.  No conclusion regarding
analysis of rice will therefore be drawn at present.

Among the wheat samples analyzed one sample in the first round was
also included in the second round. The results are included in
Figure 1, marked with numbers 1 and 2. As can be seen, AAS at
Karolinska Institute gave 77-5 ng Cd/g in the first round and
30.7 ng Cd/g in the second round. The same wheat batch harvested
in 1971 had been analyzed with Swedish AAS in connection with
another study and gave 42.5 ng/g, 50 ng/g,  51.5 ng/g and 56 ng/g
(duplicate samples) (5). Neutron activation analysis gave values
of 73 ng/g and 44.5 ng/g respectively on the same sample in the
two rounds, Keio University, Japan, reporting 170 ng/g and 120 ng/g,
and the Institute of Public Health 160 ng/g and 139 ng/g respec-
tively. These consistent differences between the two sampling times
indicate a real difference.in cadmium concentration.  This difference

-------
                                2211
is distressing as the samples were taken from the same batch. On
the other hand in the actual comparison between laboratories the
subsamples were taken from samples on which cleaning and mixing
had taken place. One way to limit the error in sampling from a
large batch would be to grind the material and homogenize it.
As our aim was to compare analysis of the original material used
for studies on daily intake, that is the grains, we felt it
would be more appropriate just to clean and mix the grains
thoroughly before subsampling.

The systematic differences between Keio University, the Institute
of Public Health and EPA, TJSA on the one hand and all other labora-
tories on the other hand which have been seen for low-level wheat
samples are not seen for the wheat sample from a polluted area
with a concentration around 2.7 ^g/g- The results range about + JO
from the average. "Standard rice' 2 jig/g" which had been analyzed
by 10 laboratories in Japan (?) giving an average of 2.33^ig/g
(variation coefficient 8.6 %) was again studied here giving
2.22 ug Cd/g at Karolinska Institute and 2.21 jig Cd/g at Insti-
tute of Public Health.

When the cadmium concentration in urine exceeds 5 jig/liter all AAS-
methods used correlated well with neutron activation analysis. The
macro-method at Karolinska Institute had systematic losses of about
^0 /£, which were statistically significant.

Methodological studies on the Japanese standard method for urine
(dithizone/chloroform extraction) (9) show a good correlation
between different laboratories. This method was also compared with
APDC/MIBK extraction with a good agreement (r = 0.996). All samples
were from a polluted area with cadmium concentrations ranging 2-26
ug/liter. The relative differences between results from different
laboratories were larger at low concentrations.

The present study shows that there are problems in analyzing
"normal" urine. None of the methods used can be looked upon as
a good reference method. The same differences as found in grains

-------
                                 2212
between Keio University and the Institute of Public Health on the
one hand and the other AAS laboratories on the other hand can be
seen in urine findings. More methodological research is needed
before cadmium content in normal urine can be analyzed with
accuracy.

Conclusions

The findings in this study emphasize the need for international
comparisons of trace metal analysis employing principally different
methods. Such studies should give full disclosure of the methods
used and any methodological studies performed earlier. A sufficient
number of samples of the same type of material should be used in
order that systematic differences can be evaluated. Efforts should
be taken to assure blind analysis and randomization of sampling
errors.
Only after such studies have been  carried out is  it meaningful  to
make comparisons e.g.  average body burden, excretion  or daily in-
take of metals in different countries.

Aknowledgements

We are grateful for  the valuable assistance of Drs. Kpbayashi,
Kubota, LaPleur, Nomiyama, Shiroishi and Vostal.
 References

 1.  Friberg,  L.,  Piscator,  M.,  Nordberg,  G.,  and Kjellstrom,  T.,
 Cadmium in the  environment,  2nd edition, Chemical Rubber Co.
 Press, Cleveland, 1974.

 2.  Ljunggren, K., Sjostrand,  B., Johnels, A.G., Olsson,  M.,
 Otterlind, G.,  and Westermark, T.,  Activation analysis of mercury
 and other environmental pollutants in water and aquatic ecosystems,
 IAEA-SM-142 a/22, p. 373-405, International Atomic Energy Agency,
 Vienna, 1971.

-------
                                2213
3. Yamagata, N., Ivrashima, K. , Kuzuhara, Y., and Yamagata, T.,
A model surveillance for cadmium pollution, Bull. Inst. Publ.
Health, 20 (3): 170-186, 1971.

4. Japanese Ministry of Health and Welfare; The method for health
examination of the inhabitants, a part of the provisional counter-
measures against environmental pollution of cadmium; Report from
the section for environmental pollution control, May 19, 1971
(in Japanese).

5. Kjellstrbm, T., Lind, B., Linnman, L., and Nordberg, G.,
A comparative study on methods for cadmium analysis of grain
with an application to pollution evaluation, Environmental
Research (in press).

6. Lehnert, G., Schaller, K.H., and Haas, T., Atomabsorptions-
spektrometrische Cadmiumbestimmung in Serum und Harn, Z. Klin.
Chera. u. klin. Biochem., Tol. 6, 174, 1968.

7. Linnman, L., Andersson, A., Nilsson, K.O., Lind, B., Kjell-
strb'm, T., and Friberg, L., Cadmium uptake by wheat from sewage
sludge used as a plant nutrient source, Arch. Env. Health, 27,
45-47, 1973.
8. Sjostrand, B., Simultaneous determination of Mercury and Arsenic
in Biological and organic materials by activation analysis, Anal.
Chem., 36, 814-8191 1964.

9. Japanese Association of Public Health, Research  about the
Standardisation  of analytical methods for cadmium poisoning,
Research Report  1970 (in Japanese).

-------
        THE DETERMINATION OF LEAD AND CADMIUM IN BLOOD
               BY ATOMIC ABSORPTION SPECTROSCOPY

                         H, T, DELVES

Hospital for Sick Children and Institute of Child Health, London
United Kingdom
ABSTRACT

     The problems of matrix interferences that are encountered
in the determination of lead and cadmium in micro-litre volumes
of blood by discrete sampling flame and flameless atomization
and atomic absorption methods are discussed.   Procedures to
overcome these interferences are given.

-------
                             2216
Introduction    The determination of lead  and  cadmium  inblood
by conventional atomic absorption spectroscopy  (AAS) requires
pre-concentration from relatively large volumes of blood
(l-5ml).  However, these determinations may be  carried out
with only micro-litre volumes of blood if highly efficient
atomization techniques such as discrete sampling flame
atomizers (l),(2), or flameless atomization systems (3) are
used.  In order to achieve accurate analyses with these
atomization techniques it is necessary to overcome matrix
interferences and interferences from non-atomic absorption
signals.
Experimental  Analyses using the Delves-cup technique were
carried out on a Perkin—Elmer '103' single beam instrument
without continuum source background correction, and analyses
with graphite tube furnace were made using a Perkin-Elmer
•303* instrument fitted with a Dg lamp background correction
accessory.  The graphite tube furnace was a modification,
by Woolley (4) of that previously described by Alexander et
al (5).
Blood-lead Analysis
       Matrix interferences  There was a 1O% enhancement of
sensitivity for blood-samples relative to aqueous standards
with the Delves-cup method for blood-lead analysis.  This
presumably resulted from the occlusion of the lead in a
volatile organic matrix.  However, with the graphite tube
furnace a depression of sensitivity was observed for the
blood samples.  In this latter technique the bulk of the
organic matrix was removed during the ashing stage prior to
atomization.  The magnitude of the depression depended upon
the power dissipated through the tube during the atomization
stage i.e. tube temperature (Figure l) and was  constant at
-16% within the power range 0.96 to 1.43kw.
       The matrix interferences were overcome, with both
techniques, by matching the matrix of the standard solutions
to those of the blood samples.  Multiple standard additions
to a series of lOjll volumes of a single control blood sample
were used with the flame sampling method, as previously

-------
                                    2217
           •30,
           .25
            .20

  Absorbance
    217.Onm

            .15
            .10
            .05
                    '/_
                    '
                                          __O O.6ngPb aqueous
                                                 0.75ngPb (blood
                              .6      .8      1.0     1.2     1.4     1.6

                             Power through tube kW
              FIGURE I     EFFECT OF H1.OOD MATRIX ON THE FLAMELESS AA

                                DETEHMINATION OF LEAD  (lul  SAMTLF.S)
              .6
    Abaorbance
       21?.Onm
fDelvea-cup method
           1O ill •ample*

 Sensitivity 4.9 x lO-11g Pb
  for IS absorption
                                      Graphite tube  furnace
                                               1 vl  sample
                                  Sensitivity 1.3 x  10-11g
                                   for 1% absorption
            20   0  20             100

                        tigPb/lOOml added
                           200
FIGURE a   CALIBRATION  CURVES FOR BLOOD-LEAD ANALYSIS

-------
                              2218
described (2).  With the graphite tube  furnacet  a  series  of
standards were prepared in a diluted blood matrix, by  diluting.
lOOv.1 of a control blood sample plus 100]il of an aqueous
standard solution to 2.OOml with water.  Calibration graphs
for both techniques are shown in Figure 2.
       Non-atomic absorption interferences arise from
radiation scatter by solid particles and/or  from molecular
absorption by species that are volatilized simultaneously
with the lead atoms. Unless these interferences  are corrected
for, the blood-lead analyses will be erronesouly high.  There
are three ways to correct for these interferences:
1.  by using a continuum source (D£ or  H% lamp)  background
    corrector,
2*  by carrying out the analysis at a wavelength that  is  free
    from this interference (this may not always  be possible)
3.  by subtracting from the absorption  signal obtained at the
    resonance line, the background absorption obtained at a
    nearby non-resonance line.
       These procedures were investigated as follows:
       Non-atomic absorption interferences with  the Delves-
cup method were investigated by making  replicate determina-
tions of lead in blood at the 283.3 and 217-Onm  resonance
lines and at nearby non-resonance lines*  Representative
results are shown in Figure 3«  Two absorption signals were
observed for each sample.  The larger signal was due to radi-
ation scatter by the combustion products of  the  sample and
the smaller, which occurred when the sample  had  finished
burning, was due to lead atoms plus any non-atomic absorption
at the lead resonance lines (283.3nm, 21?*0nm) and due to
non-atomic absorption alone at the other non-resonance lines*
There was no molecular absorption/light scatter  at 220nm,
which suggested that these interferences were also absent
at 217.0 nm.  There was however a variable non-atomic  absorp-
tion signal from 280 - 287nm, which indicated molecular
absorption rather than light scatter.   The absorption  signals
due to species other than lead at 280, 282 and 2&7nm were
found by replicate analysis to be equivalent to  blood-lead
concentrations at 283,3nm, of 6.3, 9.1  and 4. 9|ig/100ml

-------
                              2219
  0.50-
  0.40-
 Absorbance
  0.30-
  O. 2O-
  O.10-
                     NAA
            21? nra
       22<
                             NAA
                       NAA
120 nm
                                  nra
    FIGURE 3
       Pb =  Pb atomic absorption  signal
      NAA =  Non-atomic absorption signal

ABSORPTION SIGNALS AT RESONANCE AND  NON-RESONANCE
          LINES IN BLOOD-LEAD ANALYSIS
respectively.   Thus  there would be a positive bias  in blood-
lead analysis by  the above technique at 283.3nm unless  this
non-atomic absorption interference was eliminated.  However,
the above experiment did not indicate which of the  three  non-
resonance lines could be used for background correction.   The
results in Figure 3  showed no detectable interference near
the 21?.Onm line  and it  was therefore concluded that any  dif-
fer en^es between  blood-lead analyses obtained at 21?.0nm  and
283.3nm would be  due to  non-atomic absorption.  49  blood
samples were analysed for lead at these two wavelengths.   The
results (Table  1)  showed that there was a positive  bias of
approximately 5v.g/100ml  at the 283.3nra line, which  indicated that
the correct non-resonance line to be used for background

-------
                               2220
                        TABLE I
       NON-ATOMIC ABSORPTION IN BLOOD-LEAD ANALYSIS

       49 blood samples with lead concentrations from
       10-38yg/100ml analysed at 217. Onm and 283. 3nm
       gave the following results:

       Regression:  ug/lOOml

           217- Onm  =  283. 3nm x 0.99 - 5.3

           Correlation coefficient  0.93
             Mean
                              S.D.
             24.2             6.4         217.0
             24.4*            6.8         283.3

                 *5v.g/100ml subtracted from each value.

correction was the 287nm line.  These conclusions are  supported
by the work of Dawson et al (6) who used this 287nm  line to
provide continuous background correction for blood-lead
analysis at 283nm by flameless atomization using a dual
wavelength AA instrument.
         Experiments with the graphite tube furnace  and AA with
continuous background correction showed that without back-
ground correction there was a significant positive error for
blood-lead analysis at 283. 3nm, but there was much less
interference at the 21 7 • Onm line.  It is however recommended
always to use background correction with flameless atomization
systems for this analysis.
         A comparison of blood-lead analysis by the  two
atomization techniques discussed is shown in Table 2.  There
was no significant difference between the results.   The
flameless technique was more sensitive (Figure 2), but the
analysis rate was slower (40 tests per hour) than the  discrete

-------
                               2221

                         TABLE II
               COMPARISON OF BLOOD-LEAD  ANALYSES
                        Tig Pb/lOOml
Cup tech.
18
18
20
23
26
27
±7
G.T.F,
23
28
22
21
25
31
46
Cup tech.
55
60
63
67
80
82
200
G.T.F.
48
50
54
85
85
86
228
          G.T.F. 8 Graphite tube-furnace AA method

 sampling flame AA  method (60 tests per hour)  which was more
 suitable to routine  analysis of large numbers of samples.
 Blood-cadmium analysis
          Solvent  extraction  and conventional  flame AA, after
 wet  ashing of 1 ml samples,  allowed determinations to be made
 down to  0.2jig Cd/lOOml.   This procedure was however time-
 consuming and the  new atomization  systems were investigated
 for  this analysis.
          It was not possible to separate the  cadmium  absorption
signals from those of  the combustion products of the  sample
when using the Delves-cup technique, because  of the relative-
ly low vapour  pressure of cadmium  and  its  salts.   Ediger and
Coleman  (?) have described a pre-ignition  procedure for the
removal  of  the matrix  without  loss of  cadmium, but it was
not  pojbsible to reproduce successfully their  conditions or
results,  in this author's laboratory.   Alternative oxidation
procedures  along the lines suggested by Cernik  (8) are
currently being investigated.

-------
                             2222
      Preliminary work with  the  flameless  atomization
system has indicated that  31*1  of blood  are sufficient  for
the determination of cadmium.  Because  of  the volatility of
cadmium and its  salts, precise control  of  the ashing  stage
and continuous background  correction were  found  to  be
essential, to resolve the  cadmium  signals  from those of  the
matrix.  This method is still  being developed.
Conclusion   Matrix and non-atomic absorption interferences
in the determination of lead and cadmium in blood are  more
severe with discrete sampling  flame and flameless AA tech-
niques than with conventional  AA methods.  However, these
interferences may be overcome  by suitable  calibration  and
background correction procedures.
References
     KAHN, H.L., PETERSON,  G.E. ,  "Atomic  absorption  micro-
     sampling with the  'sampling  boat1  technique", Atomic
     Absorption Newsletter,  7,  35,(1968).

     DELVES, H.T., "A micro-sampling method for the  rapid
     determination of lead  in blood by  atomic-absorption
     spectrophotometry," Analyst,  95, 431  (1970).

     KIRKBRIGHT, G.F., "The  application  of  non-flame  atom
     cells in atomic-absorption and atomic-fluorescence
     spectroscopy,"  Analyst. 96,  609  (1971).

     WOOLLEY, J.F., "The determination of iron  and  copper in
     high purity glasses by  flameless AAS," Paper presented
     at the 4th International Conference on Atomic Spectros-
     c°Pyi Toronto,  Canada,  November, 1973*

-------
                            2223
5   ALEXANDER, F .W. , DELVES,  H.T.,  REESON,  R.B., "The
    application of  atomic  spectroscopy to the analysis of
    biological materials,"   3rd  International Congress of
    Atomic Absorption and  Fluorescence Spectrometry, Paris,
    1971» publ. Adam Hilger,  London,  p.44

6   DAWSON, J.B., ELLIS, D.J., FISHER, G.W.,  "A dual wave-
    length AA spectrometer  for the  determination of lead in
    blood," Paper presented at the  17th Colloquium Spectros-
    copicum Internationale,  Florence, Italy,  September, 1973«

7   EDIGER, R.D., COLEMAN,  R.L.,  "Determination of cadmium
    in blood by a Delves cup technique,"  Atomic Absorption
    Newsletter, 12, 3,(1973).

8   CERNIK, A.A., "A preliminary procedure for the deter-
    mination of cadmium in  blood,"   Atomic Absorption
    Newsletter, 12, 163,(1973>.

-------
                              2225
       MESURE DU PLOMB DANS LES OS HUMAINS PROVENANT DE
                     LA REGION PARISIENNE

         L, JEANMAIRE, F, PATH/ N, CIALELLA, R, GROS

Commissariat SL 1'Energie Atomique, DSpartement de Protection,
Service de Radiotoxicologie et Ecologie, Fontenay-aux-Roses,
France
RESUME

     Cette £tude a pour but I'obtention de donneee eyet^matiquee
our la teneur en plomb dee OB de personnes habitant la region
parieienne.   Lee prelevemente sont faite pour obtenir dea in~
format-Cone eur deux typee d'os (cftee et vertebres)t our lee
diveraes claeaea d'&gee et BUT lee deux eexee.   Apr&s m-ineva-
lieation de I'oe et concentration chimique le plomb eet doeS par
fluorescence X.   Lee r&eultata pr6sent&e correspondent d une
centaine d'Schantillone.   On note generalement une concentration
plus forte dane lee vertdbres que dane lea cdtee et une augmen-
tation en fonction de I'age.   II eat poeaible que lea concentra-
tions eoient plue faiblee chez lea femmea que chez lee hommea,
maie I'Schantillonnage actuel ne permet pae de I'affirmer.   Ce
travail n'eet qu'une &tape interme'diaire et doit Stre complete
par de nouvellee meeuree pour Stre rSellement eignificatif de la
contamination du plomb de la population parieienne.
ABSTRACT

     The aim of thie etudy wae to obtain systematic data on the
lead content of the bones of people living in the Paris area.
Samples were taken to obtain information about tvo typee of
"bones (ribs and vertebrae), various age categories and both

-------
                              2226
sexes.   After mineralisation of the bone and chemical concentra-
tion the lead was determined on the basis of X-ray fluorescence.
The results obtained were for about a hundred samples.   Generally
speaking concentrations were higher in the vertebrae than in the
ribs and increased with age.   Concentrations may be lower in
women than in ment but no proof of this was revealed in the sam-
pling.   This study only represents an intermediate stage in the
investigation and must be completed by further measurements be-
fore it can be considered a really significant indication of the
lead contamination of the population of Paris.

-------
                                   2227
I. INTRODUCTION

                    Depuis 1962, le strontium 90  et le  strontium stable
des os humalns sont dose's sur des pre*levements  provenant de  la region
parisienne  ; 1' e'chantillonnage  semble e"galement adapte*  a I'e'tude d'autres
elements  ;  c'est pourquoi nous  avons I1intention  d'ltabllr un  bilan
pour les  plombs stables et radioactifs. Ce document ne  pre'sente  que des
re*sultats partlels concernant le plorab stable.

2. METHODS

                    Les e*chantlllons sont des c8tes et  des vertebres
pre'leve'es sur des sujets des deux sexes dont 1'fige est  oomprls entre 10
et 90 ans.  Le mode de vie, la profession et la  cause du de*ces  sont  in-
connus.

                    Apres dessiccatlon a I'e'tuve  & 200", les os  sont
calcines  au four en e'levant progressivement la  temperature de  400 i
700°. Chaque e"chantillon analyst est oonstltu6  par les  cendres provenant
de 1 a 5  pre'levements. Le plomb est entrain^ sur  un pre'ciplte'  d'oxalate,
pre"cipit^ avec un hydroxyde puls copre'clplte* sur  un sulfate  de strontium;
le plomb  contenu dans ce sulfate est alors mesur6 par fluorescence  X.
Des essals  ont montre" qu'll n'y avalt ni perte  de plomb, nl  contamina-
tion des  e"chantillons durant la mineralisation.

                    Pour des teneurs en plomb superieures a  10 ug par g
de oendre,  les essals de reproductibillt^s donnent des  hearts  type  com-
pris entre  6 et 8 #,

3. RESULTAT3

                    La tableau  I, indique les r^sultats en nlcrograimies
de Pb par gramme de cendre pour environ 120 e'chantlllons group^s en
classes d'fige par de'cadres de 10 a 90 ans. Les  remarques suivantes  peu-
vent §tre faitea.

3*1. Olobalement les r^sultats  sont extremement disperses pulsqu'ils
vont de 5 ^ 150 ug.

3*2. En classant les r^sultats  par nature, on constate  que la  teneur
en plomb  des vertebres est ge*ne*ralement suprfrieure a celle des cdtes.
La valeur moyenne du rapport vertebres sur odtes  est ^gale a 1,2 environ.

3.3' Lorsque les r^sultats sont groupe*s par decade et par nature, 11
apparait  une augmentation de la teneur en plomb en fonction  de 1'tge,
Jusqu'k 70  ou 80 ans. Par centre la classe la plus ag^e semble relative-
ment molns  contamin^e. Meme avec oe type de regroupement, les  r^sultats
de chaque olasse sont tres disperse"s, 1'^cart type sur  un r^sultat  indl-
viduel ^taftt fr^quemment de 50  % ou plus. Entre 10 et 80 ans les droites
repr^sentant les concentrations en fonction de  1'fige (t) exprlmtf en
ann^es, ont les formules suivantes, lorsqU'on les fait  passer  arbltrai-
rement par  0 a 1'orlglne : C -  0,87 t pour les  odtes et V -  1,03 t  pour

-------
2228
TABLEAU I - Teneurs en Pb par g de cendre d'os
ANS
10 - 19




20-29

30 - 39


4o - 49



50 - 59


60-69
•

70 - 79



80 - 89
,t

all
O £ 10 >ig/g
C 2 P 11
C 1 M 6
V 2 M 7
C 1 M 12
VIP 11
C 2 M 25
C 3 M 13
C3 P 25
V 2 M 39
C 2 M 21
C 2 M 5
V 3 P 69
C 2 F 29
V 2 M 64
C3 P 37
V 2 M 24
C 1 M 67
C 3 M 42
V 4 M 73
C 3 M 60
C 5 P 28
V3 M 67
V 3 M 84
V 4 M 71
C 2 P 30
V 4 P 31
VIM 99
GIF 30
1
1 £
8 Z> w jxglg
V 2 P 17
VIM 6
C 2 F 13
VIM 9
C 2 M 7
V 2 M 34
C 1 M 12
C 2 F 17
V 1 F 22
V 2 M 82
V 2 M 14
C 2 P 27
V 2 F 33

V3 P 51
V3 M 32
VIM 95
C3 P 23
C 2 M 120
V3 M 59
V 5 F 39
c 3 M 57
C 3 M 147
V3 M 79
V 2 F 34
C 2 M 28
VIP 72

Q % 3> pg/g
C 1 M 10
C 1 M 10
V 2 P 14
C 1 M 8
V 2 M 11
C 2 M 44
C3 M 9
V 2 P 26
C 1 M 18
C 2 F 24
C 2 M 83
C 2 M 65
C 3 M 27

C 2 M 13
C 3 M 54
C 2 M 57
V 2 M 41
V 2 M 107

C 4 F 62
C 2 M 33
C 4 F 91
C 4 M 68
c 3 M 33
V 2 M 40
GIF 46

Jl „
GIF 8
VIM 10
C l F 7
VIM 14
C 1 M 16
V 2 M 52
V 3 M 20
C 2 P 37
VIM 19
V 2 F 36
V 2 M 122
C 3 M 101
V 3 M 22

V 2 M 44
C 4 M 43
V 2 M 81
C 2 M 40
C 1 M 93

V 4 F 74
V 2 M 44
C3 M 54
C 3 M i 61
V3 M 43
C 2 P 30
GIF 54

<5 II
V 1 F 12
C 2 M 8
VIP 12
GIF 6
VIM 17
V3 M 12

C 2 M 45
GIF 15

C 3 F 36
V 2 M 38
C 2 M 42

C 2 M 20
V 4 M 42
V3 P 75
c 4 M 57
VIM 93

C 3 M 56
V 3 M 83
C 3 M 40

C 4 P 26
C 1 M 65
C 1 F 105

Legende os C « c8te V - Vertebra
NB - Nombre de prelevements dans 1'tfchantillon.

-------
                                   2229
les vertebras. Ces valeurs sont supeVieures & celles donne"es par
Holtzraan at al. (l), Schroeder et Tipton (2), Horluchi et al. (3) qui
trouvent une pente de 1'ordre de 0,5 & 0*6.

                     L*interpretation de valeurs tree disperses est
toujours delicate et doit raster prudente ; une analyse statistique ra§roe
tres complete, ne comble pas les laounes provenant d'un e*ohantlllonnage
insuffisant.

                     En consideVant les re*sultats de 10 a 80 ans on note
une augmentation de la teneur en plomb en fonotion de 1'fige ; cet aocrois-
sement continu indique que I'e'tat d'e*quilibre n'est pas encore attelnt
a, 80 ans ; dans ce cas, entre 80 et 90 ans, les valeurs InfeVieures, ne
peuvent s'expliquer que si I'e'chantillonnage de cette classe d'Sge est
insuffisatanent repre"sentatif ou different de celui des classes pre'ce'-
dentes.

                     En ne prenant que les re" suit at s de 40 a 90 ans, on
peut admettre soit un passage par un maximum k 70-80 ans (cas identique
au pr^ce\ient), soit I1existence d'un plateau k 50 ug environ, qui serait
mal deTini par suite d'un tenantillonnage insuffisant j dans ce cas
I'e'tat d'e'qullibre aveo le milieu serait atteint.

3.4. Dans chaque olasse d'fige, les valeurs moyennes trouv^es chez les
femnea sont plutdt inf^rieures k celles trouv^es chez les homes.
         70


         60


         50
         30


         20


         10
Fig.l
                10    20    30   40   50-   60    70   80   90 Ans
        Evolution de  la teneur moyenne en Pb dana  les  vertebras ^  et dans
        les  cdtes   t   en fonctlon  de  1'Sge.  Les  ^ohantillons  sont group^s
        par  ddcade. Les barres d'erreur  correspondent  a.l'icart type sur la

        valeur moyenne.

-------
                                    2230
4. CONCLUSION

                     La dispersion des valeurs trouve"es, les differences
note'es en fonctlon de la nature de 1'os, de 1'fige des sujets et de leur
sexe, sont telles qu'll y a lieu de continuer l'4tude entreprise pour
lui dormer tine assise sollde, sans ambigulte".
 Rernereiernents t   Nous remercions Madame M.Oaroet, et Mademoiselle
                  J. Laporte de leur collaboration technique efflcace.
                        REFERENCES
(1) HOLTZMAN, R.B., LUCAS, H.F. Jr, ILCEWICZ, P.H. - The concentration
    of lead in human bone, Argonne National Laboratory report ANL -
    76 15 (1968) 4?-*9.

(2) SCHROEDER, H.A., TIPTON, I.H., The human body burden of lead,
    Arch. Environ. Health 17, 965-968 (1968)

(3) HORIUCHI, K. HORIGUCHT, S. SUEKANE, M. - Studies on the industrial
    lead poisoning - the lead contents in organ tissues of the normal
    Japanese. Osaka City Med. Journal 5, (l), 41-70  (1959)•

-------
                              2231
    ZUR BLEI- UND CADMIUMANALYTIK  IN BIOLOGISCHEN MATRICES

            M, STOEPPLER+, F, BACKHAUS+,  R,  DAHL+,
         M, DUMONT"1"*", H, HAGEDORN-G6TZ*,  K,  HILPERT+,
     P, KLAHRE*, H, Rl)TZEL+, P, VALENTA*,  H, W,  NURNBERG*

•f  Zentralinstitut ftir Analytische Chemie der  Kernforschungs-
   anlage Jiilich GmbH, BRD
++ Ecole des Hautes Etudes Industrielles,  Lille, France
KURZFASSUNG

     Es wird ttber Arbeitstechniken  und Erfahrungen  bei  der Probe-
nahme sowie ttbev Aufaohlussmethoden berichtet.    Wetter wevden
die eingesetzten Verfahren:  flammenloBe Atomabeovptioneepektvo-
metrie (auoh mit automati-eoTier Probenei.nga.be),  Pulee-Polaro-
graphie und Isotopenverditnnunge-Maaaenapektrometri.e beachrieben.
Abachlieseend werden Messergebniaae ftir Pb  und  Cd in Bovine
Liveri Blut-t Urin- und Haarproben  -  teilveiae  auoh im  Methoden-
vergleioh - mitgeteilt und diakutiert.
ABSTRACT

     The paper deale with sampling techniques and experienoet
and decomposition methods.   It also describes the processes used
in flameleas atomic absorption spectormetry  (including the method
ueing automatic insertion of samples)3 pulse polarography and
ieotope dilution mass spectrometry.   Finallyf the results of
lead and cadmium measurements in bovine liver, blood, urine and
hair samples are reported and discussed - with a comparison of
methods in some cases.

-------
                                   2232
EIKLFJTUNG

Da eich das Ausmass von Intoxikationen durch Elemente wie Blei,  Cadmium
und Quecksilber ebenso aus klinischen Befunden wie aue spurenanalytischen
Daten ergibt, let die Frage richtiger Analysenwerte hier sehr wichtig.

Kan Weiss jedoch aus einer Reihe von Vergleichsprogrammen, dass  bei
Spurenanalyeen erhebliche Abweichungen vom wahren Wert auftreten (l,2,3)|
und dass kritische Studien der Analysenmethoden mit dem Ziel einfachere,
standard!sierte Verfahren mit gro'sserer Zuverlassigkeit zu bekommen,
notwendig 1st.
In der Kernforschungsanlage Julich werden im Rahmen des von der  Bundes-
regierung geforderten Umweltprogramms der Grossforschungsanlagen der
Bundesrepublik Deutschland solche Studien durchgefiihrt• Wir untersuchen
fiir eine Reihe von Elementen die Probenvorbereitung, Aufschluss- und Be-
BtimmungBmethoden sowie Koglichkeiten zur Automatisierung der flammenlosen
Atomabsorptionsspektrometrie und der Pulse-Polerographie. Ueber  bisherige
Erfahrungen bei der BeBtimmung von Blei und Cadmium in Blut, Urin, Organ-
proben und Haaren soil hier zusammenfaseend berichtet werden. An anderer
Stelle wurden klirzlich Meeswerte fiir Blei in Biomatrices (4) sowie von
Blei, Cd und As in Oberflachenwasser, Kineralwasserj Keerwasser  und einer
Reihe von Nahrungs- und Genussmitteln (5) mitgeteilt.

E3(PERIMENTELLES

Probenahme und Probenvorbereitung
Bei Spureribestimmungen ist die Probenahme und die Probenvorbereitung sehr
viel sorgfaltiger als bei der klassischen Analyse durchzufuhren. Man muss
daher ebenso auf ausreichende Einwaage der weitgehend homogenisierten
Probe wie auf Vermeidung von Einechleppungen der gesuchten Elemente
(Kontamination) und Verluste durch Adsorption an Gefasswande etc. bei der
Probenvorbereitung achten.
Bei Blutproben verwenden wir zur Entnahme BtetB auf Pb bzw. Cd geprlifte
Behaltnisse, die mit Antikoagulantien (EDTA, Heparin, Citrat) vereehen
sind.
Aliquots der Blutproben werden entweder direkt nach der Entnahme in ein

-------
                                   2233
                            TABRLLE I

     AUFSCHLUSSKETHODEW ZUR SPURENELEKEMTBESTIMMUNQ MIT PER
     FLAKMENLOSEW AAS. PER PULSE-POLAROGRAPHIE UND PER ISO-
     TOPRNVERDUENNUNGSMASSENSPSKTROMETRIE
MF.THODE
max Menge
Zeitdauer
Bemerkungen
Muffeloefen
Spezialoefen
  10 g
   1 kg
 bis lOOh
 bis lOOh
Kikrowellen-
veraschung
  50 g
 bis lOOh
HNO, unter
Drufik
                  bis 4h
Einfach, aber
zeitraubend, im
ppb-Bereich mit
Vorbehalt

Aufwendig, aber bei
NahrungBmitteln bis-
weilen unumganglich.
Nicht fUr alle Ele-
ment e und im ppb-
Bereich gtinstig

Sehr schonend, da
max. Temp bei 200°C
Kontaminationsvor-
sorge moglich. Nahe-
zu vollBtandige Mi-
neral isierungi daher
beeonders fur Pulse-
polarographie und MS
geeignet
Trotz hoher Gerate-
kosten bei grossen
Probenzahlen glinstig

Zahlreiche Varianten
moglich, mit Ein-
waagen von 1 rag bis
1 g.
Relativ rasoh und
wegen des geringen
SSurebedarfs auch im
allgemeinen kaum
Kontaminationgefahr.
Varianten zur halbauto-
matischen Probenvor-
bereitung moglich.
Trotz klarer LBeung
keine vollBtandige
Kineralisierung, da-
her kaum fiir P.P. und
MS brauchbar

-------
                                   2234
TABELLE I (Portsetzung)
METHODE

HNO, in der
Gasphase
                   max. Menge
                       1  S
                                    Zeitdauer
                                     "bis 30 min
Fe
                        1 kg
                                     bis 20 h
 Nassauf schltisse :
                       bis 5 g
                                      bis 5
 H2S°4/HN°
 HC104/HNO
                                                     Bemerkungen
Sehr rasch und fast
frei von Kontamina-
tion. Aufwendig und
fur Serienbestimmun-
gen in der derz. Form
ungeeignet.
Trotz klarer Lb'sung
keine vollstandige
Mineral!sierung,
da-her nicht fur P.P.
und MS geeignet

Fur begrenzte Proben-
typen hervorragend
geeignet.  Kontamina-
tion im allgemeinen
gering

im  allgemeinen  bei
mittleren Einwaagen
und flu'ssigen Proben
giinstig.  Vor  all em
fur die Pulee-Polaro-
 graphie brauchbar.
 Konteminntion nicht
vernachlassigbar
 Bei Verwendung von
 Perchlorsaure sind
 Spezialabziige und
 besondere Sicher-
 heitsvorkehrungen
 notig
 HC10,  is-t fur die
 flammenlose AAS un-
 geeignet

-------
                                   2235
Aufschlussgefass uberfiihrt oder - fur die flammenlose AAS-in Polyole-
fingefassen (z.B. Eppendorf-Reaktionsgefassen) mit penta-destilliertem,
praktisch Bchwermetallfreiem Wasser hamolyBiert. Unter diesen Bedingungen
ist eine Kontamination durch Blei und Cadmium aus dem Gefassmaterial
minimal und in der Regel vernachlassigbar.
Urinproben werden in mit HNO, oder HC1 gereinigten und auf Pb gepriiften
Polyathylenflaschchen gesammelt und zur direkten Analyse entweder in
ebenfalls vorgereinigte Eppendorfgefasse uberfuhrt und angesauert oder
in eine polarographische Zelle eingebracht. Auch hierbei ist eine merk-
liche Kontamination nicht zu beobachten. Bei Organ- und Haarproben
liegen Pb- und Cd-Gehalte im ppm-Bereich vor, so dass im allgemeinen bei
Eineatz der beschriebenen Gefasse und ausreichender Einwaage die Konta-
minationsgefahr gering ist. Bei Haaren 1st der Waschvorgang vor der
BeBtimmung problematisch. Auf die umfangreichen Arbeiten hierzu wurde
an anderer Stelle hingewiesen (6,7)»

PROBENAUFSCHLUS5
Unsere bieherigen Erfahrungen mit verechiedenartigen klassischen und
moderneren AufschluBsmethoden Bind in Tabelle 1  zusanunengefaest. Ein
detaillierter Bericht hieriiber, in dem auch eine von uns entwickelte
Mikrovariante des HNO^-DruckaufsohlusBes beschrieben ist, wird zur
Zeit vorbereitet (8).

KDNTAKINATIONSPROBLEME
EB wird, falls erforderlich, in sog. clean benches oder unter streng
kontrollierten Bedingungen und mit hochreinen (z.B. Suprapur ^—')
Reagentien gearbeitet, urn eine Kontamination von eussen zu vermindern.
Als gunstigste Materialien fiir AufschlusB- und Bestimmungsge-fasse *er-
wiesen sich epanabhebend bearbeitetes Teflon und eynthetischer Quarz
(z.B. tSuprasil^), die nach Behandlung mit hochreinen Sauren sehr
niedrige Blindwerte ergaben.

METHODEN UND GERAETF.
ATOMABSORPTIONSSPEKTROKETRIE (AAS)
Bei unseren Untersuchungen wurde (Tberwiegend die erst eeit 1970 kommer-

-------
                                  2236
TABELLE  II
FLAMMENLOSE ATOMABSORPTIONSSPEKTROMETRIE

GERXTEDATEN;
Fisher- Jarrell-Ash 810, Zweikanal, Vierstrahl

Perkin-Elmer 300, Einstrahl mit Deuteriumkompensator

Perkin-Elmer GraphitkUvetten : HGA 70/72/74 + Rossimat
ANALYSENLINIEN: Pb 283.3 nm, Cd 228.8 nm
NACHWEISGRENZEN (fUr 50 ul Probeneingabe)
Pb; w&Brig L10"11g (0.2 pg/L)  Blut/AufschluBlsgg; 1-10*
    Urin:  5'10~10g


               "12
Cd; wftflrig 1-10"12g (0.02 pg/L) Blut/AufschluBlsgg: 1*10"
MATRIXPROGRAMME;
Pb in hfimolysiertem Blut;
            bis
                              Blei in Aufsdlufllsgg./Urin
Trocknen
Zersetzen I
Gleitprogramm
Atomisieren
Abktlhlen
 95®C
250°C 60
60 sec
    «
                640°C 72  "
               2400C
       5
      30
Trocknen
Zersetzen
Atomisieren
AbkUhlen
 bis 95C
bis 300°C
   24oo°C
60 sec
30  »
 f\  II
30  »
Cd in hSmolysiertem Blut:
Trocknen    bis
Zersetzen I
Gleitprogramm
 Atomisieren
AbkUhlen
                 95°C 60 sec
                250°C 30  w
                400^C 180 "
               1800°C  5  tt
                      30  n
              Cd in AufschluBlsgg. etc.
              Trocknen     bis  95°C  60 sec
              Zersetzen        300°C  30  "
              Atomisieren     1800°C   5  "
              AbkUhlen

-------
                               2237
  T A B E L L E IIJ.
  DATEN ZUR PULSE-POLAROGRAPHIE
  Methode; Anodic Stripping (Inverspolarographie), Elektro-
  lysedauer 2 min
  Gera't; PAR (Princeton Applied Research) 174
  NACHWEISGRENZEN;  (minimales Volumen 4 ml)
  Element      in wflflrigem Medium   in AufschluBlbsungen/Urin

  Cadmium      ca 0.2 ppb           ca 0.5 - 1 ppb
  Blei         ca 0.2 ppb           ca 0.5 - 1 ppb

  Relative Standardabweichung 2-1096 (matrixabhangig)
  Probenvorbereitung; Aufier beim Urin ist in alien Fallen
  von biologischen Proben eln AufschluB notwendig. Mit
  gutem Erfolg ist der MikrowellenaufschluB und auch der
  nasse AufschluB mit HpSO^/HClO^ anwendbar.
  Kontatninationsgefahr durch die Aufschlufigefa'fle und ggf.
  durch Leitsalze
 TABELLE  IV

 ISOTOPENVERDUNNUNGS-MASSENSPEKTROMETRIE
 GERATEDATEN;
 Varian-MAT CH-5 mit Thermionenquelle
 Isotopenverhaltnis: 204Pb (Spike)/natUrliche Isotope
 NACHWEISGRENZEN.  FEHLERBEREICH:
 absolute Nachweisgrenze ca 5«10~°g
 relativ abhangig von der Probenmenge
 Rel.  SAandardabweichung im gUnstigsten Bereich <1?6f
 an der Nachweisgrenze im Prozentbereich
 PROBENVORBEREITUNG:
 Aufschlufl (giinstig: mit Mikrowellen)
 Losen,  ggf.  Abtrennung von Matrixbestandteilen
'Anodische Abscheidung des Pb
 Auflosen in  wenig HN03,  Aufbringen auf mit
 Phosphorsaure  impragniertes Verdampferband
 Kontaminationseefahr bei alien Schritten

-------
                                   2238
ziell verftigbare flammenlose Variants eingesetzt, da sie gegenuber der AAS
mit Flamrae den Vorteil der grosseren Empfindlichkeit und gegeniiber der
Pulse-Polarographie den der hb'heren Analysenfrequenz besitzt.
Derzeit wird ein von Perkin-Elmer, Deutschland, nach Arbeiten von Pick-
ford und Rossi (9) gebauter Prototyp zur automatisierten Probeneingabe
in Graphitkuvetten (Rossimat) in Jiilich erprobt.
Wir hoffen, damit neben einer weiteren Steigerung der Analysenfrequenz
eine im Vergleich zur Handdosierung ura mindestens Faktor 2 bessere
Reproduzierbarkeit zu erreichen.
Tabelle II fasst die wichtigsten Daten der eingesetzten Geratekombination
und die Temperaturprogramme zur direkten Bestimmung von Pb und Cd in Blut
und Urin sowie in Aufschlusslosungen von Korperorganen und Haaren
zusajnmen.

PULSE-POLAROGRAPHIE (P.P.)
Diese, 1958 von Barker erstmals beschriebene Methode (10) erzielt vor
allem in Korabination mit dem sog, anodic-stripping-Prinzip (inverspolaro-
graphie) wegen ihres giinstigen Signal-Rausch-Verhaltnisses sehr niedrige
Nachweisgrenzen.
Ausser Urin, der direkt analysiert werden kannt ist bei den ubrigen
Matrices stets ein Aufschlusa erforderlich (vgl. Tab. l).
Vorarbeiten zu einer Teilautomatiaierung haben ebenfalls in Jiilich be-
gonnen*
Tabelle III zeigt die Daten dieser in zunehtoendem Umfang in der Umwelt-
analytik gebrauchlichen Methode.

ISOTOPEN-VERDUENNUNGS-KASSENSPEKTRCTIETRIE
Zur Beurteilung der Richtigkeit von Spurenanalysen ist ea wiinechenswert,
fiir jedes interessierende Element noch eine dritte Wethode einsetzen zu
konnen. Vor allem bei dem mit nuklearen Kethoden im extremen Spurenbe-
reich praktisch nicht bestimmberen Blei bietet sich die Isotopen-Ver-
diinnungs^assenspektrometrie als zusatzliche HSglickeit neben AAS und
P.P. an.
Tabelle IV bringt Daten dieser - gegenwartig in Jiilich in der Enderpro-
bung befindlichen Kethode.

-------
                              2239
TABELLE  V.
Bestimmung von Pb und Cd in NBS Bovine Liver
NBS-WERTE:
Pb
Cd
                       0.34 + 0.08 ppm
                       0.27 + 0.04 ppm
KFA/ZAC:
Pb rait AAS

Pb mit Pulse-Pol.
0.32 + 0.09
0.33 + 0.08
0.38 + 0.04
                                              n = 20
                                              n = 22
                                              n =  6
                       0.25 + 0.06 ppm'
                       0.21 + 0.04 ppm
Cd mit AAS
Cd Bit Pulse-Pol
                       n = 10
                       n =  6
BEMERKUNGEN:
                       1'DruckaufschluB, Einw.>200 mg
                       2 }
                        'Mikroaufschlufl, Einw. ca 10 mg
                       ^'Pb-Kontamination beobachtet
                       4}
                        'Kontamination nicht unmb'glich
 TABELLE VI
 Blei im Blut (Werte in jig/100ml Vollblut)
 Kontrollgruppe   Manner
Frauen
                 Mittelwert
                                                      Range
A (z=64)
B (z=65)
C (z=5)
D (z=40)
E
-------
                              2240
T A B E L L E VII
Vergleichsmessungen zwischen flammenloser AAS und
fuise-Foia
BLUTPROBEN
AAS
205
75
365
145
400
184
695
230
80
145
345
261
T A B E
rograpnie ^anoaic

Polarographie
270
28
405
125 pp
415 TKS = 1-08
285
720
225
70
65
485
281
L L E VIII
stripping;
URINPROBEN
AAS
115
20
20
50
70
55
20
20



46

Werte

in ppb|

Polarographie
110
47
86
65
65
60
95
25



69




^3£)
A A Q








  GERATEVERGLEICH DURCH MESSUNG VON 6 BLUTPROBEN
  (Werte in ppb)
  Probe   I
II
III   IV   Mittelwert
1
2
3
4
5
6
^ittel
370 240 370 270
240 180 190 190
130 110 130 150
480 330 370 280
200 110 180 180
630 610 640 630
340 263 310 283
312+67 (2196)
200+27 (1396)
130+16 (1296)
365+53 (1596)
167±37 (22%}
627+13 ( 296)
Gesamt-0 299+33 (1196)
Erlauterungen :
I Jarrell-Ash 810/1 am 05.04.74 mit HGA 72
II Perkin-Elmer 300 am 08,04.74 mit HGA 74
III Jarrell-Ash 810/1 am 11.04.74 mit HGA 72
IV Jarrell-Ash 810/11 " "

-------
                                   2241
Bei bisweilen betrachtlichen Abweichungen in Einzelfallen liegen die
Kittelwerte der Pulse-Polarographie - eventuell kontaminationsbedingt -
bei Blutblei nur unwesentlioh, bei der direkten Bleibestimmung in Urin
allerdings merklich hoher. Im letzteren Fall konnte einerseits die
Grenze der Mb'glichkeiten der direkten flammenlosen AAS erreicht sein,
andererseits eine Stfa'rung duroh Sn bei der Polarographie vorliegen.
Wenn man davon ausgeht, dass zumindest im Blut die direkte BeBtimmung
mit der flammenlosen AAS brauchbare Kittelwerte liefert, so tiberraschen
doch beim Vergleich von Einzelproben zwischen AAS und Polarographie und
auch zwischen Labors, die die flammenlose AAS anwenden, Werte, die nicht
selten urn mehr als den Faktor 2 differieren.
Wir haben daher versucht, dies durch Vergleichemessungen unter Routine-
bedingungen zu studieren. Dazu wurden identische Blutproben an drei ver-
schiedenen Geraten sowie eine Blutprobe ttber einen langeren Zeitraum am
gleichen Gerat geraessen.
Tabelle VIII faest die Pb-Messungen in den Blutproben zusammen. Die
Unterschiede zwischen den einzelnen Qeraten erreichen bei Einzelwerten
bisweilen fast den Paktor 2, wahrend sich die Mittelwerte meist nicht so
betrM-chtlich unterscheiden. Dennoch sind auch hier die Schwankungen
merklich.
Im Langzeittest mit einem Jarrell-Ash 810 wurde in vier Blutproben
im Verlauf von 11 Tagen arbeitetaglich der Pb-Gehalt nach jeweils neuer
Entnahme eines 50-100  ^1-Aliquots und Hamolyse bestirant, wobei jeweile
n«5 Parellelmessungen durchgefiihrt wurden*
Aus Tabelle IX ist zu erkennen, daes die Langzeitstandardabweichungen
betr§chtliche Werte erreichten.
Urn zwischen geratebedingtenMesswertschwankungen und Einfliissen der Blut-
matrix differenzieren zu konnent wurden 100 im gleichen Zeitraum durch-
geftihr^e Kontrollmessungen mit einer schwach sauren 0.02 ppm-Pb-EichlbBung
zusatzlich ausgewertet. Die Standardabweichung des Hittelwertes dieser
Messungen lag bei 10$, BO dass man einen Einfluss der Matrix nicht aus-
schliessen kPJin. Dennoch deuten die betrachtlichen Abweichungen bereits
bei w&esrigen EichlSsungen - Tag zu Tag-Abweichungen sollten dabei im
allgemeinen J?o nicht uberateigen - auf Instabilitaten hin, die entweder

-------
                                2242
  TABELLEIX
  Langzeitmessungen von vier Blutproben, Werte In ppb
  GERAT: Fisher-Jarrell-Ash 810 mit Graph!tkUvette HGA 72
  Tag
Probe 1   Probe 2   Probe 3   Probe 4   Tagesmittel
Mo
Di
Mi
Do
Fr
Mo
Di
Mi
Do
Mittel

275
230
220
200
180
340
210
270
230
240+50
"(2196)
320
310
240
310
270
350
290
320
260
300+40
"(13.
340
310
270
350
300
360
160
220
250
280+70
396) "(25%)
370
410
520
430
350
500
360
410
390
420+60
326
y\-j
312
322
275
3*7
255
305
282
308+36
"(14.3%) "(11.796)
T A B E L L E   *•;
Bleibestimmungen in Haaren nach MikrodruckaufschluB
Methode; flammenlose AAS, Eingabe in 0.5 M HNO,
Probandenzahl 30, unbelastet, Einwaage 10-20 rag
mannlich (z=15) Mittelwert 8.8 ppm. Range 2.2 - 39 ppm
weiblich (z=15) Mittelwert 6.2 ppm. Range 0.2 - 15 ppm
 TABELLE   XI
 Cadmiumbestimmungen ira Blut, Direkteingabe von
 1+15 mit pentadest. Wasser hSmolysiertem Blut in die
 HGA 72, kombiniert mit J.A. 810. Programm siehe Tab.2
 Probandenzahl; 37, keine besondere Belastung bekannt,
 keine Differenzierung zwischen Mannemund Frauen
 Mittelwert; 9.5 ppb  Range 4-19 ppb

-------
                                   2243
MESSERGEBNISSE. DISKUSSION

Die mit Blut, Urin, Bovine Liver und Haaren durchgefuhrten Messungen
zur Ermittlung von Pb— und Cd—Gehalten dienten zunachst vor allem dem
internen und externen Kethodenvergleich, urn zu einer ersten Abschatzung
der Richtigkeit dieser Bestiromungen zu korranen*
Tabelle V fasst dieErgebnisse fur Pb und Cd in MBS 1577 Bovine Liver
zusammen. Der gegenuber der AAS hohere Wert der Pulse-Polarographie fur
Pb konnte auf Kontamination beruhen, der recht niedrige fur Cd ist iiber-
raschend, aber durch hinreichend viele Messungen gesichert; er soil trotz-
dem noch einmal uberpriift werden.
Zur Ermittlung des Blutbleigehalts unbelasteter Personen im Raume
Aachen/Jiilich wurden 1973/74 zwei Kollektive mit der flammenlosen AAS
untersucht. Es ergaben sich - im Verbaltnis zu bisher angenommenen
Mittelwerten - sehr niedrige Werte, die jedoch durch neuere Arbeiten
grossenordnungsmassig recht gut bestatigt werden (11,12,13,14)* An-
echliessend wurden im Austausch mit Laboratorien in Europa und Uebersee
Blutproben von belasteten und unbelasteten Pereonen untersucht, wobei
sich signifikant hohere Werte als bei unseren beiden ersten Kollektiven
ergaben. Beim Datenvergleich mit den anderen Laboratorien ergaben sich
haufig gut ubereinstimmende Mittelwerte, es wurden bisweilen aber auch
erhebliche sy&tematische Abweichungen beobachtet, deren Ursachen noch
studiert werden mussen.
Tabelle VI zeigt einige dieser Krgebnisse. In alien Fallen zeigte sich
der auch von anderen Autoren (11,13) beobachtete signifikant niedrigere
Blutbleigehalt weiblicher Probanden. Derzeit kann noch nicht fiindeutig
geklSrt werden, ob der Unterschied zwischen dem Mittelwert von Kollek-
tiv A und dem von Kollektiv B zufallig oder relevant ist, da durchaus
mit geratebingten Schwankungen gerechnet werden muss. Aufgrund techni-
scher Schwierigkeiten beim Probenaufschluss war es bisher nicht mo'glich,
alle Vergleichsblutproben auch pulsepolerographisch zu uberpriifen. Die
bisher erhaltenen Werte - so vor all em beim Blut und auch bei Urin - be-
statigen jedoch die Grb'ssenordnung der routinemassig erhaltenen AAS-Werte.
In Tabelle VII sind Vergleichswerte zwischen AAS und Pulse-Polarographie
zusamraengestellt •

-------
                                    2244
typ- Oder  gerS.tebed.ingt  sind. Wir untersuchen derzeit  dieses Phanoraen
unter Heranziehung weiterer AAS-Gerate.
Die Messungen von  Pb  in  Haarproben konnten  nach Aufschluss mit HNO, ohne
Schwierigkeiten -  Bestimmung durch standard addition - mit guter Reprodu-
zierbarkeit durchgefuhrt werden. In Tabelle X Bind  die Werte zusammenge-
fasst, die aue Haarproben von 30 nicht belasteten Pereonen aller Alters-
stufen stammen.
Auffallend ist die grosse Streubreite unabhangig vora Alter der Versuchs-
per8onent  die wohl durch die geringe Einwaage - es  wurde  zu Testzwecken
vorwiegend in der  Kikroapparatur aufgeschlossen - mit  bedingt sein diirfte.
Schliesslich enthalt  Tabelle XI einige Reeultate von mit  der flammenlosen
AAS nach dem in Tabelle  II angegebenen Zersetzungsprogramm durchgefUhrten
jJd-Beetimmungen in stark verdunntem, hamolyeiertem  Blut.  Die Werte sind
verglichen mit der Literatur verhaltnismassig hoch  (15) und konnten bis-
her noch nicht durch  Vergleichsmessungen mit  der Polarographie gesichert
werden. Auch kann  eine Kontamination derzeit  noch nicht ausgeschloesen
werden.
Aufgrund unserer bisherigen Studien sehen wir zukiinftig gute Koglichkeiten
ebenso in  einer Teilautomatisierung von Auechluse-  und Beetimmungsmethoden
wie in der Einftihrung weiterer Technikent die Kultieelementbestimmungen
erlauben.  Der Trend wird dabei eicher dahin gehen,  aus Zeit- und Kosten-
grfinden mit den minimal mb'glichen Einwaagen auszukommen.
Durch gute zeitliche  Nutzung aller Cerate und zuverlassiger Datenbanken
in Kombination mit relevanten Modellen ftir  den Spurenmetallfluss in Um-
weltkompartimenten sollte es moglich sein,  die absolute Zahl der Anslysen
auf ein vernUnftiges  Kass zu begrenzen.
Wir danken dem Bundesministerium ftir Forschung und  Technologie der BED,
Bonn, flir  die grossztigige Untersttitzung, Perkin-Elmer, Deutschland, ftir
die Ueberlassung des  Prototypautomaten, den Kollegen Dr.  Delves, London;
Dr. Kubasik, Rochester, N.Y., USA; Dr. Kilroe-Smith, Johannesburg, Sild-
afrika , so wie dem britischen Arbeit sministerium fiir Vergleichsblutproben,

-------
                                     2245
                            LITKRATURANGAEEN
     KEPPLER, J.F.; KAXFIF1D, K.E.; MOSS. W.D. ; TIETGEN, G.; LICH, A.L.
     Amer. Ind.Hyg.Assoc.J. _3l, 417 (1970)

     BERLIN, A.; DEL CASTILHO, P.; SKEETS, J., Eur 5004 d-e-f, 1973.
     p. 1033

[ 3J  BF.RLIN, A.; LAUWERYS, R. ; BUCKET, J.P.; ROELS, H. J DEL CASTILHO, P. J
     SKEETS, J., Dieee Konferen2, Beitrag 141

[|  STOEPPLER, K.j KILPERT, K. ; VALENTA, P.; NUERNBERG, H.W. ,
     Z.Anal.Chem. im Druck

[ |  NUERNBERG, H.W. ;  STOEPPLER, K.; VALENTA, P.,
     Thalassia Jugoelavica, im Druck

[6]  HAGEDOEH-GOETZ, HV; STOFPPLER,K. ,
     Arch. Toxi col. , im Druck

[7]  BRANDT, K. ; HAGEDORN-GOETZ.H. ; STOEPPLER, K.f
     Ber.d.KPA Jiilich, ira Druck

[8]  STOEPPLER, ^.; BACKHAUS, P.; KLAHRE P.
     Z.Anal.Chem., in Vorbereitung

[9!  PICKFORD, C.J.; ROSSI, G. ,
L -1  Analyst, .21, 647 (1972)

[l6l  BARKER, G.C.; GARDNER, A.W.,
     AERE-Report C/R 2297, Harwell 1958

[ll]  SCHMIDT, D.; SANSONI, B.; KRACKE, W. ; DIETL, P.; BAUCHINGER, M.j
     STICK,  W.,
     Hunchener med. Wochenschr. Ijl^, 1961 (1972)

[12]  HAAS, Th.; WIF.DE, A.G.j SCHALLER, K.H.,
     Zbt.Bakt.Hyg.J.Att.Orig.B. 155. 341 (1972)

|J3]  STUIK,  E.J.} ZIELHUIS, P.L. ,
     Diese Konferenz,  Beitrag 57
  4J  HOWER,  J.{  PRINZ,  B. j  GONO, E.; REUSMANN, G. ,
           Konferenz,  Beitrag 61
[15]  GFXDMACHER,  V.;  KALLINCKRODT, M.| OPITZ, D. ,
     Arbeit smedizin,  Sozialmedizin, Arbeitshygiene, 10. 276, (1968)

-------
PANEL DISCUSSION

-------
                                2249
                      SUMMARY OF DISCUSSION
SZADKOWSKI (B.R.D.)
     Das Thema der Sitzung sind Gewebsmessungen, und wer sich
rait dieser Materie befasst hat weiss, dass hier erhebliche
Schwierigkeiten auftreten kOnnen, etwa gegenttber Messungen in
wSssriger LSsung.  Ursachlich daftir verantworlich zu machen
sind die sehr zahlreichen anderen Substanzen, die in biologischen
Materialien vorhanden sind und die unter Umstanden stttren
kttnnen.  Insbesondere mSchte ich hier Eiweisskttrper nennen.
Wenn wir hier (Iber Gewebsmessungen sprechen, dann meinen wir
dabei auch Blut und Harn.  Es wird allerdings auch gesprochen
werden tlber Messungen in Leber, Knochen und auch in einigen
Nahrungsmitteln.  Fiir die im Rahmen dieser Sitzung interessierenden
Schadstoffe, namlich Schwermetalle, sind in den vergangenen
Jahren eine ganze Reihe brauchbarer Methoden entwickelt worden.
     Im Rahmen der weltweit mit Intensitat angegangenen Probleme
des Umweltschutzes sind nun internationale Vergleiche unumgMnglich
notwendig.  Es ergaben sich hierbei jedoch neue Schwierigkeiten/
die unter anderem in der unterschiedlichen Methodik, in der
verschiedenen Probenahme und auch in der Aufbewahrung dieser
Proben zu suchen sind.  Es wird sich im Laufe dieser Sitzung
zeigen, wie wichtig, aber auch wie schwierig durchzufuhren de-
rartige Vergleichsprogramme sind.  Auf einem ganz anderen Sektor,
namlich auf dem der klinischen Labordaten, werden seit kurzem
in der Bundesrepublik Deutschland Qualitatskontrollen von der
zustandigen Bundesarztekammer verbindlich vorgeschrieben.  Sie
umfassen neben den regelmassig, d.h. bei jeder Analysen-Serie
durchzufuhrenden Prazisionskontrollen auch Ueberprtifungen der
Richtigkeit, d.h. der "accuracy".  Die Vorschriften fur die
Durchfdhrung dieser Qualitatskontrollen sind sehr detailliert

-------
                                 2250
und prSzise.  Unrichtige Ergebnisse  ktinnen  fiir die  betreffenden
Laboratorien finanzielle Einbussen zur Folge  haben.   Ich mSchte
nun nicht hier dafUr plSdieren, derartige verbindliche  Richtlinien
auch fiir Messungen  im Bereich der toxikologischen Umweltbelastung
elnzuftihren.  Es ware international  verbindlich  gar  nicht mSglich,
es wtirde ausserdem  die Gefahr einer  Stagnierung  der  entsprechenden
Forschungssektoren  bedeuten. Ich wollte  aber  demonstrieren, dass
auch administrativ  die Notwendigkeit vergleichbarer  Resultate
erkannt worden ist. Nun ist die Problematik vergleichbarer Resultate
in der Analytik umweltrelevanter Stoffe  ungleich differenzierter
als im Bereich des  Klinischen Labors.

     Hierftir sind insbesondere die sehr  viel  niedrigeren Kon-
zentrationen in den biologischen Materialien  verantwortlich zu
machen.  Es ist daher nur zu begriissen,  wenn  auf internationaler
Ebene Vergleichsuntersuchungen gestartet werden  und  zwar nicht
nur im Bereich der  europSischen Gemeinschaft  sondern auch liber
Atlantik und Pazifik hinweg.  Ebenso bedeutsam ist das  Bemuhen
urn eine verfeinerte und verbesserte  Methodik, die insbesondere
abzielt auf die Verwendung iiraner kleinerer  Probenmengen.  Dies
ist ein Punkt,der besonders wichtig  ist  fur die  breite  Durch-
flihrung von epidemiologischen Untersuchungen.
     The subject of this session is measurements in tissue, and
anyone who has worked with this matter will know that consider-
able difficulties can arise, as compared for example with measur-
ements in aqueous solution.  Blame for this can be laid primarily
on those numerous other substances present in biological
materials which under certain circumstances can result in
interference.  I should particularly like to mention proteins
among these.  When we talk here of measurements in tissue we
also include blood and urine.  Furthermore, measurements in
liver, bones and some foodstuffs are also included.  In recent
years a whole series of useful methods has been developed for the
toxic substances which concern this session, i.e. heavy metals.

     In connection with the world-wide problems of environmental
protection which are being earnestly tackled, international
comparisons are now unavoidably necessary.  This gives rise to
new difficulties, however, which include differing methodology,
varied sampling and the preservation of the samples.  During

-------
                                2251
 the course of  this  session it became dear  nou  important  it  is
 to carry out such comparison programmes,  but also  how much
 difficulty is  involved.   In a quite different field, namely  that
 of clinical  laboratory data, quality control by the competent
 Federal medical Association has recently  been made compulsory
 in the Federal Republic of Germany.  In addition to the regular
 precision controls  carried out in every test series this  also
 includes checks on  accuracy.  The test rules for these quality
 controls are very detailed and precise.   Incorrect results may
 mean financial losses for the laboratories  concerned.  I  am  not
 suggesting we  introduce compulsory rules  of this kind for
 monitoring in  environmental toxicology.   These  would not  be
 enforceable on a world-wide scale and might in  any case create
 a danger of stagnation in the sectors of  research  concerned.
 What I wanted  to show was that administrative circles have also
 recognised the need for comparable results.  But the question
 of comparable  results from the analysis of  environmental  substances
 entails more widely differing nuances than  clinical laboratory
 experiments.   The far lower concentrations  in the  biological
 substances are particularly responsible for this.  It is  thus
 an excellent thing  that comparative research has begun not only
 within the European Community but also across oceans and
 continents.  Equally significant are efforts towards improved,
 more sophisticated  methods using ever smaller samples.  This
 point is especially important for the execution of wide epidemic-
 logical studies.
                            DISCUSSION
TATI (Japan)

     In cadmium determination, Dr. KjellstrSm showed the
systematic difference between Japanese and Karolinska studies,
and also showed the different recovery rates between the two
countries.  What do you think about the correlation between
the different values and the recovery rates of these two
countries?

-------
                               2252
KJELLSTRttM  (Sweden)
     Of course, this study has only shown that there is a
difference and the deviation between duplicate samples in each
laboratory is rather small so there must be some kind of syste-
matic difference.  Whether this means that the atomic absorption
and neutron activation analyses done in Sweden have a lower
recovery or that the atomic absorption analysis performed in
some of the Japanese laboratories for some reason gives too
high values we cannot exactly say.  However, the Swedish atomic
absorption method has been checked with the addition of radio-
active cadmium to grains and we found that the recovery was 95%.
There may be reasons to believe that at this rather low level of
cadmium concentration in grains, there may be an exaggeration
of the cadmium levels when using the Japanese methods.  May I
also say that there have been four different analytical
laboratories participating from Japen, but unfortunately two
of them only analysed five of the samples.  The analyses from
these two laboratories correlated very well with the neutron
activation and the Swedish atomic absorption analysis.
MOORE  (U.K.)
     The 7OO°C Dr. Jeanmaire quoted for dry ashing of bone would
appear to be too high.  Work by our laboratories and by Hislop
at Harwell indicated a loss of lead from bone at a temperature
as high as this.  We use a temperature no higher than 450°C and
Hislop, 6OO C.  Could you comment on your temperature of 70O C?
JEANMAIRE  (France)
     Le proble'me de la temperature de mineralisation depend
certainement beaucoup des conditions operatoires et de la nature
de 1'echantillon.  La question s'est dejet posee en radioactivite
par example avec le cesium 137 et surtout le polonium 210 qu'on
retrouve dans les os meme apres mineralisation a 7OO°, alors

-------
                                2253


qu'il est extremement volatil.  La mineralisation est effectuee

par paliers & 200°, puis a 400° et ensuite la temperature est

amenee progressivement a 700°.  Lorsgue le poids de 1'echantillon

a mineraliser est faible, la temperature finale joue un role

tres important pour les pertes.  Lorsque le poids de 1'echantillon

et celui du residu sont grands, si I1elimination de la majority

de la matiere organigue est effectuee a une temperature assez

basse pour eviter les pertes, je crois gue 1'on peut chauffer

plus fortement ensuite, sans risgue.  C'est ainsi gue nous avons

porte jusgu'a 800  un phosphate de Ca contenant du plomb, sans

mettre en evidence de perte appreciable en cet element.


     The quest-ion of mineralization temperature certainly
depends to a large extent on operating conditions and the
nature of the sample.  The question has already arisen in
radioactivity,  for instance with caesium IS? and more par-
ticularly with polonium 210 which occurs in the bones even
after mineralization at 700°Ct then 400°C, the temperature
thereafter being brought gradually up to ?00°C.  When the
sample is light in weight, the final temperature has a very
considerable bearing on the losses.  When the sample and residue
are heavy, and most of the organic matter is elimnated at a
low enough temperature to avoid losses, I think there is no
risk in stepping up the temperature afterwards.  Thus we heated
to 800°C a Ca phosphate containing lead without appreciable
lead loss.
PFANNHAUSER  (Austria)

     Ich mCchte die vorherige Diskussionsbemerkung unterstUtzen,

Nach Untersuchungen in unserem Institut in Wien waren bei

Trockenveraachungen tlber 45O°C Verluste zu beobachten.


     As a result of research undertaken in our Institute in
Vienna, losses were observed in dry ashing at temperatures over
4BO°C.  Perhaps Dr. Kjellstr&m oah tell us about his experience
with dry ashing?

-------
                                 2254
KJELLSTROM  (Sweden)
     The temperature of dry-ashing is 450°C and it is done in a
special muffle oven, which is constructed to keep the temperature
within ± 10°C.  The sample is ashed during 15 hours.  This
procedure is done twice with the position of the crucibles reversed
within the oven between the ashing procedures.  We have studied
the losses in each different step and found that the total loss
is 5% and most of this is in the dry-ashing step.  If we do
wet-ashing and extraction in an organic solvent of grain samples
we get a loss of 15% with most of this in the extraction
procedure.  After studying the losses in each step of the procedure
we concluded that with this type of oven and at this temperature
dry-ashing of wheat samples is possible to use for cadmium
analysis.
CERNIK (U.K.)
     Has Dr. Berlin considered the use of EDTA for the homogeneous
preservation of lead in blood, to prevent the variability of
results due to non-representative sampling?  The use of heparin
can be a disadvantage with postal delayed samples because of
viscosity changes and possible clotting.
BERLIN (C.E.C)
     In the first instance we allowed each of the laboratories
to use the techniques of their choice and most of them preferred
heparin  for preservation.  We were aware of your work and in
view of the rather unsatisfactory results which were obtained,
I feel we should examine again the possibilities of using EDTA.
We had some problems with the containers for cadmium in urine
and for the aqueous mercury solutions.  We are considering using
in future programmes quartz which should certainly reduce
absorption and contamination problems.

-------
                                2255
Truffert  (France)
     J'ai constate des pertes de plomb  lorsque  1'incineration
se faisait a une temperature superieure a  5OO°C,  mSme  sur  les
os.  Cependant, pour ces derniers,  la perte  etait moins  importante
gue pour des denrees alimentaires beaucoup moins  riches  en calcium,
qui paralt retenir le plomb.  La volatilisation des  toxiques
mineraux depend beaucoup de la nature du milieu traite.  C'est
ainsi qu'en ajoutant de 1'acide nitrique aux cendres,  on peut
accelerer la mineralisation sans perte  de  plomb.  Mais alors
les pertes de cadmium sont considerables,  ce qui  n'est plus le
cas si 1'on ajoute de 1'acide sulfurique,  les sulfates de  ces
metaux n'etant guere volatiles.
     Personnellement, j'effectue des mineralisations a basse
temperature (au voisinage de 360°C) dans un  incinerateur sous
courant d'oxygene filtre , mis au point avec Mme  Girard-Walton.

     J have observed lead losses when the  incineration
temperature was over SOOoC, even in bones.   Even  so, the loss
was not eo great as for foodstuffs more deficient in calcium
which appears to retain lead.  The volatilization of toxic  min-
erals depends to a large extent on the  type  of  medium  treated.
Thus, if nitric acid is added to ash, mineralization can be
accelerated without lead loss.   Cadmium losses  are then  con~
eiderable, however, but this is not the case if sulphuric  acid
is added, since sulphates of these metals  are not very volatile.
     Personally, I conduct low-temperature mineralizations
(about S60°C)  in an incinerator with a  stream of  filtered
oxygen,  developed with Mrs. Girard-Walton's  help.
BRAETTER  (B.R.D.)
     Dir Frage richtet sich an Dr. Jeanmaire.  Haben Sie bei
Ihren Untersuchungen den vollstSndigen Wirbel bzw. die Rippe zur
Analyse eingesetzt oder nur Teilproben?  Bei systematischer
Durchmusterung von menschlichen Skeletten haben wir die ortliche
Verteilung von 20 - 30 Spurenelementen mit Hilfe der Neutronen-
aktivierungsanalyse untersucht.  Es zeigte sich, dass innerhalb

-------
                                2256
enger Bereiche bei verschiedenen Spurenelementen  Konzentrations-

Snderungen bis zu zwei Zehnerpotenzen auftreten,  die mit

Besonderheiten der Knochenphysiologie im  Zusammenhang  stehen.

Deshalb sollte bei Knochenanalysen dem Ort der  Probenahme  sehr

grosse Aufmerksamkeit geachenkt werden,


     I would like to ask Dr. Jeanmaire if he used the  entire
vertebra or rib for the analysis or only  sections of them?
In a systematic examination of human skeletons  we determined
the local distribution of 20 to SO trace  elements using neutron
activation analysis.  This showed that within very limited areast
concentration variations of up to two decades occur for several
trace elements.  These are related to certain features of  the
osteophysiology.  Great attention should  be paid,  therefore^
to the location of the bone sample taken.
JEANMAIRE  (France)

     Les cotes sont en general des cotes flottantes; nous avons
la totalite de 1'os qui est mineralise puis broye pour obtenir un
echantillon moyen.  Les vertebras sont entieres egalement.
        ^ ribs are usually floating ribs; the entire bone  is
mineralised and crushed to obtain an average specimen.  The
vertebrae are also entire.
SZADKOWSKI  (B.R.D.)

     Bin  wesentliches Anliegen dieser Sitzung war ein

Erfahrungsbericht zu geben Uber die Durchftihrung internationaler
Vergleichsprograrrane.  Wir haben gehtSrt und auf den Diapositiven
auch gesehen, dass die Ergebnisse eigentlich noch lange nicht

so sind wie wir sie uns wtinschen.  Es stellt sich die Prage ob

derartige Vergleichsprogranune Uberhaupt ndtzlich sind und wenn

ja wie sie durchgeftihrt werden kfinnen, wie sie verbessert werden
kSnnen?
     A fundamental concern of this session was to present a
report on the  experiences made in the implementation of inter-
comparison programmes.  We have seent that the results are far
from what we desired to have.  The question now arises if such
comparison programmes serve any useful purpose and if sot in
what ways can they be improved?

-------
                                2257
KJELLSTRtJM  (Sweden)
     I think it is very important when you do intercomparison
studies to take sufficient numbers of samples so that you can
make conclusions regarding systematic differences between the
laboratories.  If you ship urine and blood all over the world
there may be, especially for low concentration samples, wall
effects etc., which make the actual concentration in the sub-
sample received by certain laboratories different from the
average.  So I think that at least 10 samples of blood or 10 of
urine should be exchanged in such a study*  It is also important
that participating laboratories are identified and that the methods
used are described in reasonable detail.
BERLIN  (C.E.C.)
     The second intercomparison programme on lead having shown
no real improvement over the first, one point must be emphasised:
we had more than doubled the number of participating laboratories
and furthermore, some of the laboratories that took part in the
first did not take part in the second.  We still strongly feel
that intercomparison programmes serve to improve the results,
and this is fundamental.  Following an intercomparison programme
we often organize a meeting of the participating laboratories
so that they may discuss the results in depth.  This does lead
to one difficulty, the need to lift the anonymity of the
laboratories during the meeting in order that full discussions
may be held.
     A further point regarding the quality assurance; since
for environmental measurements the concentrations are very low
and thus the analyses difficult, national and international
authorities when they contract for research in this field
should assure themselves of the quality of the analyses which will
be performed by the laboratories.  One should really have results
at hand of which one can be sure that they are objective and not
questionable, at least from the analytical point of view.  One

-------
                                2258
will always be able to discuss other aspects of the results but
the long discussions on the validity of the analyses  should be
curtailed.
DELVES  (U.K.)
     I would agree with Dr. Berlin about removing any  'anonymity*
from the discussions after the results have been disclosed and
I think this should be agreed before the analyses are  carried
out.   But one of the big difficulties in carrying out inter-
laboratory comparative analyses is that one ends up with a
whole series of results and there still remains the question which
is the right answer?  A procedure that might overcome  this problem
would be for the reference laboratory to use at least  three
different analytical techniques to carry out the analysis of the
stock sample.  These techniques should be based on different
physical or chemical concepts, for example, anodic stripping
voltammetry, colorimetry, atomic absorption, and then  as the
National Bureau of Standards in Washington does, the reference
laboratory could issue a certified value together with a range
of acceptable values.  If any laboratory did not report analyses
within this range,  that laboratory would be in error  and one
could at least start to discuss the reasons for this.
     Secondly, I would like to see at least on spiked  sample
used for the intercomparison study.  I am aware that the metal
added will not be in the same form as that originally  present
in the sample, but one would know exactly the difference in
concentrations between the spiked and unspiked sample  and those
laboratories that did not obtain this difference would be in
error.
     In my opinion the use of spiked samples together  with samples
that have been given a range of 'acceptable concentration' by
the reference laboratory would help to solve some of the problems
of interlaboratory comparative analyses.

-------
                                2259
STOPPLER  (B.R.D.)

     Ich m6chte mich den VortrSgen der belden Vorredner anschliessen

und meine, dass wir im Augenblick einen Stand erreicht haben,

bei dem es eigentlich nicht so sehr darauf ankommt, dass eine

sehr grosse Zahl von Laboratorien an Tests dieser Art teilnehmen,

sondern dass in einem verhaitnisraSssig kleinen, aber sehr

gualifizierten Kreis diese Dinge noch etwas intensiver studiert
werden sollten.


     In my opinion we have now reached a stage at which it is
not necessary any more that a very large number of  laboratories
take part in these tests, but rather that a relatively small
number of highly qualified experts should make a more intensive
study of these problems.
SZADKOWSKI  (B.R.D.)

     Ich darf vielleicht noch folgende Anregung geben.  Auch
bei wissenschaftlichen Verfiffentlichungen, die sich nicht direkt
rait methodischen Problemen befassen, sollte doch die Methode
angegeben werden, und es sollte zumindest darauf hingewiesen
werden, wo man liber die ZuverlSssigkeit dieser Methode nachlesen
kann, damit bei einer kritischen Auswertung der Literatur wie
wir sie ktirzlich in Zusanunenarbeit mit europSischen Gemeinschaften
versucht haben, den einzelnen Autoren eine bessere Uebersicht
mtiglich ist.  Im tlbringen danke ich ftir die sehr rege Diskussion.


     In closing a suggestion should be made.  Even in scientific
publications which do not deal directly with methodological
problems the methods should nevertheless be described and at
least an indication should be given of the relevant literature
on the reliability of these methods.  This would give a much
clearer picture to those who have to evaluate critically this
literature^ as we have done recently in cooperation with the
European Communities.

-------
                       GEWEBSMESSUNGEN
                     TISSUE MEASUREMENTS
           MESURES RELATIVES AUX TISSUS BIOLOGIQUES
                MISURE NEI TESSUTI BIOLOGICI
               METINGEN VAN BIOLOGISCH WEEFSEL
                        (Continued)
                            Panel

Vorsitzender - Chairman - President - Prea-idente - Voorzitter

                  O.A. WEBER (Jugoslavija)

-------
                              2263
        THE CONCENTRATIONS OF COPPER, IRON, MANGANESE,
         ZINC AND CADMIUM IN HUMAN HAIR AS A POSSIBLE
           INDICATOR OF THEIR TISSUE CONCENTRATIONS

         E, VUORI, A, HUUNAN-SEPPA'Lfl AND J, 0, KILPld

Department of Public Health Science, University of Helskini,
Finland
ABSTRACT

     Samples from lung, muscle, liver, kidney and hair were
collected from 20 accidentally dead persons.   The tissue con-
centrations of copper, iron* manganese, zinc and cadmium were
determined by atomic absorption spectrophotometry using both
flame- and flameless atomizing systems.   Linear coefficient of
correlation was calculated between different tissues for each
element measured.

     The amounts of the trace elements measured in tissues in
the present study were approximately the same order as those
reported in literature for Finnish population.   However, in our
material the values of copper in tissues were about two times
higher than those reported earlier.

     In the present study no significant correlation between the
contents of trace elements in hair and other tissues was found.
Almost significant correlations were found between the cadmium
contents of hair and  lung and between the zinc contents of hair
and muscle.   It seems to us that the concentrations of trace
elements in human hair hardly can be used as an indicator of
their concentrations in other tissues as far as the trace ele-
ments included, in this study are concerned and the concentrations
of these trace elements fall into a  "normal" range.

-------
                             2264
^1 n t, r_gidI u c jji o n
     There has been a great number of studies in litterature
concerning the amount of various trace elements in human tiss-
ues.  The tissue concentrations of trace elements are consid-
ered to be of great importance since measuring only e.g. air,
water or food does not give reliable information about the
status of trace elements in human body and about their acute
and chronic effects on tissue metabolism.  The specimen
collection for the determination of trace element is rather
easy as far as autopsy material is concerned.  The opposite is
true of healthy human beings whose tissue concentrations have
been difficult to study mainly due to the problems of getting
biopsy material.  It is concidered that serum as a study
material has many important disadvantages (Strain et al, (11).
Hammer et al. (6), Hambidge (5)).  Therefore it seems to be
quite natural that scalp hair has been considered as a "prom-
ising" subject for trace element determinations for years.
Taking a hair biopsy is readily atraumatically performed and
it is easy to collect even large quantities of this material.
     On one hand it has been supposed that the concentrations
of some trace elements in human hair can be used as an indi-
cator of the contents of these elements in tissues (Goldblum
et al. (2), McDonald and Warren (8), Strain et al. (11)), on
the other hand it has been warned that the contents of trace
elements in hair are not very indicative of their tissue
concentrations and interpretation of analytical data requires
caution (Schroeder and Nason (10), Hambidge (5)).  Therefore
a controlled study about this problem seemed to us necessary
to perform as a part of the trace element project carried out
in our laboratory.
     This preliminary report reveals some of our main find-
ings concerning the trace element contents of hair and other
tissues.  Tho whole material will be oublished later in
detail.
Materials and methods
     Samples from lung, muscle, liver,  kidney and hair were
collected from 20 accidentally died nersons.  For analysis  the

-------
                             2265
soft tissue samples were dried overnight at 100 C and then
muffle-furnace ashed at 450 C in quartz crucibles. Hair
samples were washed before ashing as follows: the samples were
first pre-washed several times with deionized water and then
ones with ethanol. After the pre-washing the hair samples were
washed in a shaker first once with n-hexane and then four
times with 1 \ sodium lauryIsulphate. Each washing in the
shaker lasted 20 min. After all these successive washings the
hair samples were finally very thoroughly washed with
deionized water and then dried overnight at 100°C. Detailed
description about the hair washing procedure and about its
effects on the trace element concentrations of hair will be
published. In this connection it can be said that the washing
procedure had actually no effect on copper, zinc and cadmium
levels of hair but the opposite was true on iron and manganese
levels.
     For hair analysis we used only a certain part of the
whole sample, that is 1-2 cm measured from the scalp. The
ashing procedure of the hair samples was similar to the tissue
ashing. The ash was dissolved in 2 ml of 4 M HC1 and diluted
to 10 ml using deionized water. The trace metal conccentra-
tions of this solution were determined by atomic absorptions
spectrophotometry using both flame- and flameless atomizing
system (Perkin Elmer 300, HGA 70).
     In all different phases of the analytical procedure cars
was taken to avoid all possible sources of contamination. All
reagents and washing agents were checked not to cause
contamination and all glass-ware used was carefully washed
and dried before use.
     About the statistical analyses a detailed description
will also be published later.
Results
     The concentrations of copper, iron, manganese, zinc and
cadmium were determinated in hair, muscle, lung, liver and
kidney. Linear coefficient of correlation was calculated
between different tissues for each element measured.

-------
                           2266
Table I     The trace element  concentrations  of  tissues
investigated
       x            3          r-         i—          7*
       01            C          C         M*          4-i'
       M-            at          3         <          a.
       1            o          TO         co          a
                    t->                    1          O
                    CD                               *<
     23.3        6.07       6.87        23.6        13,9
     18.5        3.2        2.4         10.9         4.5
    (6.25J6     (<2.8)19    (3.4)19     (18.3)      (7.9)19
Fe
77.3
84
1.56
1.23
128
25
O.B419
1.36
1148
464
1.35
1.08
549
217
5.11
1.46
398
193
4.99
0.79
Mn

    (1.36)3     [<0.4)18    (<0,6)M    (3.8)12     (2.26)10

Zn   199         230        67.2        197         194
      64          52        17.5         66          38
    (107)6      (232)19     (53.O)19    (298)       C173)19

Cd
0.35
0.27
<0.27)6
0.2016
0.13
(<1.6)19
1.B616
1.30
(1.14)19
4.15
3.68
(3.8)
73,2
38.2
(40.7
     The numbers denote  fig/g  in dry weight.  The  first
number in each group denotes mean, the  second sd  and  the
number in brackets denotes value which  has  been reported
earlier for Finnish population. If not  otherwise  informed
the number of cases is 20.

-------
                             2267
Table II The correlations of trace  elements  measured
         between the tissues in  question
            03



Cd

hair
muscle
lung
liver
1-1
u
m
3
E
ns




bO
c
3
rH
+
ns


(_,
o
^
•H
(H
ns
*
ns

to
c
•a
•H
.*
ns
+ *
*
4
                    Cu
                    hair
                    muscle
                    lung
                    liver
                                              m            >,
                                              •-H        tl   ffi
                                              O   bO    0)   C
                                              m   c    >   TO
                                              3   3   i-l   -H
                                              E   rH   rl   JC
                     ns  ns   ns
                         ns   ns
                              44
                                                          ns
   Fe
   hair
   muscle
   lung
   liver
            o
            i—i
            u
            m
bo
c
0)
c
T)
ns  ns  ns  ns
    ns  *;   ns
        ns  * +
            ns
                    Mn
                    hair
                    muscle
                    lung
                    liver
o   bo    o
n   c    >
3   3   -H
£   rH   ^
                                               0)
                                               c
                                               TJ
                     ns  ns   ns   ns
                         ns   ns   ns
                              ns   ns
                                  ns



Zn
hair
muscle
lung
liver
l-H
u
n
3
*




bo
c
3
ns
ns


t,
0
>
•rt
ns
ns
*

 0.05)
 *    denotes almost significant  (O.Q5> p> 0.01)
 +*   denotes  significant  (0.01> p> 0.001)
 	   denotes negatively correlated

-------
                             2268
     Table I shows the trace element concentrations of tissues
investigated and Table II shows the correlations    of trace
elements measured between the tissues in question. As a whole
it can be seen in these tables that the concentrations of
trace elements investigated do not generally correlate with
the amounts of these elements in other tissues used in this
study. However, cadmium correlates significally between muscle
and kidney and almost significally between hair and lung,
muscle and liver, lung and kidney, liver and kidney. Copper
correlates significally between muscle and kidney, lung and
liver, lung and kidney and almost significally between liver
and kidney. Iron correlates significally between lung and
kidney and almost significally but negatively between muscle
and liver. Manganese shows no correlation between tissues
investigated. Zinc correlates almost significally between
hair and muscle, lung and liver.
Discussion
     It has been shown that sex, age and hair colour have
influence on hair trace element concentrations (Schroedar and
Nason (10), Petering et al. (9)).Furthermore Hambidge (5) has
shown that copper concentrations even depend on sampling
technique  so that the distal parts of hair contain more copper
than the proximal parts. Our material consisted of 20
accidentally died persons of whom 14 were men and 6 women
(mean age 34.5 years, range 13-76 years). All of them had
natural hair colour. Only the very proximal part of hair was
used in the present study. Muscle samples had been taken from
m. pectoralis major, liver and lung samples represented
parenchyma and kidney samples both cortex and medulla.
     Hambidge (4) has pointed out that the hair analysis of
chromium as an indicator of the nutritional status is valuable
if the correlation between chromium in hair and other organs
is determinated. There is no reason to suppose that this
concept is not valid as far as other trace elements are
concerned, too. However, studies where human hair trace element

-------
                              2269
concentrations have been correlated with those of other
tissues are quits few (Goldblum et al.  (2),  McDonald and
Warren (8)).
     The amounts of the trace elements  measured in tissues
in the present study were  approximately the  same order as
those reported in  litterature for Finnish population.  The only
exception was copper: in our material the values of copper in
tissues were about two times higher than those reported
earlier by ForssSn (1).
     In the present study no significant correlation between
the contents of trace elements in hair  and  other tissues was
found, Almost significant correlations  were  found between
the cadmium contents of hair and lung and between the zinc
contents of hair and muscle. On the basis of these results
the earlier suspicions of tha value of  hair  as a biopsy
material in this respect have received  more  attention.  This
is solid as far as the trace elements included in this  study
are concerned and the concentrations of these trace elements
fall into a "normal" range. The opposite might be true  when
specimens are taken from persons suffering from a disease
that is known to cause disturbance in trace  element mstabolim
[Hambidge et al. (3), McDonald and Warren (8)1. The situation
might be the same for the nonessential  trace elements (e.g.
arsenium, cadmium, lead, mercury). In these  cases human  hair
has been used to indicate the degree of exposition to these
elements (Hammer et al. (6), Kopito et  al. (7)).  The biopsies
used in this study were taken from subject representing
"normal" population with no apparent exposition.

-------
                              2270
 References
        f
 1 FORSSEN, A., "Occurance of Ba, Br, Ca, Cd, Cs  Cu, K, Mn,
   Ni, Sn, Sr,  Y and Zn in the human body", Ann .Med.Exp.Bio.
   Fenniae, 5Q, 99 (1972)
 2 GOLDBLUM, R.W., DERBY, S., LERNER, A.B., "The metal content
   of skin, nails and hair", J.Invest.Dermatol., 20,  13 (1953)
 3 HAMBIDGE, K.M., RODGERSON, D.O., O'BRIEN, D.. "Concentra-
   tion of chromium in the hair of normal children and chil-
   dren with juvenile diabetes mellitus". Diabetes, 17, 517
   (1968)
 4 HAMBIDGE, K.M., BAUN, J.O., "Hair chromium concentrations
   of human newborn and changes during infancy", Am.J.Clin.
   Nutr., 25,  376 (1972)
 5 HAMBIDGE, K.M., "Increase in hair copper concentration with
   increasing  distance from the scalp", Ann, jk Clin .Nutr., 26,
   1212 (1973)
 6 HAMMER, D.I., FINKLEA, J.F.,  HENDRICKS, R.H., SHY, C.M.,
   HORTON, R.J.M., "Hair trace metal lev/els and environmental
   exposure".  Am.J.Epiodemiol..  93, 84 (1971)
 7 KQPITO, L.,  BRILEY, A.M., SCHWACHMAN,  H., "Chronic plumbism
   in children: Diagnoses by hair analysis", JAMA, 209, 243
   (1969)
 6 MACDONALD,  I., WARREN, P.3,,  "The copper content of the
   liver and hair of African children with kwashiorkor", Brit^
   3.Nutr., 15, 593 (1966)
 9 PETERING, H,G., YEAGER, D.W.,  WITHERUP, S.O., "Trace metal
   content of  hair 1. zinc and copper content of human hair
   in relation  to age and sex".  Arch.Environ.Health., 23, 202
   (1971)
10 SCHRDEDER,  H.A., NASON, A.P.,  "Trace metals in human hair",
   J. Invest.J3erma to 1^, 53, 71 (1969)
11 STRAIN, W.H., STEADMAN. L.T.,  LANKAU.  C.A. JR., BERLINER,
   W.P., PORIES, W.J., "Analysis  of zinc levels in hair for the
   diagnosis of zinc defiency in  man",  .1. Lab .Clin .Med ., 68,
   244  (1966)

-------
                              2271
                TRACE ELEMENTS IN LUNG AND HAIR

                 G, W. GIBBS AND E, BOGDANOVIC

Department of Epidemiology and Health, McGill University, Mon-
treal, Canada
ABSTRACT

     In order to determine the health effects of airborne inor-
ganic particulates on mant it is first necessary to establish
reliable methods of measuring exposure.   Lung burden might be
a useful index, but there is little information on it's relation-
ship to measured concentrations of airborne particulates and
rates of removal of air pollutants from the lung.   Studies to
clarify these relationships and associations between the concen-
trations of trace elements in lungs and disease in man are often
hampered by the difficulty of obtaining whole lungs for analysis.
An alternative method is to use a portion of lung, but this re-
quires "knowledge of the way in which particulates are distributed
within the lung.

     Sixteen right lungs and four left lungs have been analysed
by bronchopulmonary segments for fourteen metals.   Preliminary
observations based on these results indicate that elements 'are
not uniformly distributed in the lung, the concentrations of
most elements analyzed  ( ,ug/g lung ashJ being highest in the
apex and lowest in the  anterior segments.   The total quantity
of several elements, in particular nickel and cobalt, increased
with age.   These and other elements present in the lung were
related to one another, copper being highly correlated with the
calcium content.

-------
                              2272
     Assuming the trace element content of the lung to be a
reasonable -index of exposure to airborne inorganic particulates
preliminary results suggest that the concentrations of trace
elements in hair are unlikely to provide a useful index of ex-
posure to inorganic atmospheric pollutants by the general popu-
lation.

-------
                                    2273
 1.      Introduction
        The mineral constituents  of  lung  and  other  human  tissues  can  be
 divided into  two  groups  depending on  their concentration (Koch et al [1]).
 First  the  elements such  as  calcium, sodium and potassium, which  are  gen-
 erally present  in high concentrations  (mg/100 g wet  tissue).  Elements may
 be  "essential elements"  involved in metabolism or  contaminants which, de-
 pending on their  chemical form and concentration,  may or may not have an
 adverse effect  on the  organism.
        The concentrations of  the major elements and  trace elements in lung
 and other  tissues have assumed importance for a number of reasons.   Large
 scale  studies of  metals  have  been undertaken to describe the distribution
 and concentration of stable elements in  the  organs and tissues of "stand-
 ard man" upon which the  International Commission on  Radiological Protec-
 tion [2] based  MFC calculations.  The mineral and  trace  element  contents
 of  the lungs of occupational  groups such as  coal workers have been studied
 for their  relationship or roles  in pneumoconiosis  (Rossiter et al [3],
 Keenan et  al  [4]).  The ability of certain metals to  act  as carcinogens in
 their  own  right or to  play  a  role in carcinogenicity by  inhibition of enz-
 yme activity  (Dixon et al [5]) has led to their study in cancerous and non-
 cancerous  tissue  (Mulay  et  al [6]).
        In  occupational studies it is often possible  to make some assess-
 ment of  particulate and  even  trace element exposure  from environmental
 measurements  (Gibbs and  LaChance [7], Gibbs  [8]),  while  for the  general
 population measurements  are rarely adequate.  In order to determine  the
 health effects  of  airborne  inorganic particulates  on man, it is  first
 necessary  to establish reliable methods  of measuring exposure.   Total lung
 burden may be a useful index  but there is little information on  its  rela-
 tionship to measured concentrations of airborne particulates and rates of
 removal  of air  pollutants from lungs.  Studies to  clarify these  relation-
 ships  and  associations between the concentrations  of trace elements  in
 lung and diseases  in man are  often hampered by the difficulty of obtaining
 whole  lungs for analysis and  analysis of portions of lung require a know-
 ledge  of the distribution of  trace metals and particulates within the lung.
       Although it has been recognized for may years that elements such as
 arsenic are concentrated in hair, only recently with improved analytical
methods have metals such as zinc, cadmium, copper, lead and mercury been
 detected.  The presence of trace elements such as lead in hair opened new
avenues of investigation as to the possible use of such a readily avail-
able tissue in a living population to measure the exposure of persons
 occupationally exposed to metals (El-Dakhakhny and El-Sadlk [9]).  As the
 exposure to metals by  the general population is much lower and as there
are sources of metal other than airborne particulates it is not known
whether  the trace metal content of hair from a non-occupational group
reflects exposure  to trace metals present in general air pollution.
       This report describes preliminary results of a study of the distri-
bution of  trace elements in segments of lung and their relationship to a
limited number of  trace elements in hair.

2.     Methods
       2.1   Collection of Samples
       Whole lungs and in most cases,  scalp and pubic hair were collected
at routine autopsies at seven centres in Montreal and neighbouring areas.
Tissues were transferred directly to polyethylene bags provided for  that
purpose and were stored in a deep freeze until analysed.   Hair samples,
cut as  close to the skin as  possible,  were placed in envelopes provided  by
the laboratory.   For each case,  age,  cause of death and place  of  residence

-------
                                    2274
were ascertained.   No  attempt was made  to  select  cases  with  specific  causes
of  death  (e.g. accidents),  or  to exclude  cases unless  the specimen was
incomplete.
        2.2   Samp_le_ Preparation
        The lung was weighed and cut  into  the  following bronchopulmonary
segments

    Left  Lung                   Right  Lung

Upper  lobe Apical           Upper lobe Apical         Middle  lobe Lateral
           Anterior                    Anterior                    Medial
           Posterior                   Posterior
           Lingular

Lower  lobe Superior Basal   Lower lobe Superior  Basal
           Anterior Basal              Medial Basal
           Lateral Basal               Lateral Basal
           Posterior  Basal

        Segments were  weighed and dried in an  oven at 110°C  to  constant
weight.   This took from one to four  days.  The specimens were  then placed
in  a furnace for ashing.  The  furnace  temperature was  raised slowly  (17
hours  to  reach 475°C) which prevented  conflagrations which  would cause
serious sample losses.  The tissue was ashed  for four  days, at the end of
which  period the ash  weight was constant.
        The ash from two lungs  were analysed semi-quantitatively using X-ray
fluorescence techniques to  identify  elements  which were present in suffic-
ient quantities for detailed analysis  by  atomic  absorption  spectrophoto-
metry.  For  this prupose the ash was treated  with a mixture of 1:1 concen-
trated nitric and  perchloric acids and made up to volume in nitric acid.
        Hair  samples were washed and  ashed using  the method  reported  by
Petering  et  al [10].

3.      Results
        3.1   Loss in Weight  on  Drying
        The loss in weight of lungs dried  in segments ranged 75.5 - 92.9%
(mean  83.6%, median 83.3%)  and of lungs dried in lobes ranged  65.2 - 86.9%
(mean  80.0%, median 81.7%).  This loss in weight was slightly  higher than
the published value of 78%  (Diem [11]).   The  difference between the  loss
on  drying of lungs in segments and lungs  dried as complete  lobes possibly
reflected the difficulty of removing water from  large  fragments of un-
macerated tissue.  The loss in weight  of  individual segments was not ana-
lysed  in  detail but appeared to be reasonably uniform  with  a few excep-
tions  where  large  differences  in loss  between lobes or segments were
detected.
        3.2   Lung Ash
        The ash from most lungs was white  but  in  some cases  ash from  diff-
erent  regions of the  lung showed marked differences in colour.  Ash  as a
percentage of wet  and dry lung tissue  weight  is  shown  in Table I.  Al-
though some  lungs  showed considerable  variation  in the percentage of ash
from individual segments, the  overall  ash weight was approximately one per-
cent of wet  tissue weight and  six percent of  dry tissue weight.  The ash
contents  of  the upper, middle  and lower lobes of 12 right lungs and  of the
upper  and lower lobes of six left lungs are shown In Table  II.  When the
ash contents were  expressed as mg/g  wet or dry tissue  weights,  the results
were similar for the  three  lobes, although there was a tendency for  the

-------
Table I
Ash content of lungs


Right
Lungs
(16)















Left
Lungs
(4)


Lung
No

85
49
73
39
77
12
78
99
100
13
23
21
17
27
33
19
Overall

4
11
64
22
Overall
Ash % Wet
Range

0.8-0.9
0.4-1.0
0.4-1.3
1.0-1.1
0.8-0.9
1.0-1.2
0.9-1.0
0.6-1.0
0.8-1.1
0.9-1.1
0.8-1.0
1.0-1.2
1.2-1.4
0.9-1.0
,0.9-1.0
1.0-1.3
0.4-1.4

0.9-1.3
0.8-1.1
1.0-1.1
0.9-0.9
0.8-1.3
analys ed
by segment
Weight Ash % Dry
M Range

0 8
\J » \J
1.0
0.4
1.0
0.8
1.1
1.0
0.9
1.0
1.0
0.9
1.1
1.2
1.0
0.9
1.1
1.0

1.0
1.0
1.1
6.9
1.0

fi R-fi T
U • U U * J
2.0-5.0
5.2-17.7
5.3-5.9
5.6-6.9
5.7-6.5
4.5-8.4
4.5-6.8
5.1-6.3
6.8-7.8
4.7-5.6
5.6-6.8
5.2-5.7
5.7-6.4
5.0-6.0
6.1-8.2
2.0-17.7

5.6-7.6
5.0-7.0
6.0-6.6
5.9-7.6
5.0-7.6
Weight
M

7 *>
I . j
4.8
6.8
5.7
6.1
6.0
5.9
6.2
5.6
7.0
5.2
6.2
5.3
6.3
5.6
7.5
6.1

6.6
5.9
6.3
7.1
6.0
Table II
Ash content of
Lung
No

Right 70
71
3
47
106
108
109
110
113
117
114
101
Overall

Left 115
43
107
112
83
116
Overall

lungs
mg Ash/g Wet
Upper Middle
Lobe

12
10
9
10
11
13
10
11
8
10
12
12
10

6
9
10
11
11
10
10

Lobe

9
10
8
10
11
12
10
11
10
10
12
11
10

_
-
_
_
_
_
_


analysed bv lobe
Weight
Lower
Lobe

9

8
10
10
10
10
11
7
7
9
11
10

9
9
9
11
9
7
9



mg Ash/g Dry Weight
Upper Middle Lower
Lobe

71
54
56
57
48
72
70
56
23
50
67
54
56

117
52
99
68
69
58
69

Lobe Lobe

40 64
54 57
50 54
52 56
43 4^
64 59
69 71
56 57
56 19
76 40
66 41
52 47
55 56

100
50
56
71
63
40
59

10
-J
Ul

-------
                                   2276
ash content  (mg/g dry wt.) to be higher in  the left upper lobes than in the
left  lower.
      3.3  Ash Content and Age
      The ash content of lung in mg/g dry lung weight showed little rela-
tion  to age  and  the linear correlation was  low and negative (-0.25).  How-
ever, the total  ash weight increased with age (r=*0.61).  This suggests
that  both tissue weight and mineral within  the lung increased with age,
but tissue weight increased more rapidly.
      3.4. Metals in Lung
      The range  and median concentrations of metals in  lungs examined  in
this  study are shown in Table III. With the exception of  zinc, which
occurred  in  lower concentration and calcium which was present in much
higher concentration, the results were in good agreement with those of
Tipton and shafer (12^ for victims of instantaneous death from nine cities
in the USA.  The  higher calcium levels in our study may  be related to the
older population (Mean age 65.! years).
      3.5  Change in Metal Content of Lung with Age
      If certain metals present in inorganic participates are inhaled by
the general  population and retained in the  lungs, the total quantity and
concentration of those metals might be expected to increase with age.
Certain endogenous metals might also increase with age.  The relation of
10 metals to age and to other metals in twenty lungs (16 right and 4 left)
are shown by the linear correlation matrix  in Table IV.  Nickel and cobalt
were  the elements most highly correlated with age (r-0.60 and r=0.50 res-
pectively).  Zinc showed no relationship to age.  Of interest were the
relatively low correlation of calcium with  age (0.33) and the extremely
high  correlation of copper with calcium (0.93).  Three  elements showing
little relation  to calcium or copper were cadmium, aluminium and chromium.
      3.6  Distribution of Metals within Lung
      The concentrations of eleven metals (ug metal/g ash) in. ten segments
of sixteen right lungs and eight segments of four left  lungs are shown in
Table V.  For the right lungs the ranges and median concentrations of the
metals Ni, Co, Pb, Cr, Mg, Ca and Cu were lower in the  second segment
(Anterior).  In  left lungs, the concentrations of Nl, Co, Pb, and Mn were
lower in segments two and four and the higher concentrations of Nl, Co, Pb,
Mn, Cr and Zn were present in the first segment (Apex).  Magnesium and
aluminium appeared to be concentrated in the fifth segments.  As the data
presented here are preliminary they have not been subjected to rigorous
statistical  analysis.   Nevertheless,  it is apparent that certain metals
are not uniformly distributed within the lung when expressed as concen-
trations per unit ash.  It seems likely that metals entering, the lung as
air pollutants are concentrated in the apices of the lung but also enter
other areas, but this interpretation is  complicated  by the inter-
relationships of the  metals.
      3.7  Trace Elements in Hair
      The concentrations of Pb,  Ni, Cu,  Zn and Cd in scalp hair,  pubic
hair and lung are shown In Table VI.   On these limited data there appeared
to be little or no relationship between these metals in lung and  concen-
trations of  these metals in hair and no conslstant relationship  between
the levtls in scalp hair and pubic hair.   Nickel was detected only in two
specimens and then In high concentration.   Zinc was present in all speci-
mens In concentrations In the range reported by Petering et a.l [10]  but to
date too few specimens have been analysed to reliably examine relationships
with age.   However the scatter of the results makes it unlikely  that hair
samples will prove useful in studies  to  determine exposure to air pollu-
tants .

-------
                                   2277
Table III
    Ranges and median concentrations  of metals  in lung  (ug/g tissue ash)
 LEFT LUNO.
       Hetal

     Aluminium
     Cadmium
     Calcium
     Cobalt
     Chromium
     Copper
     Iron
     Magnesium
     Manganese
     Nickel
     Lead
     Potassium
     Zinc
     Sodium
   Current series
  ug/g Tissue Ash
   range
  304-7945
   10-214
 6591-91805
    5-50
    5-198
   32-243
 5009-15099
 2689-91805
   14-284
    6-86
    8-204
 8185-215025 33997
   16-1584     167
94122-561694 211452
 Published data<10)
jig/g Tissue Ash
median
1925
54
26522
20
28
95
18080
8482
133
30
36
range
170-) 4600
< 50-270
400-29000
< 2-140
0.5-120
50-250
—
3800-16000
< 10-170
< 5-800
5-550
median
2000
<50
10000
<2
13
120
—
8600
10
<5
47
660-3100
1300
 RIGHT LUNG.
  Current series
  g/g Tissue Ash
Metal
Aluminium
Cadmium
Calcium
Cobalt
Chromium
Copper
Magnesium
Manganese
Nickel
Lead
Zinc
range
80-7946
3-639
4037-215,300
5-103
8-81*3
38-34?
1056-107,200
!0"-468
6-126
5-M6
36-20,806
median
1080
55
29.945
21
47
107
8813
116
27
50
855

-------
Table IV





                       Correlation matrix - age and  the aedian. lung content, of  10  metala

Age
Ca
Al
Zn
Ni
Co
Cr
Mn
Pb
Cd
Cu
Age
1.000
0.331
0.302
-0.004
0.602
0.500
0.409
0.308
0.284
0.114
0.388
Ca
0.331
1.000
-0.045
0.452
0.538
0.367
0.095
0.655
0.322
-0.178
0.928
Al
0.302
-0.045
1.000
-0.094
0.294
0.227
0.207
0.374
0.151
0.231
0.016
Zn
-0.004
0.452
-0.094
1.000
0.044
0.030
-0.160
0.205
-0.037
-0.111
0.452
Ni
0.602
0.538
0.294
0.044
1.000
0.369
0.477
0.565
0.293
-.0134
0.597
Co
0.500
0.367
0.227
0.030
0.369
1.000
0.693
0.654
0.569
0.247
0.416
Cr
0.409
0.095
0.207
-0.160
0.477
0.693
1.000
0.398
0.643
0.134
0.144
Mn
0.308
0.655
0.374
0.205
0.565
0.654
0.398
1.000
0.543
0.251
0.630
Pb
0.284
0.322
0.151
-0.037
0.293
0.569
0.643
0.543
1.000
0.315
0.264
Cd
0.114
-0.178
0.231
-0.111
-0.134
0.247
0.134
0.251
0.315
1.000
0.024
Cu
0.388
0.928
0.016
0.452
0.597
0.416
0.144
0.630
0.264
0.024
1.000
                                                                                                                  CO

-------
Table V

Lung
S egment
(Right)
1
2
3
4
5
6
7
8
9
10
Overall
(Left)
1
2
3
4
5
6
7
8
Overall
Ni

Range

9-126
6-29
8-63
8-88
11-59
8-57
10-67
9-111
11-98
11-45
6-126

27-86
8-29
29-44
6-25
19-61
27-81
22-40
19-24
6-86


M

43
18
31
27
29
21
27
27
23
30
27

70
13
30
14
33
31
23
22
30
Co

Range

10-103
5-21
6-67
7-68
13-58
10-38
8-52
9-52
9-37
7-46
5-103

15-72
6-14
14-50
5-10
16-29
13-37
8-28
6-27
5-50


M

40
9
15
21
21
17
24
25
13
17
21

28
8
17
8
26
23
14
9
20
Pb

Range

34-275
7-70
25-416
11-181
5-234
20-150
18-244
19-156
20-116
18-193
5-416

13-204
9-29
23-132
8-27
38-78
24-85
18-76
19-72
8-204


M

99
32
48
61
43
52
67
67
41
48
50

45
16
30
15
42
44
28
19
36
Mn

Range

64-329
10-196
27-317
14-280
23-468
14-364
45-279
15-314
13-201
29-333
10-468

133-284
18-109
99-183
23-143
122-151
80-129
44-144
14-168
14-284


M

145
115
126
117
124
81
122
108
106
108
116

170
52
147
84
133
127
105
144
133
Cr

Range

24-258
8-78
10-843
10-170
19-146
19-94
16-132
18-130
10-92
11-115
8-843

25-198
6-25
14-124
5-45
24-73
15-80
11-71
12-67
5-198
C.A

M

84
21
33
41
53
48
66
53
33
46
47

112
16
26
22
31
38
20
20
28

Range

8-327
3-235
5-383
5-639
6-482
9-327
5-319
9-268
11-218
4-342
3-639

36-120
13-94
33-106
10-130
15-91
15-132
15-79
13-214
10-214

M

53
32
24
65
56
72
66
58
58
53
55

61
48
84
'41
46
35
48
91
54
Al
Range
250-7946
192-4650
197-4833
139-4205
170-2468
135-2687
166-4028
148-4073
80-2664
116-3847
80-7946
597-4702
490-1428
707-7945
304-3522
2215-3953
891-2777
569-3685
1079-2697
304-7945

M
2001
1438
1316
1102
714
858
634
836
1094
915
1080
1519
1150
2179
2012
2748
1208
756
1818
1925
                                                                                                                                N>
                                                                                                                                N>

-------
Table V (Con'd)
 Right
 Left
Lung
Segment
1
2
3
4
5
6
7
8
9
10
Overall
1
2
3
4
5
6
7
8
Overall
Cu
Range
44-344
23-141
43-437
47-227
57-234
59-359
36-265
36-217
40-308
38-347
23-437
64-106
54-124
94-186
32-102
39-107
43-148
38-110
36-243
32-243

M
137
93
120
103
180
108
131
121
113
107
116
99
92
128
80
70
103
88
85
95
Zn
Range
74-20,806
40-4,081
51-7,438
35-5,330
93-2,505
36-9,548
63-2,725
65-6,444
66-12,139
69-8,864
36-20,806
30-1,039
32-195
41-1,584
16-1,037
33-998
44-1,095
31-950
44-515
16-1,584

M
897
795
798
885
878
822
885
866
614
862
885
821
195
421
92
140
71
67
73
167

Rai
1,557-
1,056-
3,332-
4,509-
5,039-
4,714
5,442-
5,303-
4,962-
1,216
1,056-
6,033
6,541
6,132
2,689
7,580
9,077-
6,953
7,014-
2,689-
                                                                         Mg
       Ca
                                                                               M
Range
M
1,557-18,814 8,588
1,056-15,279 7,694
3,332-17,117 8,436
4,509-18,699 8,776
5,039-19,019 9,544
4,714-84,949 8,918
5,442-107,200 8,655
5,303-23,368 8,559
4,962-18,975 9,108
1,216-19,230 9,144
1,056-107,200 8,865
6,033-8,314 6,949
6,541-7,556 7,141
6,132-91,805 7,339
2,689-23,924 10,572
7,580-19,549 10,830
9,077-17,426 8,930
6,953-18,776 8,034
7,014-21,968 7,114
2,689-91,805 8,482
11,383-116,883
^,037-132,069
12,175-146,224
15,524-88,261
11,856-166,143
15,309-69,438
12,368-66,496
13,499-162,962
16,470-215,300
15,475-148,032
4,037-215,300
12,893-19,745
16,146-53,040
12,925-91,805
6,591-34,501
19,549-42,565
15,452-31,320
17,268-46,982
15,196-40,717
6,591-91,805
31,123
21>18
23,815
39,490
32,426
23,197
36,745
22,041
30,210
31,643
29,945
14,267
37,358
17,826
22,363
24,841
23,258
28,204
29,969
26,522
                                                                                                                      N)
                                                                                                                      00
                                                                                                                      O

-------
 Table VI
Metal
              Concentration of trace elements in scalp hair, pubic hair and


Sample Location

Pb
Cu
Zn
Cd
Hi

Scalp Hair (ug /g hair)
Pubic Hair (jjg /g hair)
Lung (jig /g tissue ash)
Scalp Hair (jjg /g hair)
Pubic Hair (fig /g hair)
Lung (/ig: /g tissue ash)
Scalp Hair (ug /g hair)
Pubic Hair (ug /g hair)
Lung (ug /g tissue ash)
Scalp Hair (jig /g hair)
Pubic Hair (jig /g hair)
Lung fyig:*/g tissue ash)
Scalp Hair (»igr/g hair)
Pubic Hair (jigr/g hair)
Lung (pgr/g ^tissue ash)
77
<16.8
31.3
80.5

-------
                                    2282
4.     Discussion
       The concentrations of metals in the ash from the bronchopulmonary
segments of the lung indicate that certain elements are not uniformly dis-
tributed and that metals in both left and right lungs with the exception of
calcium, magnesium and cadmium were to varying extents, present in greater
concentration in the apical tissue.  This agrees with the findings of
Molokhia and Smith [13] that trace elements are present in higher concen-
trations in apical tissue. Nevertheless this  should be  interpreted  with
caution.

       Trace element concentrations in lung may be expressed in several
ways.  The total weight of an element might be misleading as the weights of
essential metals present (Ca, Na, K, etc.)  might be expected to vary with
lung size and contaminating metals likely to be present in larger amount
in lungs of greater capacity.  If the metal content is expressed as percen-
tage wet weight, errors arise due to oedema and blood clots.  Concentra-
tions expressed as a percentage of dry lung weight require uniform methods
of drying and local disease (e.g. emphysema or fibrosis) might lead to
articifially high or low dry lung weights.   While it is still our inten-
tion to examine our data in the above manner, in this study we chose to
describe the concentrations of metals in lung as ug metal/g lung ash.  The
results Indicate that concentrations expressed in this way must also be
interpreted with caution, as aging and calcification of diseased areas of
lung can produce higher weights of lung ash from certain regions of the
lung lowering the concentration of other elements.  This problem probably
exists even when whole lungs are used for analysis.
       Preliminary data presented in this report indicate that metals can
be detected in the hair of the general population, but the extent of the
scatter of the results obtained to date suggest that hair is unlikely to
serve as a suitable method of surveying the general population for exposure
to trace elements present in air pollutants.

Acknowledgements
       We  acknowledge with  thanks  the  technical assistance of Miss Janet
Falser.  This study was undertaken with  the  help  of a grant from  the
Quebec Department of Municipal Affairs.


REFERENCES
      KOCH,  H.J.Jr.,  SMITH,  E.R.,  SHIMP,  N.F.,  CONNOR, J.,   "Analysis of
      elements in human tissues",  Cancer,  9,  499  (1956).

      Report: of Comittee II ^on permigsib^ dose for  internal radiation,
      ICRP Publication, (1959), Pergamon  Press, London (1960).

      ROSSITER, C., RIVERS,  D., BERGMAN,  I.,  CASSWELL, C., NAGELSCHMIDT,
      H.,  "Dust content, radiology and  pathology  in  simple pneumoconiosis
      of coal workers"   (Fourth Report),  Inhaled  Particles and Vapours,
      ed.  C.N. Davles,  419,  Pergamen Press, Oxford,  England  (1967).

      KEENAN, R.G., CRABLE,  J.V.,  SMALLWOOD,  A.W., CARLBERG, J.R.,
      "Chemical composition  of the coal miner's lung", American  Ind.
      Hyg. Aasoc. J.. 392 (1971).

-------
                                      2283
 5     DIXON, J.R., LOWE, D.B., RICHARDS, D.E., CRALLEY, L.J.,  STOKINGER,
       H.E., "The role of trace metals in chemical carcinogenesis:
       Asbestos cancers", Cancer Research, 30, 1068 (1970),

 6     MULAY, I.L., ROY, R., KNOX, B.E., SUHR, H.N., DELANEY, W.E.,
       "Trace-metal analysis of cancerous and non-cancerous human tissues",
       J. Nat. Cancer Inst.. 47, 1 (1971).

 7     GIBBS, G.W., LACHANCE, M., "Dust exposure in the chrysotile asbestos
       mines and mills of Quebec", Arch Environ Health, 24, 189 (1972).

 8     GIBBS, G.W.  "Qualitative aspects of dust exposure in the Quebec
       asbestos mining industry", Inhaled Particles III. Ed. H.A. Walton,
       Vol. II, 783, Unwin Bros. Ltd., London, England (1971).

 9     EL-DAKHUKHNY, A.A., EL SADIK,  Y.M., "Lead in hair among  exposed
       workers", Arner Ind. Hyg^ Assoc. J.. 33, 31 (1972).

10     PETERING, H.G., YEAGER, D.W.,  WITHERUP, S.O., "Trace metal content
       of hair I. zinc and copper content of human hair in relation to age
       and sex", Arch. Environ Health. 23, 202 (1971).

11     DIEM, K., Documents Geigy. scientific tables. 6th ed., Geigy
       Pharmaceutical Co., Ltd., Manchester, England (1965).

12     TIPTON, T.H., SHAFER, J.J., "Statistical Analysis of lung trace
       element levels", Arch Environ Health, 8, 58 (1964).

13     MOLOKHIA, M.M., SMITH, H.  "Trace elements in the lung", Arch.
       Environ Health. 15, 745 (1967).

-------
                              2285
          TRACE METAL CONTENT IN HAIR AND THE LEVELS
     OF 5-HIAA IN THE URINE OF POPULATION EXPOSED TO LEAD

             B, STANKOVI6/ M, STANKOVIC, S, MILlC,
          Z, KORI6ANAC, LJ, MILOVANOVIC, M, DUGANDZlC

Institute of Occupational and Radiological Health, Belgrade,
Yugoslavia and Institute of Inorganic and Analytical Chemistry,
Faculty of Pharmacy, Belgrade, Yugoslavia
ABSTRACT

     The aim of this paper ie to present our first results, as
a part of a long-term project, on trace metal content of hair
in human population exposed to different pollutants* including
our observations of 5-HIAA excretion in lead exposure.

     The study population consisted of four groups, representing
four levels of lead exposure:  without exposure, urban level of
exposure - low level, intermediate exposure of people living
around lead plant, and high level of exposure of lead workers.
Absorption of lead in hair was very marked and gradually inc-
reased with the degree of exposure, from 3.76 to 71.90 ,ug/g.
Lead and cadmium in hair, as non-essential elements, of unexposed
and exposed people showed rather good degree of association of
the metals.   Our results show that the mean hair Pb level and
also Cd level did reflect community exposure and might be a use-
ful biotoxicological index of a prolonged exposure.   The con-
tents of the two essential trace elements, Cu and Zn, did not
differ so significantly between farmers, city residents, the
subjects living near lead plant and lead exposure workers.

     The study of 5-HIAA, as the end metabolite of minor pathway
of tryptophan, shows the increased levels of the excretion in

-------
                              2286
the group of 120 exposed inhabitants.   The results of our study
demonstrated that the determination of S-HIAA in the urine can
be used as a useful and additional test for better assessment of
the environmental lead exposure.

-------
                             2287
1.  Introduction
Epidemiological data suggested that, in the inhabitants  li-
ving in the vicinity of a non-ferrous metals smeltery and re-
finery, the exposure to a mixture of heavy metals is  quite
possible.
Recently, attention is paid to the determination of  various
trace metals in hair as an index of a prolonged exposure. Ha-
ir has long been recognised as a metal-containing  metabolic
end product /!/. Also hair, by some authors A,2,3 and V, be-
tter reflect  the total body pool of some elements than eit-
her blood or urine? and this tissue may prove a practical do-
simeter for metallic environmental pollutants.
In this study, hair analysis has been applied to two classes
of elements: the essential nutrients, such as zinc and copper,
and elements such as lead and cadmium for which no evidence
of essentiality has been found.
Our findings of the significant contents of lead in the hair
of the inhabitants living near a lead-zinc plant suggested
the examination of eventual  derangements in the metabolism
of some amino acids. Prom that reason the urinary excretion
of 5-hydroxyindolacetic acid /5-HIAA/ has been studied in a
group of people from that region.
2.  Materials and Methods
The study population for trace metals in the hair consisted
of male and female of three different groups, and one  group
of workers mostly exposed to lead:
  I/ a group of peasants far from any pollution /30 cases/,
  £/ a group of residents of Belgrade /40 cases/,
  3/ a group of inhabitants living at least 10 years in  the
     vicinity of a lead smelting plant /50 cases/, and
  *»•/ a group of workers from mentioned lead plant/50 cases/.
As a sensitive, accurate and relatively interference-free te-
chnique, atomic absorption allows rapid analysis for trace e-
lements. Therefore, in our study hair analysis were  done by
atomic absorption spectrometry,using a Unicam SP 90A, Ser.2.
Virtually, all of the samples were collected during the fall
of 1973 and the last two weeks of February 1974. Hair samples
were collected in the small plastic bags, numbered and  bro-
ught to the Institute of Occupational Health Lab. for analyses.
The special precautions and washing with 1# detergent soluti-
on, warm deionized water and 1# HNO, were taken to avoid trace
elements contamination. Hair samples were prepared in 2% ni-
tric acid for atomic absorption determination of metals after

-------
                             2288
wet acid-digestion with nitric acid and hydrogen peroxide.
The concentrations of 5-HIAA were determined in the fresh u-
rine samples by a spectrofluorometric method /5A From the
same urine samples the determination of delta-aminolevulinic
acid /ALA/ has been also carried out by Grabecki method /6/«

3.  jtesults and Comments
Studied population consisted of four groups, actually ranking
four levels of lead exposure:  1st group - without  exposure,
2nd group - low, urban level of exposure,  3rd group - inter-
mediate exposure in the settlements surrounding lead plant,
and 4th group - high exposure of lead workers.
The results of trace metal analyses in hair are summarized in
table I. The arithmetic means with standard deviations of le-
ad, cadmium, copper and zinc were presented for each group.

 Table  I. - Trace metal content in hair, jutg/g
             The mean values with standard deviations
*
Group

1 1st
2nd
3rd
4th
Lead

mean s
3.76 2.30
7.16 4.41
33.13 17.84
71.90 30.80
Cadmium

mean s
0.22 0.17
0.30 0.22
0.33 0.14
0.78 0.82
Copper

mean s
9.5* 3.15
9.28 2.76
7.71 2.90
10.70 3.52
Zinc

mean s
153 51.5
137 41.1
154 49.8
158 39.6
     - For the groups see the chapter: Materials and Methods
  s  - Standard deviation

The mean values for the two non-essential and toxic trace el-
ements, Cd and especially Pb, shown differences between gro-
ups, and were in accord with exposure rankings for Pb and Cd.
On the other hand, means for the two essential trace elements,
Cu and Zn, did not differ significantlly between people  in
village, city, around lead plant,and the lead workers.

-------
                             2289
Uptake of lead in hair was very marked and gradually increa-
sed with the degree of exposure. From that reason the data of
lead in hair are presented separately in table II.

  Table  II. -  Lead in hair,  jug/g
Group

1st
2nd
3rd
4th
Number
of
cases
n
30
40
50
50
Range

min
1.2
1.7
5.2
20.0

max
10.0
18.5
78.0
163.0
Arithmetic
mean
X
3.76
7.16
33.13
71.90
Standard
deviation
s
2.30
4.41
17.84
30.80
The mean values of lead* in hair of unexposed population were
3.76; than 7.16 of urban population; 33.13 of inhabitants li-
ving near lead smelting plant and ?1.90jug/g of lead exposed
workers. The calculation of t-test of the arithmetic mean va-
lues between all groups revealed the highly significant dif-
ferences /P<.001/, except between the 1st group/farmers/ and
the 2nd group /urban population/, where it was only signifi-
cant /t = 2.50; P< ,05/.
Lead and cadmium in the hair of unexposed and exposed people
showed rather good degree of association of the metals. The
levels of cadmium in all groups and their means were  lower
than those previously reported /I, 7» 8/.
Scalp hair has several of the characteristics of an ideal ti-
ssue for epidemiologic study and it is painlessly removed,no-
rmally discarded and easily collected. Hammer and co-workers
/7/ in their study minimized possible effects of age,sex, ha-
ir color, varying hair length and personal chemical  treat-
ments. Our study shown, that despite of some problems,the me-
an hair lead level and also cadmium level did reflect commu-
nity exposure and therefore might be an useful biotoxicolo-
gical index of a prolonged exposure.

-------
                             2290
The results of metal contents in hair obtained in a group of
inhabitants living in the vicinity of a lead smelting plant,
and the concentrations of lead in air /I - 80jug per nr/ of
that environment, suggested further investigation. The studi-
es of 5-HIAA excretion as the end metabolite of minor pathway
of tryptophan, parallel to the ALA excretion in the urine/as
an indicator of lead absorption/ were carried out. A larger,
homogeneous group of 120 persons from that area has been exa-
mined. As the control, a group of 30 citizens without  known
lead exposure has been also examined. The results are  shown
in table III.
 Table  III.  -  Excretion  of  5-HIAA  and ALA
Biological
parameters
5-HIAA, mg/1
ALA, mg/1
Exposed group
MQ« stand.
mean deviation
7.35 2.32
9.20 8.65
Control group
stand.
mean deviation
2.86 1.21
5.21 2.48
The increased levels of 5-HIAA in the exposed persons were
evident. The difference between the arithmetic means of uri-
nary 5-HIAA of exposed and unexposed population was statisti-
cally significant.
5-HIAA has been found in the urine presumable as the end-pro-
duct of the action of monoamine oxidase /9/ on 5-hydroxytry-
ptamine /serotonin/, which plays a role as an important phy-
siological agent. In the urine of normal human subjects the
5-HIAA is present in small amounta, a few miligrams per day.
The results of our study demonstrated that lead interferes
/possibly/ in some manner with the enzymatic degradation of
serotonin - in the long run of tryptophan. Consequently, it
seems that 5-HIAA excretion can be used as an additional and
simple biochemical test for obtaining a more complete pictu-
re in favour of the environmental lead exposure.

-------
                              2291
                          References
1. Schroeder,H.A., Nason,A.P.:"Trace metals in human hair",
   J. Inveat. Dermatol.,  53,  7! /1969/
2. Yurachek,J.P., Cemena,G.G., Harrison,W.W.: "Analysis  of
   human hair by spark source  mass spectrometry",
   Anal. Chem.,  41, 1666 A969/
3. Kyle,R.A., Pease,G,L.: "Hematologic aspects of arsenic
   intoxication", New Eng. J.  Med.,  273. 18 /1966/
4. Kopito,L., Byers,R.K., Schwachman,H.: "Lead in hair  of
   children with chronic lead  poisoning",
   New Eng. J. Med.,  276, 949 /1967/
5. Kimura,Y., Arai,Y., Shimatavi,J., Hirona,H.:  "Determina-
   tion of 5-hydroxyindolacetic acid in urine",
   Arerugi,  16, 463 A967/
6. Grabecki,J., Haduch,T., Urbanowicz,H.: "Die einfachen
   Bestimmungsmethoden der Aminolavulinsaure im Earn11,
   Int. Arch. Gewerbepath. Gewerbehyg.,  2^, 226 /1967/
7. Hammer,J.D., J.F.Finklea, R.H.Hendricks, C.M.Shy, R.J.M.
   Horton: "Hair trace metal levels and environmental expo-
   sure",  Amer. J. Epidem.,   °/£, 84 /1971/
8. Petering,H.G., Yeager,D.W., Witherup,S.O.:  "Trace metal
   content of hair",  Arch. Environ. Health, 27* 327 /1971/
9. Sjoerdsma,A., Smith,T.E., Stevenson,T.D., Udenfriend,S.:
   Proc. Soc. Exp. Biol.and Med.,  89* 36 /1955/

-------
PANEL DISCUSSION

-------
                               2295
                 SUMMARY OF DISCUSSION
WEBER (Yugoslavia)
     The unifying themes in this session will be singled out as
follows: hair, lungs and to a minor extent other tissues
as well as methods of analysis with special reference to atomic
absorption and spectrometry but we will limit the discussion to
hair and lungs.  We proposed the following order:
Hair;
   a)  can hair be used for the measurements of trace elements,
       exposure, or absorption?  what are its advantages and
       disadvantages?
   b)  How do elements in other tissues correlate with the
       levels in hair?
   c)  How are trace elements distributed in hair, for example
       along its length?
       What is the value of analysing lungs for trace elements?
       Do they reflect trace element exposures?
   b)  How are trace elements distributed in the lungs?
   c)  How should the results of element analysis be expressed,
       for instance, yg/g, ash; u9/9» w®t tissue; or some other
       dimension?

     Turning now to the first question, can hair be used for
the measurement of trace element exposure or absorption?  What
are its advantages and disadvantages?

-------
                               2296
                           DISCUSSION
VUORI (Finland)
     Everyone knows the advantages of hair as biopsy material
but I am sure that we do not know all the disadvantages that this
material has.  For instance it was not until at the end of last
year that Hambidge showed that the copper concentration of hair-
depends even on sampling technique, so that the distal parts of
hair had more copper than the proximal parts.  Further an unsolved
problem is the treatment of hair before analysis, dozens of
different procedures are published in literature.  In our case
we had to pre-wash our hair samples, because in some cases they
were badly contaminated by blood  (samples taken from persons
having died of head fractures) and this was a phase that we had
to add to those previous methods.  We prefer both organic and
detergent agents for washing hair samples before ashing procedure.
GIBBS  (Canada)
     Dr. Vuori has drawn attention to the problems of sampling
and washing hair before analysis.  The turnover rate of hair is
also likely to limit its usefulness for assessing loner-term
exposure.  For many years the analysis of hair and nails has
been accepted as a method of demonstrating the ingestion of
arsenic compounds.  It is not yet clear how the other trace
elements we are discussing enter the body or become concentrated
in hair.  Elements may be absorbed through the gastrointestinal
tract, or through the lungs, or through both, depending on the
element in question.  In addition, metals might be adsorbed to
the outer surface of the hair.  These factors must be taken into
account if hair is to be useful for the evaluation of either
exposure or absorption.

-------
                                2297
BENINSON  (Argentina)
     Hair measurements could be indicators of  intake over  a
given period, even if concentrations  in hair do not correlate  with
concentrations in tissues.  This can  be due to different retention
functions and residence times, and different characteristic
integrating times of each integrator.
GIBBS  (Canada)
     This is a very valid point and Dr. Stankovic has  shown
quite clearly that the concentrations of  lead  in hair  can  be
used for such purposes.  In our very preliminary report we
indicated that trace element concentrations in hair did not
appear to be related to the concentrations in  lung.  This  may
certainly have been because elements had  been  removed  from the
lung or because the elements present in the hair entered the
body by a route other than through the lung.
PISCATOR  (Sweden)
1.   In the case of cadmium, it is impossible to distinguish
between endogenous and exogenous cadmium in hair.  Cadmium
from air will be as firmly bound as cadmium from the body.

2.   Metals such as Cd deposited in hair will be absorbed at
different rates dependant on the pH of the hair.
WEBER  (Yugoslavia)
     This worry does not apply to cadmium only, it applies to
lead as well.  Perhaps we have an extreme case in occupational
health problems, but lead is ubiquitous and one can just by
washing hair, or just walking on the street get some lead
deposited on the hair and it is a question whether it can be

-------
                                2298
removed by ordinary washing or not.  Another point that  I want
to make is: what does washing of hair before analysis, really
mean?  How much lead or cadmium or any other trace elements are
removed.  Some of the metals are built into the structure of
hair, hair being a protein of the keratine type, where sediments
in general are quite strongly bound to different groups  in the
structure of the protein.  On the other hand some people wash
hair before the analysis in 1% nitric acid, and I am quite sure
that under these conditions part of the absorbed metals will be
removed.
BERNSTEIN (Canada)
     In testing native Canadian Indians for mercury levels, we
found they were often more hesitant to give up a strand of hair
than to allow a specimen of blood to be withdrawn.
     Has anyone had experience in the determination of hair
selenium levels?
CLEMENTE (Italy)
     We measured in Italy the Se hair concentration in three
different population groups exposed to quite different Hg con-
tamination.  Any significant difference in the Se hair concen-
tration was observed as a function of the Hg level in hair.
The average Se concentration in the hair of the three population
groups was about O.I - O.3 ppm.  These data on Se will be
published in the near future.
HINE  (U.S.A.)
     The skill of the analyst in determining small concentrations
of metals in the hair has led to certain problems in diagnosis
and therapy.  The findings of elevated levels of metals in the

-------
                                 2299
hair has resulted, on the part of some physicians not skilled
in environmental and occupational mechanisms, in the employment
of chelate therapy in an attempt to reduce the presumed increased
body burden of that metal.  The analyst should lend his skills
to a proper interpretation of the significance of the findings.
Certainly therapy should not be administered unless there are
significantly elevated concentrations in the blood and clinical
signs and symptoms confirmatory of a diagnosis of metal intoxication,
KJELLSTROM (Sweden)
     It is very important in comparisons of hair levels of
metals with the levels in other tissues to define what type of
relationship one is studying.  As Dr. Beninson Indicated you have
to define if you are trying to evaluate the hair levels as an
indicator of recent exposure, concentration in critical organ
or total body burden.  In the case of cadmium I believe the
reports at this Symposium and earlier reports show that hair is
not useful as an indicator of any of these variables.  Nordberg
has shown with radioactive cadmium no correlation between cadmium
in blood and hair, in mice.  Watanabe in Japan has not found hair
to be useful in evaluating exposure level in an area where water
but not air was polluted.
KREUZER  (8.R.D.)
     Wenn Haare Schwermetalle anreichern  (z.B. Pb, Cd) ware es
im Interesse der Interpretation und Vergleichbarkeit der erhaltenen
Ergenbnisse wichtig in etwa gleich alle Haare zu untersuchen.
Das dQrfte in der Praxis auf gewisse Schwierigkeiten stossen, da
selbst eng benachbarte Haare recht unterschiedlichen Alters
sein konnten.  Dieses Problem liesse sich durch Entnahme grosserer
Proben Haare losen, doch lauft dies wieder den Bemuhungen einer
Entnahme moglichst kleiner Proben zuwider.

-------
                                2300
     When hair is burdened with heavy metals  (e.g. Pi, Cd) it
would be of advantage for the interpretation and comparison of
the results obtained if all the hair was examined equally; this
would be difficult in practice as even neighbouring hairs can
vary considerably in age.  This problem could be solved by taking
greater samples of hair, nevertheless this would be contrary to
the effort to keep the samples as small as possible.
WEBER (Yugoslavia)

     The speakers refer to the question, how are trace elements
distributed in hair, for example along its length; now we have
heard that also there could be some differences between two
adjacent hairs.  What about a single hair along its length.
Recently I received a communication from Columbia University in
New York where they told me that there is a maximum concentration
at about 7 cm from the scalp regardless of the age or of any-
thing else, and another peak if I remember correctly at 23 cm.
I just could not interpret that and I wonder if anyone has some
experience in this respect.
TRUFFERT (France)

     La fixation des oligo-elements dans les cheveux semble se

faire sur les groupements-SH liberes par la reduction de la
cystine en cysteine.  On peut realiser une telle fixation

in vitro et apre*s reoxydation qui regene're la keratine, on

ne peut distinguer 1'oligo-element ainsi fix! de celui aui

existait in vivo.  On peut done penser que la fixation des

oligo-elements dans les cheveux se fait avec la croissance
des cheveux, en fonction de leur teneur dans 1'organisme a

ce moment.


    Trace elements appear to be fixed in the hair in the SH-
groups released by the reduction of cystine to cyeteine. This
fixation can be achieved in vitro and, after the keratin has
been regenerated by reoxidationt it is impossible^ to distinguish
a trace element fixed in this way from one existing in vivo.
It may therefore be assumed that the fixation of trace
elements in  the  hair is concurrent with the growth of  the

-------
                                2301
hair and proportionate to the levels of trace elements in the
body at a given moment.
WEBER  (Yugoslavia)
     This is quite correct, metals can be fixed either through
metabolic processes or from the outside.  It is the same thing
as in dying wool, if you like.  But the fact that in different
subjects there is always a peak at 7 and 23 cm starting from the
scalp intrigued me.
BRAETTER  (B.R.D.)
     Bei unseren noch laufenden Untersuchungen von Haaren mehrerer
peruanischer Mumien, haben wir ebenfalls ein signifikantes
Maximum der Spurenelementkonzentration im Abstand von 6 bis 8 cm
von der Haarwurzel festgestellt.  Eine Erklarung dieses Effektes
haben wir auch noch nicht gefunden.

     We are at present carrying out researches on the hair of
several mummies from Peru.  We too have found a significant
maximum of the trace elements concentration at a distance of
6 to 8 cm. from the hair root.  We have not yet found an
explanation for this effect.
WEBER (Yugoslavia)
     Having exhausted the question of hair we move on to the
lungs.  What is the value of analysing lungs for trace elements
Do they reflect trace element exposures?
GIBBS  (Canada)
     The analysis of lungs for trace elements Is time consuming
and the reasons for carrying out such analyses should be clearly
defined.  We know that airborne particulates containing trace

-------
                                2302
elements enter the lungs, some are retained, some dissolve and
are absorbed, and other particulates may be removed by other
mechanisms.  For certain insoluble particulates penetrating
deeply into the lung, the lung might act as a collector and
there may be some instances where trace element analyses of
lung may be useful.
     In order to study specific problems such as the relation-
ships between trace elements and lung cancer, there may be some
value in measuring the trace element distribution within the
lungs of cases and suitably matched controls.  The analysis of
lung to monitor the trace element exposure of the general
population is questionable, and I suspect that such analyses
will not prove to be very useful.  We are currently studying
this problem and hope to examine the relationship between trace
elements in lung and community air.  The likelihood of finding
a strong association is slight because of the removal processes
mentioned above, and because of the mobility of the population.
WEBER (Yugoslavia)
     For epidemiological studies there is an additional problem
of biopsy.
GIBBS (Canada)
     Yes, this can be a serious problem.  It is necessary to
rely on the collaboration of the pathologist and pathology
technician, and often whole lungs are not available because
segments are needed by the hospital for other purposes   When
the specimen is taken, it is important to avoid contamination
by talc, etc., from surgical gloves, washing of the specimen
and fixing in formalin.  These two latter processes may add or
remove trace elements.  Many of the problems facing the
investigator are not related to the analytical methods but to
the collection and handling of the specimen.  The investigator

-------
                                2303
is able to supervise closely the analytical stages but often has
to take specimen collection on trust.
WEBER  (Yugoslavia)
     Touching on the last question, how should the results of
trace element analysis be expressed, for example, pg/g ash,
yg/g wet tissue?  This applies not only to lung but to other
tissues as well, but what troubles me is: what does it really
mean to have sophisticated analytical procedures working hard
to try to get results with 3 or 4 significant figures when the
actual concept of the tissue as such is not defined.  Is it
better to express it as dry or wet tissue no matter how wet or
dry it is, if we express it per gram of ash has this tissue
been calcified or are some other problems involved in the tissue
depending on the height and size of the person in question, for
instance?
KJELLSTROM  (Sweden)
    If space is available I think that both wet weight, dry
weight and ash weight should be reported in order that studies
should be comparable.
PITTWELL  (U.K.)
     It is my experience when analysing other body tissues than
those discussed  (lymph glands, muscles and body fat) for trace
elements, that there are problems of water loss which start
the moment the tissue is removed from the body.  If as in the
case of my own study the trace elements may be present as
inorganic inclusions, it is essential to know the relation to
the organic content of the tissue, then the only safe way of
returning results is to determine with respect to wet, dry and
ash weights.  This can be done by quoting the drying loss as

-------
                                2304
percentage, the ash percentage and the analysis of the ash  for
trace elements.  In some instances a relation to carbon content
can be used as well, but this is harder to determine.

-------
            UNTERSUCHUNG DER GESUNDHEITL1CHEN WIRKUNGEN
                     HEALTH EFFECTS STUDIES
                 ETUDES DES EFFETS SUR LA SANTE
                STUDI DEGLI EFFETTI SULLA SALUTE
         ONDERZOEKINGEN NAAR GEVOLGEN VOOR DE GEZONDHEID
Voraitzender - Chairman - President - Preeidente - Voorzitter

                       H.E. GRIFFIN (U.S.A.)

Stellvertretender Voreitsender - Vice-Chairman - Viee-pr&eident
                Vioepreeidente - Viae-VoavaSttev

                       A.E. MARTIN (U.K.)

-------
                               2307
         HEALTH EFFECT STUDIES - NOW AND IN THE FUTURE

                         LARS FRIBERG

Department of Environmental Hygiene, The Karolinska Institute,
and the National Swedish Environment Protection Board, Stockholm
Sweden
ABSTRACT

     Health effects, panging from alight annoyance reactions at
the one extreme and death at the other, can be studied toxicolo-
gieally and epidemiologically.   Toxioologiaal studies must elu-
cidate what metabolites are formed by a given substance and what
mechanisms of action it has in nature and in the body.   The ul-
timate aim of such studies is to obtain a dose-response curve,
which is often difficult.   Much valuable epidemiologioal and
toxicologioal data are lost to the researcher simply because in-
dustrial health officers do not use their resources for the col-
lection and compilation of such data or, if they do, they must
often keep the results confidential.   The TLV values for indus-
trial exposure must be reevaluated since they do not always pro-
tect the worker.   Differences in prevalence of disease among
countries both from environmental and industrial agents have not
yet been satisfactorily accounted for.   Epidemiologioal base
registers should be initiated or, when they exist, expanded, for
example through material collected from questionnaires.   factors
of self-selection must be given due regard when data on voluntary
exposure to harmful agents, e.g. cigarettes, are to be interpreted,
Studies on twins which are discordant as to the exposure to a
certain agent may be fruitful in this respect.    Vital statistics,
cancer registries and hospital records should be given more en-
trance into epidemiological studies.   A plea is made for more
extensive training of epidemiologists and toxioologists.

-------
                              2308
Health effects cover a wide range of biological responses.
Death and disease represent one extreme of the whole spectrum
of response. At the other extreme will be found physiological
and other changes of uncertain significance, e.g.  increased
body burdens of pollutants as well as annoyance reactions due
to bad odors or noise. Health effect studies usually involve
humans or animals but may also be carried out in vitro using
different cell systems. They are as a rule of multidisciplinary
nature, utilizing medicine, chemistry, physics or behavioral
sciences.

The two major approaches are toxicology and epidemiology. Toxi-
cological studies use animals of different species, Such stud-
ies may range from more conventional determinations of acute
and chronic toxicity to sophisticated studies of metabolism,
mutagenic, carcinogenic, and teratogenic effects.  Epidemiologi-
cal studies on the other hand aim at mapping biological effects
as they occur in reality, meaning that primarily studies on
humans, often large groups, have to be embarked upon.

When new substances are intended to be brought to market, the
first step will be toxicological studies. Even if such studies
are carried out, using all available techniques, data from
animal experiments cannot easily be extrapolated to humans.
Epidemiological studies are therefore often imperative in order
to ascertain that the results from toxicological studies on a
given substance hold true. Furthermore, epidemiological studies
are indispensable as a form of health surveillance of trends in
incidence of disease and mortality in order to detect at a very
early stage health effects where no suspicion towards a speci-
fic axposure has arisen.

Scientific and practical requirements differ for different
types of health effects. The basis for study is, however, al-
ways a need for understanding the mechanism behind possible
effects and the documentation of the dose-response relation-
ships.  Several problems will come up which often will motivate

-------
                              2309
massive efforts, the magnitude of which will depend on inter
alia the risks the community is willing to take and the number
of people who may possibly be exposed.

There are many examples of the difficulties in carrying out
health effect studies. Here may be referred to the fact that
despite overwhelming evidence of a causal association between
cigarette smoking and lung cancer, this effect has not been
possible to reproduce unequivocally in animals.

It goes without saying that even if governmental and private
support for toxicological and epidemiological evaluations
should grow dramatically, only a small part of the ever-in-
creasing numbers of chemical substances brought to market can
be studied in a satisfactory way. This will be all the more
obvious when possibls interactions between pollutants, often
with potentiating effects, are considered. The toxicity, not
only of the substance itself, but also of its metabolites,
must be investigated. Metabolism may vary within a species and
among different species. This, together with different toxicity
or different exposure routes, complicates further any toxico-
logical evaluation.

Mercury gives a good example of the complexity of the toxico-
logical situation. The tragic incidences  of mercury poisoning
via  fish in Minamata and Niigata in Japan and recently, on a
large scale via a seed dressing in Iraq,  are well known. Those
mass outbreaks were caused by methyl mercury, an organic mer-
cury compound which is one of the most toxic mercury compounds
known, also involving a high risk for the fetus. It would be a
serious error, however, to conclude that  other organic mercur-
ial  compounds,  like phenyl mercury and mathoxyethylmercury,
will also have a similar toxicity. Unlike methyl mercury, these
 organic  compounds  are  readily  broken  down within  the  body,
 forming  mercuric  ion,  and  the  toxicity  is lower  and  more
 similar  to  the  one  experienced  after  exposure to  mercuric  mer-
 cury.  Metallic  mercury,  on  the  other  hand,  has  a  toxicity  which

-------
                               2310
differs from that of methyl mercury, other organic mercury
compounds and mercuric mercury salts. After absorption, it
exists for a short time as physically dissolved mercury vapor
in the blood, and in this form it easily penetrates the blood-
brain barrier and the placenta. As for accumulation and excre-
tion of absorbed mercury, methyl mercury has a biological half-
time in all organs of on an average 75 days. The biological
half-time of inorganic mercury is only a couple of months in
the blood, while, when it has reached the brain, the half-time
will be in the order of years. As regards the toxicological
evaluation of mercury, the possibilities of methylation in
nature, meaning that mercury can be transformed into methyl
mercury, must also be borne in mind.

When it comes to the relation between dose and response (Dose-
response curves) the magnitude of our efforts will depend on
the risks we are willing to take as basically it will be vir-
tually impossible to show a true no-response dose. Not even
for cigarette smoking, with all the millions of people studied,
do we know the exact relation when it comes to a low exposure.
We know that lung cancer is strongly and causally associated
with smoking but we cannot quantify the risks, if any, of
smoking say one or two cigarettes a day. It could be argued
that this is of minor importance as from the health point of
view a proper decision taken by administrators could be to
prohibit smoking. Apart from the fact that this for several
reasons is not possible, and. could lead to several very unde-
sirable effects due to the addictive nature of the smoking
habit, the question has an obvious bearing in relation to
passive smoking, meaning the tobacco smoke that nonsmokers
are exposed to. We know that concentrations of particulate
matter in rooms where several smokers are together easily will
reach concentrations in the mg range, far above concentrations
of particulate matter which by the WHO have been considered to
give rise to health effects.  One problem is that a combination
of particulate matter and sulfur oxides was considered in the
WHO criteria.  Tobacco smoke consists of particulate 'matter
together not with sulfur oxides but with a large number of

-------
                              2311

other gases.  There are some data from England which are highly
suggestive of respiratory effects in young children due to
passive smoking,  More extensive data are needed before defin-
ite conclusions can be drawn, however. To come up with dose-
response relations for passive smoking not only for the average
adult but also for children and people with pre-existing dis-
ease would involve large effort, taking into account among
other things that there are reasons to believe that tobacco
smoke as related to passive smoking would differ from freshly
produced tobacco smoke.

I sha.ll now turn to a few questions in connection with industr-
ial exposure to chemical substances. It is customary in health
effect studies to consider separately studies which are based
on industrial exposure, exposure via food, and exposure within
the general environment, e.g. ambient air and water. Admittedly
there are large differences between such different exposure
situations both in regard to the magnitude and the route of
exposure. Furthermore, the populations exposed may differ con-
siderably in relation to time of exposure and sensitivity to
the different agents.

On the other hand, it is obvious that very often our best op-
portunities to get information on effects of toxic substances
are found within  industries, particularly as the exposures
often are so much higher there.  It  is also obvious that in the
extensive criteria documents by  the US Environmental Protection
Agency much attention has been paid to results from industrial
exposure. It is certainly most unfortunate that  data from in-
dustrial  exposures are so relatively scarce as they are con-
sidering  the number of people who are exposed to different
substances. It is the more unfortunate in view of  the fact that
the possibilities of making  good studies within  the general
scope of  the industrial health services are so enormous. The
lack of data lending themselves  to  dose-response evaluations
was pointed out by a Task Group  on  Metal Accumulation at a
meeting in September  1972, for metals, but seems to be valid
also for  almost any other substances. It was then  said that

-------
                              2312

there is a lack of epidemic logical data from populations with
industrial exposure. This lack is even more striking in rela-
tion to the growing number of such studies on populations with
general environmental exposure, which is usually at a much
lower level.  There is a need in industrially exposed popula-
tions for standardized, wherever possible collaborative, epi-
demiological studies, where cohorts can be followed in time
and where groups can be related to each other.

The lack of published data from industrial exposure certainly
is due to several reasons. Very often studies are not carried
out at all within industries, responsible officers relying on
recommended TLV-values, which, however, are notoriously unre-
liable and should be completely reevaluated. One recent tragic
example concerns vinyl chloride, for which the TLV is definite-
ly too high.  It is set by the American Conference of Govern-
mental Industrial Hygieniata at 200 ppm (in Sweden set at 500
ppm) despite the fact that in the accompanying documentation
for this value there are data which indicate toxic effects on
the liver at concentrations even lower than 200 ppm. Now expo-
sure to vinyl chloride is associated with liver cancer in
humans.

In cases where industrial health studies are carried out there
are examples showing that management is unwilling to release
data which later can be used against them in connection with
legal procedures. Furthermore, very often the number of people
exposed within a given industry for a given substance may be
small. This means that statistically significant effects, when
comparisons are made against a control group, may not be de-
tected even if such effects in reality exist. In order to show
with reasonable assurance that a certain effect, also a very
severe one, does not occur in an increased prevalence in an
exposed group compared with a control group the study popula-
tions often have to be large. This could be accomplished if
collaborative studies between industries were carried out even
on a rather limited scale. To me, the question of increasing
the number of studies, improving their quality and providing

-------
                              2313
for their free publication has for the time being the greatest
potential for epidemiological evaluations of toxic substances.
This potential is valid not only for those who have the res-
ponsibility for protecting the industrial worker but also for
other health agencies. Collaboration on all levels is necessary

Collaborative efforts are also desirable on the international
level. There are several examples where an effect in one coun-
try may not be reproduced in another country despite apparently
similar exposure situations. This has been shown in connection
with international studies of lung cancer and of cardiovascular
disease. The reasons for differences in prevalence are not
clear but cannot simply be explained by the occurrence of
commonly recognized risk factors. There are other examples
which show the opposite. I would like to refer to recent stud-
ies into the effect of aircraft noise reported at this symposi-
um by a Swedish group. Despite rather large differences in cul-
tural settings it seems as though when the proper exposure and
effect parameters are used annoyance reactions due to aircraft
noise are very similar for a given dose in Sweden, France,
England and Japan.

For most epidemiological studies the aim will be to study
individuals with known exposure situations and not only to
carry out macroBpidemiological studies monitoring exposures
and effects without clear relation to each other. I would like
to advertize here the possibility of using mail questionaires
to a larger extent than hitherto. Certainly already by means
of this simple tool a considerable amount of exposure data can
be made available. This has been shown among other things in
Swedish and US studies on twins .and English, Norwegian, and
US studies on immigrants. The Swedish and US data on twins in-
cluded data on smoking habits, drinking habits, food and drug
consumption, personality factors, occupation and exposure to
air pollution. Such data may form the nucleus for an "epidemio-
logical base register" as it is often too late to get such in-
formation at a later stage when one wants to correlate effects,

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                              2314
e.g.  disease,  death,  or annoyance reactions,  with exposure
data. It would be possible, on a global basis and for reason-
able costs, to carry out such studies which would give much
better possibilities in the future to interpret differences
between different populations in morbidity and mortality
rates.

For certain types of health effects studied group comparabil-
ities between exposed group and controls are particularly
crucial. This refers to comparison of groups who to some ex-
tent are self-selected. I am thinking of studies where smokers
are compared with nonsmokers. There are good reasons to believe
that a smoker does not become a smoker by chance only but that
smokers and nonsmokers differ considerably in many other as-
pects than smoking habits. For example, it has been shown in
Swedish studies that nonsmokers are registered in a nationwide
alcohol registry to  10%, those who smoke between 1 and 10
cig/day to about 20% and those who smoke more than 10 cig/day
to about 30%. Under  such circumstances studies on twins dis-
cordant for the agent to ba studied may be of particular val-
us. Studies involving discordant monozygotic twins would keep
genetic factors under complete control and it has also been
seen that  such pairs are more similar in relation to several
confounding background factors.

Although it is obvious that epidemiological studies if possible
should be  of  the prospective type where exposure and effects
are studied on the individual basis other forms of health sur-
veillance  may also be of considerable value. I am thinking
particularly  of using data from vital statistics, cancer reg-
istries, hospital records. Such data will never be conclusive
in themselves but may serve as early warning systems for health
effects. Ideally such data should be linked with data from more
conventional  studies.

A question of particular importance refers to the training in
toxicology and epidemiology. With very few exceptions there  is

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                             2315


a lack of toxicologists and epidemiologists. There is an ex-

treme need for improving and expanding the training facilities

both in individual countries and on an international scale.

Not only are trained toxicologists and epidemiologists needed

for carrying out toxicological and epidemiological studies;

administrative bodies also need the advice of such personnel.

The evaluation of scientific data from the literature, meaning

working out criteria documents, is an extremely qualified task

and can only be carried out properly by scientists who them-

selves are familiar with the research methods used.
                           DISCUSSION
LEHTO (Finland)

     You mentioned that "management IB unwilling to release data,
which later can be used against them In connection.with legal
procedures".

     How do you see scientist's role in this situation?  Is it
enough that we see ourselves only as specialist who answer only
questions which are asked or put by administrators?


FAIBERG  (Sweden)

     This is first of all a question of education of management.
They must learn that in the long run it is for their own benefit
that data are published/ as such information must be the basis for
protective measures not only in the industries with which they
are concerned but also in other industries.  Scientists have
an extremely important role to play in such an education.


MAGE (Denmark)

     In view of the past history of industry, as in the case of
asbestos where the company doctors knew of asbestosis and meso-
thelioma in asbestos workers and did not disclose it, do you
think that labor unions should put into their contracts that
epidemiological studies shall be undertaken and published?

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                              2316
FRIBERG (Sweden)

     This might well be a recommendable way.  Again, however,
both management and labor unions should learn to understand that
the publishing of data and the proper execution of epidemiological
studies are for the benefit of both parts.  Labor unions should
also make sure that they have at their disposal highly competent
scientists in both epidemiology and toxicology.  This is often
not the situation at present.


RAMACIOTTI (Switzerland)

     Since there is a kind of automatic selection of industrial
workers, i.e. the most threatened persons are eliminated, do
you not think it dangerous to use results of epidemiological
studies carried out in industry to draw up standards for the
entire population?


FRIBERG (Sweden)

     I agree completely.  Due attention must be paid to differences
in sensitivity.  Workers who for one reason or another have
terminated their employment at a given industry should not be
omitted in epidemiological studies.  They may indeed form the most
important subgroup.  Moreover, the general population does not
consist solely of healthy people and encompasses both age extremes.
The exposure time, further, is quite different from what is ex-
perienced in industry.

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                               2317
   THE CARCINOGENIC RISK FOR MAN OF ENVIRONMENTAL CHEMICALS

                          L, TOMATIS

International Agency for Research on Cancer, Lyon, Prance
ABSTRACT

     The hypothesis that environmental factors play an essential
vole in the aetiology of human oanoer is receiving wide support
and it has been repeatedly asserted that over 80% of hitman can-
aers are due to environmental factors.

     Environmental factors may act directly on individuals exposed
and may affect pregnant women and therefore reveal an effect on a
subsequent generation.

     A direct effect of environmental factors in producing cancer
in man has been shown by:  (1) the unequivocal evidence of the
chemical origin of occupational cancer^ as in the oases of urin-
ary bladder tumours in workers exposed to aromatic amines^ lung
cancers in workers exposed to bis(chloromethyl)ethert eto.s
(2) the well-documented cases of iatrogenic cancers;  (S) the
positive correlation between cigarette smoking and lung cancers
(4) the different cancer incidences in urban and rural populations
and the possible role of air pollution;  and (S) the results of
studies in migrants showing that for some cancers migrants acquire
incidences similar to those of the host countries.   Results ob-
tained in experimental carcinogenesis are in keeping with a direct
carcinogenic effect of chemicals.

     The evidence that exposure of pregnant women to a carcinogen
may result in an increased risk of cancer in the progeny has been

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                              2318
provided by the reports on the incidences of vaginal cancer in
daughters of women exposed to stilboestrol during pregnancy,
The role of prenatal exposure in determining cancer risks in the
progeny is fully documented in experimental carcinogenesis.

     The possibilities of removing or preventing carcinogenic
hazards to man in the environment will be discussed.

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                                   2319
     It has been stated repeatedly that 80% or more of all human cancers
nay be attributed to environmental factors.  This statement, discussed,
but never seriously challenged, has been widely quoted and in practice
accepted.  This certainly has not made prevention of cancer any easier.
     Whilst it seems futile to dispute the real proportion of environ-
mentally-related cancers in the absence of unequivocal evidence, or at
least sufficient supporting data, this presentation will be focused on
situations in which evidence of the role of environmental factors is well
documented.  The purpose is to stimulate efforts towards the ultimate goal
of our activity, that is, the prevention of cancer.  In the case of
environmentally-related cancers, prevention can be achieved by the
identification and then the removal, or at least substantial decrease, of
causative agents.  Whilst it seems quite justifiable to concentrate part
of our efforts on the search for new environmental hazards, it would be
totally unjustified not to operate on hazards for which abundant evidence
already exists.
     A direct effect of environmental factors in producing cancer in man
has been shown by:  (1) the unequivocal evidence of the chemical origin
of occupational cancer, as in the cases of urinary bladder tumours in
workers exposed to aromatic amines, lung cancers in workers exposed to
bis(chloronethyl)ether, etc.;  (2) the well-documented cases of iatrogenic
cancers;  (3) the positive correlation between cigarette smoking and lung
cancer;  (4) the different cancer incidences in urban and rural popula-
tions and the possible role of air pollution;  and (5) the results of
studies on migrants showing that for some cancers migrants acquire
incidences similar to those of the host countries.  Results obtained in
experimental carcincgenesis are in keeping with a direct carcinogenic
effect of chemicals.
     Evidence that certain types of cancer were related to a particular
occupation goes back to the 17th century, when the first observation of a
high frequency of lung cancer among workers of a uranium mine were
reported.  A century later cams the observation of Percival Potts on the
high incidence of scrotal cancer in chimney sweeps, and a little more
than a century later,  high incidences of urinary bladder cancer were
observed in workers employed in dye factories.  Successively, exposure to
a number of other organic and inorganic substances, such as asbestos,

-------
                                  2320
chromates, nickel, mustard gas, etc. have been shewn to result in a high
cancer risk.
     Two preliminary conclusions can be immediately drawn:   (1) the
interval between the first observation of a causal relationship of
exposure to an environmental factor and cancer, and the adoption of
preventive measures is unjustifiably long;  and (2) to the majority of
chemicals recognized as being human carcinogens, man is exposed because
of his occupation.
     In relation to the first conclusion, it is perhaps worth mentioning
that the carcinogenic risk present in dye factories was first reported in
1896 and confirmed in 1907  (Hueper [1]), but this observation of a
carcinogenic effect in man was not judged sufficient and experimental
evidence was required.  In 1938 Hueper succeeded in inducing urinary
bladder cancer in dogs with B-naphthylamine.  It was only 20 years later,
however, that some measures were taken to prevent massive exposure of
workers to 3-naphthylamine and this was done only after the masterly work
of Case et al. [2,3] had again demonstrated beyond any doubt that both
naphthylamine and benzidine were carcinogenic to man.  In spite of this,
there were factories in seme countries in which only very recently even
elementary measures of precaution were ignored.
     As an example of a carcinogenic drug, I would like to mention
chlornaphazine, an aromatic amine pertaining to the same chemical group
as several other known carcinogens.  The fact that an aromatic amine has
been used in human therapy, ignoring its chemical parenthood to known
human carcinogens, is an indication, among others, of the fragility of
the present prevention system and of the limited diffusion given to
information on occupational cancer.
     Evidence of a carcinogenic effect in man of cigarette smoking has
     *
been abundantly demonstrated.  The risk of lung cancer in heavy cigarette
smokers is high but decreases if exposure to cigarette smoke is discon-
tinued, but still remains higher than in people who have never smoked.
According to Doll [4] "the effect of stopping exposure to a carcinogen
after a prolonged period of exposure is likely to vary with the nature of
the carcinogen".  It also depends, for a given carcinogen,on the different
target organs, as was the case of the persistence of the risk of
developing nasal cancer and the decreasing risk of developing lung cancer

-------
                                  2321
40 years after cessation of exposure to nickel carbonyl  (Doll et al. [5 3).
     Cancer incidence varies markedly between urban and rural areas.  Hie
differences are perhaps more evident for lung cancer but are certainly
not limited to one cancer site.  The incidence of colon cancer, for
instance, is higher in urban than in rural areas, while rates for stonach
cancer show a reverse trend.  A higher mortality rate from lung cancer in
urban areas than rural areas has been observed in Norway  (Kreyberg  [6]),
in the United States (Carnow and Meier [7 ], Particulate Polycyclic
Organic Matter [8]), and in the UK  (Lung Cancer in Western Europe [9]).
These differences persist even when rates for smoking habits are stan-
dardized.  In general, there is a positive correlation between mortality
rates for cancer of the lung and population density and urbanization
index.
     The role of environmental factors in altering cancer risks has
received further confirmation fron studies on migrants.  Assuming that
migrants are representative samples of the population in hone countries,
the differences observed in their cancer experience after their settle-
ment in a host country could be attributed to changes in environmental
conditions.
     Studies on Japanese migrants to the USA have shown that:  (1) morta-
lity from colon cancer, which is rather low in Japan, has undergone a
sharp upward shift in Hawaiian born Japanese and to a lesser extent in
Japanese migrants in whom the magnitude of the risk was related to the
period of residence in the host country (Haenszel and Rurihara [10],
Haenszel et al, [11]);  (2) mortality from stomach cancer, which is very
high in Japan, was lower in Japanese migrants and decreased markedly in
Japanese born in the USA;  and (3) mortality from breast cancer,  which is
very low in Japan, increased considerably in Japanese born in the USA
(Buell [12]).  This last observation is based on a study involving a
limited number of cases, and contrasts with previous observations
indicating the persistence of a low incidence of breast cancer in US-born
Japanese.  It is worth noting that the apparent persistence of low rates,
even after two generations, was taken as a demonstration that for breast
cancer, host factors or genetic factors played a preponderant role (Muir
[13]).  However,  Buell's report [12] indicated that environmental factors
also play a role on the incidence of breast cancer, which is possibly a

-------
                                  2322
good example of an interaction between host genetic and environmental
influences.
     A comprehensive review of migrant studies published recently  (Kmet
[14]) lists the factors which may be suspected to cause the change in the
cancer experience of migrants.  In some instances, and for sane types of
cancer, the available data indicate that events occurring in adult life
may change the cancer risk at a specific site within two decades
(Haenszel et at, [11]).  This is the case for incidence of bowel cancer
in Polish migrants to Australia and to the USA  (Staszewski and Haenszel
115], Staszewski et al,  [16]).  On the other hand, studies of migrants
from the UK to New Zealand {Eastcott [17]) and to South Africa and
Australia  (Dean [18, 19]) seem to indicate that exposure to pollution
early in life may produce persistent effects (Camow and Meier [7]).
     Studies of within-country migration offer good possibilities for
evaluating the inpact of a changing environment.  In a recent study,
Cutler and Devesa [20] reviewed studies in cancer incidence and mortality
in the USA for white and black populations.  Incidence of mortality from
cancer of the oesophagus, colon, prostate, lung and pancreas have sharply
increased among the black population, and in general the rate of increase
has been much higher than in whites for cancer of the lung, pancreas and
colon, even though a considerable increase has also been seen in whites.
A dramatic increase among blacks was observed for oesophageal cancer,
which is on the decrease among whites.  Although better medical care may
play a limited role, the dramatic upward shift in blacks appears more
likely to be attributable to the environmental changes the black popula-
tion have undergone in the last decades and, in particular, migration
from rural areas to the cities, and changes in occupation and in
standards of living.  Mortality and incidence of stomach cancer have
decreased in both whites and blacks, but in the latter at a slower pace
than in whites.
Possibility of prevention
     One may dispute the real percentage of all cancers related to the
environment and, in particular, to chemical agents.  What is indisputable,
however, is that at present,  primary prevention, that is the prevention
of exposure to aetiological factors, offers the best perspective for a
successful anti-cancer campaign.  It also seems the most reasonable

-------
                                    2323
approach if one expects to look at cancer as one among others and not the
only disease which may be environmentally related.  The urban and
industrial environments and air pollution are, in fact, not only respon-
sible for an increase in cancer incidence but in the incidence of other
chronic diseases, such as chronic bronchitis.  Toxic contaminants which
may be present in larger quantities in rural than in urban areas are
probably responsible for a variety of diseases other than cancer as, for
instance, seems to be the case of mycotoxins and aflatoxins in particular
 (IARC Monographs on the Evaluation of Carcinogenic Risk of Chemicals to
Man [21]).  In addition, a number of environmental contaminants may be
carcinogenic, rnutagenic or teratogenic or they may well have all three
characteristics.  Efforts towards a primary prevention of cancer can
therefore be inscribed within a broad approach towards a healthier, or at
least less deadly, environment.
     The removal, or at least drastic reduction, in occupational exposure
to hazardous chemicals may be obtained in the majority of cases by an
improvement in manufacturing procedures.  A good example of the success
in prevention which can be obtained in this way is the disappearance of
the excess risk for nasal sinuses and lung cancers din nickel workers in
the UK and Norway who began their work after an industrial process had
undergone major changes in order to eliminate exposure of workers to dust
and fumes (Doll et al. [5], Pedersen et al. [22].  The case of recognised
chemical carcinogens is, or at least should be, clear, and everybody
surely agrees that human exposure to carcinogens must be avoided.
     A major problem is the evaluation of the possible carcinogenic effect
of a chemical to man in the absence of epidemiological studies or case
reports.  As there are no objective criteria to extrapolate from experi-
mental data to man, no scientific prediction can be made.  Present
regulations, however, recamend a zero tolerance in food for chemicals
for which experimental evidence of carcinogenicity exists.  In so far as
food additives are concerned, therefore, an extrapolation for public
health purposes is carried out systematically with a procedure which
obviously implies that experimental evidence of carcinogenicity is
sufficient per se to suspect carcinogenicity in man (Tcraatis et al.  123]).
This cautious procedure, with which most people agree, has certainly not
been applied to other situations,  as for instance, chemicals present in

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                                   2324
industry and  in  the general environment.
     Tlie validity and limitations of experimental studies in preventing a
human hazard  could be illustrated by the examples of stilboestrol,
bis (chloromethyl)ether, vinyl chloride and DDT.  lhe experimental evidence
of the carcinogenicity of stilboestrol existed already in the fifties
when stilboestrol was used for human therapy in relatively high doses and
was given in  particular to pregnant women.  It was only at the end of
1970 after the report of an increased risk of vaginal cancer in daughters
of woman exposed to stilboestrol during pregnancy that some restrictions
on its use were  introduced.  Experimental evidence of the carcinogenicity
of bis (chloromethyl) ether existed in 1968 and was confirmed in 1969 but it
was only in 1973 after a retrospective epidemiological study showed an
excess of lung cancer in workers exposed to bis (chloromethyl) ether that
measures to reduce exposure in the working environment were adopted.
Similarly, for vinyl chloride, experimental evidence of carcinogenicity
existed already  in 1970 (Viola [24]) and was long preceded by the evidence
of toxic effects of the same type in man and experimental animals.  It was
only in 1974, however, that measures for drastically reducing its
acceptable concentration were adopted and only after a report on the
occurrence of a  rare type of cancer (angiosarcoma) of the liver among
workers exposed  to vinyl chloride.
     In these three cases a more cautious attitude would have resulted in
                                     i                                 ,
a successful  prevention.  We should perhaps not limit our role to that of
a sort of necrophore, waiting for  a  rare type of cancer to appear or for
a sufficiently high number of people to be killed by a less rare cancer
before being  ready to recommend preventive measures.
     The case of DDT is particularly complex because:  (1) it has already
been used for 30 years and not withstanding any measures taken now, it will
be in the environment for many years to come;  (2) its usefulness in
disease control  is beyond any doubt:  in particular, it has made the
malaria campaign a success which has resulted in saving millions of lives;
(3) its acute toxicity is relatively low and certainly lower than some of
its proposed  substitutes.  Its chronic toxicity is probably also low,
although a number of minor diseases may certainly go undetected;  (4) its
carcinogenicity was shown and confirmed in the mouse and was limited to
the liver.  A borderline carcinogenic effect was reported, but not

-------
                                  2325
confirmed, in the rat and a lack of carcinogenic effect was reported in
the hamster (Tomatis et al. [25], Turusov et at. [26], Thorpe and Walker
[27], Fitzhugh and Nelson  [28], Napalkov [29], IAKC Annual Report [30],
Agthe et al. [31]).  Measures to restrict and reduce the use of DDT have
been taken more because of its inpact on the environment at large than
because of the experimental evidence of carcinogenicity.  DDT is also an
example of a chemical for which a risk versus benefit evaluation could be
made (Safe Use of Pesticides [32]), with a procedure a posteviovi, which
cannot, however, be reccmnended.  In fact, whilst there is no obvious
evidence of a carcinogenic effect of DDT in nan, it is still not possible
at present to exclude it, as no proper epidemiological study on DDT-
exposed people has yet been reported.
     A risk versus benefit evaluation is currently being made when the use
of drugs employed in the therapy of cancer has to be decided.  As is well
known, most of these drugs are per ae carcinogenic.  Drugs for which
experimental evidence of carcinogenicity exists should not be used except
for life saving measures and in no case should a decision on its use be
made by an individual.
     The prevention of lung cancer related to cigarette smoking, although
theoretically easier to implement, has encountered serious setbacks and
there is no evidence at present that the sale of cigarettes has decreased.
There is some indication, however, that the introduction of the filter has
contributed to a slight decrease in risk (Wynder et al. [33]).
     Lung cancer is the greatest cause of cancer deaths in man.  Its
incidence has considerably increased for at least thirty years and is now
showing a strong upward shift in women.  This high uicidenoe cannot be
explained solely by cigarette smoking and must be, at least partly,
related to «jr pollution.  The incidence in urban areas is, in fact,
approximately double that observed in rural areas.  It has been estimated
that a 5% increase in the rate of death from pulmonary cancer can be
related to each increment of pollution by one microgram of benzo(a)pyrene
per 1000 cubic metres of air (Carnow and Meier  [7], Particulate Polycyclic
Organic Matter  [8]).  Conversely, it has been postulated that reduction of
air pollution would result in a substantial reduction in lung cancer death
rates.
     Studies on migrants may offer a clue for the identification of agents

-------
                                    2326
involved in the causation of human cancers.  The study of Haenszel et at.
[11] on the incidence of colon cancer in Japanese migrants is an example

of the possibility such studies may offer for the identification of
possible environmental factors.  On the other hand, it must not be
forgotten that the environment in its extended meaning includes social

status and that evidence exists that certain types of cancer occur with a

higher frequency among members of the lower social classes (Clemmesen,
Nielsen et al. J34]).
References

 1.   HUEPER,  W.C.,  "Occupational and environmental cancer of the urinary
     system",  Yale  University Press, New Haven and London,(1969).

 2.   CASE,  R.A.M.,  HOSKER,  M.E.,  McDONALD, D.B.,  PEARSON, J.T.  "TUTtOUTS
     of the urinary bladder in workmen engaged in the manufacture and use
     of certain dyestuff intermediates in the British chemical industry",
     Brit.  J.  Industr.  Med.,  11, 75-104 (1954).

 3.   CASE,  R.A.M.,  HOSKER,  M.E., "Tumours of the urinary bladder as an
     occupational disease in the rubber industry in England and Wales",
     Brit.  J.  Prev. Soc. Med., 8, 39-5O (1954).

 4.   DOLL,  R.  "Age", in: Host Environment Interactions in the Etiology of
     Cancer in Man, ed. R.Doll and I.Vodopija, IARC Lyon 1973, pp. 39 - 48.

 5.   DOLL,  R., MORGAN,  L.G., SPEIZER, F.E.,  "Cancers of the lung and nasal
     sinuses in nickel workers", Brit. J. Cancer,  24, 623-632 (1970).

 6.   KREYBEFG, L.,  "Aetiology of lung cancer", Universitetsforlaget, Oslo (1969)

 7.   CARNOW, B.., MEIER, P. "Air pollution and pulmonary cancer", Arch.
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 8.   Particular Polycyclic Organic Matter, National Academy of Sciences,
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 9.   Lung Cancer in Western Europe, Council of Europe, Strasbourg,  (1969).

10.   HAENSZEL, W.,  KURIHARA, M., "Studies of Japanese migrants. I. Mortality
     from cancer and other diseases among Japanese in the United States".
     J. Natl. Cancer Inst.. 40, 43-68 (1968).

11.   HAENSZEL, W.,  BERG, J.W., SEGI, M., KURIHARA, M., LOCKE, F.B., "Large-
     bowel cancer in Hawaiian Japanese", J. Natl. Cancer Inst.,  51, 1765-
     1779  (1973).

12.   BUELL, P., "Changing incidence of breast cancer in Japanese-American
     women", J. Natl. Cancer Inst.,  51, 1479-1483  (1973).

13.   MUIR, C.S., "Geographical differences in cancer patterns", in: Host
     Environment Interactions in the Etiology of Cancer in Man, ed. R.Doll
     and I. Vodopija,  IARC Lyon 1973, pp. 1-13.

14.   KMET, J., "The role of migrant population in studies of selected cancer
     sites: a review", J. Chron. Pis.,  23, 305-324  (1970).

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                                    2327
 15.    STASZEWSKI,  J.,  HAENSZEL,  VI.,  "Cancer mortality among the Polish-born
       in the United States.   J.  Natl. Cancer Inst., 35, 291-297 (1965).

 16.   STASZEWSKI, J., McCALL, M.G.,  STENHOUSE N.S.,  "Cancer mortality in
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      599-610 (1971).

 17   EASTCOTT,  D.F.,  "The epidemiology of lung cancer in New Zealand"
      Lancet,  l, 37-39  (1956).

 18.   DEAN, G.,  "Lung cancer  among white South Africans",  Brit.  Med.  J.,
      2,  852-857 (1959).                                   	

 19.   DEAN, G.,  "Lung cancer  in South  Africans and British inmigrants",
      Proc. R. Soc.  Med..   57, 984-987 (1964).

 20.   CUTLER,  S.J.,  DEVESA, S.S.,  "Trends in cancer incidence and mortality
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      Cancer in  Man, ed. R.Doll and I.Vodopija,  IARC Lyon  1973,  pp. 15-34.

 21.   IARC Monographs on the  Evaluation of Carcinogenic Risk of  Chemicals
      to  Man, Volume 1, IARC  Lyon  (1972).

 22.   PEDERSEN,  E.,  H0GEIVEIT, A.C., ANDERSEN, A.,  "Cancer of respiratory
      organs among workers  at a nickel refinery  in Norway",  Int.  J. Cancer,
      12, 32-41  (1973).                                      	

 23.   TCMATIS, L., PARTENSKY, C., MONTESANO,  R.,  "The predictive value of
      mouse liver tumour induction in  carcinogenicity testing -  a literature
      survey". Int.  J. Cancer,  12, 1-20  (1973).

 24.   VIOLA, P.L., "Cancerogenic effect of vinyl chloride",  X Int. Cancer
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 25.   TOMATIS, L., TURUSCV, V., TERRACINI, B., DAY, N., BARTHEL, W.F.,
      CHARLES, R.T., COLLINS, G.B., BOICCCHI, M.,  "Storage  levels Of DDT
     metabolites in mouse tissues following  long term exposure to technical
      DDT", Tumori,  57, 377-396 (1971).

 26.   TURUSCV, V.S., DAY, N.E., TCMATIS, L., GATI, E., CHARLES, R.T. "Tumors
      in CF-1 mice exposed for six consecutive generations to DDT", J. Natl.
     Cancer Inst., 51, 983-997 (1973).                             	

27.  THORPE, E., VRLKER, A.I., "The toxicology of dieldrin  (HEOD). II.
     Cccrparative long-term oral toxicity studies in mice with dieldrin.
     DDT, phenobarbitone, B-BHC and y-BHC",  Food Cosmet. Toxicol., 11,
      433-442  (1973).                         ™	

28.  FITZHUGH, O.G., NELSON,  A,.A., "Chronic oral toxicity of DDT", J.
     Pharmacol.  Exp. Ther..  89,  18-30  (1947).                     ~

29.  NAPALKDV, N.P. Personal ccmnunication.

30.  International Agency for Research on Cancer, Annual Report, 1972-1973,
     Lyon 1973.

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                              2328


31.  AGTHE, C., GARCIA,  H., SHUBIK, P., TOMATIS, L., WENYON, E., "Study of
    the potential carcinogeniclty of DDT in the Syrian Golden Hamster".
    Proc. Soc. exo. Biol.  (N.Y.), 134, 113-116 (1970).

32.  Safe Use of Pesticides, Wld. Hlth. Org. techn. Rep. Ser., No.  513  (1973) .

33.  WYNDER, E.L., MABUCHI, K.,  HEATTIE, E.J.,  "The epidemiology of lung
    cancer - recent trends", J. Amer. Mad. Assoc., 213, 2221-2228 (1970).

34.  CLEMMESEN, J., NIEISEN, A., "The social distribution of cancer in
    Copenhagen, 1943-1947", Brit. J. Cancer, 5, 159-171 (1951).

                          DISCUSSION


 SCHLIPKOTER  (Federal Republic of Germany)

      The latent period of  malignant tumors is frequently
 very long.  Tumors  caused  by asbestos, for example, may not
 appear for 20-30 years  Since we cannot wait as long  as
 this, I consider that  great importance should be attached
 to experiments on animals  to determine the carcinogenic
 effects of environmental insults until any chance of
 carcinogenesis in humans can be excluded.   Recent results
 obtained by our Institute  indicate that thin glass fibres
 are carcinogenic to rats and humans.  Should we not therefore
 pay attention to fibrous dusts of this kind now instead of
 waiting 20-30 years?


 TOMATIS (W.H.O.)

      I fully  agree  with you that more attention should  be
 paid to reliable experimental results and that experimental
 evidence of a carcinogenic effect of a substance may  well
 predict a similar effect in man.  The latency period  for
 cancer in man is often,  but not always, as long as 20-30
 years.  In the survivors of Hiroshima, for instance,  it
 was about 6 years,  and in  a few cases of urinary bladder
 cancer following chlornaphazine therapy it was 4-6 years.
 I was not aware that glass fibres were definitely proved to
 be carcinogenic to  rats and,  what is more important,  to
 humans.  If this is so,  action must be taken immediately
 to prevent human exposure.


 KJELLSTROM  (Sweden)

      The problem of the latency period may be more disastrous
 than here indicated in the way that when the carcinogenic
 effect is found in  epidemiological studies this is only
 the start of  an increasing trend in the incidence.  Even if
 the exposure  is stopped this may occur.  In two iron  mines
 in North Sweden the incidence of lung cancer has increased
 considerably during the last years.  The workers are  exposed
 to exhaust gases from  diesel engines and radon and this
 exposure started only  some years ago.  It will be very  important
 to follow the trends of the lung cancer incidence.  Do  you
 know of any data where the incidence has been followed  through
 the years after the initial finding?

-------
                             2329
      Regarding the cost-benefit analysis of profit of chemicals
 versus the costs of cancer it must be very important to engage
 the people exposed in the evaluation.  The workers in industry
 must get all the information available about the risks of
 their exposure and participate in the decision procedure.  Is
 it not the responsibility of scientists to activelv take
 his role of informer and protector of the health of the
 workers?
 TOMATIS  (W.H.O.)

      You  have  touched  a very  important  problem.   It  is  often
 said  that occupational cancers  represent  about  1% of all
 cancers.   I  have  not found  any  sound  data on which such a
 statement could be based.   What does  happen in  fact  when a
 chemical  is  found to be carcinogenic  to man?  I mentioned
 in my talk the case of /}-naphthylamine  and the  fight which
 lasted for more than half a century to  prevent  exposure to
 it, but what happens nowadays with other  chemicals?   It
 seems to  me  that  following  a  few papers or case reports
 referring to the  carcinogenic effect  of a chemical,  verv
 little information is  later collected.  Ve do not know,"for
 instance,  how  many cases of urinary bladder, skin or  lung
 cancer are due to occupational  exposure.   Sone work  has
 been  done to follow the trends  of urinary bladder cancers
 due to aromatic amines  and  of lung and  nasal cavitv  cancer
 due to exposure to nickel,  following  initial findings,  but
 there  is  certainly more to  be done in this direction.

      I agree with you that  a risk-benefit  analysis should
be done with the participation of the people who  run the
 risk  and  not only the people who evaluate  the benefits.  I
also  agree with you that scientists have  a definite
responsibility towards  society in disseminating information
which may be useful in  the prevention of  cancer.

-------
                             2331
           INTERACTION OF ENVIRONMENTAL CHEMICALS,
     ENZYMATIC CHANGES AND THEIR POSSIBLE EFFECTS ON HEALTH

                         HANS L, FALK

National Institute of Environmental Health Sciences, Research
Triangle Park, N.C., USA
(temporarily with V.H.O. Geneva)
ABSTRACT

     The interactions to be discussed here are responsible for
increased or decreased toxicity due to one environmental factor
by prior or concommitant exposure to another one.   Among the
many recorded interactions those were singled out that involve
enzyme induction or inhibition, as they may lend themselves to
easier interpretation.   Nonetheless predictions regarding the
effects of such interactions of environmental factors are not as
easy as the explanations of the results.

     The requirements for enzyme induction throw much light on
the complexity of the process.   There are differences in this
respect between species^ strains and individuals and also between
organs or tissues of the same species.   Chemicals that are lipid
soluble often are effective induoers, but other factors are in-
volved too, so that chemicals can be graded from potent to inac-
tive as enzyme induoers.   Variations in the increase in certain
enzyme activities by different chemicals suggests that multiple
enzymes are involved.

     Different effects could be produced by chemicals that act
as induoers or inhibitors depending on whether the other chemical
was toxic but could be deactivated by the induced enzyme or was
a precursor of a toxicant and would be activated by the enzyme.

-------
                               2332
Particularly complicated are situations of interactions where
multiple enzyme pathways are involved leading to inactive or
toxic products as a function of differential enzyme induction.

     Although most data oame from experiments on laboratory ani-
mals enzyme induction is also active in man and has been utilized
to good advantage.   For many chemicals the effective levels used
in laboratory experiments are not much higher than those encoun-
tered by man in his daily life.

     Several types of interactions have been discussed at a pre-
vious session of this symposium and will not be dealt with here*
but the emphasis will be on interactions which involve stimulation
or inhibition of inducible or non-induced enzyme activity.   Al-
though the mechanism of action for many of these situations ap-
pears well understoodf it is difficult to predict the outcome of
these interactions which gives the topic its importance.

-------
                                    2333
     Several typer of Interactions have oeen discussed at. a previous
session of this symposium and will not be dealt with here, but the
emphasis will be on Interactions which involve stimulation or inhibition
of induclble or non-Induced enzyme activity.  Although the mechanism of
action for many of these situations appears well understood, it is
difficult to predict the outcome of these interactions which gives the
topic its importance.
1.  Requirements for enzyme induction.
    One must start with a discussion of salient features of enzyme
Induction to appreciate the many modifying factors at work.  The
capability to respond by enzyme induction is a genetic property
possessed by all species that have been studied £_ !_/ and where the
genes are missing survival is Jeopardized.  For the perpetuation of the
species enzyme induction has played an important role whether we deal
with bacteria, insects or mammals.  However, the capacity for induction
is not the same for all species and varies greatly for species, strains
and individuals.  Comparing hepatic microsomal activity it can be seen
that man is quite low in hydroxylating activity compared to other species,
Measured by aminopyrine demethylation, mouse liver was 20 times more
efficient, rabbit, guinea pig and rat liver, 10 times, and cat liver, 5
times better than human liver.  Cytochrome P^cn baaed on mg liver
protein was 10 times higher in rodent liver than human^ but the f^o
content could not explain the superior activity of mouse liver compared
to other rodent livers ^ 2_/.
    Although enzyme induction In human liver Is far less effective than
is that of rodents, nonetheless it has recently been utilized in a case
of familial unconjugated non-hemolytic Jaundice to cure the patient by
the administration of 1.5mgAg DDT for 6 months.  It reduced the plasma
bilirubin to normal levels where it remained apparently indefinitely,
even after DDT was no longer given.  His plasma "DDT11 level rose from
5 to 1330 ng/ml during treatment and the fat content reached 200 jig/g.
After treatment plasma "DDT" levels declined slowly /"?_/.
    Effective reduction of "DDT" storage was observed in patients on
long-term antlconvulsant treatment.  In patients on phenytoin or
phenytoln and  phenobarbltal the blood DDE level was less than 2ppb,

-------
                                    2334
compared to a mean of 4.5 ppb in controls and the level in fat was 0
compared to a mean of 5-5 ppm in controls.  Phenobarbltal alone was less
effective /~V/.
    There are many variables which cause differences in enzyme Induction.
It may depend on the sex and age of the animal.  Newborn animals are
quite low in microsomal enzyme activity, but the young rat can respond
with a 10-fold increase in induction compared to a 2-fold increase in
the adult rat {_ 5_A  It also depends on the nutritional status of the
animal, on the nature of the inducer and the type of enzyme activity
measured as will be discussed below.
    Inducers are lipid-soluble and non-toxic for the tissue in which
they must accumulate to a certain level for enzyme induction.  The
compound administered need not be the inducer, but a metabolite of the
compound may be effective, such as DDE.  Those chemicals that remain
hydrophobic and are stored in specific organs will be far better
inducers than more hydrophilic ones or those that can be readily
metabolized and conjugated for rapid elimination, but even such
compounds like ethanol may act as Inducers if the concentration is high.
    Elimination of the compound may, however, not always complete the
story.  Enzymes of microbial origin may hydrolyze the conjugates or, by
enzyme induction, cause the formation of different products which on
reabsorption may show enzyme induction or, even worse,  may exert a toxic
effect.  A case in point is the enzyme Induction in fecal bacteria of
rats exposed to cyclamate, which led to increased absorption of
cyclohexylamine.  The ability to convert cyclamate to cyclohexylamine
was readily lost when cyclaraate was no longer fed to the animals [_ 6_/.
    Although most studies on enzyme induction have focused on hepatic
microsomes,  it should be mentioned that certain inducible enzymes like
glucuronyl transferase may be associated only partly with the microsome
fraction.
    Other tissues and organs are capable of enzyme induction.  It was
demonstrated for the lung, adrenal and mammary gland of rats, / ?__/
rat and rabbit kidney / 8_/, and the skin of mice, {_ 9_/.  Aryl
hydroxylase activity was demonstrated for the duodenal  mucosa of
laboratory animals and man ^~10_/.  In comparative studies on isolated

-------
                                  2335
perfused rabbit liver and lung preparations enzyme induction of
p-hydroxylatlon of N-methylaniline by phenobarbital was studied.  The
lung was half as active as the liver, but based on organ weight the
lung was more active than the liver / 8 _/.  Comparing enzyme induction
in liver and kidney of the rabbit by phenobarbital, liver was found to
have the higher rate for ring hydroxylation and N-dealkylation of
N-methylaniline than had the kidney.  Other tissues examined In this
study were barely active .  The increased enzyme activity was correlatable
with increases in yields of microsomes in liver and kidney and an
increased liver weight {_ 8 _/.  It should be noted that N-hydroxylation
of N-methylanlline was not affected by enzyme Induction, an important
observation since this step often leads to toxic products.
    A few induclble enzymes have already been mentioned, but the list
should contain N-, 0-, and S-oxidative demethylases, o- and p-aromatic
ring hydroxylases, N-hydroxylase, azo reductase, omega-oxidase and
glucuronide transf erase to mention the most important ^11 _/.
    Conditions for maximal enzyme induction may not always be
encountered .  In the absence of inducers microsomal hydroxylating
enzyme activity was at very low levels when a protein-free diet was
fed to rats, i.e. it was 1/7 the level on a normal diet.  In the
presence of an inducer, DDT, but still on the protein-free diet the
activity could be stimulated to reach near normal levels, but DDT and
a normal diet produced the expected large increase in enzyme aotivity^lg/
    On raising the protein content to yfc, i.e. still a low protein
diet, the induced enzyme activity rose to jyfa of normal as did cytochrome
p^  .  On 6$ protein in the diet the induced enzyme activity was 50$ of
normal giving a good indication for dietary protein requirements for
adequate enzyme induction.  On the protein free diet, rats showed
reduced hepatic N-demethylase activity, but increased N-oxidation of
dime thy lani line ^13, ,_/.  On a low choline diet low enzyme induction
was observed, suggesting the need for choline as a component of the
phospholipid structure of the enzyme complex,
    Riboflavin is also a required component for effective microsomal
enzyme activity.  Azo reductase is a flavoprotein which was Instrumental
in the 1940s to give early clues to investigators of carcinogenesis on

-------
                                  2336
factors that would protect animals against carcinogenic azo dyes on the
basis of'dietary supplements / l4,15_/.  Similarly, on a vitamin E
deficient diet N-demethylation was significantly reduced while
N-oxygenated products were increased.  The addition of 2 mg vitamin E/g
diet for 4 days increased N-demethylation and reduced W-hydroxylation
of N-dimethylaniline /~l6_/.
    These bits of information fit together to describe the requirements
for effective enzyme induction if the Inducer reaches the required level.
The process requires DNA-dependent RNA synthesis and protein synthesis.
One of the first enzymes synthesized is delta-aminolevulinic acid
synthetase which is required for cytochrome Phcn synthesis.  These
proteins are rapidly Increased, but their half life is very short.  On
Induction RNAase activities are Inhibited so that more enzyme can be
synthesized.  By the use of puromycln it could be shown that some of
the induced enzyme activity was not the result of de novo synthesis
but may result from activation of enzyme precursors {_ 17>l8_/.
    Some chemicals which act as enzyme inducers have already been
mentioned.  Of the various classes of chemicals, the-polycycllc aromatic
hydrocarbons (PAH) have been studied In detail.  Not all were equally
effective.  Using rat liver homogenate and measuring certain polar
7, 12-dlmethylbenz(a) anthracene (DMBA) metabolites £ 19_/ and
determlng 8-hydroxybenzo(a)pyrene formation in rat placenta, comparisons
of a number of PAH as enzyme inducers are possible {_ 20_/.   20-
Methyleholanthrene (MC) and benz(a)anthracene (BA) were most effective.
Benzo(a)pyrene and (BaP) and chrysene were also quite effective in both
systems, while anthracene and fluoranthene were weak.  Pyrene, perylene,
phenanthrene,  fluorene and naphthalene required large doses for a weak
response or were entirely Inactive. / 21_/.
    Organochlorine insecticides have been tested and were generally
found to be very effective inducers /_ 22,2£/ Chlordane,  methoxychlor,
DDT, DEE, DDD,  endrin, aldrln,  dleldrin, hexachlorocyclohexanes,
heptachlor and Its epoxide were all active.  Chlordane and DDT were
each given to rats twice daily (25mgAs) for 10 days and their liver
microsomes were incubated with testosterone,  estradiol-lTbeta,
progesterone or desoxycorticosterone (TOO mjimoles) in the presence of

-------
                                  2337
a TPNH-generating system / 24_/.  Metabolism of these steroid hormones to
more polar products was greatly enhanced, an effect which has been
considered the cause of the poor survival of some species of birds of prey.
     Steroid hormones on their part cause hepatic mlcrosomal enzyme
induction.  Progesterone has been studied in particular and was found to
give rise to a 5beta metabolite, which caused increased hepatic smooth
endoplasmic retlculum, delta-aminolevulinlc acid eyntnetase and
oytochrome P^   activity.  It was thought capable of aggravating
conditions of porphyria.  This compound is also present In human milk
and by inhibiting glucuronyl transferase appears to be responsible for
prolonged unoonjugated hyperbilirubinemia in neonates during breast
feeding ^~25_/.
     The extent to which synthetic progestogens can act as enzyme
Inducers and as competitive Inhibitors of these enzymes is In need of
study.
     Not mentioned so far as potent enzyme inducers, because they do not
belong into the picture of environmental pollutants, have been the
pharmaceutical agents, particularly analgesics, hypnotics, hypoglycemlcs,
antihistamlnes, anticonvulsants and anti-inflammatory drugs {_ 26_/.
     The observation that cigarette smoking Increased the BaP hydroxylase
and 3'-methyl-4-monomethylamlnoazobenzene N-demethylase activity of
human placentas is of great Interest.  In non-smokers no such activity
was detected.  There was, however, considerable variation in the activity
of the 2 enzymes which could not be explained by the extent of smoking.
Aromatic polyoyclic hydrocarbons were suggested as responsible for the
induction {_ 20_/.  Nicotine, however, is more likely to be the inducer
when considering quantitative requirements.  That nicotine is indeed
a good Inducer could be shown on mice given orally 2.28 mg/kg/ day of
nicotine and measuring the ataxla produced by l.p. Injection of
200 mgAg meprobamate.  The period of ataxla was significantly decreased
on nicotine treatment.  When nicotine administration was stopped the
duration of ataxla returned to normal levels In 4 days.  In vitro studies
confirmed the faster metabolic degradation by mouse liver of
meprobamate in animals treated for 2 days with nicotine*  Ethlonine
prevented the increase ^~27_/.  These observations are of great interest

-------
                                   2338
because of the widespread Intake of nicotine which belongs to a class
of compounds, the alkaloids, which have not been considered to be
effective enzyme inducers.
    Chemicals belonging to other classes have been studied sporadically
as enzyme Inducers.  Among them are dietary components, present as
anutrlents, food additives or residues.  Obtaining data on enzyme
inducers of that type Is difficult as a semi-synthetic diet is
inadequate as baseline for enzyme induction.  However, a number of
flavones were shown to induce BaP hydroxylase activity in rat liver
and lung, including tangeretln and nobiletin besides flavone itself^2§/.
Safrole was shown to induce blphenyl hydroxylase, nltro reductase,
glucuronyl transferase and cytochrome P^f-n *n ra^ liver /~2g/.  It is
of interest to note that safrole and many related methylenedioxyphenyl
derivatives are effective Inhibitors of mlcrosomal oxidative enzymes,
but become capable Inducers after they have been metabolized.  Among
the pesticides, besides the organochlorine insecticides, the urea
herbicides were shown to be active inducers.  The most effective ones
were dluron and herban causing increased 0-demethylase activity and
EPN (0-p-nitrophenyl phenylphosphonothloate) detoxification in rat
liver when 10-300 mg/kg/day had been administered for 5 days £_ 30_/.
    Among the food additives butylated hydroxytoluene (EOT) Is of
Importance.  When fed to rats above the level of 0*1$ in the diet the
liver enlargement paralleling the stimulation of mlcrosomal enzyme
systems was not considered physiological but a toxicologlc effect.  On
day 2 of BHT feeding, enzyme activity was already enhanced and
continued rising as feeding was continued.  The level of BUT in fat
stabilized at 100 ppm in that study.  BHA (butylated hydroxyanlsole)
was a less effective enzyme inducer, but also caused liver enlargement
In rats / 31_/«  Both chemicals have been shown to be radiooensitizers
                                       i
in drosophila melanogaster and murlne leukemia and lymphoma cells,
causing 1.6 times as many sex-linked recessive lethal mutations In
frultflies on exposure to 1.2 Krad gamma-radiation as did radiation
alone.  The mechanism of this symergism has not been clarified / 32_/.
2.  Interactions Involving enzyme Induction.
    The variables affecting mlorosomal enzyme induction discussed above

-------
                                    2339
will enable us to understand the interactions involving enzyme induction
that much better.  Thus feeding rats a low protein diet reduced the
hepatic mlcrosomal hydroxylase activity and dramatically reduced carbon
tetrachloride toxicity.  The LD   of CC1. was reached only at 14.7 mlAg
on the protein-free diet compared to 6.4 ml on a normal diet.  When the
rats were pre treated with phenobarbital the UV,-, was reached at 0.5 ml/kg.
Microsomal enzymes are responsible for forming a OC1  free radical, the
toxic metabolite
    Rats on a protein-free diet were also more resistant to the lethal
effects of dimethylnitrosamine (DMN).  The ID   was at 79 mgAg on the
protein deficient diet compared to the 45 mg/kg on the regular diet.
Hepatic mlcrosomal enzyme activity dropped considerably In 4 days on
the protein deficiency, reflected in very low metabolism of DMN.  The
metabolism of DMN by kidney slices, however, was not affected.  When
a large dose of DMN (60 mgAg) was given to rats i.p. while kept on
this protein -free diet no liver tumors were found, but kidney tumors
developed in all animals In 8 to 12 months {_ 24_/.  Similarly, the
mutagenlc activity of nitrosamines was shown to depend on the level of
protein In the diet.  On a protein-free diet the mutagenicity of DMN
for Salmonella typhimurium in the host-mediated assay was decreased,
while that of N-methyl-N-nitrosourea which requires no metabolic
activation was increased ,  In mice on a pure casein diet the opposite
effects were observed with these nitrosamines, which were explained on
the basis of dietary modification of hepatic mlcrosomal enzyme activity,
essential for activation of DMN and effective In the detoxification of
N-methyl-N-nitrosourea / >5_/ ',
    Rats fed 4-dlmethylamlnoazobenzene (DAB) had far fewer liver tumors
when the diet was supplemented with 200 jig ribo flavin and 2 g casein.
This supplement protected racs also against other carcinogenic azo
dyes.  The discovery of an hepatic mlcrosomal enzyme that cleaved DAB
Into aniline and dimethyl aniline - a flavoprotein - helped to explain
the findings.  Azo reductase activity could be Increased by incubating
liver slices with riboflavln /~36_/.
    The carcinogenic response to MC oould be halved If the mice were fed
a diet containing wheatgerm oil   37«  Feeding 30 - 100 times the

-------
                                  2340
dietary  requirement for alpha-tocopheol to C57 leaden mice produced
only half as many sarcomas as were seen in animals on a normal diet
given the same B.C. dose of MC in mineral oil ^ jB /.
    Several studies showed alterations in tumor incidence resulting
from interactions for which enzyme induction and enhanced detoxification
was the most likely explanation.  Simultaneous feeding to rats of
jS'-methyl-'l-dimethyl-aminoazobenzene and MC produced a far lower
incidence of hepatomas and bileduct carcinomas than feeding the a7.0
dye alone / 3gA  In further studies th« temporal relationship of feeding
the 2 compounds was of significance.  When MC was added to the azo
dye diet in the first 6 weeks hepatoma formation was prevented.  When
MC was added from the 6th to 10th week It was only partly successful
In its protection.  However, adding the hydrocarbon only after 10 weeks
of azo dye feeding was non-protective.  The temporal increase in
hepatic N-demethylase activity in the MC-fed animals which can
inactivate the carcinogen could explain the findings {_ 40_/.  With DAB
and barbltal the reduced hepatoma incidence was similarly explained
on the basis of enhanced enzyme induction /_ ^l_/«
    Using fluorenyl acetamide (FAA) as precursor of a liver carcinogen
and MC as inducer reduced the Incidence of mammary tumors, hepatomas
and earduct tumors in rats indicating that the precursor was
metabolized preferentially by aromatic ring hydroxylation instead of
by N-hydroxylatlon, thus producing inactive compounds instead of the
proximate carcinogen £ 21_/.
    However, an explanation of the results of feeding rats
N,N'-(2,7-fluorenylene)-bisacetamide with EMBA as enzyme inducer and
carcinogen is more difficult to give.  The EMBA was placed into an
artificial divertlculum in the glandular stomach where it produced few
tumors on its own.  The aromatic amine alone produced hepatomas,
cholangioflbrosls, earduot tumors and Intestinal cancers.  These
pathological conditions were markedly reduced when both chemicals
were administered, but instead, a high incidence of forestomach
papillomas, basal cell carcinomas, squamous cell carcinomas, carcinomas
of the glandular stomach as well as mammary, uterine and skin cancers
were observed / 42_/.  Explaining these findings may be possible but

-------
                                   2341
predicting them is another matter.
    Interactions of these types have also been reported in the area of
teratology.  Thalidomide (150 mg/kg) given to rabbits from day 7 to 12
of gestation produced no malformations in the offsprings unless on day
6 60mg/kg of CCl^ was administered orally.  Evidence of liver damage
was seen in the maternal animals f_ ^H>_/ •
    Other chemicals that can alter maternal hepatic microsomal enzyme
activity can affect the incidence of malformations in the fetuses as
was shown for phenobarbltal (50 or 100 mg/kg) given to mice from day
8 to 10 of gestation and the teratogenic eyclophosphamide (20 mg/kg)
given on day 10 only.  Malformations, as seen with cyclophosphamide
alone were nearly completely abolished except for the incidence of
open eyes and polydactyly which persisted /_ 44_y.
3,  Interactions Involving enzyme inhibition.
    The second part of the experiment discussed above described the
doubling of the incidence of malformations due to cyclophosphamide by
giving an enzyme inhibitor, i.e. beta-dlethylaminoethyl diphenyl-
propylacetate (SKF-525A) (32 mg/kg) to pregnant mice from day 8 to 10.
    Inhibition of microsomal enzyme activity can occur at the normal
level as well as in the Induced state.  In comparison to enzyme
induction which may last for several weeks following a single
administration of an inducer, enzyme inhibition is comparatively short
lived and reversible.
    Pesticide synergists which found widespread use as a household
insecticide combine plperonyl butoxide or a related compound with a
pyrethrum insecticide.  They produce their lethal effect at 1/10 the
normal dose of insecticide.  They function by temporarily inhibiting
detoxifying enzyme systems In insects.  They also prevent enzymatic
dehydroohlorination of DDT in soil organisms, epoxldatlon of aldrin
or heptachlor and ring hydroxylation of polycyclic hydrocarbons in
mammals, etc.  Their activity as detoxification inhibitors oould be
considered a potential health hazard for man were it not for the
limited production of these pesticide synergists because of sparce
supplies of the starting material and the interesting fact that these
compounds on being metabolized themselves give rise to enzyme inducers.

-------
                                   2342
However It should be mentioned again that methylenedloxyphenyl derivatives
are also present as anutrients In sesame oil, sasafras oil, etc {_ ^5_/«
    Ozone was found to be a good BaP hydroxylase inhibitor whether the
enzyme was in the normal or induced state.  Significant inhibition was
observed when hamsters were exposed for 3 hours to 0,75, 3 or 10 ppm
ozone and sacrificed 30 minutes later.  There was no effect on hepatic
microsomal enzyme activity £_ h6_/.
    Another air pollutant, carbon monoxide, is a potent inhibitor of
microsomal enzymes as it will bind to cytochrome Pj.,-n inhibiting Its
function.  Inhibition of the metabolism of azo dyes, drugs and
steroid hormones depend on the ratio of CO to 0_ and it was noted
that the ratio of CO to 0_ for 50# inhibition of hydroxylation of
testosterone was different dependent on which position in the molecule
was hydroxylated.  It was 0.93, 1.5^ and 2.36 for l6alpha-, 6 beta-, and
7alpha-hydroxylation of testosterone {_ ^7_/,
    Inhibition of the metabolism of estradiol-lTbeta and estrone in
Immature female rats by CC1, has been reported.  In vitro metabolism
of estrone was ?0# inhibited by 0.06 mgAg CCl^.  In in vivo studies
CC1. Interfered with estrogen metabolism, potentiated the uterotroplc
action of the compounds and increased the concentration of the estrogens
in the uterus.  Tetrachloroethylene by contrast produced none of these
effects / Jl8_/.  In other studies CC1, administered to rats increased
the sleeping time evoked by hexobarbltal, reduced the In vitro
metabolism of hexobarbital, impaired the oxidation of aminopyrine and
the reduction of p-nitrobenzolo acid.  The activity was reduced to
    of normal within 8 hours following administration and remained at
that level for 24 hours after administration of CC1. .  In contrast
chloroform had little effect and methylene chloride was ineffective as
an inhibitor /"/t9_/.
    The skin tumor incidence due to ethyl carbamate could be reduced
to only 10$ by the administration of an equlmolar amount of butyl
carbamate to mice. / 50_/,  The explanation was given that the uretnane
activating enzyme could be inhibited by a structural analog.^ 51_/«
Similarly a toxic dose of FAA (O.OJjC) could be safely administered in
the diet to rats if acetanilide at a level of 0.8# was also given.  No

-------
                                  2343

hepatomas were seen, only some hepatic hyperplasla.  Halving the dose of
acetanilide increased only the degree of hyperplasia, but produced no
hepatomas.  Since FAA requires metabolic activation to become a
hepatocarcinogen acetanilide apparently pre-empts N-hydroxylatlon of
FAA by being Itself preferentially metabolized /~52_/'
    A 50-fold potentiation of toxlcity of malathlon by another
organophosphorus insecticide (EPN) was reported in 1957«  A metabolite
of EPN was found to possess greater affinity for aliesterases than
cholinesterase and inhibition of the aliesterase prevented normal
detoxification of malathlon thus leading to the enhanced toxlcity.
This effect however is only realized at high dose levels when the
enzyme systems become overloaded./ 53_/«
                                     "~                  p
    Not quite the same results were obtained with Delnav , another
organophosphorus insecticide preparation consisting of isomers of
2J3-p-dioxanedithiol-S,S-bis-(0J,0-diethyl phosphorodlthioate) which
would only show additive toxicity when simultaneously administered
with malathion.  However, if administered 4 days prior to malathion
to rats it would enhance the expected toxicity considerably /~5\y.
    EPN not only enhanced the toxlcity of malathlon, but that of
dimethoate by blocking amidase activity as could be shown in mice and
guinea pigs.  Houseflies metabolize organophosphorus insecticides
preferentially by means of phosphatase.  Therefore inhibition of
metabolism of malathion or dimethoate by these compounds does not
occur to any extent and the synergism is not observed in houseflies/ 55_y.
    As a last example I should mention the deliberate use of monamine
oxldase (MAO) inhibitors which have found widespread use, but were
responsible for many cases of paroxysmal hypertensive attacks.  Most
of the patients on specific MAO inhibitors had eaten unknowingly high
doses of certain amines with their food, particularly tyramine, which
is present in large quantities in herring, cheeses like Cheddar or
Oruyere, but also in yeast, beer, wine, etc.  This toxlcity can also
be elicited from other amines which cannot be detoxified In persons on
MAO inhibitor drugs /"56_/.
    The Interactions discussed here have produced 10 to 50 fold
aggravation or amelioration of toxlcity and should help all of us

-------
                                   2344
concerned with environmental pollution problems to keep such interactions

in mind when considering potential health hazards from individual

pollutants alone, even though they occur in a milieu of multiple

exposures to many environmental chemicals.

BIBLIOGRAPHY
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 2.   Remitter, H. "Induction of drug metabolising enzymes in different
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 }.   Thompson, R.P.H., et al.  "Treatment of unconjugated Jaundice with
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11.   Gelboin, H.V.,  "Carcinogens,  enzyme induction and  gene action".
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-------
                                   2345
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18.   Selfert,  J., Vacha, J., and  Reamer,  H.,  "Phenobarbital  and liver
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19*   Levin, W. and Conney, A.H.,  "Stimulatory effect of polycyclic
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20.   Welch, R.M. et al., "Stimulatory effect  of cigarette smoking on
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21.   Miller, E.G. et al.  "On the protective  action of certain
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22.   Fouts, J.R.  "Factors influencing the metabolism of drugs in
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23.   Hart, L.G. and Pouts, J.R. "Further  studies  on the stimulation
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24.   Conney, A.H. et al.  "Effects of pesticides  on drug and steroid
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-------
                                  2346
25-   Adlercreut?., H. and Tenhunen, B.  "Some aspects of the
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27.   Wenzel, D.G. and Broadie, L.L.  "Stimulatory effect of nicotine on
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28.   Wattenberg, L.W., Page, M.A. and Leong, J.L.  "Induction of
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29.   Parke, D.V. and Rahman, H.  "The induction of hepatic microsomal
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30.   Kinoshite, F.K. and DuBois, K.P.  "Induction of hepatic microsomal
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31.   Gilbert, D. and Golberg, L.  "Liver response tests.III.  Liver
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32.   Prasad, O.M. and Kamra, O.P.  "Radiosensitizing property of
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33-   Slater, T.F.  "Necrogenic action of carbon tetrachloride in the
      rat.  A speculative mechanism based on activation".  Nature 209,
      36 (1966).

34.   Swann, P.P. and McLean, A.E.M.  "The effect of diet on the toxic
      and carcinogenic action of dimethylnitrosamine".  Biochem.J. 107,
      11 (1968).

35.   Zeiger, E. "Dietary effects on the  mutagenicity of N-nitroso
      compounds in the host mediated assay".  First International
      Conference on Environmental Mutagens,  Pacific Grove,  Calif.,
      USA, p 15 (1973).

36.   Kensler, C.J. et al.  "Partial protection of rats by riboflavin
      with casein against liver cancer caused by dimethylaminoazobenezene",
      Science 9J>, 308 (1941).

37.   Jaffe, W.G.  "The influence of wheat germ oil on the production of
    .  tumors in rats by methylcholanthrene".  Exp. Med. Surg. 4,
      278 (19^6).

38.   Haber, L. and Wissler, R.W.  "Effect of vitamin E on
      carcinogenicity of methylcholanthrene".  Proc. Soe. exp. Blol.  Ill,
      774 (1962).

-------
                                   2347
39.   Richardson, H.L., Stier, A.H. and Borsos-Nachtnebel, E.   "Liver
      tumor Inhibition and adrenal hlstological responses in rats  to
      which 3'-methyl-4-dimethylaminoazobenzene and 20-methyloholanthrene
      were simultaneously administered".  Cancer Res.  12,  356  (1952).

40.   Meechan, R.A., McCafferty, D.E. and Jones, R.S.   "3-Methyleholan-
      threne as an inhibitor of hepatic cancer Induced by 3'-methyl-4-
      d line thy lamlnoazobenzene in the diet of the rat.   A determination
      of the time relationship".  Cancer Res. 13, 802  (1953).

41.   Ishidate, M., Wattanabe, M. and Odashima, S.  "Effect  of barbital
      on carcinogenic action and metabolism of 4-dimethylaminoazo-
      benzene".  Gann 58, 26? (196?).

42.   Odashima, S.  "Combined effect of carcinogens with different
      action.  IV.  Effect of 7,12-dimethylbenz(a)anthracene placed
      in an artificial divertlculum of the glandular stomach combined
      with feeding of N,N'-(2,7-fluorenylene)-bisacetamlde to  rats".
      Gann 2, 8l (1968).

43.   Heine, W. et al.  "Thalldomld-Embryopathle bei Kaninchen nach
      passagerer LeberschMdigung der Muttertiere durch Tetrachlor-
      kohlenstoff".  Klin. Wschr. 42, 592 (1964).

44.   Gibson, J.A. and Becker, B.A.  "Effects of phenobarbital and
      SKF 525-A on the teratogenlclty of cyclophosphamide in mice".
      Teratol. 1, 393 (1968).

45.   Falk, H.L. and Kotln, P.  "Pesticide synergists  and their
      metabolites:  Potential hazards".  Ann.New York  Acad.  Sc. 160,
      299 (1969).

46.   Palmer, M.S., Swanson, D.H. and Coffin, D.L.  "Effect  of ozone on
      benzpyrene hydroxylase activity in the Syrian golden hamster".
      Cancer Res. 31, 730 (1971).

47.   Conney, A.H. et al.  "Inhibitory effect of carbon monoxide on the
      hydroxylation of testosterone by rat liver mlcrosomes".   J.Blol.
      Chem. 243, 2912 (1968).

48.   T«vin, W., Welch, R.M. and Conney, A.H.  "Effect of carbon
      tetrachlori'ie and other inhibitors of drug metabolism  on the
      metabolism and action of estradiol-lTbeta and estrone  in the
      rat".  J.pharmacol & exper. therap. 173, 247 (1970).

49.   Dlngell, J.V. and Helmberg, M.  "The effects of  aliphatic
      halogensted hydrocarbons on hepatic drug metabolism".  Blochem.
      Pharmacol. 17, 1269 (1968).

50.   Garcia, H. "Inhibition ofttumorigenic action of  urethane by  butyl
      carbamate".  Blologlca 34, 11 (1963).

-------
                                 2348
  51.   Kaye, A.M.  "A study of the relationship between the  rate of ethyl
        carbamate (urethan) catabollsm and urethan carclnogenesls".
        Cancer Res. 20, 2J7 (i960).


  52.   Yamamoto, R.S. et al. "inhibition of the toxicity and
        carcinogenicity of N-2-fluorenylacetamide by acetanillde".
        Toxicol. appl. Pharmacol lj>,  108. (1968).


  53.   Frawley, J.P. et al. "Marked  potentiation in mammalian toxicity
        from simultaneous administration of two anticholinesterase
        compounds".  J. Pharmacol <5b exper. therap. 121, 96 (1957).

                                                             T3
  54.   Frawley, J.P. et al. "Toxlcologic investigations on Delnav ".
        Toxicol. appl. Pharmacol. 5,  605 (196.?).


  55.   Uchida, T., Zschintzsch, J. and O'Brien, R.D.  "Relation between
        synergism and metabolism of Dlr.ethoate in mammals and insects".
        Toxicol. appl. Pharmacol. 8,  259 (1964).


  56.   Levy, J. and Michel-Ber, E.  "Difficulties and complications
        caused in man by monoamine oxidase (MAO) inhibitors, with
        special reference to their specific and secondary pharmacological
        effects", in "Toxicity and side-effects of psychotropic drugs",
        Proc. Europ. Soc. Study Drug  Toxicity, 9, 22.5 (1968K
                             DISCUSSION
ZIELHUIS (Ketherlands)
     Should one take  into account  enzyne induction or  inhibition
in setting acceptable daily intakes (ADT)  for food additives
or in  setting permissible limits  for occupational and  ambient
exposure?   Should  enzyme induction or inhibition at relevant
dosage range be regarded as inacceptable?.
FALK (W.H.O.)

      At this  stage we are  not adequately equipped  to make mean-
ingful recommendations.  In  rodents the  level of dietary intake
of  inducers has  not been adequately  documented.   The lowest
level of 2-3  week feeding  of DDT resulting in barely detectable
enzyme Induction was 1 ppm (1), 2.5 ppm  (2)  and 3.3  ppm (3).
For lindane as  inducer the dietary feeding level was O.5 ppm at
which induction  could already be observed (4).  For  most other
compounds, we do not have  such data, which are needed to be able
to  evaluate the  total picture, i.e. for  the human  we need to
know the level of Inducer  in the diet  and its relative potency.
With this information in hand, ADIs could be set,  but one would
have to determine their signigicance if  most people  are reasonably
weakly "induction-responsive".

-------
                            2349
     On the other hand, attention should be paid to workers
exposed to high doses of inducers and patients on treatment
with drugs which are potent inducers.

References

     1.  Kinoshita, F. K. et al
         Quantitative measurement of induction of hepatic
         microsomal enzymes by various dietary levels of
         DDT and toxaphene in rats,Toxicol & Appl.  Pharmacol.
         9_, 505-513, 1966.

     2.  Gillett J. W.
         "No-effect" level of DDT in induction of microsomal
         epoxidation J. Agr. Food. Chem., 16, 295-7, 1968

     3.  Hoffman, D. G. et al

         Stimulation of hepatic drug-metabolizing enzymes by
         chlorophenothane  (DDT); the relationship to liver
         enlargement and hepato-toxicity in the rat.
         Toxicol. & Appl. Pharmacol., 16, 171-8, 1970.

     4.  Kolmodin-Hedman,B. et al.
         Effect of exposure to lindane on drug metabolism:
         decreased hexobarbital sleeping time and increased
         antipyrine disappearance rate in rats.
         Toxicol. & Appl. Pharmacol., 20, 299-307, 1971.


SANOTSKY (USSR)

     What are the criteria for differentation between harmful
and not harmful changes of enzyme activity in relation to the
general state of health?


FALK (W.H.O.)

     Changes accompanying hepatic enzyme induction are
usually reversible and therefore considered of a physiological
and, by itself, harmless nature.  In the state of hyperplasia,
the liver may be prone to toxic action by other environmental
chemicals.  I mentioned CC14 as an example.

     When the enzyme-inducer  produces hyperplastic changes
which do not regress, a state of damage may be considered
permanent which could be expected in rodents, at least, to
lead to abnormal nodule formation.

     Whether enzyme induction, even only of a transitory
nature, is beneficial or harmful, will depend on the other
chemicals encountered by the organism which may be detoxified
or transformed to a more toxic agent.

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                            2350
PHILP   (U.K.)

     Dr. Falk referred to the use of semi-synthetic diets in
testing of food additives.

     This type of diet is extremely useful in such tests and
is preferrable for better control of the ingrediants.  What
was the basis for his comment in his paper that "a semi-
synthetic diet is inadequate as a base line for enzymatic
production" suggesting that enzyme induction would be difficult
to detect if such a diet was used.
FALK  (W.H.O.)

      In studies of interactions which are long term studies
in the majority of cases a completely synthetic diet has never
been  used, because of the great expense.  When these interaction
studies used well monitored diets they would not get information
on all enzyme inducers present as addition, contaminants or
anentriant food components.

      Oftentimes little attention was paid to that matter in
the past, since enzyme induction was not taken into
consideration.

      On enzyme induction studies a semisynthetic diet may
also  present problems if the various oils are not adequately
identified and studied for contaminant, antioxidants and
other components.


OLOFFS (Canada)

      Dr.  Falk, you said that the degree of enzyme induction
in humans is quite variable, and has been reported to range
from  130% (of normal) to as high as 450%.  Is it known whether
or not magnitude of induction is sex-linked in humans, as has
been  shown in animals?  Were enzymes in female subjects
induced to a higher degree than in male subjects?

FALK  (W.H.O.)

      The inducibility of microsomal enzymes has been studied on
human leucocytes in vitro.(1)They came from 353 healthy human
subjects.  Before exposure to 20 methyl cholanthrene as inducer
the enzyme activity measured by aryl hydrocarbon hydroxylatim
was less and showed little variability.

     After induction it was shown to vary from 130 - 450%.  The
lower range was more frequent than the upper range.  No informal
was given on sex.

Reference (l)Kellermann G. et al Genetic Variation of aryl
hydrocarbon hydroxylae in human lymphocytes.  Am. J. Human
Genet. 25_:  327 - 331, 1973.

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                               2351
         PERSPECTIVES IN THE ENVIRONMENTAL TOXICOLOGY
                        OF TRACE METALS

                      KENZABURO TSUCHIYA

Department of Occupational Medicine and Epidemiology, Keio Uni-
versity School of Medicine, Tokyo, Japan
ABSTRACT

     The term "environmental toxicology" has come 'into uee -In
recent years, since environmental pollutions of trace metals and
their effects on human health have been recognised.   Environmen-
tal toxicology differs from industrial toxicology in that, not
only does the former often include the latters but is concerned
usually with exposure to trace metals in much smaller doses by
larger number of the general population^ including children, the
aged and the sick.

     The use of epidemiological approaches is very important in
order to detect the insidious health effects of trace metals on
the general population.   In light of the significance of these
approaches^ the following five aspects of environmental toxico-
logy have been chosen for discussion in this paper* with refer-
ences to a number of reports on each aspect available up to the
present time.   They are:

2.   Dynamic pathways of trace metals in the ecological environ-
ment
2.   Accumulation of trace elements in human beings
S.   Dose-response relationship
4.   Detoxication of trace metals
5.   Evaluation of unspecific responses in individuals and in
groups.   Recommendations for further studies which still need
to be done concerning each of these aspects are also included.

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                                 2352






Introduction





     The term "environmental toxicology has come into common usage in the




scientific comminity in recent years as environmental pollutants have been



recognized and their effects on the health of man and animals have become




the concern of numberous scientists, many of whom are also interested in




industrial toxicology.  Recent stidies and reviews of environmental toxicology




are well represented by such monographs as "Lead, Airborne lead in perspec-




tive," published by the National Academy of Sciences, Washington, D.C., 1972,




and Cadmium in the Environment, by Friberg and Associates, 1971.  This pre-




sentation will be concerned mainly with epidemiologocal methodology evaluation




of the results obtained by epidemiological studies.




     The methodology of assessing biological effects of trace metals is




classified into three categories, namely animal experiments, experimental




studies on humans, and epidemiological studies.  Although the methodology of




research in the field of metal toxicology has remained unchanged since the




beginning of medical science, implication, significance, and meaning of "bio-




logical response" have been altered in recent years, particularly after the



recognition of health effects on the general pollution due to insidious




environmental pollution.  The reasons for this change can be ascribed to the




following facts: 1) advances in the biological sciences, particularly In bio-




chemistry, molecular biology, and physiology, which have enabled toxlcologisy




to detect minor effects of trace metals on organisms, and 2) exposure of the




general population to environmental pollutants, including toxic metals in




general, may be much smaller in amount than those having been observed among




industrial workers.  These factors have contributed significantly to the




rising importance of epidemiological studies in metal toxicology.

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                                     2353
     In this presentation I would like to point out several characteristic



features of metal toxicology which have gained prominence over the recent



years placing special emphasis on epidemiological approaches.  The topics



to be discussed are outlined in the table below.








                   Epidemiological Approaches to



                      Environmental Toxicology



    1.  Dynamic pathways of trace metals in the ecological environment.



    2.  Accumulation of trace elements in human being



    3.  Dose-response relationship.



    4.  Detoxication of trace metals.



    5.  Evaluation of unspecific responses in individuals and in groups.








1.  Dynamic pathways of trace metals In the ecological environment.



         For detecting the entity of a particular disease, physicians In



clinical practice have played an important role.  It was they who recognized



and reported the first cases of both methyl mercury poisoning (also called



Mlnamata disease) and of Itai-itai disease in Japan.  Epidemiological studies



which follow such clinical case reports aim to find and confirm, if possible,



a factor or factors directly or Indirectly associated with the cases of the



disease.  However, in many instances, the detection of the causative agent



has not been as easy as has been the case in well-known communicable disease.



It took more than six years before the source and agent of Minamata disease



was finally discovered (Kumamoto University, 1968).

-------
                                      2354
           figure 1 Pathway of Methyl Mercury in Ecosystem
              I Source ot *j«nt)          [EnlraaMnt]
                     Industry

            Hetellc Mercury ee Catelyet—» Hetellc Mercury

                                  iferthly Converted Methyl
                                  Mercury

            Comrelon to Methyl Mercury      *


                                Biologic Aapllfleetlon In
                                Icolotlcel loTironuent

                                       AccuBulttloe In
                                rteb                	> UCle« leblt Md Cuta*
 As shown in figure 1,  the mechanism of  the pathway of metyl mercury appears

 to be simple when seen In retrospeccLveThls has brought us important  know-

 ledge in environmental toxicology which had never been described  in classical

 toxicology, namely 1)  that metallic mercury produces methyl mercury in the

 process of catalytic  reaction in the reaction tower of acetoaldehyde  as well

 as by bacteria  in the environment, and  2)  that methyl mercury is  amplified

 in environmental  aquatic organisms, resulting in high concentrations  of the

 metal in fish eaten by nan.  As has been clearly shown in the case of methyl

 mercury, dynamic  pathways of other trace metals in the ecological environment,

 particularly lead,  cadmium, zinc, manganese,  arsenic and vanadium, should be

 more rigorously studied in the future.  No such studies have been performed  -

 thus far.  Future studies should include the identification of the types of

 compounds found in water, soil, plants* animals, and in man.



 2.   Accumulation of  trace metals in man


          Reports  on the accumulation and distribution of trace metals in

"normal" men on  an international scale have been rather limited.   There do

exist reports on various metals by Schroeder and Balassa (1961), Tipton and

Cook (1963), Tipton, Schroeder, Perry, and  Cook (1965) of the U.S.A.,  on

-------
                                    2355


cadmium by Tsuchiya, Seki, and Sugita (1972), also by kimura, Sumino and

Kanatanl (1970), and on lead by Horiuchi (1970) from Japan.  Prospective

epidemiclogical studies on the accumulation of trace metals in the general

population would provide us with two vital kinds of information: 1) general

background information on the Increase or decrease of those trace metals

over long periods time (years, decades, etc.) and 2) basic information needed

in order to derive biological half times of those metals in man.

     It is  important to obtain such background Information on trace metals

in environmental toxicology.  If successive observations are made according

to lapse of time, an Increase of accumulation of lead, for example, in man,

may be detected.  This would make it possible to anticipate probable effects

due to the metal prior to the development of actual health hazards.  In order

to obtain accurate and precise information on the subject, a sufficient number

of autpsies of, if possible, sudden death (since diseases over long periods

of time may change the distributions of trace elements) with proper sampling,

proper analytical methods and evaluations would have to be performed.
       £&—£    .  Total Body Burdon of Cadmium by Ag«(NM*)
                             Ttaartful MM of total body bur*n
                                 10    4*SI
                                YMIV of Af«

-------
                                     2356
     As shown in Figure 2, Tsuchiya, Sugita, and  Seki  (1972)  first  utilized


accumulation curves of cadmium in various organs  by age  for deriving Biological


half times of the metal for the whole body and for some  organs.   The reported


experimental studies on animals and on man have indicated  great  discrepancies


In the biological half times of the metal.  In our study on humans,  we  found


•uch longer biological half times of cadmium than in those on animal experiments.


We believed then, as now, that most of the cadmium absorbed would probably be


combined with metallothioneln to  forn "slow components"  if the uptake of



cadmium is low  in amount.  Since  the biological half  times of other environmental


metals have not yet been determined, more studies using  this  kind of approach


should be made  In the future for  that purpose.


3«   Dose-response relationship
          Since information on dose-response  relationship  of  metal  gathered


from animal experiments cannot be directly applied  to man.  epidemlological


studies are required, specifically concerning low doses of  exposure over  a


long period of time.

                Fig. 3
                                          Corr«»ponding Mean Air
              t.n
                                      a,
                                  A A™.* *.,»«, *+
                                                    / TtirffciyA 4T*/.
                                                         '
                                                         *t IL*,
                                                                   i
                                                                   \

                                                                   '
                  a«"
                 •
                                          I.*
                                 X«ad in Air

-------
                                    2357
Figure 3 is a combined figure of studies by Tsuchlya et al.  (1974) and Tapper


(1972), showing Che relationship between lead concentration  in the *ir and

blood lead concentration.  The two studies seem to indicate  a fairly good

agreement in their results.  However, if you look at the dot marks by Tapper,


there Is obviously no correlation between the level of lead  in the air and the


blood lead concentration in man, whereas if you look at the  cross marks In


Tsuchlya's study, they seem to show a correlation within very small ranges of


the concentration of lead in the air and of blood lead level.  In fact, the


correlation was significant with a coefficient of 0.7.  In Tsuchiya's study

                                                               3
there was no higher lead concentration In the air than 1.5 jig/m , while the


highest lead concentration in the air in Tepper's study was  nearly 4.0 jig/m  .


The correration which was observed in the Tsuchiya study may be correct or

Incorrect, due to the fact that in that study the subjects were approximately


2,300 policemen who might have been exposed to higher concentrations of lead


for longer hours per day than the housewives of the U.S.A. In the Tepper study.


Furthermore, it is also possible that the correlation of the two highest points


was strengthened merely by coincidence.  Also, the difference of lead concent-


ration in blood between the highest and lowest points in the Tsuchiya study was


only 1,5 pg/lOOg.  At the present time, therefore, we cannot draw the conclusion


that the lead concentration of air between nearly 0 up to 4 ug/m  would signi-


ficantly Increase lead concentration of the blood


     In areas of low exposures one must be very earful in drawing conclusions


about dose-response relationship before numberous other observations have been

made.

-------
                                      2358
s
I
                   Figure  4  Irrationality of Extrapolation


                                    High doia and high reeponae
                                            High correlation
                          *
                          *
                        f A
                       x  * * ^
                        * A *
  Significant correlation by extrapolating
  the reault in a high exposure to a low
  exposure

Ho correlation

Low do»a and low expoaure
                              DOM
Generally,  as shown in Figure 4,  in "normal"  exposura there is little or no


relationship because of no  response in "normal"  or a little higher  exposure,


and because of analytical errors  in measuring both and response.  On the other

hand, in higher exposures one may obtain a very  close correlation.   As shown

in the figure, when high and  low  exposures are combined, a straight linear


regression  is obtained.  From these facts 1 would like to point out here that


one should  never extrapolate  an observation obtained in the higher  or lower


exposure ranges to evaluate dose-response relationship.

                        Figur* 5  Blood LMd and   ALA In Orliw
                     40*
                     90
                    j"
                    1
                     10
                        —DOM-r«*ponMt relationship in
                         l*v*ls of exposure —
                                       A
                             to     40     M    to    100

                                 •*-•

-------
                                    2359



    An example of such a situation demonstrated by avalable data Is shown In

Figure 5 (Sakural, Suglta and Tsuchiya, 1974), which shows a dose-response

relationship between blood lead   and  ALA In the urine in a wide area ranging

from low dose to high dose.  As Indicated in the figure, the response which is

ALA concentration In urine (In this case) is constant until lead concentration

In blood rises up to about 40-50 ug/lOOg.  This means that there  IB no correlation

in lower doses of lead.  In higher exposures, however, a high correlation is

observed.   In conclusion,  here again,  to  extrapolate would be irrational,  as is ;

the case in all  biological sciences.




4.   Detoxication of  trace metals.
           In metal  toxicology  detoxication or adaptation mechanisms have never

been resported expect  for  observations  on cadmium exposure in which matallo-

thionein  Is produced in man and  in  experimental animals.   Hetallothlonein of

cadmium has been  considered to be a detoxlcating  agent which prevents toxic

effects of cadmium  on  the  cells.  Recently a study by Kimura et al. (1974)

demonstrated pertinent evidence  to  show that cadmium tnetallothioneln has an

activity  of detoxlcating the effects  of cadmium.


           Figure 6  Kelationship between production  of lt  25-(OH)
                     TD.. and Cd Metal loth ionsin an well ?.B  Cd
                  -                       (Kiraura, M.  at al., 1974)
                  I   f

                  i
                    "
                 m

                 I
                    -
                     |. Ml
                                         * » »
                             *.M* t.*l  t.lll t.OI t.l «.l «.| Ml)
                        (J COM. H W* t
                                           tl

-------
                                    2360
Figure 6 shows a result of one of his animal experiments.  In the figure it is
clear that vitamin D. activity is reduced with the increased ionic cadmium
concentration in the kidneys of animals, whereas the vitamin activity remains
normal in spite of the high concentration of cadmium which is combined with
metallothionein.  This kind of detoxication mechanism has never been demon-
strated for other metals.  However, there have been a few reports which also
suggest the existence of detoxication for lead.  Tsuchiya in 1954 performed
animal experiments in which resistance against lead toxicity was shown in
animals pre-treated with lead,  tore recently, in 1972, a similar observation
was made by Yoshikawa.  In conclusion I would like to say that more epidemi-
ological studies should be made on industrial workers as well as on the general
population exposed to excessive amount of environmental metals in order to
 discover whether  detoxication or adaptation occur.

 5.    Evaluation of  unspecific responses in individuals and  in groups.

           Since exposures  to  environmental metals  in  the general  population
 are low over  a long period of time,  the effects of  these metals are  in  general
 insidious  and are detected only by observations of  groups,  not of individuals.
 Furthermore,  very early signs of toxic  effects of metals are usually unspecific
 For instance, one early sign  of cadmium Intoxication  is  proteinuria,  but
 proteinuria due to  a number of cases is also  observed in the so-called  "normal"
 population with increased  prevalence according to  age.   Very often medical
 treatment  is  not  required  for those  people with proteinuria which is not very
 high in quantity  and not attributable to just one  specific  disease.   In soQe
 areas in Japan which are known to  be cadmium  polluted, higher prevalences of
 proteinuria have  been observed than  in  control areas. Furthermore,  protelnucil

-------
                                  2361
with low molecular proteins was observed among older  people,  parti-
cularly among females and those over seventy, in non-polluted areas.
From such observations one can state that some effects  of cadmium,
particularly  on the kidneys, exist  in the polluted areas  as affecting
a group of  people. Nevertheless,  if one person with proteinuria and
low molecular proteins from that  group was isolated,  it could not be
determined  whether that particular  individual was affected by cadmium.
Ineuch a case,  only cadmium concentration in the urine  could  be
ferred to for differential diagnosis.
u

40

10
2t
1*
t
M
Ml*
40


J
ffd
- M M
11.*
[
7
//
I

JO
M
1*

mil*


t
14.*
7.1
\


tt - -MUM-
Omtral 1 1
ATMnle Hiiw VJ/7X (IiHl*«i«f MoapnioMl mfumn to tto Mttl U •!••»
      Figure 7  Prevalence of Abnormal EHG Findings  in Arsenic Mine
               and Control Area (Talcahashi and. Kakamura 1974)
     Takahashl  (1974), as sho%m in Figure 7,  observed higher percentages  of
abnormality in  the EMG among people older than 50 years of age in a conmnlty
located near an arsenic mine in comparison with  those people living in a
control areast   The mine had been closed down more  than 20 years preceding
the study and some environmental pollution by arsenic was detected in the
water, soil, agricultural products, and also In the drinking water in the
wells of a few  homes.  This was because the sludge  of a nearby hill along
which a river flows was found to contain a very  high concentration of arsenic.
Biological samples from people In this area did  not show eleveted concen-
trations of arsenic at the tine the study was made.  Furthermore, skin ab-
normalities which are characteristic of arsenic  poisoning were not found  In

-------
                                     2362

any individual.  This situation is very similar to that  mentioned  above  for
cadmium and proteinuria.  As shown in Figure 7, percentages  of  abnormal  EMG
are  higher in the polluted areas than in the control area.   It can  be said
that the area had been polluted and minor effects of arsenic on the  health of
the inhabitants exist.  However, if one individual with  an abnormal  EMG, that
is to toy, unspecific response, were to be pointed out from  that group,  it
would not be possible to make the diagnosis that that person is suffering from
arsenic poisoning.
Thus,  the above  mentioned  two cases of  cadmium and  arsenic strongly
suggest that  toxic metals  accelerate the general aging  process of  man.
Conclusion


     In conclution,  the recent  studies of  environmental toxicology have demon-
strated various methodological  approaches  and  detected many minor effects of
environmental  metals on health  in comparison with calssical studies in the
field of  industrial  toxicology.  In  environmental toxicology epidemiological
approaches become more  and more important,  and the evaluation of the results,
the most  crucial  aspect, becomes more and  more complex.   The primary task of

 environmental toxicology is to provide the Information  needed  in  order  to
 establish environmental standards for toxic metals.  To obtain basic infor-
 mation for this  purpose we need not  only  studies of  the toxicitles of metals
 on animals and human beings, but especially, more studies to provide the
 general background of trace metals in the environment and in man.  The  ulti-
 mate goal of environmental toxicology is  a clean environment free from  pollu-
 tion by toxic metals.  This goal cannot be attained  without collaborative
 studies on an scale which would furnish more precise information  on formal
 pollution by toxic metals in the past,  present,  and  future, and in particular,
 on the effects they have on the health of the  human  race.

-------
                                 2363
                                References

Committee on Biologic Effects of Atomospheric Pollutants, Division of
Medical Sciences, National Research Council: Lead, Airbone lead, in
perspective., National Academy of Sciences, Washington, D.C., 1972.

Friberg, L., Piscator, M., and Nordberg, G., Cadmium in the Environment,
The Chemical Rubber Co., Cleveland, Ohio 44128, 1971.

Horiuchi, K., Lead in the Environment and its Effect on Man in Japan,
Osaka City Medical J., 16, 1, 1970.

Kimura, M. et al., Relationship between productions of 1, 25-[OH]2~D_ and
Cd Metallothionein as well as Cd  ., F.E.B.S. Letters, Accepted for
publication, 1974.

Kitamura, S., Sumino, K., and Kamatani, N., Cadmium concentrations in livers,
kidneys and bones of human bodies, Jap. J. Publ. Health, 17, 761, 1970  (In
Japanese).

Kumamoto University, Minamata Disease, Study Group of Minamata Disease,
Kumamoto University, Japan, 1968.

Sakurai, H., Sugita, M., and Tsuchiya, K., Biological Responses and Subjective
Symptoms in Low Level Lead Exposure.  Accepted for publication in Arch. Environ.
Health. 1974.

Schroeder, H. A. and Balassa, J. J., Abnormal Trace Metals in Man: Cadmium,
J. Chron. Pis.. 14, 236, 1961.

Takahashi, K. and Nakamura, H., Neurological Examination of Inhabitants in
Sasagadani Area of Arsenic Mine, Kankyohoken Report, 32, 1, 1974 (In Japanese).

Tepper, L., and Levin, L. S., Asurvey of Air and Pollution Lead Levels in
Selected American Communities, Department of Environmental Health, College
of Medicine, University of Cincinnati, Ohio, 1972.

Tipton, I. H., and Cook, M. J., Trace Elements in Human Tissue, II, Adult
Subjects from the United States, Health Phy.. 9, 103, 1963.

Tipton, I. H., Schroeder, H. A., Perry, H. M., and Cook, M. J., Trace Elements
in Human Tissue, III, Subjects from Africa, The Near East and Far Eastern
Europe, Health Phys., 11, 403, 1965.

Tsuchiya, K., and Kuwaki, H. A., A Study on Tolerance of Animals to Lead
Poisoning, J. of Science of Labor. 31, 291, 1955 (In Japanese).

Tsuchiya, K., Sugita, M., A Mathematical Model for Deriving the Biological
Half-Life of Chemicals, Sartryck ur Nordisk Higienisk Tidskrift. Band LIII,
sid. 105, 1971.

-------
                               2364
Tsuchiya, K., Seki, Y.,  and Sugita, M., Biological Criteria for Exposures
to Lead and Cadmium, presented In the 17th International Congress of Occu-
pational Health, Buenos  Aires, Sept. 17-23, 1972.

Tsuchiya, K., Sugita, M., and Sekl, Y.. A Mathematical Approach to Deriving
Biological Half Times of Cadmium in Some Organs — Calculations from
observed accumulations of the metal in organs, presented in the 17th
International Congress on Occupational Health, Buenos Aires, Sept. 17-23,
1972.

Tsuchiya, K., Sugita, M., Seki, Y., Kobayashi, Y., Hori, M., and Park, C. B.,
Study of Lead Concentration in Atomosphere and Pollution in Japan, Accepted
for publication in J. of Environ. Quality and Safety, 1974

Yoshikawa, H., Preventive Effect of Pretreatment with Low Dose of Metals on
the Acute Toxiclty of Metals in Mice, Ind. Health. 8, 184, 1970.
                           DISCUSSION
MAGE  (Denmark)
     In view of the very large standard deviation  in  a spatial
lead distribution in  the atmosphere  shown by Edwards in his
Paper, it is not reasonable to expect zero  correlation at low
Pb  levels, since the  blood lead level is produced then primarily
by  food and liquid intake.  At high  airborne lead levels, the
effect of food and drink is submerged and the correlation shines
through like the sun  through a cloudy sky.

     The high correlation data then correctly shows as you even
Indicate on figure 4, that in the limit as airborne  lead goes
to  zero, the response also goes to zero or a  (food-liquid) thres-
hold  which is finite but small.

     Therefore since the combined model is physically significant,
why can't we state that the extrapolation to zero is valid.


TSUCHIYA  (Japan)

     I did not mean to imply that there is never any  correlation
In  lower exposures.   I simply wish to point out that one cannot
extrapolate the results obtained  in higher exposures which were
actually observed to  lower exposures without the  evidence of
actual observations.   As is well known, in general,  the dose-
response relationship In radiation exposure, both  dose and res-
ponse will reach zero  but in heavy metal  exposures  there is no
such  linear correlation.

-------
                              2365
JACOBSEN  (U.K.)

     In your figure 5 there are two types of symbols implying
that the data are derived from two studies.  I agree that if
only the high-dose data were available then it would be unrea-
sonable to extrapolate linearly and thus infer zero response
at moderately high exposures.  Conversely, if the low-dose data
why were available then it would be equally unreasonable to
extrapolate linearly and thereby infer a slope close to zero.

     Would Dr. Tsuchiya agree, however, that if the data had
been obtained in the same study, or if the studies are suffi-
ciently similar in methodology to justify pooling the data,
then it would not be unreasonable to hypothesise a sigmoid dose-
response relationship from the result shown?


TSUCHIYA  (Japan)

     I am in complete agreement with your .comments.


KJELLSTROM  (Sweden)

     Looking at your data on relation between EMG abberations
and arsenic exposure and your discussion about proteinuria and
cadmium, it must be concluded that in environmental science
conclusions about cause-effect relationships must be drawn from
population studies.

     Do you agree that it is most unscientific to try to refer
unspecific response in an individual to a specific exposure
which is sometimes done?
TSUCHIYA  (Japan)

     I agree that it is most unscientific to try to refer un-
specific response in an individual to a specific exposure as Is
sometimes done.  If the incidence of an unspecific response is
rather high among an exposed population, it is very crucial to
confirm whether the unspecific response in an individual sel-
ected from the exposed population is actually caused by the
pollutant in question.

-------
                              2367
   ASSESSMENT OF THE INFLUENCES OF ENVIRONMENTAL POLLUTANTS
             ON CANCER AND OTHER CHRONIC DISEASES

                         S, S, EPSTEIN

Environmental Health Programs, Case Western Reserve University,
Medical School, Cleveland, Ohio, USA
ABSTRACT

     There is now little doubt that many chronic diseases, hith-
erto regarded as spontaneous, particularly cancer, are caused by
environmental pollutants.   This realization is heightened by the
exponential increase in exposure of the general population to cur-
rently used and new synthetic chemicals - and their degradation
products in air, water and soil - which, in general, are inade-
quately characterized toxicologically and ecologically.   These
considerations apply with even greater emphasis to relatively
uncontrolled occupational exposure to a wide range of "known chem-
ical carcinogens, in addition to thousands of other toxicologic-
ally uncharacterized or inadequately characterized synthetic
chemicals.

     Current toxicological techniques are relatively insensitive
and limited in their ability to detect toxic agents, particularly
carcinogens teratogens and mutagens, individually and in various
combinations or mixtures realistically reflecting low or ambient
levels and patterns of environmental exposure.   Similarly, it
is generally considered that epidemiologiaal techniques are un-
likely to detect environmental pollutants, such as weak carcino-
gens, unless there are sharp differentials in exposure of the
general population, as with cigarette smoking.   For widely dis-
persed agents to which the population-at-large is generally and

-------
                              2368
ubiquitously exposed, such as unintentional or accidental food
additives, human experience is unlikely to provide any meaning-
ful indication of safety or hazard.   Similarly, it is not pos-
sible to develop valid inferences concerning the safety of occu-
pational exposure to particular chemicals or mixtures of chemic-
als in the absence of an adequate population sample which has
been adequately followed up for several decades.

     Scientific considerations apartt there are critical defici-
encies in legislative and regulatory approaches to environmental
pollutants, including conflicts of interest in the generation and
evaluation of data, restrictions on open access to data, and lack
of qualified representation of a wide range of concerned view-
points and interest in decision making processes.

-------
                                 2369
1.   Introduction
     Although the term pollutant is often pejoratively restricted to
synthetic industrial chemicals, there is a wide range  of other chemical
pollutants.  These fit into four broad categories.   The first group con-
sists of natural chemicals in excess, such as nitrates and nitrites,
which are normal dietary components.  Additionally,  these particular
chemicals can interact with amines, natural dietary  constituents) yield-
ing nitrosamines, which may be carcinogenic,  mutagenic and teratogenic
at trace levels (Lijinsky & Epstein [1]  ).  Natural  fungal or plant
toxins in crops comprise the second group of chemical  pollutants, of
which aflatoxins and cycasins are notable exBmples.  The yields of these
toxins can generally be influenced by technological  factors,  such as
conditions of harvesting, storage and processing.  The third  group con-
sists of complex organic and inorganic mixtures, such  as community air
and water pollutants and occupational pollutants,such  as coke tar pitch
volatiles, which comprise a wide range of undefined  as well as partially
defined components.  Finally, there is the group of  synthetic chemicals-
agricultural chemicals, notably pesticides and fertilizers; food addi-
tives, which may be intentional, such as antioxidants  and dyes, or acci-
dental, such as pesticides, heavy metals and plasticizers; fuel additives;
household chemicals, and industrial chemicals.  Most of these chemicals
are petroleum-based, petroleum now being the basic stock for  synthesis
of the great majority of all organic chemicals.
     Pollutants may induce a wide range of adverse biological effects
in man, which are generically and collectively termed  toxicity.  Acute
or chronic toxicity per se_ may be expressed in fetal,  neonatal, peri-
natal, childhood or adult life, in effects ranging from impairment of
health and fitness to mortality.  More specific manifestations of
chronic toxicity include carcinogenicity, teratogenicity and  mutageni-
city.  The possibility that chronic toxicity is also manifest in immu-
ne logical impairment or in psycho-behavioral "disorders has yet been
barely explored.  Some pollutants may induce one or  more of these types
of toxicity.  Pollutants or their chemical precursors  may also interact
invitro and in vivo to produce otherwise unanticipated synergiatic
toxicity.  Synergiatic effects can also result from  interactions between
particular pollutants and otherwise harmless and common environmental
chemicals.

-------
                                 2370
     The need to use many synthetic industrial chemicals makes it essen-
tial to recognize and estimate the human and environmental hazards they
pose and their societal acceptability with regard to the real or alleged
matching benefits they confer.  Hazards from a particular synthetic
chemical,whether in consumer products or in the workplace, need not
necessarily be accepted even when matching benefits appear high, as
equally efficacious but nonhazardous alternatives are usually available.
Imposition of a mandatory criterion of efficacy prior to the introduc-
tion of synthetic chemicals into commerce may well simplify such equa-
tions.
2.   Cause For Concern
     There is now little doubt that many diseases hitherto regarded as
spontaneous, including cancer, birth defects and mental deficiency are
caused by environmental pollutants.  This realization is heightened by
the exponential increase in human exposure to new synthetic chemicals
—and their degradation and pyrolytic products in air, water and soil-
which, in general, are inadequately characterized toxicologically and
environmentally (Epstein [ 2 ] ).
     Recognition is now growing that the great majority of human cancers
are probably due to chemical carcinogens in the general or working en-
vironments, and that they are hence ultimately preventable (Dunham &
Bailar [ 3 ] ; Higginson [4 ] ).  There is also growing interest in the
role of chemical carcinogens in activating oncogenic viruses.  Epide-
miological studies have revealed wide geographical variations in the in-
cidence of cancer of various organs in the general population; in some
instances these studies have incriminated local environmental pollutants.
     The first evidence that environmental pollutants may influence the
genetic constitution of future populations resulted some four decades
ago from the discovery that high energy radiation induces mutations.
The subsequent development of nuclear energy added a new dimension and
enhanced awareness of the problem of genetic hazards.  Safeguards have
been accordingly developed to minimize radiation exposure.  Once radia-
tion-induced mutagenesis was discovered, there were reasons to suspect
that some chemicals would act similarly, but proof of this was delayed
until World War II when mustard gas was shown to induce mutations in
fruit flies.  Many and varied types of chemicals have subsequently been
shown to be mutagenic.  The likelihood that some highly rautagenic chemi-

-------
                                  2371
cdls may come into wide use, or indeed may already be in wide use,  is now
causing serious concern.  No nation has yet, however, promulgated regu-
latory requirements for mutagenicity testing.
     There is also growing recognition of the importance of environmental
pollutants in the causation of birth defects.  National incidences  of
congenital malformations are unknown in the absence of   comprehensive
registries; it has been variously estimated as ranging from 3 to 4  per-
cent of total live births.  Three major categories of human teratogens
have so far been identified: viral infections, X-irradiation and chemi-
cals, such as mercurials and thalidomide.  Although the teratogenicity of
various chemicals had been experimentally recognized for several decades,
it was only after the thalidomide disaster of 1962 that legislative re-
quirements for three-generation reproductive tests were established.
A substantive body of data is now establishing clear relationships  be-
tween exposure to a wide range of environmental and occupational pollu-
tants and delayed neurotoxic effects, ranging from mild alterations in
personality and behavior to advanced dementia.  Illustrative, are the
effects of lead, which at exposure levels and body burdens considered
to be relatively low  induce  disturbances in personality and neuro-
muscular co-ordination in workers (Morgan & Repko [5]; Seppalaninen [6]  )
and hyper kinesis and learning disorders in young children (Environmental
Protection Agency [7]  ).
3.   Toxicity Testing
     Pollutants to which humans are or may be exposed must be tested for
acute and chronic toxicity per se_, and also for the more specific effects
of carcinogen icily, teratogenicity and mutagenicity.  Toxicity testing
must not be confined to the test agents per se,  but should extend to
their chemical and metabolic derivatives, pyrolytic and degradation pro-
ducts and contaminants.  These considerations are further accentuated
when the various derivatives or degradation products are of known toxi-
cological or ecological consequence.  Test agents must be administered
acutely, subacutely and chronically to reflect the role of hepatic
microsomal enzyme function in activation and detoxification.  It may also
be necessary to test for effects of concomitantly administered and  other-
wise nontoxic chemicals that may induce or inhibit microsomal enzyme
function.  Experiments must be designed to reflect the role of possible
interactions between test agents —administered by any route —and  be-

-------
                                  2372
tween dietary factors and other chemicals, such as accidental and inten-
tional food additives, drugs and air pollutants.  Routes of test adminis-
tration inter alia should reflect human exposure.  While inhalation is
the obvious route for testing of air pollutants, the importance of this
route for other pollutants has been generally underestimated.  Respira-
tory exposure is of particular human significance for pesticide aerosols
and vapors, besides other aerosols.  Surprisingly, there are virtually
no data in the pesticide literature on chronic inhalation tests.  Ideally
and minimally, two mammalian species should be tested for toxicity per se,
carcinogenicity, mutagenicity and tera to genie ity.  In certain circum-
stances when there is specific information that the rodent metabolism of
the chemical pollutant in question is qualitatively markedly different
from that in humans, other more appropriate species, such as pigs and
subhuman primates, may also be tested.  Reliance on small numbers of pigs
or primates is no substitute for conventional rodent tests and may even
mislead.  In particular circumstances special considerations may dictate
the use of less common species.
     For carcinogenicity, teratogenicity and mutagenicity pollutants must
be tested at higher levels than those of general human exposure; irre-
spective of route of administration, maximally tolerated doses are re-
commended for this purpose as the highest dose in dose-response studies.
Testing at high doses is essential to the attempt to reduce the gross
insensitivity imposed on animal tests by the small size of samples rou-
tinely tested, such as fifty or so rats or mice per dose level per chemi-
cal, compared with the millions of humans at presumptive risk.  To illus-
trate, assume that man is as sensitive to a particular carcinogen or
teratogen as the rat or mouse; assume further that this particular agent
will produce cancer or teratogenic effects in 1/10,000 humans exposed.
Then the chances of detecting this in groups of fifty rats or mice tested
at ambient human exposure levels would be very low.  Indeed, samples of
10,000 rats or mice would be required to yield one cancer or teratogenic
event, over and above any spontaneous occurrences; for significance, per-
haps 30,000 rodents would be needed (Epstein! 2 ] ).  Of course, in any
particular instance, humans may be less or more sensitive than rodents to
the chemical in question; there is consequently no valid basis for the
prediction of the relative sensitivities of test animals and man.  Apart
from the gross insensitivity of animal test systems and the impossibility

-------
                                2373
of gauging human sensitivity from animal tests, ample data on inter-
actions between carcinogens further confirm that it is not possible  to
predict safe levels of carcinogens based on an arbitrary fraction of the
lowest effective animal dose in a particular experimental situation.
Such considerations underlie the 1958 Delaney Amendment (P.L. 85-929) to
the Federal Food, Drug and Cosmetic Act, which imposes zero tolerances
for carcinogenic food additives.  The amendment states: "...no additive
shall be deemed to be safe if it is found,  after tests which are  appro-
priate for the evaluation of the safety of food additives, to induce
cancer in man or animal. The concept of zero exposure work standard
has been accepted, but for only 14 occupational carcinogens, in regu-
lations promulgated in the Federal Register on February 11, 1974.  How-
ever, these regulations do not contain provisionsto ensure effective
implementation of the standards (Epstein [ 8 ] ).
     It must also be emphasized that testing at high dosages does not
produce false positive carcinogenic results.  There is no basis whatso-
ever for the frequent  contention, particularly  by  industrial toxi-
cologists and consultants, that all chemicals are carcinogenic, tera-
togenic, or mutagenic at high doses.  To illustrate, in the recent
Bionetics study, sponsored by the National Cancer Institute, about 140
pesticides were tested orally in mice of both sexes and strains at maxi-
mally tolerated doses from the first week of life until sacrifice at
eighteen months; less than 10 percent of these pesticides were found to
be carcinogenic (Innes et^ al [9] ).
4.   Monitoring  and Epidemiological Surveillance
     Persistent chemicals, chemical and metabolic derivatives of less
persistent chemicals and their reaction and pyrolytic products should
be detected and monitored in the environment - air, water, soil and
food - and in body fluids or tissues of plants, animals and man.
Selectively, only those chemicals or degradation products with known or
presumed toxicological relevance should be monitored.  Even with well-
planned and well-executed toxicologic testing, it is likely that un-
expected adverse effects from pollutants will occur, reflecting the  in-
sensitivity or inappropriateness of the test systems.  Epidemiological
surveys of human and animal populations may provide post ho£ information
on geographical or temporal clusters of unusual types or frequencies of
adverse effects — Including cancer birth defects, and mutations —after

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                                2374
exposure to undetected or untested pollutants in the environment.  Such
surveys are complicated by the long interval which may elapse between
exposure and subsequent adverse effects.  This may be measured in de-
cades for cancer and in generations for mutations.
     Epidmiologic techniques serve to detect trends or fluctuations in
mortality, morbidity or disease patterns.  Provided that clear differ-
entials in exposure levels to pollutants exist in the general population,
epidemiology may then correlate particular toxic effects with particular
pollutants; the association  between heavy cigarette smoking and lung
cancer is a classic example of such a relationship.  However, these
relationships are more difficult to establish when exposure differen-
tials are minimal, as with a food additive consumed at more or less
similar levels by the general population.  Additionally, logistic con-
siderations, quite apart from inadequate current surveillance systems,
may limit the utility of epidemiological approaches even when temporal
or spatial clusters of adverse effects have developed.  Disquietingly,
no major known human teratogen— X-rays, German measles, mercury or
thalidomide— has been epidemiclogically identified, even in indus-
trialized countries with good medical facilities.
5,   Contrast Between Environmental and Occupational Health Standards
     A fundamental dichotomy, both scientific and moral, exists between
current approaches to standard-setting for the working population and
for the population-at-large (Epstein [ 8 ] ).  In spite of their major
inadequacies, standards that have been developed for the protection of
the population-at-large against adverse effects from exposure to a wide
range of chemical pollutants and products are generally predicated on
the availability of an adequate data base.  Necessary information in-
cludes chemical composition of such products, labelling and disclosure
of ingredient identity, and on the testing of such products in animals
for acute and chronic toxic effects prior to their release into
commerce.  For agents producing acute and chronic toxicity per se_ in
animals threshold or no-effect levels are determined and standards are
then developed, generally, based on 100 - fold safety margins.  Agents
which induce carcinogenic effects in appropriate animal tests or which
are known to be carcinogenic to man are generally banned from commerce,
as no level of exposure can be considered safe.

-------
                                 2375

     These general requirements are in striking contrast to the  situa-
tion for occupational health standards.  Standards exist for only a
small fraction, about 450, of chemicals to which workers are exposed.
Illustratively, a "Toxic Substances" publication of the  National
                •
Institute of Occupational Safety and Hygiene (NIOSH) listed approxi-
mately 8,000 known chemicals used in industry in 1971 and approximately
25,000 in 1973; these figures clearly underestimate the  numbers  of
chemicals to which occupational exposure can occur.  Most Federal U.S.
standards are based on approximately 450 threshold limit value (TLV)
standards developed by the industrially-oriented, if not dominated,
American Conference of Government Industrial Hygienists  (ACGIH); these
TLV standards are often referred to as "proprietary", reflecting their
narrowly focused "trade" origin.
     The concept of adequate safety margins is scarcely, if at all,
reflected in occupational, in contrast with general environmental
standards.  This is well exemplified by reference to current occupatio-
                               o
nal standards on lead, 150 ug/m , in contrast with proposed general
environmental standards in California, which are 100-fold lower
(Epstein [8] ).  Furthermore, once occupational standards have been
developed and promulgated there is currently no effective method for
implementing them.  The tacit reliance on voluntary compliance is in
part, perforce, an expression of imposed fiscal, personnel, and grade
ceilings in the Occupational Health and Safety Administration (OSHA)t
and in part an expression of the philosophy of the dual standard.
     Finally, there are no current requirements for "pre-testing" or
screening chemicals prior to their manufacture and use by industry,  nor
are there even general requirements for open disclosure  of the identity
of chemical agents to which workers are exposed.  This is clearly con-
trary to the intent of the Occupational Safety and Health Act, effec-
tive April 28, 1971, which mandated the provision of a safe and healthy
working environment.  No such assurances can possibly be made in the
absence of information as to the chemical nature and possible biological
effects of these exposures.  In the absence of "pre-testing", the
worker is unwittingly used as an involuntary test subject, to whom test
data are not made available, if indeed they are ever collected and
analyzed.  Recognition of adverse effects in retrospective epidemiologi-
cal studies, is post hoc.  These human experiments are unnecessary and

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                                  2376
amoral.  Illustratively, the carcinogenic effects of bis-chloromethyl
ether and vinyl chloride, and the neuropathic effects of organic solvents,
such as methylbutylketone, could easily have been determined by simple
and standard animal tests, rather than by human experimentation.
6.   Evaluation of the Benefit-Risk Calculus
     Since World War II, there has been an exponential and,  largely,  un-
regulated increase in the numbers and quantities of synthetic organic
chemicals manufactured and used in industrialized countries.  The claimed
needs to use increasing numbers of new synthetic chemicals makes it
essential to recognize and critically evaluate carcinogenic  and other
human and environmental hazards with regard to the real or alleged match-
ing benefits they confer.  Such costing must be weighted by  factors in*-
clading the persistence and environmental mobility of the chemical, the
size of the population exposed, and the reversibility of the adverse
effect.  Total national monetary costs in the U.S., both direct and in-
direct, from cancer are estimated to be approximately $15 billion annu-
ally (National Cancer Program [10] ); these costs have hitherto been
largely externalized or discounted.  As the majority of human cancers,
both in the general population and in occupational groups, are now con-
sidered to be due to chemical carcinogens and hence preventable, there
should be clear economic,besides other, incentives to reduce the
environmental and occupational burden of chemical carcinogens.
     Carcinogenic hazards from a particular synthetic chemical need not
necessarily be accepted even when matching benefits appear high, as
equally efficacious but nonhazardous alternatives are usually available.
The mandatory criterion of efficacy, once extended from therapeutic
drugs to other synthetic chemicals, such as deliberate and accidental
food additives, feed additives and pesticides, may well simplify such
equations, especially for hazards from synthetic chemicals with no
demonstrable benefits for the general population.  The imposition of a
requirement for broad social utility may even further simplify the bene-
fit-hazard equation.  Such concepts have been recently emphasized with
regard to food additives by a leading industrial representative who
recommended that additives be excluded from products unless  they either
significantly improve the quality or nutritive value of the  food or
lower its costs as well as being safe (Kendall [11] ).  Claims that
occupational carcinogens serve industrially unique purposes, must be

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                                 2377
examined critically by economically disinterested experts  with particu-
lar recognition of the attendant and generally externalized human costs
and the lack of economic incentives to develop similarly efficacious and
non-hazardous alternatives.  In the absence of such alternatives, con-
sideration must be directed to the possible banning of the manufacture
and use of the carcinogen or to restrict its use to closed systems which
are continuously monitored with instrumentation of maximal sensitivity,
and with automated and visible read-outs.
     Inherent in toxicological and regulatory philosophy and practice is
lip service to the concept of balancing benefit, and benefit to the
public not to industry, against risk, and risk to public health or en-
vironmental integrity and not economic risk to industry.  If the chemical
in question does not serve a broad socially and economically useful pur-
pose for the general population, why introduce it and force the public-
at-large to accept potential hazards without general matching benefits?
Such questions should be vigorously directed to carcinogenic, and other-
wise hazardous, cosmetic food colouring agents, in particular, and to
all food additives, in general*  Claims have recently been made
(Gehring ejt a_l [12] ) that requirements for pre-testing of chemicals
prior to their introduction to commerce are acting as disincentives to
industrial innovation.  These claims have been particularly directed to
the manufacture of pesticides (Naegele [13] ).  Apart from the fact that
such claims are predicated on the legitimacy of externalizing public
health hazards and costs, they do not bear critical scrutiny even from
narrowly defined economic viewpoints.  A telling critique  of such claims
has been expressed by a leading industry spokesman who stated that he —
"emphatically (takes issue with the line of reasoning that) escalating
regulatory demands have made the cost of research and development pro-
hibitive, thus drying up any incentive to go develop new agricultural
chemicals...
"... In the first place (argued Dr. Sutherland), new regulations imposed
since the creating of EPA affording better protection to fish and wild-
life were overdue.  More important is the changing aspect of the market-
place, particularly in the pesticide area.  Growers now have available
to them many first-rate products... many of these are quite inexpensive.
What the chemical people are really telling you is that while research
costs continue to rise, to come with still better compounds costing no

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                                  2378
more money than what's already being sold  is a tough proposition...;
the companies with weak research organization, a shaky financial posi-
tion, are dropping out.  They would rather have FDA and EPA take the rap
rather than acknowledge the overall problem" (Sutherland [14]  ).
In any event, information in 1973 suggests that the profitability of
pesticides and their development is again rising (Bennett [ 15 ] ).
     It has now become axiomatic that there are major defects in decision
making processes in regulatory practices. It is clear that the demo-
cratic system of checks and balances is largely absent from current
regulatory practice (Epstein [2 ] ).  Apart from limited post hoc re-
course, the citizen, consumer and working person, and those who repre-
sent his or her interests, scientifically and legally, are virtually
excluded from anticipatory involvement in decisions vitally affecting
them.  The concept of matching benefits against risk has been generally
applied to maximise short-term benefits to industry, even though this
may entail minimal benefits and maximal risk to the consumer.   While
such an approach is of course detrimental to the consumer, it is also
often detrimental to the long-term interests of Industry,  which may
suffer major economic dislocation when hazardous products, to which it
has improperly developed major commitments, are belatedly banned from
commerce.  Such problems are in large measure attributable to crippling
constraints which have developed and which still dominate the  decision
making process within regulatory agencies.  Responsibility for these
constraints must be shared with regulatory agencies, by the legislature,
by the scientific community, and by citizens, consumers and workpersons
who have not yet developed adequate mechanisms for protecting  their own
vital rights.
     It is perhaps no coincidence  that the attacks on the Delaney Amend-
ment are mounting at a time when the food chemical industry is poised
for a major expansion.  The chemical industry predicts that sales of
chemical additives are expected to grow from $485 million in 1970 to
$750 million by 1980.  In providing a framework for evaluating potential
hazards of these additives, the Delaney Clause simply ratifies the pre-
vailing expert opinion in the National Cancer Institute and in other
professionally qualified groups that there is no practical method to
determine safe dietary levels for a carcinogen (Saffiotti [16];  Epstein
[17] ).  Changing the Delaney Clause to allow regulatory  discretion to

-------
                                2379
 set  tolerances  for  carcinogens  is, therefore, not only scientifically
 inappropriate,  but,  administratively  foolhardy.
      Conflicts  between crucial  social goals, such as reduction in the
 incidence of human  cancer due to environmental and occupational car-
 cinogens, and powerful concentrated economic interests are often joined
 on supposedly scientific grounds.  Illustrative, are the current U.S.
 cancellation hearings on Aldrin/Dieldrin which have largely focused on
 the  significance of carcinogenicity tests in rodents as a basis for
 risk extrapolation  to humans (Epstein [18]).  Industry and its consul-
 tants, generally toxicologists  without primary expertise in carcino-
 genesis and  pathology, have at  varying times advanced the following
 illustrative mythologies:
  1.  The mouse hepatoma is not a true neoplasm but a regenerative
      nodule.
  2.  Hepatoraas induced by mice, illustratively by persistent organo-
      chlorine  pesticides, are  "compound-dependent" and will regress
      following cessation of test exposure.
  3.  The mouse hepatoma is a benign  neoplasm and that agents inducing
      it should be classified as "tumorigens" and not carcinogens.  It
      is thus argued that standard regulatory practices for carcinogens
      are inappropriate for "tumorigens".
  4.  Transplantability of the mouse hepatoma does not necessarily
      establish its ne op las tic nature.
  5.  Dieldrin  is a "species-specific" carcinogen for the mouse and
      the mouse is endowed with a unique hypersensitivity to chemical
      carcinogens.   It is thus argued that data from mouse carcino-
      genicity tests have little or no human relevance.
  6.  Human experience, based on 826 workers of whom only 35 had been
      followed-up for over a decade,  has proven that Dieldrin is not
      carcinogenic to humans.
      In fact, the published literature clearly establishes that Dieldrin
 is carcinogenic in several strains of mice,  at the lowest dose yet test-
ed,  O.I ppm, producing raetastasizing hepatocellular carcinomas in addi-
tion to a variety of neoplasms, at other sites (Epstein [18]  ).  Addi-
tionally, more limited studies clearly establish the  carcinogenicity of
Dieldrin in the rat, in which hepatocellular carcinomas in addition to
carcinomas of other organs have been demonstrated.  It must be empha-

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                                 2380
sized that Dieldrin is highly stable and persistent, that human lip id
levels range from 0.5 to 2.9 ppm, that tolerances for Dieldrin,
petitioned by Shell, in various animal food products range from 0.1 to
0.3 ppm, and that on the basis of such petitions and existing tolerances
on raw agricultural commodities, Dieldrin levels in a standard diet have
been calculated to be 0.04 ppm.
     It is our clear professional responsibility to expose the unscien-
tific nature of the industrial mythology on toxicology, in general,and
on carcino gene sis, in particular, typified in the Aldrin hearings.
Embattled agencies, such as The Environmental Protection Agency, and
public interest groups can not be expected to unaidedly bear the onerous
burden of protecting the public health.  Pressures on agencies can sub-
vert implementation of standards and of the total regulatory process.
This has been well recognized in statements such as the following:
"It is the daily machine-gun like impact on both agency and its staff
of industry that makes for industry orientation on the part of many
honest and capable members, as well as agency staffs" (Landis [19] ).
Nevertheless, appropriate reforms in agency-industry relationships have
yet to be developed.  Reforms apart, it is clear that decisions on the
use of toxic agents,such as carcinogenic chemicals in consumer products
and in the workplace,must be made in the open political arena and  on the
basis of the evaluation of scientific data that is both expert and un-
biased.  Industry must be encouraged to avoid preoccupation with short-
term economic interests and the development of premature commitments to
products and processes which have not been adequately tested by com-
petent and independent investigators.  Such approaches will minimize or
preclude The possibility of economic dislocation which would otherwise
ensue when subsequent challenges necessitate the belated withdrawal of
the product or process from commerce and the workplace.   Such approaches
also reflect recognition of the consonance of long-term industrial In-
terests and broadly-based societal goals and values.
Acknowledgements
      Section 6 of this paper is largely based on an M.S.  "Environ-
mental Determinants of Human Cancer."  Epstein, S.S.  Cancer Res,  in
press.

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                                 2381
 References
 1    LIJINSKY,  W.  and EPSTEIN,  S.S.,  "Nitrosamines as  Environmental
      Carcinogens."  Nature,  225,  21 (1970).
 2    EPSTEIN,  S.S. "Environmental Pathology: A  Review."  Am. J. Path.,
      66 , 352,  (1972).
 3    DUNHAM, L.J.  and BAILAR,  J.C.,  "World Maps of Cancer Mortality
      Rates and Frequency Ratios."  .J. Nat. Cancer Inst., 41, 155 (1968)
 4    HIGGINSON,  J.,  "Present trends in Cancer Epidemiology," Proc. 8th
      Canadian  Cancer Res.  Conf.,  1969.
 5    MORGAN, B.  AND REPKO, J.D.   National  Institute  of Occupational
      Safety and Hygiene  (NIOSH) Contract Report,  HSM-99-72-123, 1974.
 6    SEPPALAINEN,  A.M.  In,  "Behavioral Toxicology:  Early Detection
      Occupational  Hazards",  eds. C.  Xintaras,  B.L.  Johnson and I. de
      Groot, NIOSH, HEW,  In Press, 1974.
 7    "EPA's Position on  the  Health  Implications of Airborne Lead",
      November  28,  1973.
 8    EPSTEIN,  S.S.  Testimony  before the U.S. House  of Representatives
      Committee on  Education  and Labor,  Select Subcommittee on  Labor,
      April 25,  1974.
 9    INNES, R.  et  aU, "Bioassay  of Pesticides  and  Industrial  Chemicals
      for Tumorigenicity  in Mice:  A Preliminary Note."  iJ. Nat. Cancer
      Inst. 42,  1101 (1969).
        1                       ti                  „
10    NATIONAL CANCER PROGRAM-   The  Strategic Plan. D.H.E.W. Publication
      No.  NIH 74-569, January,  1973.
11    KENDALL,  D.M.  "A Summary of Panel Recommendations."  Report of a
      Panel on  Food Safety to the  White House Conference  on Food
      Nutrition and Health, (19),22  November,  1969.
12    GEHRING,  P.J., ROWE,  V.K., & McCOLLISTER,  S.B.  (Dow Chemical Co.)
      "Toxicology:   Cost/Time", Fd_.  Cosmet. Toxicol.  11,  1097 (1973)
13    NAEGELE,  J. (Dow Chemical Co.). Testimony  to U.S.,  Congress, House,
      Committee on  Agriculture, Hearings on Federal Environmental
      Pesticide Control Act of 1971, 92nd Congress,  1st session, Wash-
      ington, D.C.:  U.S. Gov.  Printing Off.,  1971.
14    SUTHERLAND, G.L. (American" Cyanamid).   "Agriculture  Is Our  Best
      Bargaining Tool," Farm  Chemicals, 135, 44  (1972).
15    BENNETT,  I.  "Preface," Pesticides Monitoring Journal,  1: no. 1,
      (June, 1967).

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                             2382
 16    SAFFIOTTI, U.,  ''Comments on The Scientific  Basic for The 'Delaney

      Clause1".  Preventive Med. 2, 125 (1973).

 17    EPSTEIN, S.S.,  "The Delaney Amendment." Preventive Med. 2, 140
      (1973).

 18    EPSTEIN, S.S.   Testimony  at Cancellation Hearings on Aldrin/

      Dieldrin (EPA & EDF vs. Shell), March, 1974.

 19    LANDIS, J.  Report to President-elect Kennedy, 1960.



                        DISCUSSION


van ROOYEN  (South  Africa)

      One  is glad that the question of decision-making  was
raised.   There  are other  aspects that need  consideration.
During the course  of this conference, we  have  had  several
papers on the effects of  environmental factors and more
particularly emissions from specific factories on  populations
at risk.  With  few exceptions no mention  was made  of the
requirements placed on them by air pollution control authorities,
Were  these emissions completely uncontrolled,  partially
controlled, or  by  the best practicable means?  Surely  before-
and-after studies  to evaluate the effect  of control measures
are imperative.

      One  also suspects that pollution is  being exported to
developing countries because of the stringent  measures in
highly developed countries.  The former must accept this as
raising of their national product to control poverty,  pal-
nutrition and  ignorance  is paramount.  It  is  thus necessary
that  these countries should know how effective control measures
are.


EPSTEIN  (U.S.A.)

      I agree with  the concepts expressed.  Developed countries
have  a clear responsibility to avoid "exporting" pollution
to developing countries.   It is hoped that the latter  will
be encouraged to appreciate the long-term economic, besides
public health air  environmental costs of  "importing"
pollution.  It  is,  however, clear tha*t the priorities
influencing decision making processes and the  degrees  of
checks and balances  in such processes will vary widely from
country to country.
van der KREEK  (Netherlands)

     Professor Epstein,  you  complemented the Swedish Government
for the ban of synthetic colours for use in food products,
but I am informed that  this  ban is not a complete ban, but a

-------
ban of some food colours in some food products and therefore
is not a decision of principle, but a decision based on
toxicological and technological principles.
EPSTEIN (U.S.A.)

     The Swedish "Ban" is conceptual and indicates future
policy that cosmetic or coloring food additives will be
disallowed in future in the absence of unqulvocal data on
safety and on critical societal efficacy.
KIELLSTROM  (Sweden)

     I believe on the contrary that the Swedish decision was
very important in principle, but perhaps not very important
toxicologically.  All colour additives to food are banned
in principle and will only be accepted after thorough
evaluation.  This is an example of the "negative proof
philosophy" that I mentioned here earlier which means that
synthetic chemical products should be deemed deleterious to
health at any concentration as long as no data are available,
EPSTEIN (U.S.A.)

     I thank the speaker for his clarification of this
important question.


SERWER   (U.N.E.P.)

     I am sympathetic to what I take to be two of your
fundamental points:  prior evaluation of efficacy and
decentralization of decision-making.  In practice, however,
there may be a contradiction between these two "desirables."
Prior evaluation would remove decisions from the market place,
where they are at  least decentralized if not well informed.
If provision were  not made for opening up decision-making
on efficacy - and  there are practical limits to doing so -
your proposal might in fact cause the situation you deplore
to deteriorate further.
EPSTEIN  (U.S.A.)

     The concept of "freedom of choice" in the market place
is a myth.  The consumer has no options but to purchase
products "accidentally" contaminated with a wide range of
chemicals, such as Dieldrin, of which he is not informed and
for which he is not informed.  Free choice implies labelling
of ingredient and contamination concentration, hazards and
benefits.

-------
                           2384
ZIELHUIS (Netherlands)

     I agree with many of the principles brought forward by
the speaker.  However, taking up some lines of reasoning
yields consequences, I should like to question Professor
Epstein about.

     Traffic accidents constitute one of the most important
public health problems.  This could have been predicated
from the start, even without animal experiments.  According
to Professor Epstein participation in automobile transport
should be regarded as an amoral act, because non-driving
human subjects are going to be killed or injured.

     I agree that there are major defects in decision-making
processes.   One example:  the very great attention paid to
the toxicology of exhaust gases and the regulatory measures
taken.  The effort appears to be relatively too great if
compared with the effort put into the prevention of traffic
accidents.   Does Professor Epstein agree with this?
EPSTEIN   (U.S.A.)

     There are two aspects to decision making.  The first
is the generation of unbiased and expert data by scientists.
The second is the development of regulatory processes
reflecting a wide range of balancing viewpoints and
interests.

     The question of traffic accidents relates to the second
aspect.  Regulatory bodies and decision making groups have
the necessary information to decide whether saving of human
life justifies application of available techniques such as
"safety packaging" and air bags.  Such decisions should
properly be made in the open political arena.
DAVOUST (France)

     Owing to the agricultural practices in the industrialized
countries, but also to those in all countries where intensive
monoculture has been introduced, plants are sick and need
fertilizers and pesticides as much as a diabetic needs insulin,

     Set on the chemical products road, and given the numerous
scientific, political and other difficulties in decision-
making which have been referred to during the symposium as
regards environmental protection measures, do you think, Dr.
Epstein, that the situation leads to more and more complex
problems,  that a choice will have to be made between health
and consumption, and that the economic arguments weighed
against the biological ones tip the balance in a direction
not difficult to determine?  Can arbitrary or authoritarian

-------
                           2385
decision-making be avoided?  Can decision be brought into
line with the growing public conscience as regards environmental
protection?


EPSTEIN  (U.S.A.)

     Extensive use of fertilizers and pesticides in mono-
cultures is often economically counter productive.  A part
from emerging of pesticide resistance, and lack of efficacy
of pesticides, contamination of food supplies and other
segments of the environment often clearly result in long-term
economic besides public health, damage and losses.


REEVES  (U.S.A.)

     I do not believe that any members of the Threshold
Limits Committee of the American Conference of Governmental
Industrial Hygienists are present at this Symposium.  If
they were, I think they would take strong exception to
the statement of Dr. EPSTEIN that they are industrially
influenced or dominated.  In fact, no person with industrial
corrections can become a voting member of A.C.G.I.H., much
less of its Threshold Limits Committee.

     The reason why there is a discrepancy between values
on the Threshold Limits List and the Air Quality Standards
is not that anyone is more callous towards the health of
industrial workers than to that of the general population.
TLV's are figured on a 8 h./day, 5 day/week basis, while
air quality standards are figured on 24 h./day, 7 day/week
basis.  The general population includes infants, invalids,
old people, and so forth, while industrial workers are
presumably healthy and in their prime. Finally, industrial
workers can be pre-screened and educated vs. the exposure
hazard while the general population can not.
                      i
     I agree with many of the statements and philosophies
expressed by Dr. Epstein.  I believe these points could be
presented more effectively if he were to refrain from all-
egations which are inflammatory and unprovable.


EPSTEIN  (U.S.A.)

     The industrial bias of the A.C.G.H. is well recognized.
Furthermore the TLV "standards" apply to only some 450
of the thousands of chemicals to which workers are exposed
and reflect acute and sub-acute toxicity and can generally
ignore long-term and chronic toxic effects.  Recognition
of the limitation of those standards is afforded by the
recent decision of NIOSH to undertake early extensive review
of the scientific basis of these "standards."

-------
                            2386
ZUNIC  (Jugoslavia)

     This study shows the side extent of pollution in
ecological and human areas.

     As we have seen, decision are of a distorted and artificial
nature.  Since Plato's famous socialization formula
 5 = n - 1 and also since Montesquieu, democracy is
enriched neither by theory nor by practice.

     If the biological management of life is the expression
of two factors in direct competition with each other, why
can't the political decision be made complete by the simul-
taneous decision of two parties:  producers and consumers.

     We have an industrial monopoly, as Dr. Epstein emphasized.
The fact that Man has no objective knowledge leads every
civilization to its downfall.  Neither society nor civilization
dies in the physical sense of the work.
EPSTEIN  (U.S.A.)

     I thank Dr. Zunic for his perceptive comments which,
in principle, I endorse.


CHANTEUR (France)

     Without wishing to deny the value of the attempts to
rationalize choices that evaluation studies of technical
options represent, I feel that one must expose wishful thinking
and stress that, in the absence of a professional opinion,
they can lead to unrealistic, if not absurd decisions.

     In the field of ionizing radiations, this point was given
prominence by the International Commission on Radiological
Protection in report no. 22 which offers an example of a
realistic approach that could profitably be extended to other
nuisances.   Particularly in the biomedical field one should
be sufficiently humble to preserve a certain empiricism.  Of
course, this does not exclude total independence of judgment,
safeguarded by a strict division of responsibilities between
the promoter organizations and those concerned with
health protection.

     These are the principles governing the supervision of
medical radiology and nuclear energy applied daily in France by
the Central Service for Protection against Ionizing Radiations
(SCPRI), a technical service responsible to the Ministry of
Public Health via the National Institute for Medical Health
and Research.

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                              2387
      THE USE OF QUANTITATIVE EEG FOR DETECTING LOW-LEVEL
               PROLONGED EXPOSURE TO PESTICIDES

                      JOHN A, SANTOLUCITO

Environmental Protection Agency, Research Triangle Park, NC, USA
ABSTRACT

     Data were obtained from IS minutes of anesthetized sleep EEG
using three scalp electrodes positioned at mid-frontal, right and
left ocoipito-parietal regions.   Interval histograms, zero pot-
ential crossover (ZPC) rate, and right-left hemisphere correspon-
dence were obtained using computer-assisted methods.   A shift
toward higher frequencies, increased ZPC rate, and an increase
in the coefficient of variation of ZPC were most pronounced in
carbaryl, 2,4~D, and toxaphene-treated animals.   Bilateral cor-
respondence tended to be increased by all pesticides tested.

     In order to facilitate application of this methodology to
occupationally exposed humans* a portable EEG was designed and
constructed.   The instrument is completely battery operated and
will accept three channels of EEG for recording on tape cassette.
The attending physician can obtain recordings before, during, and
following an agricultural season, in the field, with minimal
interruption of work schedules.   The tape cassettes are mailed
to the laboratory, played back through a physiological data ac-
quisition unit for paper write-out and computer processing.  The
instrument will be described and preliminary findings will be
discussed.

-------
                                   2388
    The electroencephalogram (EEC) is easily obtained, non-destructive,
and non-invasive.  Consequently, it is particularly suitable as a
biological monitor.  The available literature is supportive of the con-
clusion that the electrical activity is one of the most sensitive indi-
cators of an altered functional state of the brain in the intact animal.
The EEG has been altered in repeated chronic exposure experiments with
DDT [1], endrin [2], and dieldrin [3] in which clinical signs of toxicity
eventually appear.  Specific anomalies in the EEG related to occupational
exposure to dieldrin, aldrin, and endrin without clinical signs of intox-
ication have also been reported [4],  Workers occupationally exposed to
organophosphate compounds repeatedly, showed EEG changes similar to those
seen after acute exposure [5]*  Non-human primates fed selected pesticides
for 18 months exhibited alterations in EEG frequency distribution and
right-left hemisphere waveform coincidence [6].
    As a result of the pioneer efforts of Walter [7] and Drohocki [8],
analyzer systems are readily available today.  Such systems are used
frequently in animal experiments for the quantification of EEG.  However
for human use, a remaining problem is its easy acquisition from large
numbers of the general population in a form accepted by automated data
processing systems.
    The objective of this research is first to establish quantitative
EEG indicators of prolonged, low-level exposure to pesticides using non-
human primates and second, to apply the methodology developed to
pesticide exposed humans.
    Squirrel monkeys (Saimiri sciureus) were placed on low level dally
intake of selected pesticides in 1968.  A contemporary control group has
also been maintained for evaluation of treatment effects.  Comparison of
EEG's between treated and control groups was chosen instead of compari-
sons of treated monkeys with their own pre-treatment control period to
avoid confusion with age-associated changes.
    The dosing regimen is summarized  in Table  I.
    Bipolar  EEG  recordings are made with three subcutaneous, platinum
needle electrodes  placed in mid-frontal (MF),  right, and  left occipito-
parietal  {RO-P and LO-P) locations.  Thus, three channels of EEG, repre-
senting RO-P x MF, LO-P x MF, and RO-P x LO-P, are obtained.  This
arrangement  optimizes the recovery of all  EEG waveforms with the fewest
number of electrodes.  It has  been shown that  placement of electrodes at

-------
                                   2389
      TABLE I.  Protocol for Squirrel  Monkeys Receiving Chronic,
                Low-Level Intake of Selected Pesticides.
                                           Daily Dose*
Pesticide
Parathion (6)**
Carbaryl (6)
Dieldrin (5)
Toxaphene (6)
2,4-D (6)
Lindane (5)
Control (7)
(mq/kq Body Weight)
0.03
0.007
0.1
1.0
0.2
0.05
—
     *Pesticide administered  per  os,  6 days/week
    **Number of animals

the front and back of the head form a lead pair almost equally sensitive
to sources anywhere in the entire brain [9], and right and left deri-
vations permit assessment of hemispherical bilateral symmetry.
    At the time of recording, monkeys are given 1.0 mg/kg Sernylan^, 1.m.,
to immobilize them.  They are then anesthetized with Surital^, l.v., to
loss of blink reflex and predominantly abdominal  breathing.  Scalp elec-
trodes are affixed and 15 minutes of EEG are taken using a Beckman, Type-R
Dynograph.  The anesthetized EEG is stable with time for a given depth of
anesthesia as has been found by others [10].  The data presented here were
obtained from on-Hne computer assisted interval  analysis of five minutes
of the recording after three years of pesticide exposure.
    It can be seen in Table II that the EEG from 2,4-D, toxaphene, and
carbaryl-treated animals shifted toward higher frequencies.  This was
generally accompanied by a decreased amplitude of the high voltage, slow
waves seen in anesthetized sleep.  Dieldrin was unique among the pesti-
cides tested in showing synchronization of the EEG, I.e., Increased am-
plitude of the high voltage, slow waves < 1.5 Hz.  No other consistent
amplitude differences were seen between treatment groups.

-------
                                     2390
      TABLE II.  The Distribution of EEG Wave Abundance in Selected
                 Frequency Classes as Per Cent of Total for Five
                 Minutes of Recording.
      Treatment            11-50 Hz       5.9-10 Hz      1.5-2.9 Hz
Mean *
Control
2,4-D
Lindane
Dieldrin
Parathion
Toxaphene
Carbaryl
26.
**45.
29.
31.
30.
*35.
**50.
1 ±
5 ±
7 *
9 ±
2 *
0 *
6 ±
S.E.
2.3
1.8
1.5
2.4
2.4
2.8
2.1
Mean
19.
*16.
*16.
18.
19.
18.
*17.
5
7
4
8
8
4
0
* S.
* 1.
* 1.
* 1.
* 1.
* 0.
* 0.
* 0.
E.
3
2
0
7
9
9
5
Mean
22.
*14.
22.
*19.
24.
*19.
**12.
6
3
6
3
3
0
1
±
i
±
±
i
±
±
i
S.E.
1.7
1.0
1.0
1.2
1.3
1.1
0.9
                    **p < 0.01
                     *p < 0.05

    Similarly, zero potential crossover (ZPC) rate/5 second epoch is in-
creased in the EEG of animals showing an increase abundance of higher
frequencies as seen in Table III.  More interesting is the increased
variability of this parameter and the patterning (as indicated by trend
analysis) of the variability.  The increase in the coefficient of varia-
tion is statistically significant for 2,4-D and carbaryl and nearly so for
toxaphene.  When one plots the counts per epoch in a bar graph, it is
apparent that the significant trend represents a sinusoidal pattern with
a period of 30-35 seconds.
    We felt sufficiently encouraged by the results obtained with the
monkeys to proceed with the second phase of the original objective, viz.,
application of the methodology to human studies.  The monkey research is
continuing with the application of other time-series analyses, e.g.,
cross and auto correlation, but I would like now to describe the portable
EEG developed and constructed in conjunction with Dr. Jacob Kline,
Director, University of Miami Medical Instrumentation Laboratory, School

^Phencycliciine hydrochloride, Parke, Davis & Company, Detroit, Michigan
^Sodium thiamylal, Parke, Davis & Company, Detroit, Michigan

-------
                                    2391
     TABLE III.  Characteristics of Zero Potential Crossover (ZPC)
                 Rate During Five Minutes of EEG Recording.
  Treatment
ZPC/Epoch
C.V.
Trend
Mean Successive
  Square Diff.
    Variance

Control
2,4-D
Lindane
Dieldrin
Parathion
Toxaphene
Carbaryl
Mean
19.7
**28.9
18.8
19.5
21.1
24.2
**33.4
± S.E.
* 1.9
* 1.6
± 0.8
* 1.4
* 1.2
* 1.6
* 1.7
Mean
23.0
*31.1
22.2
22.4
22.8
26.3
*28.6
* S.E.
* 2.0
i 2.3
* 1.1
* 1.2
* 1.5
* 1.7
* 0,8
Mean
2.0
*1.5
2.2
2,0
1.9
1.6
*1.3
* S.E.
* 0.2
* 0.1
* 0.1
* 0.1
* 0.1
* 0.2
* 0.1
            **p < 0.01
             *p < 0.05

of Medicine, Miami, Florida, U.S.A.  The recording unit has a self-
contained battery pack, measures 40 x 40 x 30 cm, and weighs 9 kg.   The
monitoring oscilloscope is a standard Tektronix 211  and the tape recorder
is a standard Sony stereo cassette recorder, TC-124.  The input stages
of the instrument are high performance differential  instrumentation
amplifiers in an integrated circuit package.  It performs balanced  dif-
ferential measurements with high common mode rejection (CMRR = 106  dB at
gain of 100 from DC - 100 Hz, with source unbalance of 1000 ohms).
    The second integrated circuit stage is a standard operational ampli-
fier designed with moderate gain to provide sufficient signal level to
the four input multiplexer integrated circuit.  Control of the multiplexer
is accomplished by a 7200 Hz timer clock pulse.  The three channels of EEG
are multiplexed on one channel of the stereo cassette tape.  The timer
clock pulse 1s simultaneously recorded on the second channel to be  used
1n demultiplexing upon playback.  This arrangement compensates for
differences in tape speed between units.  A demultiplexing playback unit
was also constructed for playing back the recorded information through the
Dynograph, into the data processing system.

-------
                                    2392
    Two recording units are in the field at the present time.  Agricul-
tural pesticide workers are being recorded before, during, and at the end
of the season.  The procedure consists of placing the subject on a cot,
affixing the scalp electrodes, and after assuring the presence of a good
trace on the oscilloscope monitor, obtaining 15 minutes of recording with
the subject relaxed and eyes closed.  Tapes are then mailed to the
laboratory for processing.
    It is too early to make comparative observations between pre- and
post-exposure EEG's.  However, Table IV shows the interval analysis
results on six human subjects obtained with this system.
    In conclusion, quantitative EEG measures have disclosed changes in
monkeys exposed to low levels of pesticides for three years.  The animals
have displayed no signs of toxicity, and have reproduced each season
bearing viable offspring at a rate comparable to our non-treated colony.
Our efforts to date have concentrated on the feasibility of the method-
ology and development of the portable system.  Effort is now being con-
centrated on application of a battery of time series analytical tech-
niques which can be performed on the three channels of EEG described.
     TABLE IV.  Awake EEG Interval Analysis of Six Subjects Recorded
                in the Field with Portable Instrument.
                                     Frequency Distribution
Zero Potential Crossover  	(% of Total Waves from 0.5-50 Hz) 	
   Waves                    BetaAlphaThetaDelta
5 Sec Epoch     C.V.      13-50 Hz    7.7-12.5 Hz   4 < 8 Hz    < 4  Hz
                              (Mean * S.E.)
72.6 * 7.2   9.5 * 1.6   64.3 * 3.7   24.6 * 2.9   9.8 ± 1.4   1.1 * 0.3

-------
                                   2393
                              References

1    DESI,  I., FARKAS, I., KEMEMY,  T.,  "Changes of central nervous  function
    in response to DDT administration", Acta Physio!. Acad.  Sci. Hung.,
    30,  275-281 (1966).

2   SPECK, L.R., MAASKE, C.A., "The effects of chronic and acute exposure
    of rats to endrin", A.M.A. Arch. Ind.  Health, 18, 268-272 (1958).

3   VAN GELDER, G.A., SANDLER, B.B., BUCK, B., MALAND, J.B., KARAS,  G.G.,
    "Behavioral and electrophysiological effects of dieldrin in sheep",
    Ind. Med. Surg., 38, 64-67 (1969).

4   HOOGEHDAM, I., VERSTEEG, J.P.O., DE VLIEGER, M., "Electroencephalo-
    grams  in insecticide toxicity", Arch.  Environ. Health, 4, 86-94
    (1962).

5   METCALF, D.R., HOLMES, J.H.. "EEG, psychological, and neurological
    alterations in humans with organophosphate exposure", Ann. N.Y.  Acad.
    Sci.,  160, 357-365 (1969).

6   SANTOLUCITO, J.A., MORRISON, G., "EEG  of Rhesus monkeys following
    prolonged low-level feeding of pesticides", Toxicol. Appl. Pharroacol.,
    19,  147-154 (1971).

7   WALTER, W.G., "An automated flow frequency analyzer", Electron.  Engg.,
    16, 9  and 236 (1943).

8   DROHOCKI, Z., "L'integrateur de 1'electroproduction cerebrale  pour
    1'electroencephalographie quantitative", Rev. Neurol., 80, 619-624,
    (1948).

9   RUSH,  S., DRISCOLL, D.A., "EEG electrode sensitivity - An application
    of reciprocity", IEEE Transations on Bio-medical Engineering,  16,
    15-22  (1969).

10  KLEMM, W.R., HALL, C.L., "Electroencephalographic pattern abnormal-
    ities  in dogs with neurologic disorders", Am. 0. Vet. Res., 33,
    2011-2025 (1972).
                             DISCUSSION
HAIDER  (Austria)
      Did  you use computer analysed evoked potentials  and slow
potential changes in  your studies?  In our studies with im-
planted electrodes in rats  we could demonstrate changes in
evoked potentials produced  by pesticides.

-------
                             2394
SANTOLUCITO (U.S.A.)

      We have used evoked potentials in animal studies.  We
have also found them useful, for example in a study with hexa-
chlorophene-treated monkeys, it was possible to demonstrate
visual evoked response changes at doses below that required to
produce the typical brain lesion.

      We are currently using evoked potential measurements in
a study of a series of carbamate pesticides.

      At present time, we have not attempted this in the field
for human studies utilizing the portable EEC.

-------
         UNTERSUCHUNG DER GESUNDHEITLICHEN WIRKUNGEN



                   HEALTH EFFECTS STUDIES



               ETUDES DES EFFETS SUR LA SANTE



              STUDI DEGLI EFFETTI SULLA SALUTE



       ONDERZOEKINGEN NAAR GEVOLGEN VOOR DE GEZONDHEID





                       (Continued)
Voreitzender - Chairman - President - Presidente - Voorzitte*





                     S. HERNBERG (Suomi)

-------
                                2397
             INTERNAL POLLUTION - OUR FIRST PRIORITY
                 A REVIEW OF THE STUDIES OF THE
            SPECIAL COMMISSION ON INTERNAL POLLUTION

    PETER BEACONSFIELD* NORMAN BORLAUG, A. CARPI DI RISMINI,
    SIR HANS KREBS, SIR RUDOLPH PETERS AND REBECCA RAINSBURY

Special Commission on Internal Pollution, BMA House, London, UK
+ Royal Free Hospital,  London, UK
ABSTRACT

     Pollution is the result of man's unwillingness or inability
to control hie present overpopulation, over-exploitation of nat-
ural resources, and over-production of many indestructible goods.

     However, attacking environmental pollution will be self-
defeating if we disregard man's continuous and progressive pollu-
tion of his own milieu interieur - this is what we have termed
internal pollution.

     Internal pollution is brought about in three ways;  by the
ingeation and inhalation of the products of the already polluted
external environments  by daily intake of chemical additives and
impurities prepacked in foods  and by the vast number of medica-
ments taken regularly, and more often than not unnecessarily.

     We resolved to consider the present use of chemicals under
four broad headings - medication, food additives and colourants,
agricultural aids, and household goods;  and to relate their res-
pective uses to the responsibility shared by the producers, the
consumer, the government regulatory agencies, and the biomedical
professions.

      In the laet year and a half we have examined the field of

-------
                                2398
medications, and started our inquiries  into  food,
     The general impression wast particularly since thalidomide,
that there is something wrong with the  tests, but our investi-
gations do not bear this out.   We find the  tester and the phy-
sician to be the guilty parties.

     Testers sucoomb to human weaknesses of  inefficiency and  lack
of conscientiousness, physicians are ill informed.

     Overprescribing and polypharmacy are the twin sins.

     Finally, the patient may have unsuspected sensitivity or
may be ignorant enough to believe that  doubling the dose must
double the benefit.

     As the whole field of medication is in  the control of experts
this is one of the aspects of internal  pollution capable of solu-
tion within the framework of our present technical and economic
possibilities.

     Pesticides, fertilizers, and other crop and livestock impro-
vers have come into almost universal use.    Concern has been  ex-
pressed about the long-term risks some  of these chemicals entrain.
When evaluating this, and always looking to  increase the benefit;
risk ratio of such agricultural .aids, it must be remembered that
their use should be discussed within the context of the nutritional
status of the population needing to be  fed.

     Preservatives, packaging aids, and so forth, are the result
in large measure of modern economic dictates concerning demand
and supply, storage and distribution.   The public needs and  des-
erves to be better informed about what is done  to the food supply, and why.
     We realize that an urgent need exists for clearer under-
standing of the medical, social, and economic benefits and risks
that accrue from the application of science  and technology and
the use of chemicals in today's world,  and better guide  lines for
decision-making.

-------
                                 2399
Introduction
     Over a single ten-day period this summer,  three separate meetings will
have been held on the subject of pollution and the environment*   There was
the one in Greece which ended a few days ago,  this present meeting,  and a
third which is due to take place next week in this very building. The pro-
grammes of all these meetings show that they conform to the current  trend
of being concerned with recording more accurately, detecting more quickly,
and discussing yet more fully the already identified agents of pollution.
No time has been set aside for defining the problems which underline pollu-
tion}  the general preoccupation has been with treating symptoms rather
than the disease, and, spectacular as these symptoms are, it is but  pallia-
tive to treat them without seeking their aetiology.  It is as if it  had
slipped everyone's memory that the patient is humanity itself and the
wounds entirely self-inflicted.
     Among the principal causes of pollution are over-population,  over-
exploitation of the world's finite natural resources, and an unwillingness
or inability to dispose of refuse and waste products.  For example,  it is
in the world's interest that Brazil should not reach its foolishly-declared
population "goal" of 200 million by the end of the century;  it ia in the
world's interest that its weather or radiation counts should not be affected
by pollution of the upper atmosphere.  The interdependence of rich and poor
nations was made quite clear at the 1973 United Nations Meeting  on the
Environment in Stockholm.  The industrialised countries talked of air and
water pollution and how to decrease it;  the developing countries talked of
their desire to increase industrialisation)  and the underdeveloped coun-
tries simply talked about how to feed themselves.
     At first sight it would seem impossible to find mutually acceptable
solutions when there is such disparity in development and requirements.  It
is evident that if the poorer countries are ever to change, then the rich
 nations need to change first)  and people who are well off are usually
 unwilling to do anything which might jeopardise their own standard  of
 living - and any change implies risk.  Nevertheless, if certain aspects
 of the developed countries' life-style continue unchecked, it can be
 predicted with some certainty that they will constitute a danger to the
 entire world, including themselves.  The industrialised nations' total
 dependence on chemicals exemplifies this.

-------
                                 2400
Our Chemical Age
     The ever-increa8ing use of chemicals in every sphere of activity in
the industrialised countries has been accepted almost unnoticed aa we move
into this last quarter of the 20th century, and ours could well be called
the "chemical age".  The use of chemicals directly touches far more people
and influences more aspects of daily life than any kind of revolution that
has preceded it, and this includes the atomic age.  It is profligate use
of chemicals which is the root cause of pollution - whether external pollu-
tion of the environment or internal pollution of our own bodies.  The
industrialised nations depend on chemicals for nearly every aspect of daily
living, and in addition to developing new chemicals we have growing popu-
lations and so manufacture greater quantities of these chemicals to serve
them.  It has become a vicious circle of more people demanding more goods,
and it can be broken only if the birth-rate is controlled, if the real need
for these chemicals is assessed, and if people are told that these compounds
entail risks as well as benefits.
     We have become accustomed to rely on chemicals for health, for homes,
for transport, work, leisure, food and drink, and even for fertility and
sterility.  Every object of the domestic environment, including the contents
of the bathroom, kitchen, and clothes cupboard, is partly or entirely
synthetic;  and nearly all our foods are packaged, processed, coloured,
preserved, kept fresh and otherwise "improved" by the addition of chemicals.
The economic - and hence political - implications of this way of life mean
that if we are really concerned to decrease pollution we have to consider
making radical alterations in it - alterations which would affect the work
forces and distribution of labour and perhaps change the whole direction of
national industries.
     In such circumstances it is clearly of limited value to single out
particular chemicals for special control, yet this is precisely what we are
doing.  Our aim should be to keep pollution at a minimum level while we re-
examine the implications of continuing along our present course.  Let's make
our goal what we can do and not an unrealistic ideal of what we would like
to see.

-------
                                 2401
Internal Pollution
     The advent of the chemical age has completely changed our attitudes
to health and disease.  We tend to think of medication in relation to the
cure and treatment of disease, but in fact only one-tenth of all drugs taken
in the industrialised world are for this purpose.   The rest go to maintain a
new concept - that of "positive health".  Thanks to the pharmacological
"explosion" of the post-war era, people expect to be well}  to be protected
from the great scourges of humanity like smallpox, tuberculosis, and malaria;
to be athletic and full of energy, yet to sleep well)  to be slim or muscular
according to their sex;  to be fertile when they wish and at all other times
sterile.  It is in order to attain this positive health that people swallow
tons of chemicals annually.  Few if any of these chemicals have ever before
been taken regularly over such long periods of time by people who are healthy
to start with, and the long-term effeots - biological, genetic, and economic-
are therefore not yet apparent or appreciatedt  Hence another new concept is
needed - that of Internal Pollution, the abuse of the milieu interieur.
     Internal pollution is brought about in three wayst  by the ingestion
and inhalation of the products of the already-polluted external environment;
by the daily intake of chemical additives and impurities in pre-packed food;
and by the vast number of medicaments taken regularly and, more often than
not, unnecessarily.  The last two ways could be minimised fairly easily,
since it is possible to control the mechanisms involved and the economic
consequences would be manageable.
The Special Commission on Internal Pollution
     A number of senior scientists from different disciplines and countries,
concerned at the tacit acceptance of this less visible (and hence under-
publicised) kind of pollution, formed the Special Commission on Internal
Pollution (SCIP) to study its consequences for a country's economy, for its
national progress, and for its health - both at present and in future genera-
tions.  The Commission differs from other study groups, working parties, and
the like which have been appointed by governments, industries, or the inter-
national agencies in that its members represent no one but themselves.  We
are not beholden to anybody, we are on nobody's payroll.  But if we have no
partisan political axe to grind it must be stressed that neither are we
politically naive;  we can foresee the upheaval implied by reoonmending
change.  At the moment, however, SCIP's main concern is with its impartial

-------
                                 2402
investigation of the functioning and effects of the chemical age, so that
more people can be better informed about it and thus more able to reach
reasoned opinions and sensible decisions on its development and control.
     SCIP resolved to consider the present uae of chemicals under four
broad headings - medication, food additives, agricultural aids, and house-
hold goods;  and to relate their respective uses to the responsibility
shared by the producers (that is, industry), the consumer, the government
regulatory agencies, whose role is to protect the consumer, and the bio-
medical professions who test and prescribe many of these chemicals.
     SCIP further decided to re-examine the whole concept of what we mean
by "safety" in the chemical age, with special reference to the benefits
risk ratio of the different classes of compounds.  With a clearer notion
of what constitutes risk and what benefit, we might then see how to improve
matters.
     To date, SCIP has already examined medication and is at present in-
vestigating food additives and colourants.  Although research is not yet
complete, data is also being amassed on agricultural aids and household
goods because, as will readily be appreciated, all four areas of research
do to some extent overlap.
Medication
     According to the pharmaceutical industry itself, if the current trend
continues almost every person in the technologically-developed countries
will within the next decade be on some type of daily drug regimen for the
improvement or maintenance of his positive health.
     We know that every drug is potentially both a healer and a killer, yet
there is no disagreement about availing ourselves of potent remedies for
treating serious disease:  the benefits more than compensate for any
attendant risks.  The known toxic effect of streptomycin on the auditory
nerve does not make any physician ready to withhold it in tuberculous
meningitis.
     Nevertheless not nearly enough attention has been given to the question
of benefit and risk.  Three separate factors need assessing:  the benefits
a compound can confer;  the dangers of taking it;  and the consequences of
withholding it.  In diseases which are invariably fatal if left untreated,
these factors are relatively straightforward to weigh up.  But with the
improvement of medical practice such extreme situations have become the
exception rather than the rule, and other aspects have to be considered.

-------
                                 2403
One is the question of the relative dangers and benefits of alternative
therapies)   another is the severity of the disease which may vary from
inexorably lethal, like canoer of the stomach, to fatal at a later and un-
certain stage, like Hodgkin'a disease and some leukaemias;  from the in-
capacitating like the arthritides to the merely inconvenient, like some
skin conditions.  Other factors include how long the disease may be expec-
ted to last if untreated;  is it acute and completely curable with the
appropriate therapy?  Is it short term chronic and curable?  Or long term
chronic and controllable rather than curable?  How much risk is acceptable
in therapy which will sufficiently control the disease to give the patient
a reasonable quality of life?  Which will end his life first - the disease
or the drug?  Is it possible that social opinion may change during the
natural history of a patient's chronic disease?
Side Effects
     These questions are still further complicated by the fact that no two
human beings are physiologically identical and hence will not react iden-
tically to the same drug.  In a sense, each new administering of medication
is an experiment.  So in view of the pharmaceutical companies' startling
prediction just quoted there is good reason for considerable concern over
the upsurge in iatrogenic conditions.  A conservative estimate has it that
some 5-10 per oent of all patients admitted to hospital in the UK and the
United States are there because of the side-effects of drugs they are taking:
and of these patients between 2 and 3 per cent die.  Some of the drugs have
been prescribed by the patients' own doctors, while others have been bought
over the counter.  Up to 5 per cent of a typical British general practition-
er's patients may consult him with iatrogenic disease, and of patients
already in hospital as many as 18 per cent suffer adverse reactions to
drugs, according to surveys in the US and Canada.  A British survey found
similar reactions in 10.2 per oent of a sample of 1,160 patients.1v/o to 3
per oent of congenital malformations are attributable to drugs and chemical
pollutants.
The Pharmaceutical Industry
     What are the reasons for this level of apparent failure in a field
supposedly as minutely researched as pharmacology?  It is fashionable,
especially in journalistic circles, to pin the blame on the pharmaceutical
industry itself, if only because evidence of price-fixing and substantial
profits induce people to look no further for a scapegoat.  This attitude
is not justified*  We have.found that the ethical pharmaceutical industry

-------
                                 2404
maintains high overall standards of manufacture and testing.  Furthermore,
our investigations found no support for the impression common since the
Thalidomide tragedy that the tests carried out on new drugs are ineffec-
tive or inefficient.  In fact, we can state categorically that the tests
are as good as our present state of knowledge allows.
     We found the problem to lie not with the tests, but with the testers.
Many testers succumb to the human weaknesses of inefficiency and lack of
conscientiousness - especially if their salaries are poor and their career-
structure unestablished.  They are no different from the society which
produces them.  In addition, we have found that many pre-olinioal tests are
not carried out by the scientist in charge of the projectj  he reads the
results brought to him by his technicians.  We have noticed this to be
particularly true of industry, where there is rigid adherence to a hier-
archical system.
     The truth is that many research scientists are, by nature of their
intense specialisation, unqualified to correlate diverse results, to cross-
transfer, or to recognise anomalies thrown up by different testing techni-
ques.  The picture is one of so many tests that no single scientist is com-
petent to make a final assessment of the drug under examination. The pharma-
ceutical industry itself complains of toxioity tests so numerous that the
very volume of work give}the regulating agencies an assurance of quality
that could be illusory.  In faot our experience has shown that the perspi-
cacity to pick the right animal and conduct the correct study in depth in a
small group of animals is far more likely to provide the desired information
than is sheer numerical weight of different tests.  This fact is well recog-
nised, yet it is still not accepted practice.
ClinicalJ^harmacplogy
     However, the testers are not the only ones at fault.  In fact, there
are twin culprits - the testers and also the doctors.  On the medical
profession's side, all is far from well.  In May 1973 it was reported that
some JO per cent of American doctors doing clinical trials of new drugs
whose work was spot-checked by the FDA were guilty of a range of unethical
practices, including giving wrong dosages and falsifying records.  In faot,
of all the reports submitted, in one-third the trial was never done at all,
in another third it did not follow the manufacturer's protocol, and only in
one-third were the results of any value.  In some oases the doctors have
been so slick that they were able to "take care" bf patients in America
while attending meetings on extended European vacations.

-------
                                 2405
     But even where fully-tested and established drugs are concerned,
most of the adverse drug reactions are the direct result of the indis-
criminate and over-indulgent prescribing habits of physicians who often
rely solely on advertisers' oopy for their information about the use of
the drugs.  Doctors are lamentably ill-instructed in clinical pharmacology,
a hardly-recognised speciality for which there is little or no formal
training.
     The medical curriculum for the past 100 years has been designed to
elicit diagnosis, for until recent times diagnosis was all we could offer
the patient apart from surgery.  Drugs were few and their mode of action,
even when curative, was ill-understood.  Now drugs are many and potent and
their method of action and interaction must be known by the presoriber.
Although this fact is recognised, few medical schools give any formal train-
ing in the speciality, and fewer still have academic departments of clinical
pharmacology.  Courses in clinical pharmacology and therapeutics running in
parallel with clinical medicine are rare, and those who design and run them
are thought of as second-olass citizens by their colleagues - whether these
are general physicians or academic pharmacologists.  The need for this
speciality is underlined by the fact that the rate of introduction of new
therapeutic and prophylactic agents during the last 30 years has almost
entirely outdated the knowledge of a doctor who graduated before then.
Polypharmaoy has now become the rule instead of the exception;  certain
hospital surveys have identified patients taking up to 30 different prepara-
tions a day.  Small wonder they develop new diseases, and who is to say
whether their symptoms are caused by the disease or the drug combination?
     Since the whole field of medication is in the hands of experts and is
therefore amenable to control, the cavalier attitude of the medical prof-
ession   is inexcusable.  In faot medication is the aspect of internal
pollution most easily dealt with in the framework of our present technical
and economic possibilities.
Food
     The use of chemicals in the food industry has had even more far-
reaching effects on Western society's economy than has the concept of
positive health.  More people eat, and they eat a greater diversity of
foods in a greater number of places than ever before.  Food is produced in
one place, processed in another, sold in a third, and retailed elsewhere
again*  No longer do the seasons dictate the menus of the developed coun-
tries.  We have what we want when we want it, and without chemical help,

-------
                                 2406
most of this would be impossible.
     The economic implications of all this are immense.  Whole new indus-
tries employing hundreds of thousands of people have been created; fanning
itself has been modified to produce a crop of maximum suitability and
convenience for harvesting and packaging.
     A distinction must be drawn here between those chemicals which are
valuable in improving the product or in preserving it better, and those
which are added purely and simply for profit.  It is one thing to add a
preservative to give a good product a better shelf life;  but it is nothing
other than sharp practice to feed a pig a chemical compound which will
cause it to retain water to make it weigh more in the market place.  SCIP
was told that one well-known manufacturer puts up to 20 different chemical
additives into his pork pies.  Nowadays this is the rule rather than the
exception.  We may well ask - why?
     At present few people realise the degree of adulteration undergone by
the food they buy.  We believe that in addition to the obligation of food
manufacturers to put dates on their packages, which has only recently become
mandatory in most countries,      all the ingredients and their proportions
should similarly be detailed.  The consumer needs to be educated to under-
stand and use this information to his best advantage*
The Green .Revolution
     Pesticides, fertilisers, and other agricultural or livestock improvers
have now come into almost universal use.  These, together with genetically.
improved crops, have produced a new concept - the farming equivalent of
"positive health" - the so-called "green revolution".  It now takes less
manpower to produce more food from a given acreage in a greater diversity
of conditions.  However, all this is only with the expenditure of more
energy for the same tonnage of production*  And just as consumers need even
greater quantities of chemicals to maintain their standard of living, so
intensely-farmed soils come to rely more and more on chemicals to make up
for their depleted structures*
     However, in all evaluations of "agribusiness" it must be remembered
that these 20th century farming aids have to be seen in the context of the
nutritional status of tho population needing to be fed*  We all know about
the dead fish in the Baltic and Lake Michigan, but what would happen if DOT
were not used in Ceylon?  Once again, the point is the same as it is in the
matter of "positive health"i  what benefits do we demand and what risks are
we prepared to run for them?

-------
                                 2407
Closing Comments
     We should like to stress again that there is practically nothing
about pollution - whether internal or external - which is not already
known.  We know what pollution isf we know how pollution takes place,
and we know who does the polluting.  SCIP feels that we should now be
concentrating more effort into uncovering the circumstances which at
present guarantee that such pollution is inevitable, and could remain so.
     In our view, safety as a concept is inaccurate, and "safe" products
do not exist.  The use of this epithet to describe any chemical is wrong
and misleading.  The World Health Organisation itself is not above criti-
cism for publishing technical papers referring to such mindless concepts
as "Safe Uae of Pesticides".  We must learn instead to talk in terms  of
benefit and freedom from risk, when it is accepted that substances like
DDT can be dangerous but that without their judicious use millions would
starve in many parts of the world.
     Up to the present, few people have understood that the comfortable
life-style of the industrialised world entails its own risks, and that
those risks may not be generally known.  A man getting into his car to
drive to work accepts - albeit subconsciously - that there is a definite
statistical chance of his becoming one of the eight million injured or
250,000 killed annually on the roads of the industrialised world.  Yet
the same man sitting down to dinner does not even consider that his pre-
cooked TV repast, bought at the local supermarket, could entail any risk.
It is wrong that he should continue in this state of uninformedness.   He
has a right to know so that he himself can decide whether any risk is
worth taking.  A separation of what can from what can't be done within our
present technological and economic capabilities has to be made*  In the
case of the pharmaceutical industry, for instance, we consider that the
tests applied to new compounds are as good as modern knowledge and techno-
logy permits, but that this does not necessarily mean they are done as well
as they could be.  It is perfectly possible to remedy this, just as it is
equally possible for the medical profession to improve its knowledge  of the
drugs it so freely and generously dispenses.  We have seen how necessary it
is for doctors to be trained to prescribe:  it is not enough to let the
advertisers' copywriters do it for them.  So far as food and agriculture
are concerned, up to the present no clear distinction has been drawn  between
chemicals used to improve quality and quantity of food production and those
used solely to increase producers' grofit margins at no price advantage to

-------
                                 2408
the consumer and at a possible risk to his health.
     The value of many goods, particularly in the domestic consumer
industry, ia overstressed by advertising}   the public is conditioned
just as effectively as any Pavlovian dog by the sheer weight  and repeti-
tiousness of the advertising copy thrust upon it.
     At present the consumer is exploited through his technical  ignorance.
SCIF realises that an urgent need exists for clearer understanding of  the
medical, social, and economic benefits and risks that accrue  from the
application of science and technology and the use of chemicals in today's
world, and better guidelines for decision-making.
     The purpose of our endeavour is to enable us to inform and  instruct
the citizen on the causes of internal pollution and what is entailed in
trying to remedy them.  In a free society, at the end of the  day it is the
citizen who finally decides what kind of life he wants.   It is our respon-
sibility as scientists to see that he has the necessary knowledge to make
the right decision.
     Unfortunately, meetings such as the three this summer do not achieve
this objective;  they satisfy the egos of a small scientific  filite, but
do nothing for the wider public of which we are also members. It is up
to us  to start a dialogue with our fellow citizens rather than  conduct
monologues among ourselves;  otherwise it will begin to look  as  if the
sheer number of speeches, like the sheer number of diverse laboratory  tests
on a new drug, will reveal this air of purposefulness to be an illusion.

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                              2409
       A COMPREHENSIVE TOXICOLOGICAL LIMITATION OF TOXIC
   SUBSTANCES IN THE ENVIRONMENT, FUNCTIONAL AND METABOLIC
                 CRITERIA OF NOXIOUS ACTION
                N,F, IZMEROV, I.V, SANOTSKY
Institute of Labor Hygiene and Occupational Diseases, Moscow
                             USSR
ABSTRACT

     Methods baaed on the determination of the adaptability
limits of the organism and on metabolioal criteria may be re-
garded as a methodological basis for determining hazardous three-
holds of chemical pollutants in the environment^ and correspon-
dingly for determining hygienic standards.  Synergiatio and
 cumulative  effects of pollutants are also considered.

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                             2410
      In solving questions concerning a comprehensive toxi-
cological limitation (hygienic standards) of chemical com-
pounds in the environment, prime attention is devoted to
threshold and safety levels of toxins that enter the organ-
ism separately. These levels may be established, proceeding
from the knowledge about the stages of chronic intoxication,
which is an interaction of the processes of physiological
adaptation and compensation of pathological phenomena (so-
called "acclimatization") with a transition to decompensa-
tion. Difficulties are encountered when differentiating ordi-
nary reactions of adaptation, including orientating reac-
tions, which cannot be considered injurious, from patho-
logical reactions (temporary sub-clinical, compensated).
      In order to determine the borderlines of genuine
adaptation, the following laws of biology were suggested:
      - the law of unity of the organism and environment,
      - the law on unity of the organism as a biological
system,
      - the law on permanent numerity of a species, and
other laws, (I.V. Sanotsky, N.G. Ivanov, N.M. Karamzina,
V.N. Pomenko  [101).
      The first presumes determining stability of organisms
subjected to chemical action, to additional physiological and
extreme stresses. The use of different pharmacological func-
tional tests, as well as physical stresses (cooling, accord-
ing to restoration of rectal temperature test; ionizing
radiation - according to lethality and others) has made it
possible to classify the state of genuine adaptation or sub-
clinical pathology under the action of  small doses and con-
centrations of mercury, lead, CCl*t benzene, dimethylform-
amide, pyridine, carbon disulfide, triphthazine and many
other compounds.

-------
                             2411
      The second presumes a comprehensive study of the
organism since one or several indices may be interpreted
erroneously. For instance, the "acclimatization" to many
irritant toxins was proven to be the phase of chronic in-
toxication. Adaptation to certain mutagenic (ethera of tri-
chlorphenoxyacidic acid) aberration test of chromosome of
the bone marrow cell is accompanied by a rise in chromosome
aberrations in the cells of the liver*
      The third - the permanent numerity of a species is
connected with the action of a substances on the reproduc-
tion processes. A study of the selectivity and active
thresholds for several dozen substances has made it possible
to establish that comparatively few substances have specific
gonadotrophic, embryotrophic, mutagenic properties, namely -
chloroprene, ethylenimine, urethane, ethyline oxide and
others.
      The criteria of hazardness should be the deviations of
the indices beyond the limits of normal physiological fluc-
tuations (2£>  from the mean or average seasonal values), but
not yet obvious signs of pathology.
      The metabolioal criteria of hazardous action many be
the following criteria:
      - the level of action at which semi-ejection of the
toxin from the organism increases as compared with lower
levels of action (I.V. Sanotaky),
      - the level of action at which the activity of enzymes
is suppressed (24  from the mean or average seasonal fluctua-
tions) ; the effect is aggravated when there is a specific
pharmacological stress on the enzyme (V.V* Kustov, L.A. Tiu-
nov  [131).
      The above-mentioned methods may be regarded as a metho-
dological basis for determining hazardous thresholds of
chemical pollutants in the environment, and correspondingly,
for determining hygienic standards*

-------
                             2412
      The safety levels of action in case of separate entry
of substances of separate environments must take into con-
sideration remote  effects - such as gonadotrophic, embryo-
tropic, mutagenic, blaetomogenic, arteriosclerotic action
and other effects* In this problem, in spite of certain
headway that has been made, there are still many unsolved
questions, in particular, extrapolation to human beings of
data obtained from experiments involving animals, and othera
      Upon simultaneous entry of chemical substances through
the gastro-intestinal tract and the respiratory organs, most
frequently there is a summation of effects at the MFC or
chronic action threshold levels, (S.U. Pavlenko [8]). The
calculations by E.I. Spinu  1113 indicate that the quantity
of pesticides entering the organism through the environment
at MFC level for each environment, when summed up exceeds
the harmless dosage. It has been proposed to establish a
harmless maximum perroissable dose of a preparation for man,
obtained as a result of summing up the maximum permissable
doses and concentrations in all environments with considera-
tion for the share of each environment. In order to calculate
the maximum harmless dose during simultaneous entry of the
substance from different environments, it has been proposed
to use the principle of hygienic standardization that is em-
ployed at the present time in respect to combined entry of
substance with the same action, proc-eeding from the additive
effect (A.I. Korbakova, N.I. Shumskaya  [4])*
      Certain difficulties are encountered when involved
in comprehensive hygienic standardization in view of sign-
ificant differences in the toxlcity and nature of the action
in respect to certain substances when toxins enter through
different channels: nitryl fluoride (Yu.N, Korshunov [53)i
bromium (A.M. Klyachkina  [6], O.A. Krylov  [121).
      In natural conditions, when these substances are
inhaled or enter through the gastro-intestinal tract, there
is a considerable difference in the regimes of action by
these chemical compounds. It has been established that the
intermittent regime of action in certain cases proves to be

-------
                                 2413
 more  harmful  for the  organism (G.P. Babanov  [2],  I.P.  Ulano-
 va  [14], T.V.  Lomanova [?] , L.N. Burkatskaya  [3]).
        At present, data has been obtained pointing to  the
 practical need for a  comprehensive approach to hygienic
 standardization of the contents of chemical compounds that
 are present simultaneously in several  environments: fluorine
 (A.A.  Petina  [9] , G.P.  Pankratova  [1] j  benz-a-pyrene  (N.Ya.
 Yanusheva  Cl6j,  L.M.  Shabad,  I.V. Sanotsky  [15]  and others).
        Besides  the evaluation  of the environment,  an im-
 portant  index  of the  sum  total action  of a chemical factor
 for a  number of  substances may be the biological value of
 the integral MPC that  has been established with  the help of
 highly sensitive exposition tests.
References

1.  O.G. AVILOVA,  E.IA. GOLUBOVITCH, N.M. MALTZEVA,  G.P. PANKRATOVA  et al.
    "Atmospheric pollution by chemical compounds! adaptation and compensation"
    Moscow 1973, 45

2.  G.P. BABANOV et al "Toxicology and hygiene of petrochemical compounds"
    1972 32-45

3.  IA. N. BUKEATZKAIA, V.I.  MATIUSHINA,  "Hygiene and toxicology of pesticides,
    clinical poisoning" Kiev 1968, 6th Edition, 698-703

4.  A.I, KORBAKOVA, I.I. SCHUMSBLYA, G.N. ZABVA, T.K. NIKITENKO, "Scientific
    basis for the current hygienic normative efforts regarding the cheroioal
    compounds present in the environment" Moscow 1971, 35-40

5,  Yu. N. KORSCHUNOV, "Toxicology of the new industrial chemical compounds"
    Moscow 1969 2nd edition,  78-85.

6.  N. Q. IVANOV, A.L. GERMANOVA, V.S. POZDHIAKOV, A.M. KLYATEHKYNA et al
   "Adaptation and compensation of the organism confronted with chemical
    pollutants" Moscow 1973,  75-91

7.  T.V, LOMANOVA, M.N. FROLOVA, M.A. GRITVSKI "Psychophysiology and hygiene
    of various work activities" 1967, 57-67

8.  S.M, PAVLENKO, Hygiene and Sanitation 1972, 1. 40-4

9,  A.A. PETINA, L.N. ELNITEHKIN, "Fluorine and its profilaxis" Leningrad
   1967, 50-5

-------
                                     2414
10.  I.V. SANOTSKY, N.G.  IVANOV, N.M,  KARAMZYNA, V.N. FOMENKO "Scientific
     basis for the current hygienic normative efforts regarding the chemical
     compounds present in the environment" Moscow 1971t  63-68

11.  E.I. SPYMJ "Hygiene, application and toxicology of pesticides; clinical
     poisoning" Kiev, 1968, 103-9

12.  O.A. KRYLOV, The Setzenova Journal 1966, vol. 52, No.  7, 906-910

13.  L.A. TIUKOV, V.V. KUSTOV.  "Methods for the determination of the toxicity
     and danger of chemical compounds" Moscow 1970» 231-43

14.  I.P. ULANOVA et al,  "Adaptation and compensation of the organism confronted
     with chemical pollutants" Moscow 1973, 64-75

15.  M.I. SCHABAN, I.V. SANOTSKI et al. Hygiene and Sanitation. 1973, 4i  78-81

16.  N. Ya. YANSCHEVA, Hygiene ami Sanitation. 1972, 7t  8?

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                               2415
             BEHAVIORAL METHODS FOR INVESTIGATING
                 ENVIRONMENTAL HEALTH EFFECTS

                         BERNARD WEISS

Department of Radiation Biology and Biophysics, University of
Rochester School of Medicine and Dentistry, Rochester, New York
USA
ABSTRACT

     One may conceive of three phases in the assessment of be-
havioral changes induced by environmental contaminants.   The
first is the non-specific, or screening phase, during which a
search is made for general CNS effects.   The methodology emp-
loyed stresses relatively simple techniques, partly because, for
some agents, any CNS activity prohibits their use and partly be-
cause one aim is to estimate the relevant range of dose levels.
These techniques encompass observational tests of unlearned be-
havior, and relatively simple learned behavior.

     The second phase focuses on the specific functions that are
affected^  for example, sensory and motor function and complex
discriminative processes.   The pertinent techniques almost all
involve highly-trained animals, and advanced instrumentation,
including computers.

     The third phase focuses on human health.   Its purview ex-
tends from laboratory studies of complex functions to what might
be called behavioral epidemiology.   Naturally, it i& the most
difficult phase.

-------
                                    2416
1.  Introduction
    Numerous environmental insults and contaminants express their actions
through alterations in behavior.  These effects range from subtle, non-
specific symptoms to overt, easily discernible behavioral disruption.
The site of action can either involve the central nervous system (CNS)
directly, as with organic mercury poisoning, or indirectly, as with
respiratory irritants that induce performance impairment by provoking
discomfort.
    Only recently has it been recognized in the United States and most of
Western Europe that behavioral measures could play a significant role in
the assessment of toxic processes.  Part of this new visibility arises
from the pressures on public agencies to set standards on the basis of
minimal interference with health, not simply on the basis of clear
mortality and morbidity.  Part is due to the recognition that new tech-
niques are now available, largely from psychopharmacology, with which to
study chemical influences on behavior.  A significant contribution also
comes from an awareness that standard setting in countries such as the
U.S.S.R. historically has been guided by evaluations of CNS function.
    I plan, in this paper, to survey some of the ways in which beha-
vioral assessments may contribute to the resolution of certain Issues
posed to toxicology in the context of environmental health.  The frame-
work I will employ is the sequence of questions to be answered with the
Introduction of a new chemical agent presumed to Involve human exposures.
Other effects on the total ecosphere are set aside for the present.
    I conceive of such an assessment as encompassing three phases.  The
first is the screening phase, which corresponds to the preliminary obser-
vations undertaken in the pharmaceutical industry when a new compound is
being scrutinized.  The second is the specific function phase, during
which, if required, detailed assessments are made of systems that seem to
be targets of undesirable actions.  Such assessments encompass complex
behavioral processes in animals and the examination of sensory and motor
deficits.  The third phase deals with human susceptibility and its
parameters, both In the field and In the laboratory.
2.  Screening Phase
    Although a relatively straightforward, somewhat standardized approach
to CNS drug screening has been evolved by the drug industry, It falls
short for environmental agents.   Weighing risks and benefits is a much
different matter for a therapeutic chemical, used under limited condi-

-------
                                        2417
 tions,  than  for an agent  that might  be dispersed widely, linger for

 many  years,  and expose large  segments of the  population.  Such consi-

 derations direct one  to a sequential  evaluation such  as that  diagrammed

 in Figure 1.   This is a flow  diagram  devised  for a forthcoming
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Figure 1.  Flow diagram illustrating  a proposed sequential testing
            protocol  for behavioral effects.
 publication  from the U.S. National Academy of  Sciences, The Evaluation

 of Chemicals  for Societal Use,  prepared for  the Environmental  Protection

 Agency [1].

     As in conventional screening, we  recommend inclusion of CSS obser-

 vations in the total acute toxicity studies.   Such observations would

 Include simple indices of locomotor impairment, neurological Indices

 such  as tremor, ptosla, and convulsions, alterations  in varioua reflexes,

 and disorders of regulation.

-------
                                   2418
     Instead of stopping with  these  observations,  however,  an  environ-
 mental agent should  be  further  examined  for delayed  effects.  We  are all
 familiar with agents capable  of producing damage,  not  immediately after
 exposure,  but following a prolonged latent period.   Carcinogens are
 notorious  for such actions, but certain  CNS poisons  are  equally treache-
 rous.   Methylmercury is an example.   A recent  mass outbreak of methyl-
 mercury poisoning in Iraq, due  to grain  treated with a methylmercury
 fungicide,  would  have been much more constricted  in  impact if the effects
 had  not been visible only several weeks  after  the tainted  bread had begun
 to be  eaten [2].
     prenatal exposure is  another danger  in certain instances.  The combi-
 nation of  an immature,  developing nervous system  with  a  lack  of adequate
 detoxification mechanisms makes the fetus and  neonate  especially  suscep-
 tible.  Sometimes, the  consequences of such exposure may remain dormant,
 at least overtly, until the organism reaches a relatively  advanced age.
 Spyker's experiments on prenatal methylmercury exposure  [3] indicated
 that behavioral teratology is a question that  embraces the total  life
 span.   An  adequate screen must  confront  this set  of  problems.
    Certain aspects of the elementary screen are amenable  to easy quan-
 tification.  Gross locomotor activity can be measured in a variety of
ways, some of which are able to distinguish small amplitude movements,
such as  those associated with tremor, from  the large amplitude movements
 involved in walking and running.  Sauerhoff and Michaelson  [4] and, also,
 Silbergeld and Goldberg [5] observed significant Increases in the loco-
motor activity of rodents exposed to lead as neonates.
    Other simple measurements can be made with the open field test,
which helps an observer gauge an organism's exploratory activity,  and
which proved useful In Spyker's experiment.  She also found swimming
 tests  to be useful for revealing deficits in coordination not discer-
nible In ordinary locomotion.   The rotarod, which is simply a rod that
 revolves at a specified rate, can also be used to test  gross coordina-
 tion.  Certain CNS agents impair the ability of rodents to maintain
 themselves on the rod.
    Although body weight often serves as an index of toxic potential,
 peculiarities in  regulatory functions may not be revealed by the usual
measures.  Suppose, for example, the agent  in question destroys hypo-
 thalmic  tissue.  Gross damage may eventually produce hyperphagia and

-------
                                  2419
obesity or adlpsia and inanition.  During the development of such a
malfunction, the sole index of regulatory impairment might be a certain
finickiness about food, such as exaggerated responses to the tastes of
quinine (bitter) and saccharin (sweet).  We are examining raethylmercury
for such effects in mice.
3.  Specific Functions
    A chemical would be explored during phase 2 only If it seemed
capable of conferring some especially useful benefits.  An assessment
in depth requires a substantial Investment in time, talent, and money.
Sometimes, of course, such an assessment is inescapable, as in the case
of methylmercury, because it is so pervasive In food.
    3.1  Sensory Function
    Research on problems of what has been called animal psychophysics
 [6] is flourishing, taking advantage of progress both in operant
behavior and in technologies such as the digital computer.  These
developments offer toxicology an important tool for gauging impairment
in sensory systems, which, for some classes of substances, are primary
 targets.
     A cogent  example of the  utility of animal  psychophysics  is provided
 by work  performed by Dr. Hugh Evans in our laboratory.   Methylmercury,
 at  least in primates,  appears to concentrate in those  areas  of the
 cerebral cortex that subserve vision.   Not  only is  tissue  destruction
 visible  in  this  area,  but poisoned humans  experience constriction of
 the  visual  field.
     Starting with the  knowledge  that ablation  of visual  cortex impairs
 discrimination  of form more  than simple discrimination of brightness,
we decided  to approach the assessment  of visual function by  training
monkeys  to  distinguish among different geometric shapes.  We also
 reasoned that the constricted visual fields might simply be due to the
poorer representation  of the peripheral visual field on the cortex (as
well  as  in  the  distribution of retinal receptors).  If this were the
case, reducing the number of receptors stimulated, especially the rods
in the peripheral field, should exaggerate whatever impairment is
present.  This parameter was investigated by varying the luminance of
the stimuli from easily visible at ordinary room illumination to barely
detectable after 10 minutes of dark adaptation.

-------
                                   2420
      Figure 2 shows the arrangement devised for this experiment.   The
                                                     >

Figure 2.   Arrangement for testing visual discrimination in monkeys.
           A mirror was 'osed to show both front and rear views.   A small
           quantity of fruit drink is delivered through the brass pipe if
           the monkey presses the key with the square.

  monkey presses the  key  on which  the square  is projected  in order to
  obtain a  juice reward.  Incorrect  responses  are  followed by a  period
  during which no  events  are programmed.  All  of these  events are con-
  trolled and recorded by an on-line digital  computer.  Computer tech-
  nology is  used throughout our  laboratory  for these  purposes; we believe
  that precise exposures  and biochemical  analyses  should not be  hampered
  by  crude  behavioral analyses.   (See Weiss  [7] for a survey of  applica-
  tions .)
      The history  of  one  monkey  (M.  speciosa)  exposed to methylmercury
  appears in Figure  3.  The  uppermost panel  represents  the brightest
  luminance. The  lowest  panel shows accuracy at  the  dimmest  luminance.
       After 4 priming doses  of methylraercury chloride, the monkey's  whole
  blood  level  of methylmercury was maintained at  about 2500 ppb  by weekly

-------
                                    2421
  oral  doses  of  0.5  rag/kg.   After approximately 10 weeks, performance at
  the lowest  level began to deteriorate, was partially regained for a few
  weeks,  then finally deteriorated quite badly.  At the same time, the
                         100
                         90
                         80
                         70
                         100
                      s
                      s
 90
 80
 70
100
 90
 80
 70
100
 90
 80-
                         70
     10H2 mtombert
     (BRIGHTEST)
10~a9m Lambert
                             10'31m Lambert
                            10  m Lambert
                             (DIMMEST)
                           2   6  10  14  18  22  26  30
                        WEEKS SINCE METHYLMERCURY BEGAN
Figure 3.  Deterioration of visual performance  in a monkey treated
           chronically with methylmercury.
  monkey was able to maintain virtually perfect  accuracy at the higher
  luminances until about the 30th week, when  the monkey was virtually
  blind.
      Similar techniques have been used In other modalities.   Stebbins [8],
  for example, has shown that ototoxlc antibiotics  can be detected by
  training monkeys, in essence, to trace  their audiograms.
      3.2  Motor Control
      Animals can be trained to make many different varieties of motor
  responses, some of rather awesome complexity.   Since behavior basically
  comprises an organism producing changes in  its environment by various
  muscular movements, evaluations of motor performance should play a
  major role in detailed studies of toxic effects.

-------
                                    2422
      An example of how one might pursue such questions is illustrated in
  Figure 4.  The picture shows a squirrel monkey (S. sciureus) inserting
  its paw into a slot that contains a lucite plate mounted on a strain
  gauge.  The monkey was trained to press the plate with a force between
  5 and 15 grams in order to obtain a 90 mg sucrose pellet from the
  feeding magazine on the right.  Using auch a paradigm, it becomes
  possible to measure variables such as proportion of response time spent
  within the specified limits, the number of excursions above and below
  the limits, and other indices of precision of motor control [9, 10].  A
  digital computer controlled the experiment and was used for analysis.
Figure 4.   Arrangement to assess fine motor control.   The monkey inserts
           its paw into the slot and ia reinforced with a sucrose pellet
           It  it maintains a force of 5 to 15 grams for 2 consecutive
           seconds.

-------
                                     2423
        Figure  5  comes  from an experiment whose  aim was  to  study the onset
    of motor incoordination after weekly doses of methylmercury chloride.
    What can be observed on the figure is  an acute disruption of perfor-
    mance soon  after the weekly oral doses  plus  a gradual deterioration
    through the total period of exposure.   This  deterioration was mani-
    fested as a high initial force,  then a  gradual slide into the desig-
    nated range instead of the precise entrance  into the band charac-
    teristic of baseline performance.
        One can use similar reinforcement  contingencies  to  measure strength.
    A recent publication by Dews and Herd  [11] examined  the effects of
    ganglionic  blockers on the pressor responses Induced by muscular con-
    traction.   The situation employed monkeys (M.  raulatta)  trained to pull
    a weighted  rod for  a food reinforcement.   In our laboratory, to examine
                                MONKEY 91

                                       ,
                                   '




           (
                                                 .  IbHaMS/21]

                                      •




                                                      .

         6/7                  [U«t/I9v.

                                                       •

                       •

Figure 5.  Changes in the ability of a methylmercury-treated monkey to
           exert a relatively constant force.  Note how the first dose
           (5/21) produced an acute effect on  5/24 .   Deterioration of
           performance on 6/7 and 6/9 was marked by inability to come
           directly into the proper range (cf. , 5/17 and 5/19).

-------
                                  2424
static strength, we have trained squirrel monkeys to maintain forces
of over 200 grams in order to avoid an electric shock to the tail.
    3.3  .Comple_x_Pr_qc_eiBBes
    Many human activities, and many toxic phenomena, involve not pure
sensory or motor function, but responses to complex relationships in
the environment.  Some of these may be approached by examining an
organism's behavior in the context of what we call "Schedules of
Reinforcement"; that is, how the properties of behavior are governed
by specified relationships between behavior and its consequences.  The
advantages of such an approach lie in the large number of variations
an experimenter can explore using the same basic technique.  It has
proved immensely useful to behavioral pharmacology, and I predict that
behavioral toxicology will see its repertoire of methods enhanced in
the same way.
    One current project in our laboratory, carried out by Dr. Tina
Levine, is directed toward the behavioral effects of carbon disulfide,
partly because of Its interesting neuropharmacologic properties:  it
appears to act as a dopamine- 0-hydroxylase inhibitor, so that it
retards the conversion of dopamine to norepinephrine.  Pigeons are the
first species we turned to because they are easy to handle and train.
    Figure 6 shows how some of our pigeon chambers are constructed.
The pigeons, usually maintained at 80 percent of their free-feeding
body weight, are trained to peck an Illuminated disk connected to a
switch.  When the pigeon is reinforced, a food magazine, loaded with
grain, is elevated into position for eating.
    Figure 7 shows how one can quantify certain effects of carbon
disulfide exposure.  The pigeons whose records appear here were  tested
in a slightly different chamber—one with two pecking keys.   If  the
pigeon pecked the left key 8 or more times before pecking  the right
key, it  received access to food.  Otherwise, the left key  counter was
reset.   Latles  [12]  found that such a schedule was  sensitive in
different ways  to a  variety of drugs.  The tracings in Figure 7  demon-
strate that with increasingly longer exposures to  carbon disulfide the
rate of  responding decreased, but at no  sacrifice  In accuracy.   The
levels required to produce such an effect are rather high  compared to

-------
                                   2425
Figure 6.
Pigeon teat chambers.  The pigeon In the middle is pecking at
the response key.  The pigeon  at  the bottom is eating from
the food magazine.
                        CS2 FCN8

            4HRS    BIRD 165

               8HRS DAY 1   jS  	/
                                      8MRS DAY 2
                                                  §200
                                                  §  100
                                                        I 35
                                                      MINUTES
                              8HRS DAY I

                                   BIRD 2965
Figure 7,
                       8 MRS DAY 2
Changes in  rate  of  pecking induced by carbon disulfide
exposure.   The records  are cumulative; each response produces
an increment  in  height, so that, with the recording paper
moving at constant  speed,  slope is equivalent to response rate.
The slashes on the  record  indicate reinforcements.  The
pigeons were  removed from  the exposure chambers for testing.

-------
                                   2426
the TLV for humans, but we currently are more interested in the relation
between the neurochemistry of carbon disulfide and behavior than in stan-
dard setting.  Later, we will be asking such questions of primates.
4.  Human Evaluations
    Although we all recognize that the assessment of health effects in
exposed human populations poses the most difficult challenges to environ-
mental health science, the problems are not insurmountable.  Particularly
for specific questions about function, behavioral toxicology can draw on
many decades of refinement in experimental psychology for techniques to
determine impairments in sensory and discriminative processes, complex
motor function, and phenomena such as those subsumed under terms like
"learning" and "memory."
    When our laboratory needed to evaluate three women who had been
occupationally exposed to mercury vapor, we arranged the system shown
in Figure 8  [13].  The patient kept her finger in a lucite trough
attached to  a strain  gauge.  Lights on a small box facing the patient
indicated whether the force exerted on the strain gauge, which is  an
isometric device, was above, below, or within prescribed limits.   (Note
how this task is analogous to the one used with the monkeys.)  We wished
to obtain measures of motor function that not only reflected overt tremor,
but that could give us an indication of fine control.
Figure 8.  Device tor measuring tine motor control in humans.  The lucite
           trough is connected to a strain gauge.  The indicator lights
           denote "start," "above" and "below."

-------
                                    2427
     One of  the patients, when she  first appeared in the laboratory, gave
  us  records  such as  the one shown in the upper section of Figure 9.  This
  patient was unable  to  keep  the  applied force between  the designated  limits
  of 10 and 40 grams. Nine months  later, with no  intervening  exposures,
  performance greatly improved (lower section of  Figure 9).
                                                                 4-9-70
5
5
^
100 q
 70:
     10
                                                                 1-14-71
                                        15SBC.-
                                                           •I
  Figure 9.  Polygraph  tracings of a patient  (see Figure 8 apparatus)
            exposed  to mercury vapor  (upper) and 9 months after cessation
            of  exposure  (lower).
      A more useful way  of tracking progress is shown in Figure 10.  The
  patient whose  record is shown in Figure 9  appears here as "Mrs. T."
  "Mr. T." is her husband, who served aa a control.  "Mrs.  N." is a co-
  worker.  The figure  shows that the proportion of time within the pre-
  scribed limits attained stable values in about two months, during which
  the plasma level of  mercury fell to about half its earlier value.
  "Mrs. N." showed a more rapid response, but her plasma levels were
  lower, and, in addition, she was treated with the chelation agent,
  n-acetylpenicillamine.
      A further parameter of quantification  appears in Figure 11.  These
  charts represent a frequency analysis of the tremor.  The earlier
  evaluation shows multiple peaks and a huge variance.  The later one
  shows a much narrower  distribution of power.
      This demonstration testifies to the utility of quantification and
  precise measures of  motor function.  Similar examples for discrimina-
  tive functions can be  extracted from the literature on carbon
  monoxide [14].

-------
                                2428
                     100 1
                     50
                      0
                     100

                                            I
                   20

                    0
                 g
                     50
  s^**-***!^    ^
 //^N/    
-------
                                   2429

                 32.0l
                 16.0-
                                               4-9-70
        r
                 l.On
0.5-
                                              1-14-71
                         t
                                       10
                               15
20
                              FREQUENCY (Hz)
Figure 11.  Power  spectral analysis of the tremor shown in Figure 9.  This
           chart  shows the distribution of power (variance) over fre-
           quency.  The effects of mercury vapor exposure were not simply
           to  increase tremor amplitude.
      Laboratory  studies such as those adduced above are, I will admit,
  projects  of  minimal vexation compared to the frustration presented by
  studies in the  field.  When exposed individuals cannot be brought into
  the laboratory  for testing, or when the symptoms do not lend themselves
  to laboratory studies, we have to turn to other techniques.  This is
  especially true when dealing with the relatively nonspecific complaints
  associated with many early intoxications.
       Mercury vapor exposure, for example, engenders a collection of
  symptoms  so  well known in the clinical literature that It even has a
  label—erethism.  Yet, the components of erethism sound Just like a
  catalog of so-called neurotic complaints:  fatigue, irritability,
  anxieties about social relationships, numerous mild somatic complaints,
  and, in general, a marked lability in behavior.  Carbon disulflde and
  lead are other  examples of substances that may produce such complaints.
       Although I would hardly minimize the obstacles to reliable measure-
  ment of this class of variables, the methodology is a standard procedure

-------
                                  2430
in psychology.  Many test and survey instruments are developed each year
according to these procedures.  Unfortunately those aimed at some of the
questions that environmental health specialists view as important are
not pertinent because they tend to focus on enduring traits, not the
assessment of change.
     4
    There are, however, enough materials available to provide a firm
beginning to the design of a useful questionnaire instrument.  Examples
such as Goldberg's General Health questionnaire [15] and the Symptom
Distress Check List  [16] can be used as guides to useful items.  Once
refined, moreover, such an instrument could be validated in more easily
quantifiable ways than psychological tests typically have access to:
body burden measurements, days of absenteeism, exposure duration, and
so on.

5.   Coriclusions
     I have surveyed  the range of what I believe behavioral  toxicology
encompasses in the broad context of environmental health.   Naturally,  I
have had to ignore many fascinating and useful aspects, and creative
workers, in order to keep this paper within bounds.  I hope, however,
that I have helped familiarize some of you from other specialties with
the ways in which certain vital issued in environmental health are
joined to behavior science.
                            Acknowledgment

    The preparation of this paper was supported, in part, by Grant
MH-11752 from the National Institute of Mental Health, Grant NS-08048
from the National Institute of Neurological Diseases and Stroke, Grant
GI-30097X from'the RANN Program of the National Science Foundation, and
in part by a contract with the U. S. Atomic Energy Commission at the
University of Rochester Atomic Energy Project and has been assigned
Report No.  UR-3490-541.

-------
                                    2431
                               References

 1    Weiss, B.,  Brozek, J., Hanson, H., Leaf, R.C., Mello, N.K.,
     Spyker, J.M.,  "Effects on behavior."   In The Evaluation of Chemicals
     for  Societal Use, National Academy of  Sciences, Washington, D.C
     (in  press).

 2    Bakir, F.,  Damlugi,  S.F., Arnin-Zehi,  L., Murtadha, M., Khalidi, A.,
     Tikriti,  S., Dhahir, H.I., Clarkson, T.W. ,  Smith, J.D.,
     Doherty,  R.A. , "Methylmercury poisoning in  Iraq:  an  inter-
     university  report,"   Science, 181, 230 (1973).

 3    Spyker, J.M.,  Sparber, S.B. , Goldberg, A.M., "Subtle  consequences
     of methylmercury exposure:  behavioral deviations in  offspring
     from treated mothers,"   Science,  177,  621 (1972).

 4    Sauerhoff,  M.W., Michaelson, I.A., "Hyperactivity and brain
     catecholamines in lead-exposed developing rats,"  Science, 182,
     1022 (1973).

 5    Silbergeld, E.K.,  Goldberg, A.M.,  "A lead-induced behavioral
     disorder,"   Life Sci.. 13, 1375  (1973).

 6    Stebbins, W.C. (Ed.), Animal PsychojahysjLcjs, Appleton-Century-Crofts,
     New  York  (1970).

 7    Weiss, B. (Ed.), Digital Computers in  the Behavioral  Laboratory,
     Appleton-Century-Crofts, New York (1973).

 8    Stebbins, W.C.,  Miller,  J.M., Johnson, L.G. , Hawkins, J.E.,
     "Ototoxic hearing loss and cochlear pathology  in  the  monkey,"
     Ann. Otol.  Rhinol.  Laryngol., 78,  1007 (1969).

 9    Falk, J.L. , "The behavioral measurement of  fine motor control:
     effects of pharmacological agents," in Thompson, T.,  Pickens, R.,
     Meisch,  R.  (Eds.),  Readings in Behavioral Pharmacology, Appleton-
     Century-Crof ts,  New York (1970).

10    Notterman,  J.M., Mintz,  D.E., Dynamics of Response, Wiley,
     New  York  (1965).

11     Dews, P.B., Herd, J.A.,  "Behavioral activities and  cardiovascular
      function:  effects of hexamethonium on cardiovascular changes
      during strong sustained  static work in rhesus  monkeys,"
     J. Pharmacol.  Exper. Therap..  189 12  (1974).

12    Laties,  V.C.,  "The modification  of drug  effects on  behavior by
      external  discriminative  stimuli," J. Pharmacol^ Exper. Therap.,
      183, 1 (1972).

-------
                               2432
 13   Wood,  R.W.,  Weiss, A.B., Weiss, B., "Hand tremor induced by
      industrial exposure to  inorganic mercury," Arch. Environ. Health,
      26, 249 (1973).

 14   Laties, V.G.,  Beard, R.R., Dinman, B.D., Schulte, J.H.,
      "Behavioral effects of  carbon monoxide poisoning."  In Efj!ects of
      Chronic Exposure to Low Levels of Carbon^Monoxide on Human Health,
      Behavior and Performance.  National Academy of Sciences,
      Washington,  D.C. (1969).

 15   Goldberg, D.P., The Detection of Psychiatric Illness by
      Questionnaire. Oxford University Press, London, England  (1972).

 16   Parloff, M.B., Kelman,  H.C. , Frank, J.D., "Comfort, effectiveness
      and self-awareness as criteria of improvement in psychotherapy,"
      Amer.  J. Psychiat. 3, 343  (1954).
                            DISCUSSION
SANOTSKY (USSR)
      In the determination  of the threshold  of  chemical action
what  is comparative sensitivity of the condition reflex tests
or EEC-tests?


WEISS (U.S.A.)

      The spontaneous EEC contains much electrical noise; that
is, random activity.  Only fairly advanced  analytical methods,
such  as Fourier analysis,  can make much sense  of such infor-
mation.  The availability  of the small digital computer, and
methods such as the Fast Fourier transform, will help.  I am
convinced, however, that the evoked potential  method because it
controls for many more extraneous phenomena, is the method of
choice  for studies of brain electrical activity.  New mathematical
methods are also being developed.

      As to the comparison  you requested behaviorial pharmacology
offers  cogent evidence for a greater sensitivity of behavioral
methods.   You can see why, in a behavioral  experiment, you can
control many extraneous variables, manufacture the behavior to
specifications, and measure exactly what you wish.


STEENSBERG (Denmark)

      May I draw attention  to what was said  by  Prof. Lawther on
the recent colloquium in Luxembourg on carbon  monoxide environ-
mental  pollution and public health organized by the Commission
of the  European Communities.  In the discussion on perception
and performance tests Lawther was giving a  warnining to the

-------
                             2433
usefulness of some of the more subtle methods of investigation.
We may register effects of carbon monoxide which are equivalent
to the effect of a cup of coffee or tea or the intake of a small
amount of alcohol.   The practical importance of such investig-
ations is limited.
WEISS (U.S.A.)

     We must not forget that the society, and the public agencies
that represent it, roust determine what are "small" effects.
Remember that the CO literature is based on young, healthy men;
and we know that older people are more susceptible.

     Furthermore, CO is only one constituent of a chemical soup
in which we live.  The ingredients interact - may have additive,
or synergistic properties.  CO may at times,  simply be the
additional load that breaks beyond the threshold.


NEEDLEMAN  (U.S.A.)

1.   Would you extend your comments on the discriminating power
of a neurological examination to the sensitivity of health
questionnaires of routine clinical examinations?

2.   Could you put your remarks into a developmental context?
That is, what are the possible effects of such interferences in
perceptual, motor or cognitive processes on a growing child's
brain, during critical- periods of neurogenesis, myclogenesis,
or other forms of maturation and learning?


WEISS (U.S.A.)

1.   Routine clinical examinations are not meant to detect subtle
functional impairment.  They are designed to elicit indications
of frank disease.  Even the addition of a skilled neurological
examination may not be enough to distinguish phenomena that
might only be expressed as vague, subjective complaints,  or
complex malfunctions discerned only with advanced technology.

2.   The developing nervous system is especially susceptible to
toxic processes.  As I pointed out in a recent article in Ped-
iatrics, some of these effects are less important in themselves
than in what they portend.  Experiments on prenatal exposure to
methylmercury indicate that many such consequences may be re-
vealed only in advance age, when other processes also contribute
their effects.

-------
  SCHLUBSITZUNG






 CLOSING SESSION






SEANCE DE CLOTURE






 SESSIONE FINALE






   SLOTZITTING

-------
                             2437
     CONCLUSIONS ET REFLEXIONS DES CONSEILLERS SCIENTIFIQUES
                       Presentees par

                   le Prof. A. LAFONTAINE
                          Belgique
     En matidre de protection de 1'nomine et de 1'environnement,
les critSres sont dSfinls comme un jugement, base sur des don-
nfies scientifiques tradulsant une relation, autant que possible
BxprimSe num6riquement, entre I1exposition d'une cible a un
facteur d'agression et le risque et/ou 1'effet dSfavorable ou
indSsirable au niveau de cette cible: I'Stablissement de critSres
est la premiSre des demarches qui, en logique cartSsienne, doit
aboutir d'abord a la fixation d'objectifs de qualite", ensuite &
1'etablissement de normes, de regies et de codes de bonne pra-
tique .

     La santfi de 1'homme et de son espfice etant sans conteste
la plus importante des preoccupations, il etait logique que
les trois organisations, responsables du Symposium aient eu pour
premier souci de rfiunir les connaissances scientifiques exis-
tantes pour progresser dans 1'etablissement des relations entre
1'exposition aux polluants et les effets directs et indirects
sur la sant£, afin d'arriver a une Evaluation aussi rationnelle
que possible des risques, et, dans un stade ultSrieur, pour
decider des programmes d'intervention et des actions en vue de
prSvenir ou reduire la pollution ou d'en limiter les rfipercus-
sions.

     L1 experience ayant h61as, montrfe combien souvent fragmen-
taires fitaient les informations et combien peu exploitables
Staient de nombreuses Etudes expSrimentales et certaines fitudes
6pid6miologlques, les promoteurs se rendaient parfaitement com-
pte des difficultfis qul se presenteraient, d1autant plus que
certains facteurs moins pertinents risquent de s1 intriguer aux
aspects strictement scientifiques.  Us savaient aussi que beau-
coup de recherches avaient 6t6 conduiteg pour des raisons dif-
£6rentes de celles de la poursuite de I'fitablissement de critS-
res. -Aussi, en plus des nouvelles donnSes qui pourraient £tre
recueillies, avaient-ils pour but:
a)  d'fitablir une meilleure planification, une motivation plus
    sp6ci£ique et une orientation plus efficace et rigoureuse-
    ment objective des recherchesj             .  -
b)  de d£couvrir la nature et I1importance des lacunes:
c)  d1identifier, autant que possible, les secteurs ou des
    realisations sont imperatives en raison de 1'urgence ou de
    la prioritfi des problSmes et ceux pour lesquels les dbnnfies
    existantes, mSme si clles sont encore incomplStes, sont
    suffisantes et valables pour passer a 1'action.

-------
                              2438
     Malgrg les difficultfis, malgrfi le caractSre inegal et par-
fois dispersfi des communications presentees, les conseillers
scientifiques estiment neanmoins le bilan coirane largement posi-
tif, si 1'on considers les divers objectifs envisages.
     Une sfirie de donnSes nouvelles ont 6t§ apport6es concernant
entr'autres la pollution atmosphe'rique urbaine, les dangers des
m6taux  lourds, les nuisances acoustiques,  les techniques experi-
mentales,  les indicateurs d'exposition, les mesures au niveau
biologique, les nouvelles approches expfirimentales des phfinomenes
du comportement. Des nouveaux problemes ont 6t6 abordes tels
ceux poses par les biphfinyls polychlorSs ou le chlorure de
vinyle.  Mais davantage se  sont pre'cise'es  les donnfies scientifi-
ques requises pour la prise de decision pour la protection de
la sant£ et ont £t£  identifiers les lacunes fondamentales dans
les connaissances, les orientations futures des reoherches et
des terrains  a explorer, pour arriver a des bases scientifiques
nficessaires pour  les processus de reflexion et de d§cision.

     Nous  manquerions toutefois d1objectivity  si  avant de pr€-
ciser certaines reflexions  et suggestions, nous ne soulignions
pas certaines faiblesses dfimontrees par les documents pr£sent£s.
Par example,  1'engouement trop unilateral  pour certains themes
comme les  me'taux  lourds ou  pour certains secteurs comme les
polluants  atmospheriques.   Les causes en sont probablement les
suivantes: enthousiasme ou  facility scientifique, orientation
politique, pression  du public.  Par centre, d'autres thdmes ont
attir£  peu 1'attention comme les d6riv€s nitres, les nitrosa-
mines,  les mycotoxines ou le manganese.  Bien qu'ils ne manquent
pas d'actualite,  certains secteurs comme ceux des rSsidus solides
et de la contamination du sol n'ont oas prSoccunfi  les auteurs,  et
toxicologie des eaux destinees a la consommation humaine pourrait
paraltre aujourd'hui pour un auditeur non  averti, comme un pro.*
bl@me rfisolu.  D'autres secteurs comme celui des medicaments ou
des additifs  alimentaires ou certaines nuisances comme celles
liees aux  radiations ionisantes et non ionisantes n'ont pas etg
consid6r6s au cours  de ce Symposium: les raisons majeures sont
liees a des cloisonnements  devenus traditionnels mais qui mSrite-
raient  d'etre leves  en vue  d'une approche  plus globale des pro-
blemes,  la  cible de ces diverses nuisances  £tant la meme, 1'honutie
et son  espece.

     Par allleurs, si les Conseillers Scientifiques regrettent
une dispersion parfois exagerfee des problemes de la sant6 de
1'environnement au niveau d'un nombre considerable d'organismes,
ils tiennent  au contraire a souligner la cooperation qui s'est
6tablie entre les trois organisations responsables du Symposium
et a les remercier pour 1'opportunity exceptionnelle qui a &t&
donnfie  a des  chercheurs europfiens et amfiricains et par la liai-
son de  1'O.M.S.,  et 3 des chercheurs  du mond entier,  de  se  ren-

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                              2439
contrer pour chercher a Stablir des crit&res de sant£ (les crite-
res restent en dehors de toute consideration socio-e'conomique ou
politique)  communs a tous les hommes de cette "seule terre" et
pour oeuvrer de mani&re aussi coordonne'e et fraternelle que pos-
sible.
     Parmi les nombreuses reflexions <3ui suggftrent les
du Symposium, nous pensons utile de retenir, outre la mise a
execution des programmes d'action concernant la liste pr§sent6e
par 1'O.M.S. de substances pour lesquelles des informations
doivent etre assemblies et des recherches poursuivies, quelqUes
propositions particuliSres:
1.   Avant tout, il importe de s'exprimer clairement et la
ngcessitS d'un glossaire dSfinissant le sens des termes utilises
en matiere d'environnement s'est confirmee.
2.   Trop souvent, on considdre ind£pendamment chaque polluant
et chaque secteur en oubliant que I'homme joue un role d'int§-
grateur des nuisances: un meme toxique peut 1'atteindre par di-
verses voies et divers agents altSragdnes, y compris ceux lie's
a certaines habitudes  (usage du tabac par exemple), peuvent
toucher le meme organe cible. Les risques Ii6s aux polluants
chimiques et physiques ne doivent pas faire oublier les risques
biologiques et microbiologiques.  La notion de charge corporelle,
d1incorporation totale doit se g^n^raliser de meme que celle  des
doses admissibles par jour, semaine, mois ou ann§e, suivant  le casp

     De plus, a la notion de concentrations dans 1'atmosphore,
dans 1'eau ou dans le sol, doit s'ajouter celle de la capacity
toxicoloaioue d'un milieu, d'une region, d'un bassin, en narti-
culier pour les substances non deqradables: il s'agit de la auan-
tit£ totale de substances polluantes oue peut supporter sans
dommages intolerables ce milieu, cette region, ce bassin.

3.   Les d€bats ont  souligne  le besoin  de developpements metho-
dologiques, notamment  en ce qui concerne  1'epid^miologie,  les
modeles animaux,  le  pretesting et  les balances avantages/desa-
vantages:
     a)  les techniques epidemiologigues  sont  fondamentales,non
seulement pour ^valuer  I1influence  directe  ou  indirecte  des
pollutions  sur la sant£, mais  aussi pour  identifier de nouveaux
risques ou  inconv£nients et pour  apprecier  1'efficacite  des
actions entreprises.   L'epide"miologie devrait  pouvoir  s ' appuyer
davantage sur  les informations apportees  par un  public pr^venu
et  forme et par un  corps de medecins praticiens  avertis:  la
clinique generale et les observations en  milieu  professionnel
pouvant etre d'une  importance  capitale, meme  si  cela  demande
une preparation du  corps medical.   La mise  en  oeuvre  de  program-
mes gpidemiologiques plurinationaux peut  etre  tres  utile pour
^valuer  1'effet concomitant d'autres facteurs  comme les  condi-
tions  d1ambiance  ou les influences geographiques,  climatiques,
etc.

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                               2440
     b)   le dSveloppement, au plan des essais toxicologiques,
de modules anlmaux pour  lea maladies humaines est  Indispensable
pour mieux comprendre  les effets observes  et pour  assurer  le
maximum d'efficacite 3 1'examen prgabable  des nouveaux produits
et des nouvelles  technologies.  Ces modules devront  etre concus
de maniSre S  serrer au plus pres les Studes 6pidemiologiques;
     c)   leprobl&ne  du "pretesting" doit etre un souci majeur
des scienti'fiqu.es: il  importe, sous peine  d'exposer  1*human!t6~
a des risques imprfivisibles, que les nouveaux produits chimiques,
les nouveaux procSde's  et les nouvelles sources d'gnergie soient,
prialablement SL leur eraploi fitendu, 1'objet d'une  Evaluation
aussi large que possible sur la nature et  1*importance des ris-
ques a court, moyen et long termes y compris la recherche des
effets mutagenes, canc6rogenes et t6ratogenes 6ventuels et de
1'etablissement d'une  balance avantages/de'savantages pour
1'homme et son espece.  Un mficanisme mgthodologique doit fitre
61aborS qui tente de prSvoir au mieux les  risques  toxicologiques
et £cologiques divers  mais pr£voie figalement les criteres de
puretfi applicables, le sort dans 1'environnement des substances
mises en  oeuvre et leur detection  (du produit pur, ainsi que
des impuretes et metabolites 6ventuels) dans les diffSrents
composants du milieu;

     d)   une balance entre les avantages et les dfisavantages
pour la santfe des produits et techniques mis en oeuvre pour
1'homme est indispensable: toutefois, les  approches  actuelles
sont souvent  inadfiquates, sur le plan conceptuel comme sur le
plan technique et il importe de repenser le probl&me de maniftre
multidisciplinaire.  Dans un meme ordre d'id§es, 11  importe de
mieux Svaluer le  coOt  de la pollution et de la lutte centre les
polluants ainsi que I'efficacitfi des moyens de lutte et de leurs
effets secondaires 6ventuels sur le plan de la sante.  Ces no--
tions seront utiles SL  c6t6 d'autres Elements socio-4conomiques
et regionaux dans le processus de decision partant des criteree
et allant vers les objectifs de qualite1 et les normes.

     A ce dernier point de vue, une attention particuliere de-
vrait $tre accord^e aux problSmes qui se posent pour  les pays
en voie de dfiveloppement: il pourrait arriver que  les besoins en
developpement 1'emportent en urgence sur les exigences de la
protection.

4.   On observe souvent a c6t6 de manifestations franches ou
du moins  assez aisfiment identifiables des  chanqements considerftp
comme Infra-pathologiques ou d1interpretation difficile comme~	
la stimulation des enzymes mitochondriales ou la modification
de la dehvdratase.de 1'acid delta aminolfivulinioue chez les DerBrtn
nes exposees au plomb.  Des recherches doivent permettre de   i^°"
donner un sens-plus prficis a ces modifications et  de savoir s'ii
s'agit de simples concomitants ou d'alterations subtilea pr6c6dant
des manifestations pathologiques: en meme  temps, il serait
de savoir si ces modifications peuvent etre employees comme
indicateurs d1exposition.

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                                2441
  5.   En relation avec le point precedent, se soulSvent des
  problSmes:

       a>   le choix des groupes critiques a prendre en consid€-
  ration au point de vue de la sant<§ et de 1'ampleur des risques
  eventuels qui pourraient etre raisonnableroent acceptables pour
  ces groupesf                                               ^

       b>   le choix des prioritgs  en matiSre de recherche scien-
  tifique bashes notamment sur les connaissances sur la nature du
  risque,  son ampleur,  sa reversibilite et sur les possibility
  d  action,  qu'il s'agisse de polluants naturels ou provenant de
  1  activity  humaine.

  6.    De meme,  il importe de considSrer differemment les risques
  observed dans  les conditions habituelles et les risoues acciden-
  ¥±2''  ?;fs  Unites acceptables dans ce dernier cas seront dgrivees
  de  critdres applique^ a I1 exposition aigue et les mesures
  d.fnj?ry!nti°n aPPr°PriSes a 1'ampleur du risque et du nombre
  d'individus menaces.

  7.    En  ce  qui concerne les valeurs  actuellement admises comme
  normes,  il  est sur que  certaines d'entreelles a la lumiSre des
  connaissances  acquises,  doivent  etre revues:  c'est particulidre-
  ment  le  cas de certaines  concentrations  maximales admissibles
  dans  les milieux  de travail.   En effet,  certains effets a long
  terme n  ont pas 6t€ suffisamment pris  en consideration parce
  qu'on n'a peut etr^ pas tenu assez compte du  fait oue  dls  Expo-
  sition non professionnelles viendraient  s'ajouter a des exposition
  professionnelles et sur tout parce au'on a parfois un peu  perdu
  de vue aue des femmes en 6tat de procr^er travaillent de plus
  en plus dans I1 Industrie: certaines valeurs doivent indiscutab-
  lement etre repensSes en tenant compte du transfert transplacen-
  taire et transmammaire et de la protection de foetus et du nour-
  risson.


      Nous avons dgfini  le terme critSre  au d^but de cet exoosfi
 dont rintlrSt' T"* d'envls^er certaines Etudes fondame^Sfes
 A* L    ?ffet SSt aPParu au cours ^s debats, attirer  1' attention
 de nos collegues sur trois points qui nous paraissent importants?
                      plutot comme des
les odeurs ou Certains inconvenients acous-
 t22tefola P°ssible de chiffrer 1' exposition
 nl      »
 oc<-   *q2   ^lfler 16S SffetS et P01^tant un jugement critrial
 eat  g6nSralement possible,  qui permettrait la mise en marche
 du processus  dficisionneli  cela ne veut pas dire qu'un effort
 technique  et  m^thodologique ne doit pas etre fai? pour arrlver
 le plus tot possible I  des  expressions numeriques.      arriver

 doiveJt ItL1^^^  dS  S0uli9ner  Par  ailleurs  que des  decisions
 doivent etre prises dans  certains cas quant  a  la fixation de
surJs L°" de rjeurs de reference ou a  la  prescription de ml-
sures reglementaires avant que  les critdres n'aient  pu  etre

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                               2442
€labor€s; 11 importe dans ce cas de rSunir le maximum de donnfies
scientifiques, meme partielles ou analogiques, S la condition
qu'elles soient suffisamment Svidentes pour Stayer ces decisions.
Elles doivent susciter le plus tot possible des confirmations
expSrimentales ou epide"miologiques.

3.   Enfin, il imports de poursuivre, des recherches sur le plan
experimental et clinique pour mieux ^valuer par exemple les ef-
fets psychologiques de certaines nuisances ou de certains stres-
ses de I1existence moderne et mieux connaltre les effets psycho-
logiques de certaines nuisances  (comme les Equivalents biologi-
ques du bruit).
     Enfin, parmi les recherches fondaraentales d'appui n€ces-
saires que ce Symposium a permis d1identifier, nous retiendrons
particuliSrement les points suivants:

1.   I16tude des facteurs d'agression, qu'ils soient d'origine
naturelle ou apport£s par les activit£s humaines doit Stre
approfondie, quant & leur mScanisme d1action, quant S leurs
interactions au niveau du milieu et des ciblesj
2.   une attention plus approfondie doit etre accorded aux ef-
fets sur le potentiel gSnfitique, sur  1'organisme foetal  (m§me
s'ils sont tr&s tardifs et n'apparaissent que lorsque le  jeune
etre est arrive1 £ I1 age adulte) , sur  les transferts transplacen-
taires et transmammairesj
3.   les transformations me'taboliques des substances xSnobioti-
ques dans le milieu, le long des cycles biologiques et au niveau
de 1'organisnie humain n'ont pas tou jours §t€  suff isamment pris
en consideration^ des recherches exp§rimentales dans ce  sens
sont indispensables;
4.   de nouvelles recherches sont  indispensables en ce qui con-
cerne 1"existence ou I1absence de  seuil pour  les effets  cancero-
gSnes: les mficanismes de reparation,  prouv^s  par la biologie
mole'culaire, oblige a repenser cette  notion;
5.   par ailleurs, n'existe-t-il pas  a 1'Sgard de  certains
facteurs d'agression des m6canismes d1adaptation ou de compen-
sation autres que la selection naturelle et qui se sont  mani-
festfis au cours de I'-histoire humaine?  Us mSriteraient  d'etre
identifies, ^tudi^s et 6valu6s;
6.   tout en 6cartant des preoccupations actuelles les critSres
6cologiques, il importe toutefois  d1identifier et  d'fivaluer les
effets indirects sur la santS de I'homme des  effets ficologiques
comme  1'atteinte de certains v6g6taux essentiels a la survie
de I'homme ou comme la pollution thermique avec ses retentis-
sements sur 1'eijvironnement et  entr'autres sur le  plan microbio-
logique et parasitaire;

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                                2443
  7.    Les  techniques  actuelles  de  mesure  dans  1'environnement
  (monitoring)  sont  parfois  encombrantes:  elles doivent  etre
  mieux  adaptees  a  1'evaluation numSrique des  expositions  et par
  ailleurs  des  nouveaux  indicateurs de  pollutions/ si  possible
  polyvalents,  doivent etre  recherches  dans le  milieu  et chez
  1'nomine pour  faciliter  les enquetes Spidemiologiques et les ren-
  dre plus  efficacesf

 8.    parmi les effets § long terme, le phenomgne de vieillis-
 sement acce"ler€ sur le  plan physique et mental n'a pas  gt€
 suffisamment pris en consideration comme peut etre aussi, certai-
 nes repercussions impreVues sur 1'ontogenese;   une approche fon-
 damentale  est souhaitable en meme temps que des enquetes €pide-
 miologiques orientees;
 9.    tout  en reconnaissant qu'il vaut  mieux prevenir que guSrir,
 et  qu'il vaut mieux eviter la pollution que la combattre, il
 n'empeche  qu'il peut etre precieux de  chercher sur le plan
 biologique et physiologique des parades et des remedes  a cer-
 taines contaminations d'autant  plus que certaines   d'oriaine
 naturelle, sont inevitables.
      En  terminant  cet expose,  je voudrais insister au nom de
 mes  collogues  et au  nom des  etres que nous voulons protSger
 sur  1'importance de  1'information centripete d'une part,  et sur
 la constitution d'une banque de  donnSes  d'autre  part.   Trop de
 donnees  disponibles  au niveau  de 1'Industrie et  de 1'agriculture
 ou au niveau de la medecine  professionnelle ou preventive,  pour
 des  raisons diverses ne sont pas communiquees aux  responsables
 des  etudes epidemiologiques.   Par ailleurs,  le medecin praticien,
 peut etre,parce qu'il n'est  pas  suffisamment prepare,  neeliee  de
 transmettre des informations qui seraient  souvent  precieuses;
 son  attention  devrait etre particulierement portee sur 1'obser-
 vation des groupes a haut risque et sur  1'interet  du "monitoring"
 biologique.  Souhaitons  que  cet  appel suffise et qu'une rgponse
 bienveillante  permette  d'eviter  d'aboutir  a  des  mesures r^gle-
 mentaires pour obtenir  ces diverses informations.

      Corre'lativement,  la constitution d'une  banque de  donn§es
 toxicologiqu.es Par collecte aleatoire des  coincidences suspectes
 nous  paralt indispensable, allant de  pair  avec la  surveillance
 reguliere d'echantillons de population representatifs  ou  cor-
 respondant a des groupes particulierement  sensibles.   Une telle
 banque de donnees  serait precieuse pour aider  1'O.M.S.  dans  ses
 efforts et constituerait un fondement precieux pour  les enquetes
 epidemiologiques que nous suggerons.

     Souhaitons que  les efforts entrepris de part  et d'autre
de 1 Atlantique se conjugent et s'amplifient dans  ce sens et que
cette realisation soit une des concretisations de  ce symposium
qui n a pu aboutir  que par une collaboration intense des trois
organismes interesses a ce symposium et grace au dynamisme
sympathique des organisateurs.

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                               2444
   CONCLUSIONS AND REFLECTIONS OF THE SCIENTIFIC ADVISERS
                        Presented by
                     Prof. A. LAFONTAINE
                           Belgium         (translation)
      In relation  to  the  protection  of man  and  the  environment,
 criteria may be defined  as  judgements based  on scientific  data,
 reflecting  a relationship  (expressed as  far  as possible  numeri-
 cally) between the exposure of  a  target  to an  agressive  factor
 and  the resulting risk and/or unfavourable or  undesirable  effect
 on this target; the  establishment of criteria  is the  first of
 the  steps which,  in  cartesian logic, should  result initially in
 the  setting of quality objectives,  followed  by the establishment
 of standards, rules  and  codes of  good practice.

      Since  man's  health, individually and  collectively,  is un-
 questionably the  most important issue at stake, it was logical
 that  the three organizations responsible for the symposium
 should be concerned  above all to  collect existing  scientific
 knowledge in order to make  progress in establishing relation-
 ships between exposure to pollutants and direct and indirect
 effects on  health, to achieve the most rational possible asses-
 sment of the risks,  and,  at  a later stage,  to decide on action
 programmes  to prevent or reduce pollution  or limit its effects.

     Experience having unfortunately demonstrated  the often frag-
mentary nature of information at hand and  the  scant use that
 could be made from many experimental studies and certain epidem-
 iological studies, the sponsors were fully aware of the diffi-
 culties which would  arise,  all  the more  so as  there is a risk
 of certain  less relevant factors becoming mixed up  with the
 strictly scientific  aspects.  They also  knew that  a great deal
of research had been carried out for other reasons  than the
desire to establish  criteria.  Therefore,  in addition to the new
data which might be collected, it was their  intention:

 a)   to provide for better planning more specific motivation,
     and more efficient and strictly objective orientation of
      research work}

b)   to discover the nature and size of  the gaps in present
     knowledge;

c)   to identify as far as possible the fields in which At is
     necessary to act,  in view of the urgency or priority nature
     of the problems, and those for which existing data,  even
     if they are still  incomplete, suffice for action to be
     taken.

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                              2A45
     Despite all the difficulties, despite the uneven and at
times disparate nature of the papers presented, the scientific
advisers nevertheless consider the result as highly positive,
in view of the diversity of the objectives envisaged.
     Mew data have come to light concerning, inter alia, urban
atmospheric pollution, the dangers of heavy metals, acousitc
nuisances, experimental techniques, indicators of exposure,
biological measurements and new experimental approaches
to behavioural phenomena.  New problems have been tackled, such
as those caused by biphenyl polychlorides or vinyl chloride.
Even greater contributions were made, however, in the matter of
the scientific data required for decision-making in health pro-
tection and the identification of fundamental gaps in our know-
ledge, the future direction of research work and the fields to
be explored in order to lay the necessary scientific foundation
for reflection and decision-making.

     We would, however, be lacking in objectivity if, before
embarking on various reflections and suggestions, we did not
draw attention to certain weaknesses in the documents submitted,
such as the one-sided obsession with certain subjects like
heavy metals or certain sectors such as atmospheric pollutants.
These could probably result from scientific enthusiasm, political
emphasis or public pressure.  On the other hand, little attention
was paid to other subjects such as nitro derivatives, nitrosamines,
mycotoxins or manganese.  Despite their topical interest, certain
sectors like solid residues and soil contamination did not pre-
occupy the authors, and today the toxicology of water intended
for human consumption might to the unaware seem like a problem
of the past.  Other sectors such as medicinal products, food
additives and certain harmful effects such as those of  ionizing  and
non-ionizing radiation were not considered during this symposium,
chiefly on account of a now traditional corapartmentalization,
which ought to be sacrificed to a more global approach to the
problems, since the target of these various nuisances is the
same, namely, man and the human race.

     However, although the scientific advisers regret the some-
times excessive tendency of environmental problems to be spread
out over a considerable number of organizations, they would also
like to draw attention to the cooperation which has come into
being between the three organizations responsible for the
symposium and to thank, them for the exceptional opportunity
given to research workers from Europe, America and, thanks to the
WHO, the whole world, to meet and seek to establish health
criteria - criteria uninfluenced by any socio-economical or
political consideration - common to all men in this "one world"
of ours, and work together in the most fraternal and cooperative
spirit.

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                              2446
     Among the many reflections inspired by the discussion held
during the symposium, we believe that apart from the implemen-
tation of action programmes concerning the list submitted by
the WHO of substances for which information must be collected
and research conducted, the following particular points deserve
to be stressed:

1.   It is of the utmost importance to express oneself clearly
and the need for4a glossary defining terms used in environmental
studies was felt yet once again.

2.   It too often happens that each pollutant and each sector
are considered independently and it is forgotten that man acts
as an integrator of nuisances; the same toxic agent may reach
him in different ways and various degenerative agents, including
those linked to certain habits (use of tobacco for example), may
affect the same target organ.  The risks linked with chemical
and physical pollutants must not make us overlook biological
and microbiological risks.  The idea of body burden, of total
incorporation/ must become general, as must the idea of allow-
able doses per day, week, month or year, depending on the case,

     Furthermore, to the concept of concentrations in the atmo-
sphere, in water or in the soil, one must add that of the toxi-
cological capacity of a medium, a region or a basin(especially
for non-degradable substances), that is the total amount of
polluting substances which this medium, region or basin can
tolerate without irreparable damage.

3.   The discussions stressed the need for methodological
developments, especially with regard to epidemiology, the use
of animals models pre-testing and the balance -sheet of adven-
tages and disadvantages:

     a)   epidemiologj^cal techniques are of fundamental impor-
tance , not only to assess the direct or indirect influence
of pollution on health, but also to identify new dangers or harm-
ful influences and to appreciate the effectiveness of the actions
undertaken.  Epidemiologists should be able to rely more exten-
sively on information supplied by an informed public and by a
body of informed practising doctors: general clinical practice
and workplace observations can be of cardinal importance, but
do require some preparation of the medical profession.  The
implementation of multinational epidemiological programmes may
be very helpful in assessing the concomitant effect of other
factors, such as ambient conditions or geographical and climatic
influences, etc.

     b)   development of the use of animals for simulating human
diseases in toxicological tests is indispensable for a better	'
understanding of the effects observed and to ensure maximum
effectiveness when pre-testing new products iind new technologies
These tests must be devised in such a way as to remain closely
related to the epidemiological studies;

-------
                              2447
     c)   the problem of pre-testing must be of major concern
to scientists;  humanity may be exposed to unforeseeable risks
unless new chemical products, new processes and new sources of
energy are subject, before being used on a large scale, to the
most comprehensive possible assessment of the nature and size
of the short-, medium- and long-term risks, including the search
for any mutagenic, carcinogenic and teratogenic effects and the
drawing up of a balance sheet of the advantages and disadvantages
for man.  A methodological system must be devised which would
attempt to forecast the various toxicoligical and ecological
risks as accurately as possible, but would also foresee the
criteria of purity applicable, the fate of the substances in-
volved in the environment, and methods of detecting them  (the
product in its pure state, as well as any impurities and meta-
bolites) in the various constituents of the environment;

     d)    a balance sheet of the advantages and disadvantages
for health of the products and techniques employed is essential;
however, present approaches are often inadequate in both con-
ception and technique, and the matter must be reconsidered on a
multidisciplinary basis.  In the same way, it is important to
assess more accurately the cost of pollution and pollution con-
trol as well as the effectiveness of control methods and their
possible secondary effects on health.  This knowledge will be
used, together with other socio-economical and regional elements,
in the process of making decisions starting from criteria and
aiming at quality objectives and standards.

     In this connection, particular attention should be paid to
the problems of developing countries thus, the need for develop-
ment might take precedence over the need for protection.

4.   Alongside clear or more-or-less easily identifiable
symptoms, changes regarded as infra-pathological or difficult
to interpret, like mitochondrial enzyme stimulation or modifi-
cations of delta-aminolevulinic acid dehydratase in persons ex-
posed to lead, are often observed.  Research is needed to enable
us to interpret these changes more accurately and to know whether
they are just concomitants or subtle impairments preceding out-
right pathological symptoms:  it would also be useful to know
whether these changes may be used as indicators of exposure.

5.   In connection with the preceding point, the following
problems arise:
     a)   the choice of critical groups to be taken into con-
sideration from the health point of view, and of the extent of
any risks which might reasonably be accepted for these groups;

     b)   the choice of priorities in scientific research, based
in particular orTknowledge of the nature of the risk, its extent
and reversibility, and of the possibilities for action, both for
natural pollutants and those originating in human activity.

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                              24A8
6.   Similarly, we must consider in a different light risks
observed in normal conditions and accidental risks; the accep-
table limits for the latter must be derived from criteria
applied to acute exposure and appropriate counter measures must
be in proportion to the extent of the danger and the number of
individuals threatened.

7.   Regarding the values currently accepted as standards there
is no doubt that some of them must be revised in the light of
present knowledge» this is particularly the case for certain
maximum allowable concentrations at work places.  Certain long-
term effects have* not been taken sufficiently into consideration,
perhaps because not enough attention has been paid to the fact
that occupational exposure is increased by non-occupational ex-
posure, and above all because it has sometimes been over looked
that more and more women capable of child-bearing  are working
in industry:  certain values must unquestionably be rethought
and account must be taken of transplacental and transmammary
transfers and of the protection of foetuses and breast-fed
children.
     We gave a definition of the term "criterion" at the begin-
ning of this paper.  Before considering certain fundamental
studies the significance of which was stressed during the dis-
cussions, we should like to draw our colleagues1 attention to
three points which seem important to use:

1.   Firstly, some exposures effects like odours or certain kinds
of noises tend to take the form of annoyances; although it is
not possible to calculate these exposures nor to quantify their
effects.  A judgement on criteria could nevertheless be made,
enabling the decision making process to be set in motion.  This
does not mean that technical and methodological efforts should
not be made to evolve numerical data as soon as possible.

2.   It is also necessary to stress that in certain cases de-
cisions must be taken regarding the setting of standards or
reference values, or the introduction of regulations, before
the criteria can be finally established; in such cases it is
necessary to collect as much scientific data as possible, and
these may even be incomplete or analogical, as long as they are
clear enough to support the decisions made.  Such data must be
confirmed experimentally or epidemiclogically as quickly as
possible.

3.   Finally, there is a need to pursue experimental and clinical
research, for example a more effective evaluation of the psycho-
logical effects of certain nuisances and stresses of modern
living, and a better knowledge of the psychological effects of
certain nuisances (equivalent to the biological effects of
noise)

-------
                              2449
     Among the fundamental back-up research which this symposium
has shown to be necessary, we would lay particular emphasis on
the following points:

1.   the study of agressive factors, whether of natural origin
or derived from human activity, must be extended/ with respect
to the manner in which they act and their interactions in the
environment and in the targets;

2.   greater attention must be paid to the effects on genetic
potential, on the foetal organism (even if they are very slow-
acting and do not appear until adult age) and on transplacental
and transmammary transfers;

3.   the metabolic transformations of xenobiotic substances in
the environment, throughout the biological cycles and at the
level of the human organism have not always been sufficiently
taken into consideration, and experimental research into these
is essential;

4.   research must also be undertaken into the question of whether
a threshold for carcinogenic effects does or does not exist:
the existence of repair mechanisms, proven by molecular biology,
demands that this concept be considered a fresh,

5.   furthermore, have not adaptation or compensation mechanisms
other than natural selection appeared with regard to certain
agressive factors during the course of human history?  These
should be identified, analysed and appraised;

6.   while excluding ecological criteria from our current preoc-
cupations, it is nevertheless important for us to identify and
assess the indirect effects on human health of-adverse ecological
influences affecting, for example, certain plants essential to
human survival, or thermal pollution, with its repercussions on
the environment and, amongst other things, on microbiological
and parasite life;

7.   current monitoring techniques are sometimes cumbersome;
they must be better adapted to the numerical evaluation of ex-
posure and new and if possible polyvalent indicators of pollution
in the environment and in man must be sought in order to facili-
tate and increase the effectiveness of epidemiological surveys;

8.   among long-term effects, the phenomenon of acceleration of
physical and mental ageing has not been sufficiently taken into
consideration, nor have, perhaps, certain unforeseen repercus-
sions on ontogeny; a fundamental approach is desirable as well
as guided epidemiological surveys;

9.   although prevention is admittedly better than cure, and it
is better to avoid than to combat pollution, nevertheless it
may be extremely profitable to search for countermeasures and
remedies at the biological and physiological level to certain
kinds of contamination, all the more so since some of them,
being of natural origin, are inevitable.

-------
                               2450
     In conclusion, I would like to stress, on befalf of my
colleagues and those whom we wish to protect, the Importance of
centripetal information flow, on the one hand, and of setting
up a data bank on the other hand.  Too many data, available in
the fields of industry or agriculture or of preventive of oc-
cupational medicine for various reasons fail to reach those
responsible for epidemiological surveys.  Moreover, the prac-
tising doctor, perhaps because he is not sufficiently prepared,
neglects to communicate information which could often be in-
valuable; his attention should be drawn in particular to the
observation of high-risk groups and to the usefulness of bio-
locical monitoring.  Let us hope, that this appeal will suffice
and that a favourable response will make it unnecessary to lay
down rules for the transmission of this information.

     Similarly, the setting up of a toxicological data bank by
the random sampling of suspicious coincidences seem essential
to us, and should go hand-in-hand with regular surveillance of
population samples, either representative or corresponding to
particularly sensitive groups.  A data bank of this type would
be of immense value to WHO and would constitute a vital
foundation for the epidemiological surveys which we are proposing,

     Let us hope that the efforts made on both sides of the
Atlantic will be united and concentrated in this direction and
that this will prove to be one of the concrete achievements of
this symposium, which was made possible only thanks to the very
close collaboration of the three organizations concerned and the
unflagging energy of the organizers.

-------
                               2451
                        CLOSING  SPEECH
                      DR,  B.H. DIETERICH
           Director  Division  of  Environmental  Health
                  World Health  Organization
     In the few minutes available for my statement, I want to
tell you how we of WHO appraise and will be able to make use of
the material presented during this symposium and of the many
interesting discussions that took place.  We have been pleased
to co-sponsor this symposium for two particular reasons, namely:
     1) To help provide the widest possible forum where scien-
tists from all countries could freely exchange their experiences,
and
     2) To demonstrate how a national regulatory agency, an
international health agency and a regional economic community
can approach problems of environmental health together.

     I will present my reviews as replies to two questions:
1)  How will we be able to use the specific information presented?
     We have obtained a great deal of specific information on
methodology.  Most important in this connection is perhaps the
information presented on the design of population studies tak-
ing into account the need for a multi-factorial approach.  We
are interested in promoting methodology which will enable scien-
tists in various countries to work together, and your papers
have provided examples as to how this can be done.

     We have learned on the one hand that some of the work done
and some of the methods applied did not always produce results
which could be related to the specific problems posed in environ-
mental health, but on the other hand that you have made use of
rational models in planning your scientific studies.  This ex-
perience will contribute to 6ur efforts to develop and promote
agreement on methods for international comparative analysis of
environmental health effects.

     Still on methodology, interesting proposals have been made
for the selection of meaningful parameters both for environmental
conditions and for measuring human response.  I believe that the
symposium helped to promote agreement amongst scientists in
respect of prediction models which might be applied in future
scientific work and this too is of great interest to us.

-------
                               2452
      There is no doubt that the symposium has confirmed the
value of both toxicological and clinical and epidemiological
methods as well as the limitations inherent in both of them.
I believe that we will have to accept that the planning and
implementation of meaningful population studies is a problem in
itself, and that it would be false to expect immediate results
which would help the legislator and administrator within just a
few months in their respective tasks  of setting and enforcing
environmental standards.  However, this should not discourage
us, and I believe that we must continue to give a major role to
human studies and we feel encouraged by the result of this sym-
posium.

     In this connection, we are grateful for the suggestions
made to improve the monitoring of individual and population ex-
posure and response for the elaboration of more meaningful stat-
istical methods of analysis, for the suggestions which will help
to plan and implement work in comparative toxicology and for the
study of mechanisms of action and inter-action.

     While the lessons learned with respect to methodology are
important, I believe that the WHO Environmental Health Criteria
Programme will greatly benefit from the data presented by you.
This programme depends on research throughout the world, and I
can state that the information presented during this week will
broaden the scientific basis of this programme, particularly as
regards the criteria documents which WHO has under preparation,
such as for oxides of nitrogen, cadmium, lead, mercury, oxides
of sulfur and resulting compounds, ozone and other oxidant and
suspended particulate matter.  The criteria documents on these
substances are at various stages of development and in all the
cases mentioned, I am sure that the specific data presented here
will be of great help.  This also applies to the papers on the
effects of PCBs, and those on other persistent substances, fib-
rous dusts and the interaction studies on which some of you have
been working.  I should like to thank all of you for making this
material available to us.

     The results of this week's deliberations are also of spe-
cific interest in the promotion and planning of an increasing
number of specific international case studies which we like to
see organized wherever suitable occasions arise.  Not only have
we been able to learn more about a great number of new studies,
but we were able to appreciate the fields of scientific interest
and the scientific capacity of many people assembled here, with
whom we were not acquainted in the past.   I believe that this
will help us in expanding contacts with the scientific community
in making proposals for new programmes and priorities and in
organizing more, and more meaningful, international comparative
research.

-------
                               2453
2)   Did the symposium validate the current approach to the
     assessment of environmental health factors^and <3*d it
     identify new directions which we should follow in the
     future?

     While I would like to make a few remarks in reply to this
question, I should first  reiterate that we believe in (i) making
scientific information an essential basis of administrative
decisions at the national or regional level, and (ii) that ad-
ministrative decisions are needed in the countries not only for
the formulation and enforcement of national standards for environ-
mental quality, but also for the allocation of more funds to
environmental health research on a continuous basis.

     Although I heard some pessimism, we have been reinforced
in our belief that the assessment of environmental health effects
of pollution and the presentation of information on dose/effect
relationships in the foim of international criteria documents
are valid approaches and thai: they can be effective decision-
making tools for use by legislators and administrators.  In this
connection, I would like to refer to a resolution adopted by the
World Health Assembly in May 1974, in which WHO was requested to
strengthen its efforts in the establishment and the promotion of
international agreement on environmental health criteria.

     Nevertheless, we acknowledge that for some time to come we
will not he entirely successful in attempting to make regulatory
action a" fully rational process.  Perhaps we must learn how best
to support but not to replace and on a temporary basis, the
scientific approach by prudent pragmatism in the planning and
implementation of regulatory control.  Although not too many
suggestions have been made in this respect during the symposium,
I believe that what has been said will help us to qtrenghten
our contribution towards making health protection the ultimate
objective of environmental protection.

     Sure enough, our symposium did not make, nor was it intended
to make, suggestions which would allow the application of prin-
ciples of systems analysis to the planning of health oriented
programmes of environmental pollution control.  However, it
provided a deeper insight into the environmental origin of some
common chronic diseases, and it brought out the need to face
the task of scientifically studying what might be the socially
accepted risk of such diseases and the comparison of such risks
with benefits derived from modern ways of life.

     I believe we all should accept this challenge and undertake
research in this respect.  Perhaps this research will also pro-
vide further insight into the age-old question of the responsib-
ility of the scientist who engages in creating new knowledge,
which he must expect to lead to new hazards to public health.

     I am not personally in favour of closing a meeting of this
nature with a call for more support to research, but it appears

-------
                               2454
 that  stronger  financial and moral support  is needed  for environ-
 mental health  effects research.  One wonders whether some re-
 allocation  is  not  called  for, keeping  in mind  the environmental
 origin of many of  the chronic diseases  for which massive research
 programmes  are now under  way.

      The symposium also showed  that we  can still improve com-
munication among us and particularly with the younger scientific
 generation,  of whom we were pleased to  meet so many  during  the
 symposium.   We must also  do more to explain science and scien-
 tific findings more effectively to the  general public  and to
 the decision makers.  In  planning future research we should
 accept the  challenge expressed*  by one  speaker  during the early
 part  of the symposium that apparently  everybody does what every-
 body  else does, but that  nobody does what  nobody else  thinks
 about.  Without making a  case for exaggerated  futurology, we
 have  learned during this  week that we  must strengthen  our efforts
 towards early  identification of potentially new hazards to
 public health.  This must become a continuing  activity involving
 international  collaboration, and I am  sure that it will not only
 influence the  planning of future research, but also  the priorities
 which we have  been following in the health assessment  of the
 effects of  environmental  pollution.

      Finally, on behalf of WHO, I would  like to  thank  all those
 who have made  this symposium what I consider a great success.
 This  includes, first and  foremost, those who have prepared  the
 papers and  have come here to present them, but also those who have
 spoken during  the  discussions.   It includes our Secretary
 General, the Organizing Committee, the Scientific Secretariat,
 the Scientific Advisors,  all of the conference staff,  the
 interpreters and all those who  have contributed to our work but
 have  not been  visible during the sessions.  I  would  like to
 thank Unesco for making such beautiful facilities available to
 us and  I would like to state that I personally consider it  a
 great success  and  an encouragement for the future, that a nat-
 ional regulatory agency,  a regional economic community and  an
 international  health agency have been  able to  organize a sym-
 posium  in  the  spirit of mutual  understanding and in  pursuit of
 a common objective, namely to create an environment  in which
 man can  live and work and enpoy himself.

-------
                              2455
                        CLOSING   SPEECH
                       A,   C,  TRAKOWSKI
    Assistant Administrator for Research and Development
        United States Environmental Protection Agency
     I wish to express  the gratitude of the United States En-
vironmental Protection  Agency (EPA)  for the privilege of co-
sponsoring and working  with the  World Health Organization and
the Commission of the European Communities to organize and pro-
duce this stimulating scientific meeting.   It has been a great
effort by all concerned,  from which we found a welcome and mem-
orable diversion last Tuesday evening.  The reception given by
the Commission of the European Communities was a unique event at
a unique place that will  long remain a treasure in our memories,
and we wish to most sincerely thank the Commission for its grac-
iousness, its generosity  and its hospitality.  Also, we wish to
extend deep appreciation  to UNESCO for their provision of these
facilities and staff support, without which this meeting could
not have been as successful.

     Hundreds of people have met here for five days to present
and to learn of advancements in  knowledge of the health effects
of environmental pollution, and we have  given plenary consideration  to
the problems of 'the use of scientific information in decisions
to protect human health.   Out of these presentations has come
useful information in many disciplines and in many problem areas
too diverse to be given comparative judgment or individual ass-
essment here.  However, in the totality of these presentations,
I believe it is possible  to observe certain trends and concepts
in the pursuit of scientific knowledge on health effects caused
by pollution and its application to pollution control.  I would,
therefore, like to put  forth our observations in this regard,
for whatever use they may be given as guides to further advance-
ment toward the knowledge we are seeking.

     First, we wish to  note and  to compliment the many contri-
butors at this symposium on the  interdisciplinary breadth and
communication in their  work.  Indeed, we have seen during this
week that the inter-disciplinary approach to investigations of
health effects can succeed where single discipline studies
have failed to reach desired research goals.  We believe the
advantages are apparent in dealing with the problems of health
effects from pollutants,  and we strongly urge intensification
of the interdisciplinary approach.

     Observation of the character of the progress reported at
this symposium and of the methods by which this progress has

-------
                               2456
been achieved suggests that, at the present time, there are lim-
itations to the scientific basis for pollution control.  Scien-
tific methods are valuable for describing natural phenomena both
qualitatively and quantitatively.  In a time when man has been
transported with exquisite precision from earth to the moon and
back, it is difficult to believe that there are any problems that
do not yield to scientific inquiry, or any decision  not subject
to rational, scientific information.

     But, man's many-sided relationship to his environment must
not be underestimated.  Man is a complex and often inscrutable
creature.  He has created an intricate societal fabric that is
now global in extent.  This society, dependent on a precarious
ecosphere, has become increasingly capable of exerting major
influences on that ecosphere.  The diverse problems already in
our view appear to exceed our capability to solve them in a
scientifically rigorous manner.  We may reasonably question our
abilities to solve even more complex environmental problems not
yet suspected.

     We probably possess the conceptual framework and techniques
with which to develop solutions to these problems, given suffici-
ent time and resources.  But, faced with the urgency of self-
evident needs for environmental control, we do not have the
luxury of unlimited time.  Additionally, research on environ-
mental needs must compete with other strong requirements for
limited resources.  The interactions between environmental con-
cerns and the complex, non-linear, social, political, economic,
health and welfare aspects of human society must be pursued with
imagination and unbiased dedication by scientists who have pre-
viously devoted themselves to more restricted fields and more
classical methods.  While they continue to direct their efforts
toward the pursuit of scientifically substantiable information,
they must also seek publicly acceptable methods for applying
available, though inadequate, information to the decision-making
processes which depend also on economic, political and geogra-
phical considerations.

     We recognize that application of the results of scientific
inquiry on the health effects of pollution may be limited in its
usefulness at the present time, but we know no other way to
acquire the needed information.  What then, can we do, that the
results of this symposium suggest, to improve our direction and
techniques in our scientific pursuits.

     We have seen increasing evidence to indicate that many
environmental pollutants probably act differently in combination
than they act separately.  We know that urban environmental
pollutants are rarely, if ever, present alone.  Should we not,
then, in order to devise an adequate control program, assess the
effects associated with pollutants according to the way pollutants
are actually found.

-------
                               2457
     It is possible that this kind of information may require
future standards for combinations of pollutants.  It would re-
quire methods to measure the constituents of the important com-
binations of pollutants simultaneously.  It would require field
networks to simultaneously monitor the constituents of important
combinations, source inventories of all constituents of the
combinations, models linking sources of all constituents of the
combinations to ambient levels, joint control plan strategies
for all constituents of the combinations, and adequate enforce-
ment plans.

     We may find that scientific information to support regulatory
strategies for control of combinations of pollutants will be
many times more difficult to obtain than information for single
pollutants.  Indeed, we may be well advised to begin now to plan
systematically for the collection and analysis of such data.

     If the harmful combinations of pollutants are known to
exist in certain regions, perhaps we can use the level of a
single pollutant of the combination as an index of the level of
the combination.  However, the accuracy of this approach will
have to be experimentally evaluated.


     We have seen this week that epidemiological studies reflect
progressively greater degrees of sophistication concerning their
design and methodologies both for assessing the pollutant expos-
ures and identifying and quantifying health effects.  He believe
that the state-of-the-art has now been sufficiently developed
to the point where studies involving common pollutants can and
should use standardized methodologies concerning both the phys-
ical/chemical monitoring and the biological effects assessment.
Such an approach will allow the development of standardized data
bases utilizing the multiplicity of studies conducted throughout
the world.  It will, therefore, expedite our attainment of know-
ledge and allow for the more efficient use of our seperate lim-
ited research resources.  Some effort of this nature is already
underway.  For example, the cooperative epidemiology studies
presented at this meeting being conducted by the CEC and the
WHO.  in the area of environmental monitoring, the WHO air pol-
lution monitoring program is using regional and many other inte-
national air monitoring sites where there has been cooperative
agreement on standardized sampling, analytical, and data manag-
ement systems.  EPA, serving as the WHO International Reference
Center forAir Pollution Control, is intimately involved in this
program as well as with the WHO Air Quality Criteria Program.
I am sure other examples could also be cited.  Such cooperative
efforts involving standardized protocols should be expanded on
both the national and international level.

     At the present time, most effort toward standardization
have concerned monitoring programs, health questionnaires, and
to a more limited degree, pulmonary function measurements.  These

-------
                               2458
should be continued and improved.  For example/ similar efforts
should be applied to many other biological parameters being used
as health indicators/ as well as to biological sampling and
analysis and the statistical treatment and presentation of the
basic data.  The degree of success achievable by any of these
efforts will depend totally upon the inclusion of an adequate
quality control program.  Standardized methodology with quality
control ean benefit all of our criteria and standard development
activities.

     Of course/ this recommendation is only intended for applied
research and not meant to disregard the obvious need for contin-
uing efforts in the development of new, or the improvement of
existing research methodologies.  Results from this area of more
basic research should, after being proved effective in the field,
be incorporated into cooperatively developed standardized prot-
ocols.

     The magnitude and difficulty of research needed to provide
the necessary information base for pollution control tells us
that we cannot afford the time or resources given to investig-
ations, that do not contribute directly to the needed information
base.  As our research becomes more difficult and our regulations,
and the society to which they apply, become more complex, it is
necessary for the scientific community to assess its status and
systematically determine its future direction.  To do otherwise/
that is to say, pursue our investigations separately according
to separate interests and techniques and without coordination
is not only wasteful of time and resources, but promises to
yield results that lack practical usefulness because of basic
differences in objectives and methods.

     A systems analysis approach to the identification and
solution of future problems could assure application of research
effort to the most critical needs.  It would also assure the
integration of the necessary interdisciplinary contributions to
each study.  It would tend to eliminate project activity of
doubtful significance and the collection of questionable data.

     One practical way to internationally unify the direction
of research is to internationalize the preparation of criteria
documents which would include the assessment of current infor-
mation and identification of gaps in knowledge which would
serve as needs to be fulfilled through research.  A practical
way to this achievement would be for all member countries to
participate in the WHO Air Quality Criteria Program.

     I would like to make a comment on the future of meetings
such as this.  This symposium provided a complete overview of
the advances to date of research on the health effects of
environmental pollutants.  It was necessarily  large and compre-
hensive.   It has served its purpose well. We now know where the

-------
                              2459
state-of-knowledge of this subject stands, and it has suggested
to us concepts by which we can guide future efforts.  It would
seem that because this symposium has so well accomplished its
purpose, consideration of future meetings in this subject area
might be in terms of specific pollutants, or classes or combin-
ations of pollutants, and their effects, or in terms of
areas of pollutant control, for example, pollutants associated
with particular industries or sources.

     Again, I wish to express deep thanks to all the organizers
and the participants in this highly succussful symposium.  It is
only through such extensive and enthusiastic cooperative part-
icipation that we can have full confidence in the results and
ideas that have come forth from this monumental effort.

-------
                              2461
                      REMARQUES FINALES
                        DR,  P,  RECHT
            Directeur de la  Protection Sanitaire,
           Commission  des Communautes europeennes
                  Luxembourg (Grand-Duch€)
     A I1issue d'un Symposium, harassant pour les organisateurs
et les participants en raison de la densite du sujet, du rythine
des sessions, du nombre des communications et de I1intensity des
discussions, il est certain que nous avons tous un grand nombre
de reflexions 3 presenter.  Ceux qui m'ont pr€c€d£ ont exprimS
de £a9on excellente les reactions et les commentaires gue meritait
une telle reunion.

      Parlant le dernier, il m'est facile de m'associer au nom
de la Commission des Communautes europeennes aux declarations
qui viennent d'etre faites par mes deux collSgues et amis de
1'Organisation Mondiale de la Sante et de 1'Agence de la Pro-
tection de 1'Environnement des Etats-Unis.  Je souscris entifire-
ment a ce qu'ils ont dit et je me permets d'ajouter que nous
pouvons nous feliciter d'avoir trouv6 dans la mise sur pied de
cette conference la clef d'une collaboration tripartie  efficace
et benfifique pour 1'avenir.

     Ce Symposium prfisentait des risques qui sont apparus au
moment oil le Comite organisateur et les conseilliere scientifiques
ont examine un nombre impressionnant de rapports et de communi-
cations et ont dQ faire un choix dans les propositions qui ont
ete envoyees.

     II est indeniable qu'il y a dans les communications que
vous avez entendues pendant cinq jours une dispersion qui indique
que les problemes de 1'environnement ont des aspects multiples
et complexes et qua des lors il 6tait indispensable de tenter la
recherche d'une methodologie commune pour les affronter.

     Malgre le caractSre positif des resultats qui ont et6 ap-
portes, nous sommes forces de reconnalisre que sur un trSs grand
nombre de sujets, il n'y a pas encore de rfiponse satisfaisante
du point du vue scientifique.  Cette remarque ne doit pas Stre
consideree comme un reproche a 1'encontre de la recherche scien-
tifique dans le domaine de 1'environnement.  C'est une constata-
tion qui doit nous conduire, cher Monsieur Dietrich, a demander
a 1'O.M.S. d'intensifier encore son action et de jouer un role
indispensable dans 1'etablissement de critSres et de normes
sanitaires communes.  II est temps que dans le domaine de 1*en-
vironnement, les problemes de la protection de 1'homme et de son
milieu prennent leur place veritable et soient trait6s par ceux
qui en ont la competence et la responsabilite.

-------
                               2462
     Nous  avons trouvfi dans  la presence de Mme Veil, Ministre
 frangais de  la Sante, un t£moignage dont  le congres peut etre
 particuliSrement fier et heureux.  Je voudrais sur ce point par-
 ticulier,  au nom du Symposium, remercier  le Gouvernement fran£ais
 de nous avoir deiSguS pour prSsider la seance d'ouverture la
 personnalite officielle qui  par vocation  et par  fonction a pour
 tache de proteger  la sante humaine.  Si,  comme j'ai eu  1'occasion
 de le dire lors de la table  ronde, il faut appliquer a  1'environ-
 nement les concepts et les mfethodes de la m€decine, cela ne
 signifie pas pour  autant que 1'environnement soit essentiellement
 un problSme  medical.  Les sciences de 1'environnement ressortis-
 sent a de  nombreuses disciplines qui cherchent actuellement &
 s'entendre et a se concerter, dans un souci d'economie  et d'ef-
 ficacite.                            »

     A cet 6gard,  le  Symposium est une rSussite  car avec la
 participation de 800 dSlSgu^s, reprSsentant aussi bien  les scien-
 ces fondamentales  que la medecine clinique, la toxicologie et
 1'ecologie,  nous avons pu jeter les bases d'un accord et degager
 des objectifs communs, malgre certaines lacunes  et imperfections
 que Monsieur Lafontaine vient de nous presenter.

     Lors  de la conference de presse de ce matin, nous 'avons eu
 1*occasion de dire que jamais la science  ne s'est trouv€e devant
 une telle  responsabilite et  que jamais le rdle moral qu'elle
 doit jouer n'a ete aussi important et lourd de consequences pour
 1'avenir de  1'homme.

     II n'est pas  possible d'etudier les  probldmes qui  sont
 pose's en oubliant  les buts auxquels nous  devons  tendre  et qui
 doivent nous servir de point d'accrochage pour nos entreprlses
 et nos reflexions.  Certes,  nous devons accepter de concSder a
 la technique la part  suffisantfi qui lui revient, en vue de la
 developper et d'en harraoniser les performances,  mais nous devons
 orienter le  dSveloppement de la technologic dans la mesure ou
 elle est indispensable SL rfealiser 1'objectif gereral de qualite
 de vie que nous nous sonunes  assign^s.

     La prise de conscience  de I1importance de 1'environnement
 est r€cente  et dans certains cas elle a 6t€ dramatique.  La
 revelation du nombre de problSmes SL rfisoudre, la quantite con-
 siderable  de mat;iSres a etudier et qui se sont entassfies ces
 dernifires  annfies devant les  chercheurs et les responsables
 politiques sont tels qu'un certain dfisarroi et un certain disor-
 dre  sont  apparus  dans les milieux qui veulent promouvoir le
 bonheur de 1'humanite et ame'liorer la qualite de la vie.

     Je crois que  le Symposium qui vient  de se terminer a permis
 d'opSrer une certaine classification dans les problemes pos6s,
 d'etablir  des priorites et surtout de donner leur veritable
 dimension  a  des preoccupations dont le monde entier est conscient
 en soulignant les questions  urgentes et en de'dramatisant des     '
problemes mal poses.

-------
                               2463
     Nous pouvons nous demander guel est 1'enjeu de la lutte
contre les pollutions.  Get enjeu est immense mais il est cer-
tain qu'il d£passe le cadre d'une generation et gue parmi les
grands problemes a rSsoudre il y a une meilleure connaissance
des effets lointains des pollutions, y compris les effets herSdi-
taires, alors qua, sauf pour les radiations ionisantes/ la plu-
part des elements polluants du milieu ne sont £tudi£s que dans
les perspectives d'une action potentielle de quelques annees.

     II y a dans 1'extraordinaire aventure collective od nous
nous engageons, deux objectifs qui paraissent contradictoires.
II y a d'une part la promotion de la qualitfi de la vie indivi-
duelle, a laquelle nous sommes tous personnellament attaches et
d'autre part, le deVeloppement d'un type de soci6t6 particuliS-
rement efficace mais qui met en danger 1'individu et ses aspi-
rations profondes.  Cette contradiction entre 1'efficacitfi de
la societ^ et la recherche du bonheur individuel est dgroutante
et doit nous conduire a nous poser une serie de questions et a
tenter de les rSsoudre.

     De tels probldmes ne sont pas uniquement du ressort de la
philosophic.  Nous devons en tant qu'homme
ministrateurs de la santfi prendre conscience de la complexity
des points d'interrogation et leur appliquer la mSthode scien-
tifique dont ce congres a demontrfi la valeur et 1'irremplacable
mission de conseil et d'inspiration politique.

     Le hasard a voulu que notre Symposium se tienne a Paris,
dans ce pays oQ sont nes la mfithode expSrimentale et le raison-
nement scientifique, et ce n'est pas sans motif que 1'un des
participants a fait allusion Si Claude Bernard et a 1'oeuvre qui
1'a immortalise.  Mais il faut que nous soyons honnetes avec
nous-memes et que nous n'attribuons pas a la science plus qu'elle
ne peut et ne doit accepter.  Si son rdle est d'aider, en ana-
lysant et en objectivant les ph£nomenes scientifiques, les
autorites qui doivent prendre des decisions, elle ne peut se
substituer a ces dernieres pour transferor sur le plan de la
rSglementation ou de la politique les rfisultats des constatations
scientifiques.

     La conference de presse que nous avons tenue ce matin et
qui a etfi1suivie avec beaucoup d'int£r§t par un grand nombre
de journalistes, indique que la presse entend jouer le rSle
qui lui revient dans 1'information objective du public et dans
la mobilisation de 1'opinion en fa^veur de la defense et de la
protection de 1'environnement.

     II est particulierement favorable que nous ayons pu, au
cours de ces cinq journ6es, en toute objectivitfi et en toute
franchise,  avec les portes largement ouvertes vers 1'ext£rieur,
presenter un bilan positif souvent et negatif parfois,de 1'fitat
de nos connaissances, et indiquer les grandes lignes et les
orientations d'une action en faveur de 1'environnement et de
la sante humaine.

-------
                              2464
     Ce Symposium a suscitS une esperance pour 1'avenir de nos
socifitfis humaines mais il a aussi prouvfi que les chercheurs et
les responsahles de la sant6 et de 1'environnement poss6dent
I'ind^pendance et la libertfi d1esprit qui sont toujours les
conditions essentielles au progrSs de 1'humanitg.

-------
                              2465
                     CONCLUDING REMARKS (translation)

                        DR.  P,  RECHT
               Director for  Health Protection,
           Commission of the European Communities
                  Luxembourg (Grand Duchy)
     Having reached the end of this symposium, a strenuous one
for organizers and participants., alike in view of the wide-ranging
subject matter, the tight schedule of sessions, the large number
of reports and papers and the intensity of the discussions, we
all no  doubt have many comments to make.  Those who have already
spoken have summed up admirably the reactions and comments which
this gathering has certainly evoked.

     As the last speaker, I should like, on behalf of the
Commission of the European Communities, to endorse the opinions
which have just been expressed by my two colleagues and friends
of the World Health Organization and of the United States Envi-
ronmental Protection Agency.  I agree wholeheartedly with their
views and should like to add that in organizing this conference
we have laid the foundation for an effective tripartite collab-
oration which will certainly prove its worth in the future.

     The difficulties inherent in this symposium became apparent
as soon as the organizing committee and the scientific advisers
began to sift the large number of reports and papers submitted,
with a view to selecting the most appropriate.

     By their very diversity the papers presented over the past
five days have clearly illustrated the many-sided and complex
character of environmental problems, and thus the need to de-
velop a common methodological approach to tackle them.

     Despite the positive results obtained, it is certain that
scientifically valid solutions have yet to be achieved in very
many fields.  This remark is not intended as a criticism of
scientific research in the environmental field, but as an encour-
agement,dear Dr. Dietrich, to ask W.H.O. to intensify its activ-
ities still further, and to play a vital role in the establishment
of common health criteria and standards.  It is time that the
protection of man and his living environment was given its proper
place in environmental studies, and be tackled by those competent
and responsible for doing so.

     We have been particularly honoured and gratified by the
presence here of Mrs Veil, the French Minister of Health.  On
behalf of all those who have attended this symposium I should
like to thank the French Government for having delegated Mrs Veil,

-------
                              2466
who both professionally and in her official capacity has the
task of protecting human health, to preside over our opening
session. Although, as I had occasion to say during the round-
table discussion, medical concepts and methods must be applied
to the environment, this does not mean that the environment is
essentially a medical problem.  Environmental sciences embrace
a wide range of disciplines, which are at present endeavouring
to work together in closer harmony in the interests of economy
and efficiency.

     In this respect the symposium has been a success because,
with 8OO delegates representing not only the fundamental sci-
ences but also clinical medicine, toxicology and ecology, we
have been able to find common ground»and establish common ob-
jectives, despite the gaps and imperfections to which Dr. Lafon-
taine has just referred.

     At this morning's press conference we said that science had
never before had to carry such a great responsibility, and that
its moral role had never been &o important and fraught with con-
sequences for man's future.  When examining the problems facing
us we must always keep clearly in mind the goals we are aiding
at, for it is these which must guide our thinking and our action.
Technology must, of course, be allowed its proper role, and
must be developed and its results harmonized, but its develop-
ment must depend on the degree to which it can help us achieve
our overall goal of improving the quality of life.

     The general awareness of the crucial importance of the
environment is recent, and in some bases has come dramatically.
The realization of the enormous number of problems to be solved
and fields to be studied by research workers and politicians
has caused a certain amount of confusion and disarray among those
who aim to promote human happiness and improve the quality of
life.

     I feel that the symposium which has just ended has enabled
us to some extent to sort out the problems facing us, to estab-
lish priorities and above all to put in their proper perspective
the problems which are troubling the world, emphasizing the
urgent ones and stripping the less important ones of their sen-
sationalist aspects.

     We must think about what is at stake in our struggle against
pollution.  There is, of course, and immense amount at stake,
but it is certainly not confined to one generation: one of the
major  problems confronting us is how to improve our knowledge
of the long-term., and hereditary effects of pollution, since,
except in the case of ionizing radiation, most pollutants have
been studied solely with a view to their possible effects over
periods of a few years.

-------
                              2467
     In the extraordinary adventure in which we are all engaged
there are two apparently contradictory objectives.  On the one
hand we wish to improve the individual's quality of life, - this
concerns us all personally - and on the other we seek to develop
a highly efficient society, which threatens the individual and
his most profound aspirations.  This contradiction between the
efficiency of society and the search for individual happiness
is disturbing, and should cause us to ask ourselves a number of
questions, and try to answer them.

     Such questions are not confined to the realms of philosophy.
As scientists and health administrators we must be aware of the
complexity of the problems facing us and apply scientific methods
in solving them.  This symposium has shown these methods to be
of irreplacable value as sources of guidance and political in-
spiration.

     Chance has ordined that our symposium should be held in
Paris, in this country which is-the home of experimental methods
and scientific reasoning, and it is significant  that one part-
icipant alluded to Claude Bernard and the work which immortalized
him.  But we should be honest with ourselves and not ask of
science what it cannot, indeed may not, give.  While those who
have to make decisions should be assisted by the objective  ana-
lysis of scientific phenomena, the results of scientific  obser-
vations must not become the all-important factors in legislative
and political decisions.

     This morning's press conference, which aroused much interest
and which was attended by a large number of journalists, indi-
cated that the press intends to fulfill its role of providing
objective information and of educating public opinion in favour
of environmental protection.

     We have been particularly fortunate over the past five days
in being able, frankly and with complete objectivity, and with
our doors wide open to the outside world, to discuss the freq-
uently positive and sometimes negative aspects of our knowledge
and to outline the programmes which will have to be introduced
to protect the environment and human health.

     This symposium has given us fresh hope in the future of
our societies but it has also proved that research workers and
those responsible for health and the environment possess the
independence and freedom of mind which are always the essential
prerequisite for human progress.

-------
    TEILNEHMERLISTE



 LIST OF PARTICIPANTS



LISTE DES PARTICIPANTS



LISTA DEI PARTECIPANTI



LUST VAN DE DEELNEMERS

-------
                              2471
                           ARGENTINA
BENINSON D.
  Gerencia de Proteccion
  Radiologica y Seguridad
  Buenos Aires 29
                           AUSTRALIA
ALLAN A.
  Mount ISA Mines Ltd.
  -Mount ISA
  Queensland   4825

JOHNSON H. M.
  Broken Hill Associated Smelters
  Box 219, Port Pirie
  South Australia   5540
MICHAUX E,
  Health Service
  21 Rokeby Terrace
  Taringa   4068
  B r i sbane, Queensland

SHEPHERD W.
  C.S.I.R.O.
  Division of Atmospheric
  Physics
  Station Street
  Aspendale, Victoria
                       BELGIE / BELGIQUE
BORMANS M. J. J.
  Fe'de'ration des Industries
  Chimiques, ECOCHEM
  49 Square Marie Louise
  1040 Bruxelles

BOUQUIAUX J. J.
  Institut d'HygiSne et
  d'Epidfimiologie
  14 rue Juliette Wytsman
  1050 Bruxelles

BRUAUX P.
  Institut d1Hygiene et
  d'EpidSmiologie
  14 rue Juliette Wytsman
  1050 Bruxelles

BUCKET J.-P.
  Unite de Toxicologie Indus-
  trielle et Mgdicale
  University Catholique de Louvain
  Avenue Chapelle aux Champs 4
  1200 Bruxelles
COFFIELD T. H.
  Ethyl S.A.
  1 rue Paul Lauters
  1050 Bruxelles
DAMS R.
  Instituut voor Nucleaire
  Wetenschappen
  Rijksuniversiteit Gent
  Proeftuinstraat 86
  9000 Gent

DEMARET Y.
  Groupement International
  des Associations Nationales
  de Fabricants de Pesticides
  28 Square Marie Louise
  1040 Bruxelles

DE MEULEMEESTER A.
  Texaco Services (Europe) Ltd,
  31 Pres. J. F. Kennedylaan
  9020 Gent

-------
                              2472
DEMUYNCK M. R. R.
  Instituut voor Nucleaire
  Wetenschappen
  Rijksuniversiteit Gent
  Proeftuinstraat 86
  9000 Gent

DRUMAUX P.
  Minist&re SantS Publique
  Administration Hygiene
  Service des Nuisances
  CitS Administrative de 1'Etat
  1010 Bruxelles

DUHAMEAU W.
  Institut National des Indus-
  tries Extractives
  rue de Che"ra 200
  4000 Liege

FAGNIART E. A.
  DSpartement Radiobiologie
  CEN/SCK
  2400 Mol

GRANDJEAN J. F. F.
  Institut Royal M6t6orologique
  de Belgique
  Avenue Circulaire 3
  1180 Bruxelles

HAWKES A.
  Ethyl S.A.
  1 rue Paul Lauters
  1050 Bruxelles

HERMAN P.
  Institut de Recherches Chimiques
  Ministere de 1'Agriculture
  5 Moslenstraat
  1980 Tervuren

IMPENS R.
  Faculte"  Sciences Agronomiques
  de  1'Etat
  5800 Gembloux

RUMMER J.
  Universite" Libre de" Bruxelles
  100 rue  Belliard
  1040 Bruxelles

KUSTERS E. M. V.
  Medische Dienst
  BASF Antwerpen NV
  Eikenlei 15
  2130 Brasschaat
LAFONTAINE A.
  Institut d1Hygiene et
  d'EpidSmiologie
  14  rue Juliette Wytsman
  1050 Bruxelles

LAUWERYS R.
  Unit§ de Toxicologie Indus-
  trielle et M6dicale
  UniversitS Catholique de
  Louvaln
  Avenue Chappelle aux Champs 4
  1200 Bruxelles

LEFEVRE M. J.
  Solvay et Co. - DCP
  33 rue Prince Albert
  1050 Bruxelles

LEGRAND M.
  Institut d'Hygiene et
  d'EpidSmiologie
  14 rue Juliette Wytsman
  1050 Bruxelles

MERCIER M.
  Universitfi de Louvain
  Sentier du Biereau 13
  1348 Louvain-la-Neuve

NEURAY M.
  Institut National des Indus-
  tries Extractives
  rue de Ch6ra 200
  4000 Liege
OVERSTEYNS M.
  MetallurgiS Hoboken Overpelt
  Fabrieksstraat 144
  3583 Overpelt

ROELS H. A. C.
  UnitS de Toxicologie Indus-
  trielle et Mfidicale
  Universitfi Catholique de
  Louvain
  4 Avenue Chapelie aux Champs
  1200 Bruxelles

RONDIA D. J, J.
  Laboratoire de Toxicologie
  Boulevard de la Constitution
  4000 Liege

-------
                                2473
  SPAAS  J,
   Coraitfi  de  Liaison  des  Indus-
   tries de MStaux  non  Ferreux
   de la CommunautS Europe'enne
   boulevard  de  Berlaimont  12
   1000 Bruxelles
  TITE M.
   D6pt.  de  Toxicologie
   R.U.G.  Belgique
   Hospitaalstraat  13
   9000 Gent
 STASSEN J.-J.
   UniversitS de LiSge
   rue des Augustins 49
   4000 Liege
 SWYNGEDOUW Y.
   Institut d1Hygiene et
   d'Epidgmiologie
   14 rue Juliette Wytsman
   1050 Bruxelles
 VAN MEIRHAEGHE A.
   Dienst Openbare Gezondheid
   Stad Gent
   23 Stationstraat
   9000 Gent

 VUYLSTEEK K.
   Institut d1Hygiene et de
   M6decine Sociale
   Institut d1Hygiene R.U.G.
   Akad. Z iekenhuis
   be Pintelaan 135
   9000 Gent
                   BUNDESREPUBLIK DEUTSCHLAND
ANGERER J.
   Zentralinstitut  fur
   Arbeitsmedlzin
   2 Hamburg 76
   Postfach 56 45

ARSOVIC H. M.
   Emissions Technisches
   Institut
   757 Baden-Baden
   Beutigweg 1

BECK E. G.
   Hygiene-Institut
   Justus-Liebig Universitfit
   63 Giessen
   Friedrichstr. 16

BEHNE D.
   Hahn-Meitner-Institut fUr
   Kernforschung Berlin GmbH
   Bereich Kernchemie und
   Reaktor
   1 Berlin 39
  Glienicker Strasse 100

BEITZ L.
  Siemens A.G,
  Bereich Mess- und Prozess-
  technik
  75 Karlsruhe
  Rheinbruckenstr.  50
BERGE H.
   Institut fur Umweltschutz
   u. Agrikulturchemie
   5628 Heiligenhaus
   Bez. Dusseldorf
   Am Vogelsang

BRAETTER P.
   Hahn-Meitner Institut fur
   Kernforschung Berlin GmbH
   1000 Berlin 39
   Glienicker Strasse 100

BRETSCHNEIDER W.
   Deutsche Forschungsgeraein-
   schaft
   53 Bonn-Bad Godesberg 1
   Kennedya1lee 40

BRyCH J.
  Med. Institut fttr Lufthygiene
  und Silikoseforschung an
  der Universit&t Dttsseldorf
   4000 Dttsseldorf
  Gurlittstr. 53

DEIMEL H.
   Institut fur Lebensmittel-,
  Wasser- u.  Luftuntersuchungen
  der Stadt K6ln
  5 Koln 1
  Eifelwall 7

-------
                                2474
 Von DEPKA J.
   Rheinische  Olefinwerke GmbH
   5047  Wesseling
   Postfach 31

 DIEHL J.  F.
   Institut fur  Strahlen-
   technologie der Bundes-
   forschungsanstalt fur
   Lebensmittelfirschhaltung
   75 Karlsruhe  1
   Engesserstr.  20

 ELLISSEN  W.
   Chemie-Mineralien K.G,
   28 Bremen
   Postfach 1804
 FISCHER A. B.
   Med.  Institut  fur Lufthygiene
   und Silikoseforschung  an
   der Universitat Dusseldorf
   4000  Diisseldorf
   Gurlittstr. 53

 FOERSTER  H. J.
   Daimler-Benz A.G.  Abt.E  6
   7000  Stuttgart-Unterturkheim
   Postfach 202
 FRIEDRICHS K.-H.
  Med.  Institut fur Lufthygiene
  und Silikoseforschung an
  der Universitat Dusseldorf
  4000 Dusseldorf
  Gurlittstr.  53

 GOERKE W.
  Bundesministerium des
   Innern
   53 Bonn
  Rheindorferstr. 198

 GRAUL E. H.
  MEDICEF
  355 Marburg/Lahn
  Kornerstr. 19

GREFEN K.
  Verein Deutscher Ingenieure
  Kommission Reinhaltung der
  Luft
  4000 Dusseldorf
  Graf-Recke-Str. 84
 GROMZIG H.
   Stadtverwaltung Duisburg
   Gesundheitsamt
   41 Duisburg 1
   Landfermannstr. 1
 GRUNSPAN M.
   Med.  Institut fur Lufthygiene
   und Silikoseforschung an
   der Universitat Dusseldorf
   4000  Dusseldorf
   Gurlittstr.  53

 GULLEMANN D.
   Universitat Bielefeld
   48 Bielefeld
   Postfach 8640

 HALBRITTER G.
   Institut fur Angewandtle
   Systemtechnik und Reaktor-
   physik
   Kernforschungs zentrum
   75 Karlsruhe
   Postfach 3640

 HARKE H.-P.
   Forschungsinstitut der
   Cigarettenindustire e.V.
   Gazellenkamp 38
   2  Hamburg 54

 ROLL K.
   Bundesministerium fur Jugend,
   Familie und  Gesundheit
   53  Bonn-Bad  Godesberg 1
   Deutschherrenstr.  87

 HOWER J.
   Universitatskinderklinik
   43 Essen
   Hufelandstr.  55

 JANSEN G.
  Universitatsklinikum
   43 Essen
  Hufelandstr.  55

KNAUP M.
  Arbeitskollektiv Zwei
   35 Kassel
  Kirchweg 77

-------
                              2475
KNAUTH P.
  Institut fiir Arbeits-
  physiologie an der
  Universitat Dortmund
  Ardeystr. 67
  Dortmund

KOPETZ K.
  Chemische Werke Hiils A.G.
  437  Marl Kreis Recklinghausen
  Postfach 1180

KREUZER W.
  Bereich  fur Hygiene und
  Technologic der Lebens-
  mittel Tierischen Ursprungs
  Universitat Miinchen
  8 Miinchen 22
  Veterinarstr. 13

LANDHAUSSER 0A
  Rheinische Olefinwerke GmbH
  5047 Wesseling
  Postfach 31

LEHMANN K.
  Deutscher Normenausschuss
  1 Berlin 30
  4-7  Burggrafenstrasse

LORKE  D.
  Institut fiir Toxikologie
  der  Bayer A.G.
  56 Wuppertal 1
  Fried. Ebertstr. 217

LOSER  E.
  Institut ftir Toxikologie
  der  Bayer A.G.
  56 Wuppertal 1
  Fried. Ebertstr. 217

MAINTZ E.
  Hamburgische Electricitats-
  Werke A.G.
  2000 Hamburg 60
  Uberseering 12

MATHEIS U.
  Institut fiir Angewandte
  Systemtechnik und
  Reaktorphys ik
  Kernforschungszentrum
  75 Karlsruhe
  Postfach 3640
MENKE J,
  Stadtverweltung Miilheim
  433 Mulheim-Ruhr
  Amt 53, Ruhrstrasse

OLTERSDORF U.
  Institut fiir Ernahrungs-
  wissenschaft
  6300 Geissen
  Wilhelmstr. 20

PIETRZOK W.
  Arbeitskollektiv Zwei
  35 Kassel
  Kirchweg 77

POETZL K.
  Institut fiir Atmospharisc'.e
  Umweltforschung
  8 Miinchen 90
  Harthauserstr. 109

PRENZEL J.
  Staatliches Gesundheitsamt
  fiir den Landkreis Wesermarsch
  2880 Brake/Utw.
  RBnnelstr. 10

PREUSSMANN R.
  Deutsches Krebsforschungs-
  zentrum
  Institut fiir Toxikologie
  und Chemotherapie
  69 Heidelberg
  Kirschnerstr.  6

PRINZ B.
  Landesanstalt  fiir Iramissions-
  und Bodennutzungsschutz des
  Landes NW
  43 Essen - Bredeney
  Wallneyer-Str. 6

RAEDE D.
  Bayer A.G.
  4150 Krefeld
  Rheinuferstr.  7-9

RAHLENBECK U.
  Fachdokumentation Reinhaltung
  der Luft, VDI
  4 Diisseldorf  1
  Graf-Recke-Str.  84

-------
                              2476
REINHARDT J.
  NiedersSchsisches Landes-
  verwaltungsaiot - Dezernat
  Arbeitsmedizin, Imraissions-
  und Strahlenschutz
  3 Hannover
  Bertastr. 4

RETHFELD H.
  Landwirtschaftliche Unter-
  suchungs- und Forschungs-
  anstalt
  Joseph-KSnig-Institut
  4 Miinster (Westf.)
  von Esmarch-Str. 12

RICHTER E.
  Institut fur Pharraakologle,
  Toxikologie und Pharmazie
  der TierSrztl. Fakultat
  8000 Munchen 22
  VeterinSrstr. 13

ROSSBACH H.
  Geschaftsstelle des Rates
  von Sachverstandigen fur
  Uraweltfragen
  62 Wiesbaden
  Statistlsches Bundesamt
  Gustav-Stresemann-Alice 11

RUDOLF W.
  Bundesstelle fur Umwelt-
  angelegenheiten
  6000 Frankfurt/M
  Feldbergstr. 45

RtJDEN H.
  Hygiene-Institut der
  UniversitSt
  53 Bonn 1
  Venusberg, KlinikgelSnde 35

SCHLIPKOTER H. W.
  Med. Institut fur Lufthygiene
  und Silikoseforschung an
  der Universitat Diisseldorf
  4000 Diisseldorf
  Gurlittstr. 53

SCHMIDT E.
  Bundesgesundheitsamt
  1 Berlin 33
  Postfach
SCHMIDT K, G.
  Institut fur exp, Toxikologie
  und Chemotherapie
  69 Heidelberg
  Kirschnerstr. 6

SCHOLZ G.
  Stadtverwaltung Mulheim
  433 Mulheim / Ruhr
  Rathaus

SEYBOLD I.
  BASF Farben + Fasern AG
  Bereich Siegle Stuttgart
  7000 Stuttgart 30
  Postfach 30 06 20

STEINMANN D.
  Ministerium fur Wirtschaft
  Mittel»stand und Verkehr
  des Landes NW
  4 Diisseldorf
  Haroldstr. 4

STOEPPLER M.
  Zentralinstitut ftir
  Analytische Chemie der
  Kernforschungsanlage
  Julich GmbH
  Postfach 365
  517 Julich

SZADKOWSKI D.
  Zentralinstitut ftir Arbeits-
  medizin
  2 Hamburg 76
  Adolph-Schonfelderstr. 5

THIEL W. R.
  Verein Deutscher Ingenieure
  Koramission Reinhaltung der
  Luft
  4 Diisseldorf 1
  Postfach 1139

VALENTA P.
  Zentralinstitut fiir
  Analytische Chemie der
  Kernforschungsanlage
  517 Julich
  Postfach 365

-------
                               2477
WEIGAND W.
  Farbwerke Hoechst A.G,
  Pharma Forschung Toxikologie
  6230 Frankfurt/M. 80

WITTENZELLNER R.
  Gesellschaft fur Strahlen-
  und Uraweltforschung mbH
  8042 Neuherberg
  Post Oberschleissheim
  Ingolstadter Landstrasse 1

WOLFERMANN H.-F.
  Max-Planck-Instltut fttr
  Landarbeit und Landtechnik
  655 Bad Kreuznach
  Am Kauzenberg
WOSING-U,
  Bundesgesundheitsamt
  1 Berlin 33
  Postfach

WUNDERER A.
  Bayer A.G.
  Abt. Zentrale Anwendungs-
  technik
  Leverkusen - Bayerwerk
                            CANADA
BATES D. V.
  University of British
  Columbia
  Dean's Office
  Faculty of Medicine
  Vancouver, B.C.

BERNSTEIN A.D.
  Medical Services Branch
  Health and Welfare Canada
  255 Argyle Avenue
  Ottawa, Ontario  K1A OL3

BUTLER G. C.
  Division of Biological
  Sciences
  National Research Council
  of Canada
  Ottawa, Ontario  K1A OR6

DENNIS C. A. R.
  Prairie Institute of Envi-
  ronmental Health
  2220 Lome Street
  Regina, Saskatchewan

FINKELSTEIN B,
  St. Joseph's Hospital
  49 Dow Avenue
  Hamilton, Ontario
GIBBS G. W.
  Department of Epidemiology
  and Health
  McGill University
  3775 University Street
  Montreal, Quebec   H3A 2B4

HAZUCHA M.
  McGill University
  Department of Physiology
  3655 Drummond Street, No 1116
  Montreal, Quebec

LAO R. C.
  Air Pollution Control
  Directorate
  Environmental Health Centre
  Tunney's Pasture
  Ottawa, Ontario   K1A OH3


LESAGE M.
  Association des Mines
  d'Amiante
  5 place Ville Marie
  Chambre 412
  Montreal, Quebec

LEVY D.
  812 King Street East
  Hamilton, Ontario

-------
                                2478
 LINDELL K.
   Institute of Occupational
   and Environmental Health
   R.R.4., Danville
   Quebec   JOA 1AO
 LISTON A. J.
   Environmental Health
   Directorate
   Department of National
   Health and Welfare
   Ottawa, Ontario   K1A OL2

 LORENZI L.
   588 Barton Street E.
   Hamilton, Ontario

 MONKMAN J. L.
   Air Pollution Control
   Directorate
   Environmental Health Centre
   Tunney's Pasture
   Ottawa, Ontario   K1A OH3

 NEWHOUSE M.
   McMaster University
   St. Joseph's Hospital
   Hamilton, Ontario

 OLOFFS P. C.
   Department of Biological
   Sciences
   Simon Fraser University
   Burnaby 2, B.C.
PICHE L.
  University de Montreal
  DSpartement de Chimie
  Montreal 101, Quebec H3C 3V1

SMITH D. M.
  Gouvernement du Canada
  MinistSre de la Sant6 et
  du Bien-Etre Social
  Services Internationaux
  d1Hygiene
  Ottawa, Ontario   K1A OK9

STOPPS G. J.
  Ministry of Health
  Province of Ontario
  15 Overlea Boulevard
  Toronto, Ontario

WIEBE A.
  Atmospheric Environment
  Service
  Department of Environment
  4905 Dufferin Street
  Downsview, Ontario  M3H 5T4

ZERBE H.
  640 Boulevard Graham
  Ville Mont Royal
  Quebec
                       CITTA' DEL VATICANO
 DUBARBE D.
   Nonciature Apostolique
   72 Avenue de Tervuren
   Bruxelles, Belgique
SATRA J.
  Nonciature Apostolique
  72 Avenue de Tervuren
  Bruxelles, Belgique
                           Ci SiS
PELECH L.
  Institute of Postgraduate
  Training for Physicians
  and Pharmacists
  Bratislava
PFEIFER I.
  Czechoslovak Research and
  Development Centre for
  Environmental Pollution
  Control
  100 42 Praha 10
  Srobafova 48

-------
                               2479
ROSIVAL L.
  Research  Institute of
  Hygiene
  Bratislava
  Ul.Cs.armady 40

SCHMIDT P.
  Institute of Hygiene and
  Epidemiology
  Centre of General and
  Environmental Hygiene
  100  42 Praba 10
  Srobafova 48
STEPANEK V.
  Czechoslovak Research and
  Development Centre for
  Environmental Pollution
  Control
  120 00 Praha 2

SYMON K.
  Institute of Hygiene and
  Epidemiology
  Centre of General and
  Environmental Hygiene
  100 42 Praha 10
  Srobafova 48
                            DANMARK
CARSTENSEN J.
  National Food Institute
  Institute of Toxicology
  Mtfrkhtfjgaard
  19 Mtfrkhtfj Bygade
  2860 Stfborg

FAKSTORP J.
  F. L. Smidth & Co. A/S
  Environmental Technology
  Division
  Gl. Koege Landevej 55
  2500 Valby

FLYGER H.
  Danish Atomic Energy
  Commission
  Research Establishment
  Rlstf
  4000 Roskilde

FUNCH-ENGBERG A,
  National Food Institute
  Institute of Toxicology
   19 MiZirkheSj  Bvaacte
   2860  Stfborg
HANSEN J. C.
  Institute of Hygiene
  University of Aarhus
  Universitetsparken
  8000 Aarhus C

MAGE D.
  Laboratoriet for Varme-
  og Klimateknik
  Danmarks Tekniske H^skole
  Bygning 402
  2800 Lyngby

STEENSBERG J.
  Ministry of the Environment
  Agency of Environmental
  Protection
  47 Store Kongensgade
  1264 Copenhagen K

UHL E.
  Sundhedsstyrelsen
  St. Kongensgade 1
  ?.264 Copenhagen K
                  DEUTSCHE DEMOKRATISCHE REPUBLIK
 HORN K.
   Ministerium fur Gesundheitswesen
   102 Berlin
   Rathausstr. 3

-------
                              2480
                             EGYPT
ATTIA M.
  Ministry of Health
  Cairo

EL-SHEBOKSHY A.
  Ministry of Health
  Cairo
FARAG S, A.
  Air Pollution Unit
  National Research Center
  Dokki, Cairo

MASSOUD A.
  Department of Industrial
  Medicine
  Cairo University
  Faculty of Medicine
  14 Gezira Street
  Zamalek Cairo
                            FRANCE
ALARY R. S. E.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

ALQUIER J. P.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

ALVINERIE R. M.
  Institut National de la
  Recherche Agronomique
  180 chemin de Tournefeuille
  31300 Toulouse

ANDREUX J.-P.
  Hopital Boucicaut
  78 rue de la Convention
  75015 Paris

ARSAC F.
  FacultS de Phannacie
  Laboratoire de Toxicologie
  rue J. B. Clement
  92290 Chatenay-Malabry

AUBERT M.
  C.E.R.B.O.M.-I.N.S.E.R.M.
  Pare de la Cote
  1 avenue Jean Lorrain
  06300 Nice
AUDRAN R.
  Institut National de
  Recherche et de Sficuritfi
  30 rue Olivier Noyer
  75680 Paris Cedex 14

BEAU P.
  Commissariat 3. 1'Energie
  Atomique
  Bolte Postale 6
  92260 Fontenay-aux-Roses
BENARIE M.
  I • R.C .H.A•
  Bolte Postale 1
  91710 Vert-le-Petit

BENET M.
  Soct£t£ Languedocienne de
  Soufre et Micron-Couleurs
  n • lr • %•• • O • A •
  Bolte Postale 79
  11102 Narbonne

BERTIN M.
  Electricite-Gaz de France
  30 avenue Wagram
  75008 Paris

BIGNON J.
  Clinique de Pneumo-
  Phtisiologie
  Hopital Laennec
  42 rue de Sevres
  75007 Paris

-------
                              2481
 BITTEL R.
   Commissariat  SL  1'Energie
   Atomique
   Departement de  Protection
   Bolte Postale 6
   92260 Fontenay-aux-Roses
BLAKE M.
  Centre Universitaire de
  Savoie
  Laboratoire de  Zoologie
  et Biologie
  Bolte Postale 143
  73011 Chamb^ry

BLANCHER G. L. G.
  Faculte" de MSdecine de
  Paris  (Quest)
  1 rue de l'Universit6
  75007 Paris

BOISVIEUX
  Socl§t6 Antar
  4 rue LSon Jost
  75017 Paris

BONNEFOUS
  Association National de
  1'Environnement
  18 rue Duphot
  Paris

BORDET F.
  Elf-Union
  12 rue Jean Micot
  75340 Paris Cedex 07

BOUDENE C..
  I.N.S.E.R.M.,  U.122
  Universitfi Paris-Sud
  rue Jean Baptiste Clement
  92290 Chatenay-Malabry

BOURDEIX J.
  74 Cours Lafayette
  69003 Lyon

BOURGEOIS R.
  La Presse Francaise
  64 rue de Rome
  75008 Paris

BOVARD P.
  Commissariat a  1'Energie
  Atomique
  D^partement de Protection
  Bolte Postale 6
  92260 Fontenay-aux-Roses
 BOYD S.
   Sierra Club
   24 avenue Charles  Floquet
   75007  Paris

 BRESSION H,
   Prefecture de Paris
   Laboratoire d'HygiSne
   1  bis  rue  des HospitaliSres
   St. Gervais
   75004  Paris
 BRESSON G.
   Commissariat a 1'Energie
   Atomique
   DSpartement de Protection
   Bolte Postale 6
   92260 Fontenay-aux-Roses

 BRILLE  D.
   Institut National de  la
   Santg et de la Recherche
   Mfidicale
   184 rue du  Faubourg St.
   Antoine
   75012 Paris

 CABANE  E.
   Ciba-Geigy  S.A.
   2-4 rue Lionel Terray
   92505 Rueil Malmaise

 CABRIDENC R.
   Institut de Recherche
   Chimique AppliquSe
   Bolte Postale  1
   91000 Vert-le-Petit

 CAILLARD L.
   Socie'te' Rhone-Poulenc
   21 rue Jean Goujon
   75008 Paris

 CASANOVA H. P.
   I.N.R.A.
   Route de St. Cyr
   78000 Versailles

 CHAMBON P.
   Laboratoire de Toxicologie
   et Hygiene  Industrielle
   Faculte Mixte  de Mfidecine
   et Pharmacie
   8 avenue Rockefeller
   69008 Lyon

CHANSON
  Socifitfi Antar
  4 rue L6on Jost
  75017 Paris

-------
                              2482
CHANTEUR J.
  Ministere de la Sante Publigue
  et de la SecuritS Sociale
  78110 Le Vesinet
CHAPOUX E.
  Union Technique de  1'Auto-
  mobile, du Motocycle et du
  Cycle  (UTAC)
  157 rue Lecourbe
  75015 Paris

CHAUVEAU J.
  Institut de Recherches
  Scientifiques sur le
  Cancer
  Bolte Postale 8
  94800 Villejuif

CHEMINAT J. C.
  Hopital Sabourin
  Botte Postale 12
  63018 Clermont-Ferrand

CHESNAIS J. C.
  I.N.E.D., C.N.R.S.
  Bureau 316
  27 rue du Commandeur
  75675 Paris-Cedex 14

CHEVRIER M.
  Regie Nationale des
  Usines Renault
  Direction des Laboratoires
  Bolte Postale 103
  92109 Boulogne Billancourt

CHOVIN P.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

CICOLELLA A.
  I.N.R.S.
  route de NeufchSteau
  54500 Vandoeuvre"

CLAUDE J. R.
  Laboratoire de la
  Prefecture de Paris
  88 rue de la Jonquiere
  75017 Paris
COHEN M.
  HygiSne Industrielle
  Esso S.A.F.
  92080 Paris la Defense
  Cedex 2

COIN L.
  Bolte Postale 42
  94220 Charenton

CONSO F,
  Hopital Fernand Widal
  Clinique Toxicologique
  200 rue Fbg St, Denis
  75010 Paris

CORTIER J.
  Ciba-Geigy S.A.
  2-4 rue Lionel Terray
  92505 Rueil Malmaise

COSSON G.
  Pharmacie de 1'Hopital
  Broussais & Paris
  110 due Pierre Brossolette
  (P. Corby)
  92320 Chatillon

COULON
  Picheney Ugine Kuhlmann
  23 rue Balzac
  75008 Paris

COURGIBET G.
  R^gie Nationale des
  Usines Renault
  Direction des Laboratoires
  Bolte Postale 103
  92109 Boulogne Billancourt

COUTEAUX H.
  SocietS Nationale des
  Pfitroles d'Aquitaine
  26 avenue des Lilas
  64000 Pau

CUMONT G.
  MinistSre de I1Agriculture
  Direction des Services
  VSterinaires
  Laboratoire Central
  d'HygiSne Alimentaire
  43 rue de Dantzig
  75015 Paris

-------
                               2483
DANA V.
  Management Science Oper-
  ations Research
  36 rue Claude Decaen
  75012 Paris

DANIEL H.
  CERCHAR
  Bolte Postale 27
  60103 Creil

DAVOUST P.
  Institut Universitaire de
  Lorlent
  Dgpartement d1Hygiene et
  de SScuritfi
  rue de Londres
  56100 Lorient

DECLOITRE F.
  Institut de Recherches
  Scientifiques sur le Cancer
  Bolte Postale 8
  94800 Villejuif

DEIT N.
  Revue Technique Inter-
  nationale de 1'Eau
  9 rue Coetlogon
  75006 Paris

DELAMAIN J.
  MinistSre de 1'Environnement
  13 rue de Bourgogne
  75007 Paris

DE LAVOUR
  I.N.R.A.
  Laboratoire de Phyto-
  pharmacie
  C.N.R.A. route de St.Cyr
  78000 Versailles

DELPLA M.
  Electricity de France
  Comite1 de Radioprotection
  3 rue de Messine
  75008 Paris

DELSEY J.
  Institut de Recherche
  des Transports
  Centre d1Evaluation et de
  Recherche des Nuisances
  109 Chemin St. Jean
  69000 Bron
DESBORDES J.
  Pr. Centre Hospitaller
  76600 Le Havre

DETRIE J. P.
  Centre Interprofessionnel
  Technique d1Etudes de la
  Pollution Atmosph£rique
  28 rue de la Source
  75016 Paris

DUBOIS G. J.
  FacultS de M£decine
  3 Allge des Piquiers
  59700 Lille

DUCHESNE M. C.
  Ministere de 1'Agriculture
  Service de la Repression
  des fraudes et contrOle de
  la qualitfi
  42 bis rue de Bourgogne
  75007 Paris

DUPUIS P. J.
  39 Pare du Val d'Or
  92210 St. Cloud

EFTHYMIOU M. L.
  Hdpital Fernand Widal
  200 rue du Faubourg St Denis
  75475 Paris-Cedex 10

ESCOULA L. G.
  Institut National de la
  Recherche Agronomique
  180 chemin de Tournefeuille
  31 Saint Martin du Touch

FABRE M.
  Laboratoire d'Hygifine de
  la Ville de Paris
  1 bis rue des HospitaliSres
  St. Gervais
  75004 Paris

FARGES L. F. P.
  Commissariat & 1'Energie
  Atomique
  Bolte Postale 6
  92260 Fontenay-aux-Roses

FAUDEMAY F.
  I.N.S.E.R.M.
  78 avenue de Suffren
  75007 Paris

-------
                                2484
 FAUGERE J. G.
   Laboratoire Municipal
   rue du Professeur Vezes
   33000 Bordeaux

 FESTY B.
   Laboratoire d1Hygiene
   1 bis rue des HospitaliSres
   St. Gervais
   75004 Paris

 FONDIMARE A.  H. P.
   Lab.  d'Analyses Path.
   76600 Le Havre

 FONTVIEILLE D.  A. J.
   Centre Universitaire  de
   Savoie
   Laboratoire de Zoologie
   et  de Chimie
   Bolte Postale 143
   73011 ChambSry

 FOURNIER E.
   Hfipital  Fernand Widal
   200 rue  Fbg St. Denis
   75010  Paris

 FROC  J.
   Institut  National de
   Recherches Agronomiques
   C.N.R.A.  Rte  de St. Cyr
   78000 Versailles

 GALTIER P.  E.
   Institut  National de la
   Recherche Agronomique
   180 Chemin de  Tournefeuille
   31300  Toulouse

GARNIER
   SPS/PE
  Dfipartement de Protection
  Centre d1Etudes NuclSaires
   Bolte Postale  6
  92260 Fontenay-aux-Roses

GARREL P. M.
  SocietS de Mgdecine du
  Travail
  37 rue du Depart
  75014 Paris
 GAUDIN G.  J.  L.
   Secretariat General de la
   Defense  Nationale
   51 bd de Latour Maubourg
   75007 Paris

 GERIN-ROZE C. M.  M.
   Laboratoire d1Hygiene de
   la Ville de Paris
   1 bis rue des  Hospitalifires
   St. Gervais
   75004 Paris

 GILLES G.
   Laboratoire des Services
   VSterinaires
   66 rue St.  S§bastien
   13252 Marseille Cedex 2

 GILLON J.  J.
   Revue le Concours M6dical
   37 rue de Bellefosse
   75009 Paris

 GIRARD-WALLON C.
   Universitg  Paris  VI.  UER
   Broussais,  Hotel  Dieu
   185 avenue  Aristide  Briand
   94230 Cachan

 GODIN J.
   I.N.S.E.R.M., U.122
   Laboratoire  de  Toxicologie
   Facultg  de  Pharmacie
   rue J. B. Clement
   92290  Chatenay-Malabry

 GRADISKI D.
   I.N.R.S.
   route  de  Neufchateau
   54500 Vandoeuvre

 GRIFFATON G.
   Institut  National de  la
   SantS et  de  la Recherche
  Medlcale
   Laboratoire de Nutrition
   Humaine
   Hopital Bichat
   170 boulevard Ney
   75018 Paris

GRISONI S.
  Laboratoire d'HygiSne de la
  Ville de Paris
  1 bis rue des Hospitalieres
  St. Gervais
  75004 Paris

-------
                                2485
 GROS R.
   Commissariat a 1'Energies
   Atomique
   10 Allee Jacques Cartier
   91430  Igny

GUERIN M.
   Soci§t6  Frangaise de PStrole
   B.P.
   10  qua!  Paul Doumer
   92  Courbevoie

GUIBKY M. T.
  14 rue Juliette Lamer
  75017 Paris

HALPERN J.
  I.N.S.E.R.M.
  Centre de Documentation
  HOpital de Bicfitre
  94270 Le Kremlin-Bicfitre

HEDDE L. M.
  Societe Octel
  8 rue Bellini
  75782 Paris Cedex 16

HENRY G.
  Association Frangaise des
  Femmes MSdecins
  65 boulevard Garibaldi
  75015 Paris

HO M. T.
   I.N.R.S.
  route de Neufchateau
   54500 Vandoeuvre

HUBERT-HABART M.  A..
  Fondation Curie
   Institut de Radium
   26  rue d'Ulm
   75005 Paris

JARRY J. J.
   Institut National de
   Recherche et de  SficuritS
   30  rue Olivier Noyer
   75680 Paris Cedex 14

JEANMAIKE  L.
   Commissariat a 1'Energie
   Atomique
   DSpartement de Protection
   Bolte Postale  6
   92260 Fontenay-aux-Roses
JUGUET B. E. E. 0.
  Laboratoire d1Hygiene
  1 bis rue des Hospitalieres
  St. Gervais
  75004 Paris

KAUFFMANN F.
  I.N.S.E.R.M., U. 68
  H&pital Saint Antoine
  75571 Paris Cedex XII

LABONDE J.
  Laboratoire d1Hygiene
  1 bis rue des HospitaliSres
  St. Gervais
  75004 Paris

LAMBOTTE J.
  Ministere de 1'Environnement
  Direction de la Prevention
  des Pollutions et Nuisances
  13 rue de Bourgogne
  75007 Paris

LAREDO N.
  ComitS Anti-Pollution
  Boues Rouges
  66 boulevard Jean JaurSs
  94000 Fresnes

LECLERC F.
  Hopital Broussais
  96 rue Didot
  75014 Paris

LECLERC J.-L.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

LECLERC M.
  HOpital Broussais
  96 rue Didot
  75014 Paris

LECLERC P.  P. M.
  Service de Pneumologie
  Hotel Dieu
  160 rue Blomet
  75015 Paris

LE COADIC Y.
  Institut  Universitaire de
  Lorient
  D6partement  Hygiene et
  Security
  rue de Londres
  56100 Lorient

-------
                               2486
LEIDNER J.
  CES Ecologie Humaine
  UER BiomSdicale
  2 rue Saint ExupSry
  92700 Colombes

LEJEUNE X.
  Ciba-Geigy S.A.
  92505 Rueil Malmaison

LE MOAN R.
  University RenS Descartes
  170 boulevard Ney
  75018 Paris

LEYNAUD G.
  Division Qua1it6 des Eaux
  PSche et Pisciculture
  C.T.G.R.E.F.
  14 avenue de St. MandS
  75012 Paris

LIEVAIN 0.
  Service de Chirurgie
  Clinique du Pare Monceau
  21 rue de Chazelles
  75017 Paris

LOWY R.
  C.N.R.S.
  Laboratoire de Nutrition
  Humaine
  HQpital Bichat
  75018 Paris

LUQUET F. M.
  Laboratoire d*Industrie
  LaitiSre - E.N.S.I.A.
  13 rue de 1'University
  59000 Douai

MACLOUF C.
  Prefecture de Police
  14 rue Bremontier
  75017 Paris

MAGADUR J. L.
  I.N.R.S.
  route de Neufchateau
  54500 Vandoeuvre

MARIEN J.
  Laboratoire d1Hygiene
  1 bis rue des HospitaliSres
  St. Gervais
  75004 Paris
MEININGER J.
   1 boulevard de Vaugirard
   75751 Paris Cedex  15

MESNIL M.
   Roussel U.C.I.A.F.
   102 route de Noisy
   Romainville

MICHON G.
   C.E.A, Saclay  (C.I.R.E.A.)
   Bolte Postale 8
   91190 Gif sur Yvette

MOLLARET P.
   Institut de MSdecine et
   d'Epidfimiologie Africaine
   Hfipital Claude Bernard
   10 avenue Porte d'Aubervillief
   75019 Paris

MONCEAUX R. H.
   Revues MSdicales
   51 rue de la Procession
   75015 Paris

MONCELON B.
   I.N.R.S.
   route de Neufchateau
   54500 Vandoeuvre

MOREAU M.
  University de Bretagne
  Occidentale
  Facultfi des Sciences
   6 av. Victor Le Gorgeu
   29283 Brest Cedex

MORICHAU-BEAUCHANT J.
   Institut d'Etude du
  Dfiveloppement Economique
  et Sociale
  5 rue des Colonnes-du-Trdne
  75012 Paris

MOUTET J.-P.
  I.N.S.E.R.M.
  44 chemin de Ronde
  78000 le V£sinet

MOUVIER G. J.
  University de Paris 7
  Laboratoire de Physico-
  Chimie Instrutnentale
  2 place Jussieu
  75221 Paris  Cedex 05

-------
                              2487
MOYEN E.
  Ministere de la QualitS
  de la Vie
  14 bd du G6n6ral Leclerc
  99000 Neuilly/Seine

MUEL B.
  Fondation Curie
  Institut du Radium
  26 rue d'Ulm
  75005 Paris

NIEF G.
  Commissariat a 1'Energie
  Atomique
  Centre d'Energie Nucleaire
  de Saclay
  Bolte Postale 2
  Jif-sur-Yvette

OPPENEAU J. C.
  Ministere de la QualitS
  de la Vie
  14 bd du General Leclerc
  92000 Neuilly/Seine

ORIOL P.
  I.N.S.E.R.M., U. 68
  HSpital Saint Antoine
  75571 Paris Cedex 12

OSTROWSKI Z.-L.
  C.I.E.
  3 Square du Rhfine
  75017 Paris

PARENTEAU J.P.
  MinistSre de 1'Environnement
  13 rue de Bourgogne
  75007 Paris

PARROTT J.
  De Pietro Press
  55 boulevard de Charonne
  75011 Paris

PATTI F.
  Commissariat a 1'Energie
  Atomique
  Bolte Postale 6
  92260 Fontenay-aux-Roses

PERDRIZET S.
  I.N.S.E.R.M.
  38 rue de Bretagne
  75003 Paris
PEYRONNET J.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

PHAM Q. T.
  I.N.S.E.R.M., U. 14
  Plateau de Brabois
  54500 Vandoeuvre Ifis Nancy

PIVA C,
  HOpital Fernand Widal
  200 rue du Fbg St.  Denis
  75474 Paris-Cedex 10

POUSSE G.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

QUINOT E.
  I.N.R.S.
  route de NeufchSteau
  54500 Vandoeuvre

RAVIART M.
  I.M.A.-I.N.S.E.R.M.
  Hopital de Bicetre
  94270 Le Kremlin-Bic§tre

REY COQUAIS B.
  SocietS Prodelec
  25 quai Paul Doumer
  92408 Courbevoie

RIBARD P.
  Centre Antipoison de Lyon
  HOpital Edouart Heriot
  69000 Lyon

RIPS R.
  I.N.S.E.R.M.
  17 rue du Per a Moulin
  75005 Paris

ROBERT E.
  Compagnie Fran?aise BASF
  DSpartement Agricole
  140 rue Jules Guesde
  Bolte Postale 118
  92303 Levallois-Perret

ROTHSTEIN G.
  13 rue de Poissy
  75005 Paris

-------
                              2488
ROTHSTEIN M.
  13 rue de Poissy
  75005 Paris

ROUGH Y.
  Mfidecine du travail et
  Hygiene industrielle
  HOtel Dieu
  31052 Toulouse

ROUIT C.
  SociStS Francaise des
  Petroles BP
  10 quai Paul Doumer
  92401 Dourbevoie

SADOUL P.
  I.N.S.E.R.M., 0. 14
  Plateau de Brabois
  54500 Vandoeuvre-lSs-Nancy

SARASIN A.
  Institut de Recherches
  Scientifiques sur le Cancer
  Centre National de
  Recherche Scientifique
  Bolte Postale 8
  94800 Villejuif

SEBAN R.
  15 rue des Marguettes
  75012 Paris
SEIBEL G. M. G.
  F.F.S.A.
  53 rue Francois 1
  75008 Paris
er
SIMON B.
  Dfipartement MSdical
  ESSO, S.A.F.
  92080 Paris La Defense
  Cedex 2

SIOU G.
  Shell Francaise
  29 rue de Berri
  75380 Paris Cedex 08

SMAGGHE G.
  Produits Chimiques
  Ugine Kuhlmann
  Bolte Postale 175
  75782 Paris Cedex 16
STUPFEL M.
  I.N.S.E.R.M., U.  123
  44 chemin de Ronde
  78110 Le VSsinet

SUCHET A.
  Fondation A. de Rothschild
  Service M6decine  Preventive
  56 avenue Mathurin Moreau
  75019 Paris

SYROTA M. J.
  MinistSre de I1 Environnement
  13 rue de Bourgogne
  75007 Paris

TESSIER J. F.
  Laboratoire d'Hygiene
  Facult6 de Me'decine
  place de la Victoire
  Bordeaux

THAL M. F.
  Commissariat £ 1'Energie
  Atomique
  DSpartement de Protection
  Bolte Postale 6
  92260 Fontenay-aux-Roses

TREMOLIERES J.
  Laboratoire de Nutrition
  Humaine
  I.N.S.E.R.M.
  HOpital Bichat
  170 boulevard Ney
  75018 Paris

TRICARD D.
  Ministfire de la Sante"
  20 rue d'Estrfies
  75007 Paris

TRUCHOT A.
  Compagnie Francaise de
  Raffinage
  39/43 quai Andre" Citroen
  75739 Paris-Cedex 15

TRUHAUT R.
  Centre de Recherches Toxi-
  cologiques de la FacultS des
  Sciences Pharmaceutiques et
  Biologiques de 1'Universite
  Rene1 Descartes
  7 avenue de 1'Observatoire
  75006 Paris

-------
                              2489
TRUFFERT L.
  7 avenue Barbes
  94100 St. Maur des Fossfis

TULLIEZ J.
  Institut National de la
  Recherche Agronomique
  C.N.R.Z.
  Laboratoire des Additifs
  Alimentaires
  78350 Jouy-en-Josas

VAREILLE J.
  Laboratoire Central de la
  Prefecture de Police
  39 bis rue de Dantzig
  75015 Paris

VEIL S.
  Ministre de la SantS
  20 rue d'Estrfies
  75007 Paris

VIAL J.
  Institut Pasteur Lyon
  69365 Lyon Cedex II
VILLERMAY D.
  Revue de 1'InfirmiSre
  9 rue de Berri
  75384 Paris Cedex 08

WARNET J. M.
  62 rue Brancion
  75015 Paris

WHIPPLE P.
  Education Nationale
  46 Avenue du GSne'ral de
  Gaulle
  Bolte Postale 132
  49400 Saumqr

YANA A.
  Ministers de la QualitS
  de la Vie
  14 boulevard du GfinSral
  Leclerc
  92521 Neuilly/Seine

ZERAFFA J.
  I.N.S.E.R.M.
  101 rue de Tolbiac
  75013 Paris
                            GREECE
ZAPHIROPOULOS M.
  Environmental Pollution
  Control Project
  Voulis 4
  Athens
                           HELVETIA
BERGEN P.
  World Wide Medical News
  Service A.G,
  6 rue du L6man
  1201 Geneve

COLLET R. A.
  Universite de GenSve
  Section de Chlmie
  30 gue Ernest-Ansermet
  1211 Geneve 4
DUPUIS G.
  Ciba-Geigy A.G,
  4002 Basel
  Rosental

HOGGER D.
  Eidg. Kommission
  Lufthygiene
  8032 Zttrich
  Kapfstr. 16
fur

-------
                               2490
MAREK  B.
  Swiss Federal  Health Office
  Division of Food Control
  Section Pesticide Residues
  and  Contaminants
  3000 Berne
  Haslerstrasse  16

MERIAN E.
  Schweiz. Arbeitsgemein-
  schaft fur Umweltforschung
  4106 Therwil
  Im Kirsgarten  22

MULLER U.
  Kantonales Laboratoriura
  3000 Bern 9
  Postfach

RAMACIOTTI D.
  Institut de MSdecine So-
  ciale et Preventive
  20 quai de 1'Ecole de
  MSdecine
  1205 Gen&ve

REY P.
  Institut de MSdecine So-
  ciale et Preventive
  20 quai de 1'Ecole de
  Mfidecine
  1205 Genfive

RINZEMA L. C.
  Dow  Chemical Europe SA
  8832 Wollerau
  Erlenstr. 46
RYLANDER R,
  Institut de MSdecine So-
  ciale et Preventive
  20 quai de 1'Ecole de
  M6decine
  1205 Geneve

SCHLATTER C.
  VeterinSr- Physiologisches
  Institut der Universitat
  Winterthurerstr. 260
  8057 Zurich

SCHLATTER-LANZ I.
  Schweiz. Toxikologisches
  Informationszentrum
  8028 Zurich
  Zurichbergstr. 8

SEILER J. P.
  Eidg. Forschungsanstalt
  fUr Obst-, Wein- und
  Gartenbau
  8820 Wadenswil

STAM J.
  Du Pont de Nemours Int.SA
  29 bis rue de Lancy
  1211 Geneve 24

VOUK L.
  6 avenue du Lignon
  Geneve
                            HUNGARY
KOVARI K.
  1012 Budapest
  I. Attila 107
                           INDONESIA
SETIADY I.
  Directorate General of
  Communicable Disease Control
  Ministry of Health
  Jl. Percetakan Negara
  Post Bos 223
  Jakarta

-------
                              2491
                              IRAN
AHMED W.
  Human Environment Division
  Department of Environmental
  Conservation
  Teheran

AREFI M.
  Human Environment Division
  Department of Environmental
  Conservation
  Teheran
ROUANI M.
  Health Affairs
  National Iranian Oil
  Teheran
Co,
                            IRELAND
CHAMBERS P.
  Department of Pharmacology
  University of Dublin
  Trinity College
  Dublin

DEAN G.
  The Medico Social Research
  Board
  73 Lower Baggot Street
  Dublin 2

DUGGAN C.
  An Taisce
  The National Trust for
  Ireland
  Warrenscourt
  Lissarda
  County Cork
GRIMES H.
  Department of Pathology
  and Bacteriology
  Regional Hospital
  Galway

HEAGNEY J. K.
  North Western Health Board
  Kintogher
  Bundoran Road
  Sligo

HICKEY N.
  Department of Social and
  Preventative Medicine
  University College
  Earlsfort Terrace
  Dublin 2

KEVANY J.
  Trinity College Medical
  School
  St. James's Hospital
  Dublin 8
                            ISRAEL
WASSERMAN M.
  Department of Occupational
  Health
  The Hebrew University
  Hadassah Medical School
  P.O. Box 1172
  Jerusalem

-------
                              2492
                            ITALIA
ALBERTONI F.
  Istituto di Tisiologia
  II Catt. Universita di
  Roma
  Via Baldassarre Peruzzi 6
  00153 Roma

ALESS10 L.
  Clinica del Lavoro
  Via S. Barnaba 8
  20122 Milano
                       10
 BARRO G.
   Regione dell'Umbria
   Via. S. Bonaventura
   06100 Perugia

 BERETTA F.
   Clinica del  Lavoro
   Via M.  Greppi 10
   20135 Milano
 BIANCO A.
   Laboratorio  Radioattivita
   Ambientale C.N.E.N.
   C.S.N.  Casaccia, C.P.  2400
   00100 Roma
 BONAZZI G.
   Universita Parma
   Laboratorio  di Ecologia
   Borgo Carissimi 10
   43100 Parma

 CACCURI S.
   Istituto di  Medicina del
   Lavoro  dell1   Universita
   Policlinico
   Piazza  Miraglia
  80136 Napoli

 CASTELLINO N.
  Medicina Lavoro
  Universita Napoli
  Via  Raffaele de Cesare 31
   80132 Napoli

CASULA D.
  Istituto di Medicina del
  Lavoro dell'Universita
  degli Studi di Cagliari
  Via S. Giorgio 12
  09100 Cagliari
 CASULE G.
   Regione Toscana
   Piazza Liberta 15
   50129 Firenze

 CERQUIGLINI-MONTERIOLO S.
   Istituto Superiors di
   Sanita
   Viale Regina Elena 299
   00161 Roma

 CLEMENTE G. F.
   Laboratorio Radioattivita
   Ambientale C.N.E.N.
   C.S.N. Casaccia
   C.P. 2400
   00100 Roma

 COLOMBINI M.
   Ministero  della Sanita
   Via  Liszt  34
   00100  Roma

 CONENNA  C.
   "Italia Nostra"
   Piazza Fontane Marose 6/5
   16123  Geneva

 COTTA-RAMUSINO F.
   Istituto Superiore di
   Sanita
   Viale Regina Elena 299
   00161 Roma

 DEL BORGHI M.
   Istituto di Scienze e Tec-
   nologie dell'Ingegneria
   Chimica
   Facolta di Ingegneria
  Via all'Opera Pia 11
   16145 Geneva

FADDA F.
  Enti locali ecologia
  urbanistica
  Regione Sarda
  09100 Cagliari

FAILLACE R.
  Regione Toscana
  c/o Dip.  Sicurezza Sociale
  Piazza della Liberta 15
  50129 Firenze

-------
                              2493
FAVRETTO L.
  Istituto di Merceologia
  Universita di Trieste
  34100 Trieste

FILASTO-NARESE M.
  Regione Toscana
  Dipartimento sicurezza
  sociale
  Piazza della Liberta 15
  50129 Firenze

GADONI S.
  Enti local! ecologia
  urbanistica
  Regione Sarda
  09100 Cagliari

GARIBALDI P.
  Snam Progetti
  20097 S.Donate Milanese
  Milano

GARLANDA T.
  Montedison S.p.A.
  Via Turati 7
  20121 Milano

GATTI G. L.
  Istituto  Superiore di
  Sanita
  Viale Regina Elena 299
  00161 Roma

GHETTI P.  F.
  Laboratorio  di Ecologia
  Universita di  Parma
  Strada Universita  12
  43100 Parma

GRASSO C.
  Universita degli Studi di
  Firenze
   Istituto di  Igiene
  Viale  G.  B.  Morgagni 48
   50134  Firenze

LA  FORGIA  M.
  Assessorato  all'Igiene e
  alia Tutela  dell'Ambiente
  Regione  Emilia-Romagna-
  Bologna
  Viale  Silvani  6
  40122  Bologna
LANZOLA E.
  Istituto di Igiene
  dell'Universita
  Viale Forlanini 1
  27100 Pavia
MAGI F.
  E.N.I.
  P.le Enrico Mattel 1
  00144 Roma

MARINI G.
  Enti locali ecologia
  urbanistica
  Regione Sarda
  09100 Cagliari

MARRACINI A.
  Enti locali ecologia
  urbanistica
  Regione Sarda
  09100 Cagliari

MONTEROSSO M.
  Regione dell'Umbria
  Via S. Bonaventura 10
  06100 Perugia

NARESE C.
  via Rondinelli  2
  50123 Firenze

PALMAS A.
  Enti  locali ecologia
  urbanistica
  Regione Sarda
  09100 Cagliari

PASQUINI R.
  Istituto d'Igiene dell1
  Universita degli  Studi
  di Perugia
  Via del Giochetto
  06100  Perugia

PECORA L.
   Istituto  di Medicina del
   Lavoro
   Universita Napoli
  Via Schipa 61
   80122 Napoli

-------
                              2494
PERTOSA M.
  Istituto d'Igiene dell1
  Universita degli Studi
  di Perugia
  Via del Giochetto
  06100 Perugia

SACCHETTI A.
  Assessorato all'Igiene e
  alia Tutela dell1Ambiente
  Regione Emilia-Romagna-
  Bologna
  Viale Silvani 6
  40122 Bologna

SANTARONI G.
  Istituto Nazionale della
  Nutrizione
  Citta Universitaria
  00161 Roma

SANTI L.
  Istituto di Oncologia
  dell'Universita di Genova
  Viale Benedetto XV
  16132 Genova

SAVING A.
  Istituto d'Igiene dell'
  Universita degli Studi
  di Perugia
  Via del Giochetto
  06100 Perugia
 SCASSELLATI SFORZOLINI G.
   Istituto d'Igiene dell1
   Universita degli Studi
   di Perugia
   Via del Giochetto
   06100 Perugia

 SEMIANI P.  L.
   E.N.I.
   Piazzale  E. Mattel  1
   00144 Roma

 SENNI  L.
   Tecneco S.p.A.
   (E.N.I.)
   C.P.  47
   Fano  (P.S.)

 SIRIGU  M.
   Enti  locali ecologia
   urbanistica
   Regione Sarda
   09100 Cagliari

 VANINI  G.
   Istituto  di Igiene
   dell'Universita Torino
   Via Santena 5 bis
   Torino

 VIVOLI  G.
   Istituto  di Igiene
   Universita di Modena
  Via Campi  287
   41100 Modena
                             JAPAN
GOTO M.
  Laboratory of Ecological
  Chemistry
  Faculty of Science
  Gakushuin University
  Toshima-Ku
  Tokyo 171

ISAO U.
  Environmental Health Sec-
  tion of Osaka Fu Gov.
  25-1 Otemae no Cho
  Higashi Ku
  Osaka
KABURAGI K.
  Mitsubishi Metal Corporation
  5-2 Ohte-Machi 1-Chome
  Chiyoda-Ku
  Tokyo 100

KAWAI K.
  National Institute of
  Industrial Health
  221 Kizuki-Sumiyoshi-Cho
  Kawasaki

-------
                              2495
TAGUCHI S.
  Department of Physiology
  School of Life Science
  Rissho Women's University
  3337 Minamiogishima
  Koshigaya-shi
  Saitama

TAKENAKA K.
  Environment Agency
  3-1-1 Kasumigaseki
  Chiyoda-ku
  Tokyo

TATI M.
  Department of Public
  Health
  Gifu University
  School of Medicine
  Tsukasa-maci 40
  Gifu
TOMITA K.
  Public Nuisance and Medical
  Research Institute, Inc.
  c/o Mitsui Mining & Smel-
  ting Co., Ltd.
  2-1-1 Nihonbashi-Muromachi
  Chuo-Ku
  Tokyo

TSUCHIYA K.
  Department of Preventive
  Medicine & Public Health
  School of Medicine
  Keio University
  35 Shinanomachi
  Shinjuku-Ku
  Tokyo

WATANABE H.
  Public Health Institute
  of Hyogo Prefecture
  2-1 Aratacho, Hyogoku
  Kobe City
                          JUGOSLAVIA
ACKO M.
  Institut za patolosko
  morfologijo
  Zaloska 4
  61 005 Ljubljana

BOGOMIL H.
  Dispanzer fur Arbeits-
  medizin
  Zdravstveni dom Celje
  Zvezna 8
  63000 Celje

BYRNE A.
  Institute "Jo2ef Stefan"
  Ljubljana
  Jamova 39

CERKEZ F.
  Institute of Hygiene
  Medical Faculty
  71 000 Sarajevo
  Bjelave 70/1
DJUKIC Z.
  Health Protection Depart-
  ment of the "Boris Kidric"
  Institute of Nuclear
  Sciences
  11 000 Beograd
  P.O.B. 522

HOCEVAR B.
  Sanitary Inspector
  Republiski Sekretariat za
  Zdravstvo in Socialno Varstvo
  Zupanciceva ul 6 - Pp 644
  Ljubljana

KORICANAC Z.
  Institute of Inorganic &
  Analytical Chemistry
  Faculty of Pharmacy
  University of Belgrade
  Dr. Subotica 8
  11 00 Beograd

MIL1C S.
  Institute of Occupational
  & Radiological Health
  11 000  Beograd
  Deligrdska 29

-------
                              2496
MILOVANOVIC L.
  Institute of  Inorganic  &
  Analytical  Chemistry
  Faculty of Pharmacy
  Dr. Subotica 8
  11 000 Beograd

ORUS&V T.
  Institut de Medecine du
  Travail
  Rudnici i Zelezarnica
  Skopje

ROSIVAL L.
  Research Institute of
  Hygiene
  Bratislava
  Ul.Cs.armady  40

STANKOVIC B.
  Institute of  Inorganic
  and Analytical Chemistry
  Faculty of  Pharmacy
  Studentska  43/10
  11 000 Beograd

STANKOVIC M.
  Institute of  Occupational
  and Radiological  Health
  11 000  Beograd
  Deligradska 29
VERHOVNIK V.
   62000 Maribor
  Kamniska 28

VERHOVNIK Vilma
   62000 Maribor
  Kamniska 28

WEBER 0.
   Institute for Medical
  Research
  Yugoslav Academy of
   Sciences and Arts
  P.O. Box 293
  Mose Pijade 158
   41 001 Zagreb

ZARKOVIC G.
   Institut za Higijenu i
   Socijalnu Medicinu
  MoSe Pijade 6
   71 000 Sarajevo

ZELENKO V.
   Institut Joseph Stephen
   Ljubljana
   Okiskega A

ZUNIC R.
   Zajednica za naucno  istra-
   zivanje zivotnih uslova
   23 ul.Cara H.  Selasija
   11 000 Beograd
                             LIBERIA
WESLEY J.
   Liberian  Embassy
   8  rue Jacques  Bingen
   75017 Paris, France
                           LUXEMBOURG
 LANGEVIN M.
   177  rue  de  Dippach
   Bertrange

-------
                              2497
                             MAROC
AZIZI A.
  Ministere de la Santfi
  Publique
  1 boulevard Jeanne d'Arc
  Rabat Agdal
          CHAUDERON J. J. M.
            Ministere du Travail
            11 rue des Courlis
            Casablanca
                           MAURITIUS
MAURICE J.
  Ministere de la Santfi
  lie-Maurice
                            MEXICO
BRAVO A, H.
  Instituto de Geofisica
  Universidad Nacional
  Autonoma de Mexico
  Torre de Ciencias - 50,
  Ciudad Universitaria
  Mexico 20
Piso
LOWE A. C.
  Instituto de Geofisica
  Universidad Nacional
  Autonoma de Mexico
  Torre de Ciencias - 50.Piso
  Ciudad Universitaria
  Mexico 20
                           NEDERLAND
BIERSTEKER K.
  Municipal Health Department
  Baan 170
  Rotterdam

den BOER M. C.
  Ministerie van Volks-
  gezondheid
  D.G. Milieuhygiene
  Dr. Reyersstraat 10
  Leidschendam

BOLIJN A. G.
  Bedrijfsgeneeskundige
  Dienst
  Hoogovens IJmuiden BV
  Medisch Centrum 3D.10
  IJmuiden
          de BRUIN A.
            Universiteit van Amsterdam
            Laboratorium van Arbeids-
            h^giene
            1  Constantijn Huygens-
            straat 20
            Amsterdam

          CANTON J. H.
            Rijks Instituut voor de
            Volksgezondheid
            Postbus 1
            Bilthoven

          del CASTILHO P.
            Laing's Nekstraat 49
            Amsterdam

-------
                              2498
EVENDIJK J. E.
  Dienst Centraal Milleu-
  beheer Rljnmond
  Stationsplein 2
  Schiedam

GOOTJES P.
  Rijks Instituut voor de
  Volksgezondheid
  Postbus 1
  Bllthoven

GUPTA S.
  Department of Toxicology
  Rijks Instituut voor de
  Volksgezondheid
  Postbus 1
  Bilthoven

HENDERSON P. T.
  Farmacologisch Instituut
  Geert Grooteplein noord 21
  Nijmegen

HERZBERG S.
  Koninklijke/Shell-
  Laboratorium
  Badhuisweg 3
  Amsterdam-Noord

JOOSTING P. E,
  TNO Research Institute for
  Environmental Hygiene
  Schoemakerstraat 97
  P.O.B. 214
  Delft

KOLK J. J.
  AKZO Research and Engin-
  eering
  Velperweg 76
  fabriek Kleefse Waard
  Arnhem

van der KREEK F. W.
  Ministerie van Volks-
  gezondheid en Milieuhygiene
  Dokter Reijersstraat 10
  Leidschendam

van der LEE C, G.
  AKZO Research and
  Engineering
  Postbus 60
  Arnhem
de MOOR A. G.
  Ministerie van Volks-
  gezondheid en Milieuhygiene
  Veterinaire Hoofdinspectie
  le van den Boschstraat  4
  Den Haag

van der NOORDAA J.
  Organization for Industrial
  Research
  Industrial Liaison Dept.
  97 Schoeniakerstraat
  Delft

NOTTEN W. R. F.
  Farmacologisch Instituut
  Geert Grooteplein noord 21
  Nijmegen

OOMS A.
  Chemisch Laboratorium TNO
  Postbus 45
  Rijswijk (Z.H.)

SCHNEIDER T.
  Rijksinstituut voor de
  Volksgezondheid
  Postbus 1
  Bilthoven

SIBINGA MULDER J.
  Bedrijfsgeneeskundige
  Dienst
  Hoogovens iJmuiden BV
  Medisch Centrum 3D.10
  IJmuiden

den TONKELAAR E.  M.
  Rijksinstituut voor de
  Volksgezondheid
  Postbus 1
  Bilthoven

VERBERK M.  M.
  Universiteit van Amsterdam
  Laboratorium van Arbeids-
  hygiene
  ie Constantijn Huygens-
  straat 20
  Amsterdam

de VRIES J.
  Organization for Industrial
  Research
  Industrial Liaison Dept.
  97 Schoemakerstraat
  Delft

-------
                              2499
de ZEEUW
  Instituut voor Toepassing
  van Atoomenergie In de
  Landbouw
  Postbus 48
  Wageningen
ZIELHUIS R. L.
  Universiteit van Amsterdam
  Laboratorium van Arbeide-
  hygiene
  ie Constant!jn Huygens-
  straat 20
  Amsterdam
                            NIGERIA
SULAIMAN A. B.
  Environmental and Occupa-
  tional Unit
  Federal Ministry of Health
  Yakubu Gowon Street
  Lagos
                             NORGE
AASETH J.
  Institute of Clinical
  Biochemistry
  University of Oslo
  Rikshospitalet
  Oslo 1

SKOGVOLD O. P.
  Norwegian Institute for
  Air Research
  P.O. Box 115
  2007 Kjeller
THRANE K. E.
  Norwegian Institute for
  Air Research
  P.O. Box 115
  2007 Kjeller

WANNAG A.
  Institute of Occupational
  Health
  Box 8149
  Oslo-Dep
  Oslo 1
                          OSTERREICH
CZEDIK-EYSENBERG P.
  Osterreichische Unilever
  Ges.M.B.H.
  Schenkenstrasse 8-10
  1010 Wien

HAIDER M.
  Institut fur Umwelthygiene
  der Universitat Wien
  Kinderspitalgasse  15
  1095 Wien
PFANNHAUSER W.
  Forschungsinstitut der
  Ernahrungswirtschaft
  Blaasstrasse 29
  1190 Wien

PLANK S.
  Ludwlg Boltzmann-Institut
  fur Umweltwissenschaften
  und Naturschutz
  Heinrichstrasse  5
  8010 Graz

-------
                              2500
PRIGLINGER R.
  Amt der 8st. Landesregierung
  Sanitatsdienst
  Harrachstr. 16A
  4020 Linz
                            POLSKA
KOTLAREK-HAUS S.
  Medical School
  Department of Haematology
  Pasteura 4
  50-367 Wroclaw

LOSZEK W.
  Ministerium der Schwere-
  industrie
  00-163 Warszawa
  Str.Karmelicka 15m44
WLODEK S.
  Universitfi Polytechnique
  Institut de Genie de I1 En-
  vironnement
  00-661 Warszawa
  Pl.Jednosci Robotniczej 1
                   REPUBLIC OF SOUTH AFRICA
van ROOYEN G. I.
  Department of Health
  Private Bag X88
  Pretoria
                   REPUBLIQUE CENTRAFRICAINE
BEDAYA NGARO 5.
  MinistSre de  la Sant6
  Publique et des Affaires
  Sociales
  Bangui
MOKEMATKENGUEMBA G.
  Soci6t4 National
  d'Habitat
  Bangui
                             SUOMI
HERNBERG S.
  Institute of Occupational
  Health
  Haartmaninkatu 1
  00290 Helsinki 29
LAAMANEN A.
  University of Kuopio
  Puistokatu 20
  70100 Kuopio 10

-------
                               2501
LEHTO V.-P.
  Environmental Protection
  Council
  Secretary of Work and
  Housing
  Environment Section
  Ministry of Internal Affairs
  Fredrikinkatu 58 D 62
  00100 Helsinki 10

OJAJARVI J.
  Department of Public Health
  Science
  Haartmaninkatu 3
  00290 Helsinki 29
SUMARl P,
  Institute of Occupational
  Health
  Haartmaninkatu 1
  00290 Helsinki 29

VIRTANEN I.
  Ill Department of Pathology
  University of Helsinki
  Kaartmaninkatu 3
  00290 Helsinki 29

VUORI E.
  Department of Public Health
  Science
  Haartmaninkatu 3
  00290 Helsinki 29
                            SVERIGE
ABDULLA M.
  Research Department 2
  University Hospital
  22185  Lund

BERGLUND U. L.
  Psychological Laboratories
  University of Stockholm
  Box 6706
  11385  Stockholm

BERLIN M.
  University of Lund
  Department of Environmental
  Health
  Box 2009
  22002  Lund 2

BJORLING M.
  Saab-Scania AB
  46100  TrollhStten

DANIELSON L.
  Swedish Environment
  Protection Board
  17120 Solna 1

FRIBERG L.
  Department of Environmental
  Hygiene
  The Karolinska Institute
  10401 Stockholm 60
GAGE J. C.
  Department of Environmental
  Health
  University of Lund
  Box 2009
  22002  Lund 2

HOLM S.
  Department of Environmental
  Health
  University of Lund
  Box 2009
  22002  Lund 2

HOLMQVIST I.
  Bolieden Aktiebolag
  Bomsk&rsverken
  93200 Skelleftehamn

KJELLSTROM T.
  Department of Environmental
  Hygiene
  The Karolinska Institute
  10401 Stockholm 60

LINDVALL T.
  Department of Environmental
  Hygiene
  The Karolinska Institute
  10401 Stockholm 60

-------
                              2502
PISCATOR M.
  Department of Environmental
  Hygiene
  The Karolinska  Institute
  10401 Stockholm 60
KINDER L.
  University of GSteborg
  40033 Goteborg

SVENSSON K.
  Saab-Scania AB
  46100  Trollhatten
                            TUNISIE
ATALLAH S.
  Service de 1'Assainissement
  Ministfire de la Santfi
  Publique
  Tunis
WILSON J. C.
  Ecole Nationale des IngS-
  nieurs de Tunis
  B.P. NO. 37
  Tunis-Belvedere
                        UNITED KINGDOM
BARLTROP D.
  Paediatric Unit
  St. Mary's Hospital
  Medical School
  Norfolk Place
  London W.2.

BARRATT R. S.
  Chemistry Department
  University of Birmingham
  P.O. Box 363
  Edgbaston
  Birmingham B15 2TT

BARRY P.
  The Associated Octel Co,
  Ellesmere Port
  Cheshire

BROOKS A. G. F.
  Medical Research Council
  Air Pollution Research Unit
  St. Bartholomew's Hospital
  Medical College
  Charterhouse Square
  London E.C.I.
BROWN P.
  Elsevier North Holland
  Scientific Publishers
  55 Benslow Rise
  Hitchin
  Hertfordshire SG4 9QY

BULCRAIG W. R.
  Pilkington Brothers Ltd.
  R and D Department
  Watson Street
  St. Helens
  Lancashire WA10 3TT

BURGESS J. E.
  Dow Chemical Company
  Huntingdon Research Centre
  Estuary Road
  King's Lynn
  Norfolk

CERNIK A. A.
  Central Reference Laboratory
  Department of Employment
  1-13 Chepstow Place
  Westbourne Grove
  London W.2.

-------
                               2503
CHAPMAN J.
  Medical Research Council
  Headquarters
  20 Park Crescent
  London WIN 4AL

CHARLTON J.
  Environmental Services
  Department
  Trafalgar House
  Paradise Circus Queensway
  Birmingham Bl 2BQ

CLARKE R. H.
  Central Electricity Gener-
  ating Board
  Berkeley Nuclear Labora-
  tories
  Berkeley
  Gloucestershire GL13 9PB

COMMINS B. T.
  Medical Research Council
  St. Bartholomew's Hospital
  Medical College
  Charterhouse Square
  London EC1M 6BQ

DANIEL J. W.
  Life Science Research
  Stock
  Essex CM4 9PE

DELVES H. T.
  Department of Chemical
  Pathology
  Institute of Child Health
  University of London
  30 Guilford Street
  London WC1N 1EH

DERRETT C. J.
  Medical Research Council
  Air Pollution Unit
  St. Bartholomew's Hospital
  Medical College
  Charterhouse Square
  London EC1M 6BQ

DUCK B. W.
  British Petroleum Company
  B.P. Research Centre
  Chertsey Road
  Sunbury-on-Thames
  Middlesex
DUNCAN K. P.
  British Steel Corporation
  33 Grosvenor Place
  London SW1X 7JG

FLOREY C. du V.
  Department of Epidemiology
  and Social Medicine
  St. Thomas's Hospital
  Medical School
  60 Sancroft Street
  London S.E.I.

GILTROW J.
  Central Unit on Environ-
  mental Pollution
  Department of the Environment
  2 Marsham Street
  London S.W.I.

GRIFFITH R. R.
  Associated Octel Co. Ltd.
  20 Berkeley Square
  London W1X 6DT

HARRISON G. F.
  Associated Octel Co. Ltd.
  20 Berkeley Square
  London W1X 6DT

HARRISON R. M.
  Imperial College of Science
  and Technology
  Public Health Engineering
  Section
  Imperial Institute Road
  London SW7 2BX

HEDGES W. V.
  Blythe Colpurs Ltd.
  Cresswell
  Stoke-on-Trent ST11 9RD

HISCOCK S.
  Lead Development Association
  34 Berkeley Square
  London W1X 6AJ

HISLOP J.
  Environmental and Medical
  Sciences Division
  A.E.R.E.
  Harwell
  Didcot
  Berkshire

-------
                               2504
 HOLLAND W. W.
   Department of Clinical
   Epidemiology and Social
   Medicine
   St. Thomas's Hospital
   Medical School
   London SE1 7EH

 HOWE D.
   Paintmakers Association
   of Great Britain
   22 Hightree Drive
   Henbury
   Macclesfield
   Cheshire

 HUGHES E.  G.
   Johnson,  Matthey &  Co.  Ltd.
   78 Hatton Garden
   London ECIP 1AE
HUNTER W. J.
  General Foods Europe
  Banbury 0X16 7QU

INGIMUNDARSON E.  V.
  National  Institute of
  Environmental Health
  Bernard Sunley House
  40-44 Evelyn Gardens
  London S.W.7.

IRWIG L.
  Department of Clinical
  Epidemiology and Social
  Medicine
  St. Thomas's Hospital
  Medical School
  London SE1 7EH

JACOBSEN M.
  Institute of Occupational
  Medicine
  8 Roxburgh Place
  Edinburgh EH8 9SU

KING E.
  National Occupational
  Hygiene Service Ltd.
  12 Brook Road
  Fallowfield
  Manchester
 LEEDER S.
   Department of Clinical
   Epidemiology and Social
   Medicine
   St. Thomas's Hospital
   Medical School
   London SEl 7EH

 LEESE W. L. B.
   Mobil Oil Corporation
   54 - 60 Victoria Street
   London S.W.I.

 LEMON R. C.
   Shell International
   Petroleum Co.
   Medical Division
   Shell Centre
   London SEl 7NA

 MCNEIL N.
   Scottish Home and Health
   Department
   St.  Andrew's House
   Edinburgh

 MARTIN A. E.
   Department of Health  and
   Social  Security
   Alexander Fleming House
   Elephant and  Castle
   London  SEl  6BY

MILLER D. S.
  Department of Community
  Health and Medicine
  Medical School
  Nottingham University
  Nottingham NG7 2RD

MILLS A.  L.
   Burmah-Castrol Company
   Castrol Research  Laboratories
   London  Road
   Bracknell
   Berkshire RG12 2UW

MOORE J. P.
  Occupational Hygienist
  Shell International
  Petroleum Co.
  Medical Division
  Shell Centre
  London SEl 7NA

-------
                              2505
MOORE M.  R.
  Department of Materia Medica
  Stobhill General Hospital
  Glasgow G21  3UW

MOORHOUSE J.
  Rio Tinto-Zinc Corporation
  6 St. James's Square
  London  SW1Y  4LD
MULLINS C.
  Instrumentation Laboratory
  (U.K.) Ltd.
  Station House
  Stamford New road
  Altrincham
  Cheshire

OLDHAM P. D.
  MRC Pneumoconiosis Unit
  Llandough Hospital
  Penarth
  Glamorgan CFG 1XW

PACKHAM R. F.
  Water Research Centre
  Medmenham Laboratory
  Ferry Lane
  Medmenham
  Marlow
  Buckinghamshire SL7 2HD

PERRY R.
  Imperial College of Science
  and Technology
  Public Health Engineering
  Section
  Imperial Institute Road
  London S.W.7.

PHILP J. McL.
  Unilever Limited
  Unilever House
  P.O. Box 68
  London EC4P 4BQ

PITTWELL L.  R.
  Department of the Environment
  Directorate General of
  Water Engineering
  2  Marsham Street
  London SW1P 3EB
PURI G.
  Liverpool Polytechnic
  Department of Biology
  Byrom Street
  Liverpool 3

RAINSBURY R.
  British Medical Journal
  BMA House
  Tavistock Square
  London W.C.I.

RATCLIFFE J. M.
  Imperial College of Science
  and Technology
  Department of Mathematics
  London S.W.7.

REED S. B.
  Greater London Council
  40 Brynmaer Road
  London SW11 4EW

SAYERS M. H. P.
  Department of Employment
  Central Reference Laboratory
  1-13 Chepstow Place
  Westbourne Grove
  London W.2.

SHERWOOD R. J.
  Honey Bottom
  Dry Sandford
  Abingdon
  Oxfordshire 0X13 6BX

SILVERMAN A. P.
  Imperial Chemical Industries
  Industrial Hygiene Research
  Laboratories
  Alderley Park
  Near Macclesfield
  Cheshire SK10 4TJ

STANLEY F.
  Medical Research Council
  Social Medicine Unit
  London School of Hygiene
  and Tropical Medicine
  Gower Street
  London W.C.I.

-------
                              2506
STOTT N. B.
  Environmental and Medical
  Sciences Division
  A.E.R.E.
  Harwell
  Didcot
  Berkshire

STREHLOW C.
  St. Mary's Hospital
  Medical School
  Paediatric Unit
  Norfolk Place
  London W.2.

SWANN P. G.
  Esso Petroleum Co. Ltd.
  Esso House
  Victoria Street
  London SW1E 5JW

TAYLOR J. F.
  Chloride Batteries Ltd.
  P.O. Box 5
  Clifton Junction
  Swinton
  Manchester M27 2LR

TWIBELL J. D.
  Home Office Central
  Research Establishment
  Aldermaston
  Near Reading
  Berkshire

VAILE H.
  Commonwealth Smelting Ltd.
  St. Andrews Road
  Avonmouth
  Bristol BS11 9HP
WAKELIN E. N.
  Environmental Services
  Department
  Trafalgar House
  Paradise Circus Queensway
  Birmingham Bl 2BQ

WALLER R. E.
  Medical Research Council
  Air Pollution Unit
  St. Bartholomew's Hospital
  Medical College
  Charterhouse Square
  London EC1M 6BQ

WILLIAMS M. K.
  Chloride Automotive
  Batteries Ltd.
  Fortyfoot Road
  Leatherhead
  Surrey

WILSON J. R.
  British Gas
  45 Oaken Lane
  Claygate
  Esher
  Surrey KT10 ORQ

WOOLLEY J. F.
  Standard Telephones Ltd.
  Harlow
  Essex

YEOMAN W. B.
  Regional Toxicology
  Laboratory
  Dudley Road Hospital
  P.O. Box 293
  Birmingham B18 7QH
                   UNITED STATES OF AMERICA
ALBERT R.
  New York University
  Medical Center
  Institute of Environmental
  Medicine
  550 First Avenue
  New York, N.Y.   10016
ALLEN C.
  Veterans Administration
  Veterans Hospital
  Madison, Wisconsin

-------
                               2507
ALLEN J. R.
  Department of Pathology
  University of Wisconsin
  Medical School
  470 North Charter Street
  Madison, Wisconsin   53706

ANDERSON J.
  Department of Biology
  Texas A & M University
  College Station
  Texas   77813

EARTH D. S.
  Environmental Protection
  Agency
  National Environmental
  Research Center
  Las Vegas, Nevada   89114

BOUHUYS A.
  Yale University Lung
  Research Center
  333 Cedar Street
  New Haven
  Connecticut   06510

BRAMAN R. S.
  University of South Florida
  Department of Chemistry
  Tampa
  Florida   33620

BURGER E. J.
  Science and Technology
  Policy Office
  National Science Foundation
  Washington D.C.   20550

BUSH B.
  New York State Department
  of Health
  Division of Laboratories
  and Research
  New Scotland Avenue
  Albany
  New York   12201

BUTLER R. C.
  E. I. du Pont de Nemours
  & Co.
  Petroleum Chemicals
  Division
  Wilmington
  Delaware   19898
CALANDRA J. C,
  Northwestern University
  and Industrial Bio-Test
  Laboratories Inc.
  1810 Frontage Road
  Northbrook
  Illinois   60062
CAPURRO P.
  Elkton
  Maryland
U.
  21921
CARNOW B. W.
  University of Illinois
  School of Public Health
  P.O. Box 6998
  Chicago
  Illinois   60680

CHAPMAN R. S.
  Environmental Protection
  Agency
  National Environmental
  Research Center
  Research Triangle Park
  North Carolina   27711

CLAYTON J. W.
  Environmental Protection
  Agency
  Health Effects Division
  Room 609 West Tower WSM
  401 M Street SW
  Washington D.C.   20460

COMMINS J.
  J. A. Commins & Associates
  7708 Orchard Way
  Philadelphia
  Pennsylvania   19118

COOPER W. C.
  Tabershaw Cooper Associates
  2180 Milvia Street
  Berkeley
  California   94704

CRAMMER M. F.
  National Center for Toxi-
  cological Research
  Jefferson
  Arkansas'

-------
                              2508
CROCKER K.
  Southern California College
  2525 Newport Boulevard
  Costa Mesa
  California   92626

CROCKER T. T.
  University of California
  Department of Community &
  Environmental Medicine
  Irvine
  California   92664

DAVID O.
  State University of New York
  Downstate Medical Center
  451 Clarkson Avenue
  Box 1195
  Brooklyn
  New York   11203

DAVISON T. H.
  Chicago & North Western
  Transportation Co.
  500 W. Madison Street
  Chicago
  Illinois   60606

DONALDSON W. T.
  Environmental Protection
  Agency
  Southeast Environmental
  Research Laboratory
  College Station Road
  Athens
  Georgia   30601

DUNCAN S.
  Institute for Research
  in Hypnosis
  315 E 77th Street
  New York City
  New York   10021
EDWARDS H. W.
  Colorado State University
  Department of Mechanical
  Engineering
  Foort Collins
  Colorado   80521

ENGEL R. E.
  Environmental Protection
  Agency
  Office of Research and
  Development
  401 M Street SW
  Washington D.C,   20460
ENGLISH T.
  Environmental Protection
  Agency
  National Environmental
  Research Center
  Research Triangle Park
  North Carolina   27711

EPSTEIN S. S.
  School of Medicine
  Department of Pharmacology
  Cast Western Reserve
  University
  Cleveland
  Ohio   44106

FISHBEIN L.
  National Center for
  Toxicological Research
  Jefferson
  Arkansas   72079

FISHER F.
  Office of Toxic Substances
  Environmental Protection
  Agency
  Room E-705-B
  401 M Street SW
  Washington D.C.   20460

FREEMAN G.
  Stanford Research
  Institute
  333 Ravenswood Avenue
  Menlo Park
  California   94025

FRIEDMAN N J,
  University of California
  Department of Psychiatry
  Irvine
  California   92664

GARDNER D. E.
  Environmental Protection
  Agency
  Experimental Biology
  Laboratory
  National Environmental
  Research Center
  Research Triangle Park
  North Carolina   27711

GOLDBERG A. M.
  Johns Hopkins University
  615 N. Wolfe Street
  Baltimore
  Maryland   21205

-------
                              2509
GOLDSMITH J. R.
  National Cancer Institute
  and California State
  Health Department
  7910 Woodmont Avenue
  Bethesda
  Maryland   20014

GOLDSTEIN I.
  Columbia University
  Division of Epidemiology
  School of Public Health
  600 W 168th Street
  New York
  New York   10032

GREEN F.
  Environmental Protection
  Agency
  Waterside Mall
  401 M Street SW
  Washington D,C.   20460

GRIFFIN H.
  Graduate School of Public
  Health
  University of Pittsburgh
  Pittsburgh
  Pennsylvania   15261

GROSSMAN A. M.
  Los Angeles County Air
  Pollution Control District
  8712 Wilshire Boulevard
  Beverly Hills
  California   90211

GUINEE V. F.
  New York City Department
  of Health
  377 Broadway  (4th floor)
  New York
  New York   10013

HANG Y.
  New York State Agricul-
  tural Expt. Station
  Food Research Laboratory
  Geneva
  New York   14456

HELLMAN R. A.
  U.S. Senate Committee on
  Public Works
  Minority Counsel
  Room 4210/Dirkson
  Senate Office Building
  Washington D.C.
HINE C. H,
  University of California
  School of Medicine
  60 King Avenue
  Piedmont
  California   94611

HUETER F. G,
  Environmental Protection
  Agency
  National Environmental
  Research Center
  Special Studies Staff
  Research Triangle Park
  North Carolina   27711

JACKSON D.
  Environmental Protection
  Agency
  Office of the Administrator
  401 M Street SW
  Washington D.C.   20460

KAMINSKI E. J.
  Northwestern University
  Medical School
  303 East Chicago Avenue
  Chicago
  Illinois   60611

KARLAN M. S.
  Beverly Hills Medical
  Society
  Environmental Pollution
  Control
  9730 Wilshire Boulevard
  Beverly Hills
  California

KEITZ E. L.
  The Mitre Corporation
  1820 Dolley Madison Bvd
  McLean
  Virginia   22101

KENNEDY G. L.
  Industrial Bio-Test
  Laboratories Inc.
  1810 Frontage Road
  Northbrook
  Illinois   60062

KLEMPERER E.
  Institute for Research
  in Hypnosis
  315 East 77th Street
  New York
  New York   10021

-------
                              2510
KNEIP T.
  Institute of Environmental
  Medicine
  New York University
  Medical Center
  550 First Avenue
  New York
  New York   10016

KNELSON J. H.
  Environmental Protection
  Agency
  University of North
  Carolina
  Mason Farm Road
  Chapel Hill
  North Carolina   27514

LAUER G.
  Rockwell International
  Science Center
  1049 Camino Dos Rios
  Thousand Oaks
  California   91360

LEUNG S.
  California Air Resources
  Board
  638 Coolidge Street
  Davis
  California   95616

LEVERE T, E.
  North Carolina State
  University
  Department of Psychology
  Raleigh
  North Carolina   27607

LIU B, Y. H,
  University of Minnesota
  Mechanical Engineering
  Department
  Minneapolis
  Minnesota   55455

 LLOYD W.  J.
  National Institute for
  Occupational Safety and
  Health
  5600 Fishers Lane
  Room 10-28
  Rockville
  Maryland   20852

LYNAM D. R.
  International Lead Zinc
  Research Organization Inc.
  292 Madison Avenue
  New York
  New York   10017
McCABE E. B,
  International Lead Zinc
  Research Organization Inc.
  1202 Frisch Road
  Madison
  Wisconsin   53711
MCCARTY D, L.
  3810 East Coast Highway
  Suite One, Corona del Mar
  California   92625

MCNEIL j.
  Texas Technical University
  School of Medicine
  Suite H, 6211 Edgemere
  El Paso
  Texas   79925

McNESBY J. R.
  National Bureau of Standards
  Washington D.C.   20234

METZGER R.
  Department of Physical
  Science
  San Diego State University
  San Diego
  California   92115

METZGER S.
  Department of Physical
  Science
  San Diego State University
  San Diego
  California   92115

MICHAELSON I. A.
  University of Cincinnati
  College of Medicine
  Department of Environmental
  Health
  3223 Eden Avenue
  Cincinnati
  Ohio   45219

MITCHELL R. I.
  Battelle Columbus Labora-
  tories
  505 King Avenue
  Columbus
  Ohio   43201

MORRIS S.
  Brookhaven National
  Laboratory
  Upton
  New York   11973

-------
                             2511
MRAK E. M.
  Environmental Protection
  Agency
  University of California
  Davis
  California

MUIR W.
  Executive Office of the
  President
  Council on Environmental
  Quality
  722 Jackson Place NW
  Washington D.C.   20006

NEEDLEMAN H.L.
  Children's Hospital
  Medical Center
  300 Longwood Avenue
  Boston
  Massachusetts   02115

NELSON N.
  New York University
  37 Detries Avenue
  North Tarry Town
  New York   10591

O'KEEFFE A. E.
  Environmental Protection
  Agency
  Chemistry and Physics
  Laboratory
  National Environmental
  Research Center
  Research Triangle  Park
  North  Carolina    27711

 OMENN G.
  Office of  the  Chairman
  B-413
  Atomic Energy  Commission
  Washington D.C.   20545

 OREL J.  V.
   Environmental  Consultant
   522 Warner Avenue
   Los Angeles
   California   90024

 PALLOTTA A.  J.
   Environmental Protection
   Agency
   401 M Street SW
   East Tower Room 401
   Washington D.C.   20460
PETERSON R. W.
  Chairman of the President's
  Council on Environmental
  Quality
  722 Jackson Place NW
  Washington D.C.   20460

RALL D.
  National Institute of
  Environmental Health Sciences
  P.O. Box 12233
  Research Triangle Park
  North Carolina   27709

REEF A.
  AMAX Inc.
  1270 Avenue of the Americas
  New York
  New York   10020

REEVES A.
  Wayne State University
  School of Medicine
  540 East Canfield
  Detroit
  Michigan   48201

REUTER L.
  Sanitary Engineer
  Research Branch
  U.S. Army Medical
  R & D Command
  Washington D.C.   20314

RUBIN  R.  J.
  Johns Hopkins University
  School  of Hygiene and
  Public  Health
  615  N Wolfe  Street
  Baltimore
  Maryland   21205

 SANTOLUCITO J.  A.
  Environmental Protection
  Agency
  Pesticide  and Toxic
   Substances  Effects  Lab.
  National Environmental
   Research Center
   Research Triangle Park
   North Carolina   27711

 SCHUCK E. A.
   Environmental Protection
   Agency
   National Environmental
   Research Center
   P.O. Box 15027
   Las Vegas
   Nevada   89114

-------
                              2512
SCHWING R. C.
  General Motors Corporation
  Technical Center
  Warren
  Michigan   48090

SILBERGELD E. K.
  Johns Hopkins University
  615 N Wolfe Street
  Baltimore
  Maryland   21205

STARA J. F.
  Environmental Protection
  Agency
  Office of Research and
  Development
  National Environmental
  Research Center
  Cincinnati
  Ohio   45268

STEELE T. D.
  Department of Interior
  Geological Survey - Water
  Resources Division
  Quality of Water Branch
  Washington D.C.

SUTHERLAND L. C.
  Wyle Laboratories
  128 Maryland Street
  El Segundo
  California   90245

TER HAAR G.
  Ethyl Corporation
  Research and Development
  Department
  1600 West Eight Mile Road
  Ferndale
  Michigan   48220

TRAKOWSKI A. C.
  Environmental Protection
  Agency
  Waterside Mall
  401 M Street SW
  Washington D.C.   20460

WALSH R. G.
  Office of  Research and
  Development
  Environmental Protection
  Agency
  Washington D.C,   20460
WARNER P. O.
  Wayne County Department
  of Health
  Air Pollution Control
  Division
  1311 East Jefferson
  Detroit
  Michigan   48207

WEAVER N. K.
  American Petroleum
  Institute
  1801 K Street NW
  Washington D.C.   20006

WEIR F. W.
  Biomedical Science Dept.
  Research Laboratories
  General Motors Corporation
  Technical Center
  Warren
  Michigan   48090

WEISS B.
  University of Rochester
  School of Medicine and
  Dentristry
  Department of Radiation
  Biology and Biophysics
  Rochester
  New York   14642

WESOLOWSKI J. J.
  Air and Industrial Hygiene
  Laboratory
  California State Depart-
  ment of Health
  2151 Berkeley Way
  Berkeley
  California   94704

WETHERILL G. W.
  Department of Planetary
  and Space Science
  University of California
  Los Angeles
  California   90024

WIERZBICKI  R.
  3745 West 56th  Street
  Chicago
  Illinois   60629

-------
                             2513
YANIV S. L.
  Environmental Protection
  Agency (HM-571)
  Washington D.C.    20460

YU M.-H.
  Huxley College of Environ-
  mental Studies
  Wast Washington State
  College
  Bellingham
  Washington   98225
ZAVON M, R,
  Ethyl Corporation
  451 Florida Avenue
  Baton Rouge
  Louisiana   70801

ZBAR J. E.
  Bellevue Hospital
  New York City
  2690 Kennedy Boulevard
  Jersey City
  New Jersey   07306
                           U.S.S.R,
SANOTSKY A.
   Institute of Industrial
   Hygiene and Occupational
   Diseases
   Academy of Medical Sciences
   31 Meyerovskij Prozd
   Moscow E 273
                           VENEZUELA
 SALAS-BERTI M.
  Direccifin de Malariologla
  y  Saneamineto Ambiental
  Ministerio  de Sanadad y
  Asist,  Social
  Apartado 2676
  Caracas 101

-------
INTERNATIONALE ORGANISATIONEN



 INTERNATIONAL ORGANIZATIONS



ORGANISATIONS INTERNATIONALES



ORGANIZZAZIONI INTERNAZIONALI



 INTERNATIONALE ORGANISATIES

-------
                              2517
   ECONOMIC AND SOCIAL COMMITTEE OF THE EUROPEAN COMMUNITIES
DE GRAVE M.
  Avenue Charles Woeste 208
  1090 Bruxelles
  (Belgique)
PEEL J. H.
  Warren Court Farm West
  Tytherley near Salisbury
  Wiltshire (United Kingdom)
                      EUROPEAN PARLIAMENT
KUSTER B.
  Commission Santfi Publique
  Centre European du Kirchberg
  Luxembourg (Grand Duch£)
               FOOD AND AGRICULTURE ORGANIZATION
GRIFFITHS R. C.
  Fishery and Marine Science
  Officer
  c/o Unesco
  7 Place de Fontenoy
  75007 Paris  (France)
              INTERNATIONAL ATOMIC ENERGY AGENCY
KOBAYASHI S.
  Radiation Biology Section
  Division of Life Sciences
  P.O. Box 645
  Kartner Ring 11
  1011 Vienna (Austria)
                  INTERNATIONAL LABOUR OFFICE
LARRUE M. G.
  Bureau de Paris de I1Office
  International du Travail
  Paris  (France)

-------
                               2518
         INTERNATIONAL ORGANIZATION FOR STANDARDIZATION
SIMONSGAARD V.
  Dansk Standardiseringsrad
  Aurehtfvej 12
  2900 Hellerup (Danmark)
            INTERNATIONAL UNION ON HEALTH EDUCATION
ORJOLLET M.
  12 rue Gustave LeBon
  75014 Paris  (France)
     ORGANIZATION FOR ECONOMIC COOPERATION AND DEVELOPMENT
ALEXANDRE A.
  2 rue Andr6 Pascal
  75775 Paris-Cedex 16
   (France)

HAINES J. A.
  2 rue AndrS Pascal
  75775 Paris-Cedex 16
   (France)

HUNTER G.
  2 rue AndrS Pascal
  75775 Paris-Cedex 16
   (France)

IDMAN M.
  2 rue Andr6 Pascal
  75775 Paris-Cedex 16
   (France)
KRAMISH A.
  U.S. Delegation to the
  O.E.C.D.
  19 rue de Franqueville
  75016 Paris (France)

NOBBS C.
  2 rue Andr€ Pascal
  75775 Paris-Cedex 16
  (France)

WYZGA R.
  Central Analysis and
  Evaluation Unit
  2 rue Andre1 Pascal
  75775 Paris-Cedex 16
  (France)
                        UNITED NATIONS
SERWER D.
  U.K. Environment Programme
  Geneva Liaison Office
  Palais des Nations
  Geneva  (Switzerland)

-------
                              2519
UNITED NATIONS EDUCATIONAL, SCIENTIFIC AND CULTURAL ORGANIZATION
BATISSE M.
  DSpartement des Sciences de
  1'Environnement et des Re-
  cherches sur les Ressources
  Naturelles
  7 Place de Fontenoy
  75700 Paris (France)

von DROSTE B.
  Division des Sciences
  Ecologiques
  7 Place de Fontenoy
  75700 Paris (France)

FREDERIKSEN M.
  Section de la Recherche
  dans le domaine des
  Sciences de I'IngSnieur
  7 Place de Fontenoy
  75700 Paris (France)
GILBRICH W.
  Division des Sciences de
  1'Eau
  7 Place de Fontenoy
  75700 Paris  (France)
MAHEU R.
  Director-General
  7 Place de Fontenoy
  75700 Paris  (France)
                   WORLD HEALTH ORGANIZATION
DIETERICH B. H.
  Division of Environmental
  Health
  Avenue Appia
  1211 Geneva 27
  (Switzerland)

FALK H.
  Avenue Appia
  1211 Geneva 27
  (Switzerland)

KttHNER A.
  Avenue Appia
  1211 Geneva 27
  Switzerland

KUMPF J.
  Regional Office for Europe
  Scherfigsvej 8
  Copenhagen 0
  Denmark
LAMBO T. A.
  Deputy Director General
  Avenue Appia
  1211 Geneva 27
  (Switzerland)

TOMATIS L.
  International Agency  for
  Research on Cancer
  Unit of Chemical Carcino-
  genesis
  150 Cour Albert Thomas
  69008 Lyon
   (France)

VOUK V. B.
  Division of  Environmental
  Health
  Avenue Appia
  1211 Geneva  27
   (Switzerland)

-------
                               2520
            COMMISSION OF THE  EUROPEAN  COMMUNITIES
AMAVIS R.
  Direction Protection
  sanitaire
  Centre Louvigny
  23 Avenue Monterey
  Luxembourg  (Grand-Duche1)

ANGELLETTI L. M.
  C.E.N., D.P.S.
  Bolte Postale  6
  92260 Fontenay-aux-Roses
   (France)

BATTI R.
  C.E.N., D.P.S.
  Bolte Postale  6
  92260 Fontenay-aux-Roses
   (France)

BERAUD R. C.
  Service Juridique
  B/ll-64
  200 rue de  la  Loi
  1040 Bruxelles
   (Belgique)

BERLIN A.
  Direction Protection
  sanitaire
  Centre Louvigny
  23 Avenue Monterey
  Luxembourg  (Grand-Duch§)

BERTINCHAMPS A.  J.
  Direction G6n6rale de la
  Recherche, Science et
  Education
  200 rue de la Loi
  1040 Bruxelles
  (Belgique)

BIGLIOCCA C.
  C.C.R.  Euratom
  21020 Ispra
  Varese (Italia)

BOURDEAU P.
  Direction GenSrale de la
  Recherche, Science et
  Education
  200 rue de la Loi
  1040 Bruxelles
  (Belgique)
CARPENTIER M.
  Service Environnement
  200 rue de la Loi
  1040 Bruxelles
   (Belgique)

DEL BINO G.
  Service Environnement
  200 rue de la Loi
  1040 Bruxelles
   (Belgique)

DEPAUS R.
  C.C.R. Euratom
  Petten
   (Netherlands)

DI FERRANTE E.
  Direction G6n6rale de la
  Recherche, Science et
  Education
  200 rue de la Loi
  1040 Bruxelles
   (Belgique)

FACCHETTI S.
  C.C.R. Euratom
  Chemistry Division
  21020 Ispra
  Varese (Italia)

GAGLIONE P.
  C.C.R. Euratom
  21020 Ispra
  Varese (Italia)

GEISS F.
  C.C.R. Euratom
  21020 Ispra
  Varese (Italia)

GIBB J. M.
  Direction GenSral de
  1'Information Scientifique
  et Technique et Gestlon de
  1'Information
  29  rue Aldringen
  Luxembourg (Grand-Duchfi)

HENTZ P.
  Direction SScuritfi et
  Mfidecine de Travail
  Centre Louvigny
  23  Avenue Monterey
  Luxembourg (Grand-Duch§)

-------
                              2521
HILLERY P. J.
  Vice-President
  200 rue de la Loi
  1040 Bruxelles
  (Belgique)

KARCHER W.
  C.C.R. Euratom
  Petten
  (Netherlands)

KARHAUSEN L.
  C.E.N., D.P.S.
  Bolte Postale 6
  92260 Fontenay-aux-Roses
  (France)

MAGNAVAL R.
  C.E.N., D.P.S.
  Bolte Postale 6
  92260 Fontenay-aux-Roses
  (France)

MULLER W.
  C.E.N., D.P.S.
  Bolte Postale 6
  92260 Fontenay-aux-Roses
  (France)

MUNTAU H.
  C.C.R. Euratom
  21020 Ispra
  Varese  (Italia)

OTT H.
  Direction General de la
  Recherche, Science et
  Education
  200 rue de la Loi
  1040 Bruxelles
  (Belgique)
RECHT P.
  Direction Protection
  sanitaire
  Centre Louvigny
  23 Avenue Monterey
  Luxembourg  (Grand-Duche")

SCHORN H.
  C.E.N., D.P.S.
  Bolte Postale 6
  92260 Fontenay-aux-Roses
   (France)

SCOPPA P.
  C.C.R. Euratom
  Gruppo di Biologia
  21020 Ispra
  Varese  (Italia)

SHANKS M.
  Directorate General for
  Social Affairs
  200 rue de la Loi
  1040 Bruxelles
   {Belgique)

SHEETS J.
  Direction Protection
  sanitaire
  Centre Louvigny
  23 Avenue Monterey
  Luxembourg  (Grand-Duche")

TREU-RICCO' G.
  Direction Protection
  sanitaire
  Centre Louvigny
  23 Avenue Monterey
  Luxembourg  (Grand-Duche*)

VAN DER VENNE M.
  Direction Protection
  sanitaire
  Centre Louvigny
  23 Avenue Monterey
  Luxembourg  (Grand Duch£)

-------
                              2522
                   ADMINISTRATIVE STAFF
Conference Officers:              D.  BLAND (EPA)
                                  A.  BONINI (CEC)
                                  E.  MILLAR (CEC)
                                  R.  MILLAR (CEC)
                                  T.  SONNEN (CEC)
Press Officers:                   G.  ANOUIL (CEC)
                                  J.  PARRISH (EPA)
                                  R.  REPOND (WHO)
Editorial Board:                  W.  BRAY (CEC)
                                  B.  O'NEILL (CEC)
Technical Services:               R.  LINSTER (CEC)
Secretariat:                      S. BRAMAN (WHO)
                                  A. DOYLE (EPA)
                                  M. PASSERINI  (EPA)
                                  L. PROOST (CEC)

-------
VERZEICHNIS






   INDEX






   INDEX






  INDICE






   INDEX

-------
                             CXIII
AASETH, J., 913
ADLER, M.W., 77.
ALBERT, R.r 1167, 2061, 2O68,
  2069
ALESSIO,  L., 1123, 1129, 1130,
  1167
ALLART-DEMUL, C., 1303
ALLEGRINI, M., 1697
ALLEN, J.R., 385, 397
ALTMAN, D.G., 269
ALTSHULER, B., 2O61
ALTSHULLER, A.P., 21O9
ANDERSON, J., 1449, 1461, 1468,
  1469
ANGERER,  J., 1317, 1327, 1328
ARHIRII,  M., 339
ARONOW, R.,  1177
ARSAC, F., 603
AOBERT, M.,  1613

BABCOCK,  L.R.,  2083
BACKHAUS, F., 2231
BAKER,  F.D., 879
BARHAD, B.,  333
BARQUET,  A., 695
BARRATT,  R.S.,  1397,  1779
BARSAN, E.T.,  1073
BARTH,  D.S.,  1875,  1877,  1921,
   1922,  1925, "~I9~2"6,  1928,  1930,
   1931,  1933,  1934,  1935,  1939,
   194O, 1950,  1959,  I960,  1962,
   1963, 2073,
BASTENIER, H.,  13O3
BATES,  D.V., 10O7,  1967,  1978,
   1979, 2001,  2003,"TT6"2
BATTI, R., 1531
BEACONSFIELD, P., 2397
BECK, E.G., 1031, 1O40, 1041
BEITZ, L., 1417, 1430
BELCHER, R., 1779
BELL, A., 1101
BENARIO, M., 2169
BENINSON, D., 845, 1878, 1924,
  1927, 1934, 1935, 1941, 1949,
  2297
BENSON, F.B., 423
BERGLUND, B., 119
BERGLUND, U., 119, 142
BERLIN, A.,  552, 611, 629, 693,
  859, 1087, 1100, 1238, 1959,
  2OO3, 2185, 2254, 2257
BERLIN, H.,  156, 491, 895, 1259
BERNA, H., 231
BERNER, A., .1729
BERNSTEIN, A.D., 1O5, 116, 117,
  1029, 2298
BERNSTEIN, D.,  431
BIANCO, A.,  1039
BIERSTEKER,  K.,  1881, 1924,  1926,
  1933, 1934, 1943, 1957, 1958,
  1959
BIGNON, J.,  1189, 1196,  1197
BINDER, R.E., 669
BITTEL, R.,  714, 1441,  1449,
  1469
BLACKBURN, C.R.B., 63
De BOECK,  R.,  1131
Den  BOER,  M.C.,  1247
BOGDANOVIC,  E.f  2271
BONNAUD,  G., 1189
BONNEFOUS, M.,  51O
 Underlined numerals denote authors
 Arabic numerals denote participants in discussions

-------
                              CXIV
BORDAS, E., 1145
del BORGHI, M., 1807
BORLAUG, N.f 2397
de BORTOLI, M., 1287
BOTZENHART, K., 1757
BOUDENE, C., 603, 612
le BOUFFANT, L., 1645, 1651
BOUHUYS, A., 669, 675
BOUQUXAUX, J., 1239, 1298
BOURBON, P., 269
BOURDEAU, Ph., 263 363, 366,
  510
BOUVILLE, A., 1531
BRADEN, M., 485
BRAETTER, P., 2255, 2301
BRAMAN, R.S., 117, 258, 1328,
  1363, 137O, 1397, 1954
BRAVO  A, H., 468, 2O91, 2160,
  2161
BREIDENBACH, A.W., 751
BRILLE, D., 269, 356, 667
BROCKHAUS, A., 781
BROMBERG, P.A., 1989
BROOKS, A.G.F., TT_
BRUAUX, P., 1131, 1143
                                CACCURI, S., 1823
                                CAGNETTI, P. , 1451
                                CALANDRA, J.C., 772
                                CANTON, J.H., 1479, 1489
                                CAPURRO, P.U., 1579
                                CARNOW, B.W., 45, 313, 353, 266.
                                  368, 588, 1129, TT6~7
                                CARPENTER, L., 1729
                                CARPI dl RISMINI, A., 2397
                                CARSTENS, L.A., 385
                                CARSTENSEN, J., 969
                                CARTER, M.H., 1399
                                del CASTILHO, P., 2185
                                CASOLA, D., 1693
                                CERNIK, A.A., 1207, 1221, 1237.
                                  2254        	
                                CERQUIGLINI-MONTERIOLO, S., 13O1
                                  1383                      """"*•
                                CHAMBERS, P., 104, 259, 397, 51O.
                                  7O2, 2045
                                CHANTEUR, J., 2386
                                CHAPMAN, R., 193, 207, 208, 21O,
                                  645, 658
                                CHARLTON, J., 112O
                                CHATTOPADHYAY, A., 1685
                                CIALELLA, N., 2225
BROCH, J., 781, 791, 1O4O, 12O6 CICOLELLAr A., 1661
de BRUIN, A., 259, 911, 2O59
BUCKET, J.P., 631, 887, 2185
BULCRAIG, W.R., 2O9
BURGER, E.J.jr.,  145,  155_,
   156,  157
BUSH,  B., 879, 885, 911,  1339   CtggfB'
                                CIGNA ROSSI, L., 1451
                                C1AEYS-THOREAU, F., 1131
                                CLAUDE, J.R., 397
                                CLAYTON, J.W.Jr., 371, 383, 383
                                            ,F., 260, 1028,
BUSTUEVA, K., 10O9
                                COFFIELD, T.H., 858
BUTLER, G.C., 1431, 1884, 1930  _ft_.  _
  1937, 1948, 1955, 1956, 1978  COIN' L- '
BUXTON, R. St.J.,  1113
BYRNE, A.R., 245,  258, 259, IW/***™'  "• • 1M
  1370
                                COLMAN, R. , 485

-------
                             cxv
COLUCCI, A.V., 1043
COOPER, W.C., 555, 568, 569,
  1196
COTE, R.W., 47_
CROCKER, K., 2068
CROSSMANN, G., 14O7
CRUZ, R., 1685
CUCU, M., 333

DAHL, R., 2231
DAMS, E., 513, 1430
DAMS, R., 409
DANIEL,  H., 1645, 1651
DANIEL,  J.W., 877/ 902, 1041
DANIELSON, L., 116, 885
DAVID,  0., 588, 1549
DAVIDOW, B., 545
DAVIES,  J.E., 695
DAVOUST, P., 2384
DEAN, G., 190, 643
DEHNEN,  W., 781
DELEANU,M., 1583
DELCARTE, E., 1675
DELVES,  H.T., 2215, 2258
DEMUYNCK, M., 409
DENNIS,  C.A.R., 1029, 1543,
  1953
DEPAUS,  R., 1341
von  DEPKA, J., 5O6
DERWENT, R.G., 1669
DESBORDES, J., 1199
DEVOTO,  G.f 1693
DIEHL,  J.F., 958, 1121
DIETERICH, B.H.,  2451
DIETL,  P., 1853
DI FERRANTE,  E.,  1956
DJURIC, D., 1829
DOBIN, D.D., 1223
DOBRYSZYCKA, W., 685
DOLGNER, R., 279
DONALDSON, W.T., 1399, 1405,
  1406
DONNIER, B., 1613
DONZELLI, A.,  231
DUBOIS, L., 1331
DUGANDZIC, M.  2285
DOMONT, M., 2231
DUNCAN, K.P.,  2175
DUNCAN, L.J.,  1241
DUPUIS, P.J.,  491, 1196,  1314

EDWARDS.  H.W., 1277,  1285,  1286
EFTHYMIOU, M.L., 1789
EGELS, W., 14O7
EISENBUD, M.,  431
ENGLISH, T.D., 401, 470,  471,
  472
EPSTEIN, S.S., 552, 569,  749,
  814, 1120, 2367, 2382,  2383,
  2384, 2385,  2386
van ESCH, G.J., 1017
ESPINOSA, M.E., 2091
EVENDIJK, J.E., 1351

FACCHETTI, S., 1287,  1298,  1299
FAGNIART, E.,  1675
FAIRWEATHER, F.A., 1113
FALK, H.L., 2331, 2348, 2349,
  2350
FAVRETTO, L.,  1511
FAVRETTO  GABRIELLI,   L.,  1511
FERRAIOLO, E.G., 18O7

-------
                             CXVI
FERRARI-BRAVO, F., 1797
FINE, P.R., 1223
FINKLEA, J., 193, 645
FISCHER, A.B., 1O31
FISHBEIN, L., 725, 749
FLOREY, C., 289
FONDIMARE, A,, 1199, 1206
FOURNIER, E., 1433, 1439, 1789
FREDERIKSON, M., 1959
FREEMAN, G., 685, 833, 844,
  10O7
FRENCH, J., 193, 645
FRIBERG, L., 2307, 2315, 2316
FRIEDMANN,  J., 507, 51O, 511
FRIEDRICHS, K.H., 715, 723,
  724
FUMAROLA,  G., 1807
FORIOSI, N.J., 833

GADDO,  P.P.,  1287
GAFFEY, W.R., 555
GAGE,  J.C., 895
GAGLIONE,  P., 1287
GARDI,  R., 1807
GARDNER,  D.E.,  7O5,  713,  714,
GARIBALDI, P.,  1287
GARNIER,  A.,  1441
GENT,  M.,  1263
GHELBERG,  N.W.,  1145,  1583
GHETTI, P.F.,  1957
GIANANI,  G.,  1697
GIBBS,  G.W.,  1197,  12O6,  2271,
   2296, 2297,  2301,  2302
GIBSON, R.J.W.,  289
von GIERKE, H.,  1249
GIOVANNINI, I.,  1797
 CLAUDE, P., 1341
GLOBUS, G., 5O7
GODIN, J., 603
GOERKE, W., 37
GOLDBERG, A.M., 793, 803
GOLDBERG, H., 193
GOLDSMITH, J.R., 62, 155, 189,
  382, 585, 675, 1889, 1923,
  1935, 1936, 1942, 1945, 195O,
  1956, 2156, 2165, 2179,
GOLDSTEIN, I., 1275
GONO,  E.,  591
GOOTJIS, P., 1316, 1951
GRAB,  B.,  339
GRADISKI,  D., 1631, 1661
de GRAEVE, J., 523
GRAOVAC-LEPOSAVIC, L.,  1829
GRASSO, C., 861,  878
GREENLAND, R.D.,  805
GREVE, P.A., 1479
GRIECO, A., 231
GRIFFIN,  H.E.,  23O5
GROLL-KNAPP,E.,  989
GROS, R.,  2225
GRUENER,  N.,  1067
GRUNSPAN,  M.J.,   773, 2OO3
GUINEE,  V.F,,  545,  551,  552,
   553, 1166

 HAAG, A., 1317
 HAASE, J., 1417
 HADDAD,  R., 451
 HAGEDORN-GOETZ, H., 2231
 HAIDER,  M., 475, 989, 999, 2393
 HARDWICK, D.F., 961
 HARKE, H.P.,  1327, 1773
 HARRISON, P.R., 1741
 HARRISON, R.M., 1111, 1285,  1349.
   1370, 1405, 1783

-------
                             CXVII
HAYES, C., 645
HAZUCHA, M., 1979, 2O01
HEBBELINCK,  D., 1303
HEM, B., 1189
HENDERSON, P.Th., 2047
BENIN, J.P., 1651
HERNBERG, S., 568, 692, 1129,
  1142, 2395
HERTZ, M.B., 1763
H 1CKEY, N., 658
BILLERY, P.J., 7_
HILPERT, K., 2231
BINE, C.H., 2O7, 260, 1O4O,
  113O, 1937, 2O29, 2298
HINTON, D.O., 1769
HISLOP, J., 959
HO, M.T., 1519
HOFMAN, B., 1017
BdGGER, D., 1247, 2157
HOLL, K., 612
HOLLAND, W.W., 39, 45
HOLM, S., 895, 902
HOLMQVIST, I, 613, 629
HORIE, Y., 2143
HOSEIN, H.R., 669
HOUCK, C.L., 879
BOWER, J., 591, 601
HOETER, F.G., 261, 352, 363,
  364
BOTCHINSON, T.C., 1685
BOTH, P., 715
HUUNAN-SEPPALA, A., 2263

ZMPENS, R., 1675
IOVENITTI, L., 1797
ZRWIG, L., 289, 360
IWANKIEWICZ, S., 685
IZMEROV, N.F., 2409
JACKSON, D.L., 161, 176, 177,
  178, 1956
JACOBSEN, M., 89, 211, 228, 229,
  1478, 1502, 2365
JACYSZYN, K., 685
JAMIN, P., 523
JANSEN, G., 513, 999, 2O44
JEANMAIRE, L., 2225, 2252, 2256
JERVIS, R.E., 1685
JOHNSON, D.L., 1363
JOOSTING, P.E.f 76, 20O5, 2029
JOST, D., 2115
JUHOS, L., 833

KAMINSKI, E.J., 551, 8O3, 1168
KARCHER, W.,  1341, 1349
KARHAUSEN, L., 367
O'KEEFFE, A.E., 21O9, 2164, 2166
  2168, 2169, 2172
KEITZ, E.L.,  1241, 1247, 1248
KELLER, M.D., 47
KEY ANY, J., 89
KILPIO, J.O., 2263
KIRCHMANN, R., 1675
KJELLSTR6M, T., 62, 1221, 1939,
  2197, 2252, 2254, 2257, 2299,
  2303, 2328, 2365, 2383
KLAHRE, ?•/ 2231
KLEINMAN, M., 431
KNAUTH, P., 2031, 2043, 2O44
KNEIP, T.J.,  431, 466, 473
KNELSON, J.H., 181, 189, 19O,
  361, 973     	
KOPPLE, J.D., 847
KORICANAC, Z., 2285
KOSTA, L., 245
KOTLAREK-HAUS, S., 685, 692, 693

-------
                            CXVIII
KRACKE, W., 1853                LLOYD, W.J., 1085
KREBS, H., 2397                 LO, Fa-Chun, 879
van der KREEK, F.W., 959, 1028, van LOON, J.,  1685,  1849
  1065, 2382                                   	   	
                                LORKE, P., 817, 830,  831
KREUZER, W., 601, 1853, 2O45,    _        	
  2299            	          LOSER, E., 817
RUMMER, J., 1303, 1314, 1315,   "WE* G'J'' 3°1' £45.
  1316, 1339                    LOWE, A.c.f 2O91
KUMPFM.J., 339, 363, 365, 1957 LYNAM, D.R., 543, 791
LACOURLY, G., 1441
LAFONTAINE, A., 382, 1131, 1942
  2437
LAMBO, T.A., 11
LANESE, R.R., 4J7
LANG, R., 659
LANGER, H., 1757
LANZOLA, E., 1697
LAO, R.C., 1331, 1339, 1340,
  1468
LAUER, G., 423, 2156, 2163
LAUWERYS, R., 542, 568, 631,
  831, 887, 1238, 2185
LEEDER, S.R.,  6_3, 76, 1111
LEFEVRE, M.J., 988, 2029
LEGRAND, M., 1131
LEHNERT, G., 1317
LEHTO, V.P., 1007, 2315
LELLOUCH, J., 269
LENGHEL, I., 1583
LEUNG, S., 1248
LEVERE, T.E., 493, 506
LEVY, D., 1263, 1275, 1276
LEVY, E.A., 1797
LIND, B., 2197
LINDVALL, T., 119, 1954
LINNMAN, L., 2197
 MAGADUR, J.L., 1631, 1661
rMAGE, D., 176, 207, 1275, 2O68
   2156, 2158, 2162, 2164, 2165,
   2168, 2170, 2174, 2176, 2178,
   2179, 2181, 2315, 2364
 MAGE, D.T., 2097
 MAGI, P., 857
 MAGNAVAL, R., 1531
 MAHEU, R., 1£
 MALONE, D.W., 1569
 MANOJLOVIC, N., 1O31
 MARCUS, A.H., 1505
 MARQUARDT, H., 16O7
 MARTIN, A.E., 1113, 112O, 1121,
   1122, 2305
 MARTIN, J., 973
 MARTIN, J.C., 1645. 1651
 MATERNE, D., 631
 McCABE, E.B., 1168
 MCDONALD, G.C., 1491
 McGUIRE, J.M., 1399
 MCNEIL, J.L., 571, 584, 586,
   588, 589
 McNESBY, J.R. , 1371, 1383
 MEININGER, J., 603
 van  MEIRHAEGE, A., 2O44
 MENENTI, M., 1797
 MERIAN, E., 929, 1396, 2175

-------
CXIX
MERLUZZI, F., 231
MICHAELSON, I.A., 805
MILIC, S., 1829, 2285
MILOVANOVIC, Lf., 2285
MIRE, B., 973
MITCHELL, C.A., 669
MITCHELL, R.I., £7, 62
MOKEMATKENGUEMBA, G., 178,
MOLDOVAN, N., 1583
MOLLARET, P.,  1938
MONCELON, B., 1661
MONCHAUX, G., 1189
MONKMAN, J.L., 1015, 1331
MOONEY, T.F., 1637
MOORE, M.R., 535, 1171, 2252
MOORE, W., 751
MOREAU, M.,  1315
MORGADE, C., 695
MORGAN, G.B., 2073
MORRESI, N., 231
MORRIS, S.C., 677, 683
MOSE, J.R.,  1617
MOULE, Y., 967
MRAK, E.M.,  1965
MURAYAMA, H., 91
MUSSENDEN, R., 833

NAGDA, N.L., 2083
NANGNIOT, P., 1675
NEEDLEMAN, H.L., 584, 1155, 4.4.
  1167, 1168, 1169, 1936T~2433
NEUBERGER, M., 989
NEWHOUSE, M., 383, 1263, 1953
NEWILL, V.A., 161
NIEUWSTRATEN, N., 1351
   NORBACK,  D.H.,  385
   NORSETH,  T.,  913
   NOTTEN, W.R.F., 2O47, 2O59, 206O
   NURNBERG, H.W. , 2231
   OLEKSYK,  E., 685
     9_61,' 1065 )'1953* 2350
   OMENN, G.S., 1563, 1952
   OREL, J.V., 2177
   ORIOL, P., 269
   OTT,  W.R., 2097
   OUW,  K.H., 11O1
   PACIGA, J., 1685
   PACKHAM, R.F., 1468
   PATTI, F., 2225
   PEAT, J.K., 6_3
   PECORA, L., 1823
   PELECH, L., 279
   PERROTEY, J., 1199
   PERRY, R., 1285, 1328, 1385,
     1405, 1783
   PETERS, R., 2397
   PETERSON, R.W., 3_
   PFANNHAUSER, W., 258, 858, 1328,
     1719, 2253
   PHAM, Q.T., 208, 470, 973, 988
   PHILP, J.M., 2350
   PICHE, L., 1314, 1468
              ., 830, 951, 958,
              , 2297
   PITTWELL, L.R. , 2303
   PIVA' C"
   POWELL, W., 833

-------
                              cxx
PRINZ, B.f 591, 1471, 1489
PTASNIK, J.A., 571

QOAGLIARDI, A., 1287
QOINOT, E., 1519

RABINOWITZ, M., 847
RAFFONELLI, A., 695
RAINSBORY, R., 2397
RALL, D.,  37
RAMACIOTTI, D., 659,  2316
REAY, J.S.S.,  1669
RECHT,  P., 157, 1894, 1928,
   1938, 1946,  1961,  2641
REEVES, A.,  724,  1637,  2385
RETHFELD,  H.,  1407,  1416
REUSMANN,  G.,  591
REUTER, L.,  1066
REY,  P., 659,  667
ROBERTS, T.M., 1685
ROELS,  H., 631,  887, 1237,
   2185
ROGGI,  C., 1697
RONDIA, D.,  523,  535, 1339
ROOSKEN, A.A., 1351
ROOTS,  L.M., 1113
van ROOYEN,  G.I.i 2382
 ROSCA,  Gh.,  1073
 ROSCA, S., 1Q73
 ROSIVAL, L., 877, 1187, 1237
 ROSSANO, A.T., 2083
 ROSSI, A», 1823
 ROBIN, R.J., 903, 911, 912
 RUDEN, H., 1757
 RUDOLF, W., 2115
 ROTENFRANZ, J.,  2031
RUTZEL, H., 2231
RYLANDER, R., 137, 477, 484,
  667

SABATINI, G.C., 1697
SADOUL, P., 973
SALAZAR, S., 2091
SANOTSKY, I.V., 1O09, 2349,
  2409, 2433
SANSONI, B., 1853
SANTARONI, G.P-., 1O3O, 1451
SANTOLUCITO, J.A., 7O2, 1051,
  2387, 2394
SAOERHOFF, M.W.,  805
SAVING, A., 931,  1749
SAYERS, M.H.P., 1207
SCASSELLATI-SFORZOLINI G.,  931,
  1749
SCHALLER,  K.H., 1O87
SCHNEIDER,  T.,  1954,  2071,  2155,
  2158,  2162,  2166,  2168,  2172,
  2174,  2175,  2178,  2182
SCHILLING,  R.S.F.,  669
SCHLATTER,  C.,  723
SCHLIPK6TER,  H.W.,  190,  228,
   369, 521, 667,  723, 771,
   2328
 SCHMIDT,  P.,  279, 354
 SCHOENBERG, J.B., 669
 SCHUCK, E.A., 2073, 2159, 2161,
   2164, 2171, 217T, 2176, 2177,
   2179, 2180, 2181, 2182
 SCROLLER, P.L., 1O17
 SCHULZ, C.O., 9O3
 SCHWING, R.C., 1491, 15O2, 1503,
   2178, 2181
 SCOPPA, M., 958
 SEBASTIEN, P., 1189
 SECCHI, G.C., 1123

-------
                             CXXI
SEIFERT, D., 1407
SERWER, D., 2383
EL-SEWEFY, A.Z., 1589
SHAHEEN, H.f 1589
SHAMS El-DEEN, A., 1589
SHAPIRO, I.M., 1155
SHAPIRO, M.A., 677
SHERWOOD, R.J., 471, 134O, 1954
SHUVAL, H.I. , 1067
SILBERGELD, E.K., 793, 814, 1O51
SILVERMAN, A.P., 1601
SKJAERASEN, J., 1233
SLATER, D.H., 1783
SHEETS, J., 1087, 2185
SMIDT, U., 1557
SMITH, R.G., 1637
SMITH, G., 1729
SORENSEN, S., 137, 477
STANKOVIC, B., 2285
STANKOVIC, M., 2285
STANKOWSKA, K., 685
STANLEY, R., 1729
STARA, J.F., 714, 751, 771, 772,
  813
STEELE, T.D., 1954, 1961, 217O
STEENSBERG, J., 472, 1O06, I960
  2432
STEPANEK, V., 1473, 1478, 1503
STEPHAN, W.I., 1779
STERN, A.C., 2143
STEVENS, L., 1O01
STEWART, H.N.M., 1669
STIDL, H.G., 989
STOEPPLER, M., 2231, 2259
STOOFF, W., 1479
STOPPS, G.J., 229
STUIK,  E.J., 537
STUPFEL, M.,  89, 77O, 844,
  1275, 1625. 1963, 2043
SUESS, M.J.,  339
SULAIMAN, A.B., 657, 683, 1952
SUTHERLAND, L.C., 484, 485, 491
SWYNGEDOUW, I., 1131
SYROTA, M.J., 399, 457, 459,
  462
SZADKOWSKI, D., 1841, 2183, 2249,
  2256, 2259
SZPERLINSKI, Z., 1711

TAKABATAKE, E., 2197
TATI, M., 2251
TAYOT, J., 1199
TER HAAR, G., 8O3, 1177
THOMAS, R.S., 1331
THOMAS, T.J., 1569
THOMPSON, J.M., 1779
TOMATIS, L., 1053, 2317,  2328,
  2329
TOMPKINS, E., 2197
den TONKELAAR, E.M., 1O17,  1O28,
  1029, 1030
TRAKOWSKI, A.C., 2455
TREMOLIERES, J., 19O2, 1921,
  1927,  1944, 1950
TRUFFERT, L., 2255,  23OO
TSUCHIYA, K.r 2197,  2351,  2364,
  2365
TULLIEZ, J., 921, 929
TOMASONIS, C.E., 879
TWIBELL, J.D.,  1385, 1397,  1398
TYTUN, A.r 545

VALENTA, P., 1416,  2231
VALJAREVIC, V.,  1829

-------
                              CXXII
VANINI, G., 1289
VEIL, S., 1, 25
VERBERK, M.M.f 20O5
VOINIER, B., 659
VOUK, V.B.,  467, 469, 473,
                                YORDANOV, D., 1815
                                ZAPHIROPOULOS, M., 1961
                                de ZEEUW, M., 1947
  1912, 1925, 1930, 1931, 1947  ZELENKO, V., 24J7
VUORI, E., 2263, 2296
WALDBOTT, G.L., 1575
WALLER, R.E., 77_, 89, 90
WANNAG, A., 1233, 1237, 1238
WARNER, P.O., 1001, 1006, 1007,
  1008
WASSERMANN, D., 1053
WASSERMANN, M., 103, 156, 396,
  683, 702, 713, 877, 1053,
  1065, 1066, 1439, 1933
WATANABE, H., 9^, 104
WEAVER, N.R., 369
WEBER, 0., 260, 2261, 2295, 2297,
  2299, 2301, 2302, 2303
WEICHERT, N., 1417
WEIR, F.W., 1989, 2001, 2O02,
  2003, 2004
WEISS, B., 2415, 2432, 2433
WESOLOWSKI, J.J., 471, 1729
WETHERILL, G.W., 847, 857, 858,
  859, 1298
WILCOX, S.L., 1241
WILLIAMS, H., 1601
WILLIAMS, M.K., 551
WILLIAMS, R., £7
WISSMATH, P., 1853
WOIDICH, H., 1719
WOOLCOCK, A.J., 63
WORTH, G., 1557
ZIELHUIS, R.L., 175, 364, 537,
  542, 543, 692, 104O, HOOT"
  1168, 2002, 2060, 2348, 2384
ZUNIC, R., 2386
ZWIERS, J.H.L., 1O17
YANIV, S.L.,  1249, 1260

-------
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