xvEPA
           United States
           Environmental Protection
           Agency
           Office of Health and
           Environmental Assessment
           Washington DC 20460
EPA-600/8-83-033F
August 1984
Final Report
           Research and Development
Revised Evaluation of
Health Effects
Associated with
Carbon Monoxide
Exposure:

An Addendum to the
1979 EPA Air Quality
Criteria Document for
Carbon Monoxide
  Final
  Report

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                            EPA-600/8-83-033F
                                August 1984
  Revised Evaluation of Health
Effects Associated with Carbon
       Monoxide Exposure:

 An Addendum to the 1979 EPA
Air Quality Criteria  Document for
         Carbon Monoxide

            Final Report
      Environmental Criteria and Assessment Office
     Office of Health and Environmental Assessment
        Office of Research and Development
        U.S. Environmental Protection Agency
      Research Triangle Park, North Carolina 27711

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                      NOTICE

This document has been reviewed in accordance with
U.S. Environmental Protection Agency policy and
approved for publication.  Mention of trade names
or commercial products does not constitute endorse-
ment or recommendation for use.
                       ii

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                                   ABSTRACT
     This addendum reevaluates  the  scientific data concerning health effects
associated with exposure  to  carbon  monoxide (CO) at ambient or near ambient
levels by providing:  (1) a concise summary of key health effects information
pertaining to relatively  low-level  CO exposure;  and (2) an  overview of the
limited volume of new evidence on the subject.  This reevaluation is performed
in light of  the  diminished  value of  studies  by  Dr.  Wilbert Aronow on human
health effects of  exposure  to low levels of CO.   These studies figured in to
the preparation of the U.S.  Environmental Protection Agency's 1979 Air Quality
Criteria Document for Carbon Monoxide and to  the  Agency's  proposed retention
of the  8-hour and revision of  the  1-hour primary standards for  CO  (45 FR
55066; August 18, 1980).
                                        i i i

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                          CONTRIBUTORS AND REVIEWERS
The following authors contributed to the writing of this addendum.

Dr. Lester D. Grant, Environmental Criteria and Assessment Office,
  U.S. Environmental Protection Agency, Research Triangle Park, NC  27711

Mr. James A. Raub, Environmental Criteria and Assessment Office,
  U.S. Environmental Protection Agency, Research Triangle Park, NC  27711

Dr. Vernon A. Benignus, Neurotoxicology Division, Health Effects Research
  Laboratory, U.S. Environmental Protection Agency, Building 224H, University
  of North Carolina, Chapel Hill, NC  27514.

Dr. David J. McKee, Strategies and Air Standards Division, Office of Air
  Quality Planning and Standards, U.S. Environmental Protection Agency,
  Durham, NC  27701


     A working draft of this addendum was circulated for preliminary peer-review
to the individuals listed below, and comments received were taken into account
in preparing the present addendum.  The views expressed in the present addendum
should not be taken, however, as representing those of any single individual or
group listed here.

Dr. Laurence D.  Fechter
Department of Environmental Health Sciences
School of Hygiene and Public Health
John Hopkins University
615 North Wolfe Street
Baltimore, MD  21205

Dr. George M. Goldstein,
Clinical Studies Branch, Health
  Effects Research Laboratory
U.S. Environmental Protection Agency
Building 224H, University of
 North Carolina
Chapel Hill, NC  27514

Dr. Jack D.  Hackney
Environmental Health Service
Rancho Los Amigos Hospital
7601 Imperial Highway
Downey, CA  90242
                                        i v

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                          CONTRIBUTORS AND REVIEWERS
Dr. Stephen M. Horvath
Institute of Environmental
University of California
Santa Barbara, CA  93106
Stress
Dr. Victor G. Laties
Environmental Health Sciences Center
University of Rochester, School of Medicine
Rochester, NY  14642
Dr. Patricia M. Mihevic
Institute of Environmental
University of California
Santa Barbara, CA  93106
Stress
Dr. John J. O'Neil
Clinical Studies Branch, Health
  Effects Research Laboratory
U.S. Environmental Protection Agency
Building 224H, University of
 North Carolina
Chapel Hill, NC  27515

Dr. David S. Sheps
Department of Cardiology
University of North Carolina
  School of Medicine
Chapel Hill, NC  27514

Dr. Jaroslav J. Vostal
Executive Department
General Motors Research Laboratories
Warren, MI  48090
Dr. Jeames A. Wagner
Institute of Environmental
University of California
Santa Barbara, CA  93106
Stress

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                    CLEAN AIR SCIENTIFIC ADVISORY COMMITTEE
     The substance  of  this  document was  independently peer-reviewed  in public
session by the Subcommittee on Air Quality Criteria for Carbon Monoxide,  Clean
Air  Scientific  Advisory Committee, and  the  Environmental  Protection Agency
Science Advisory  Board.   The Committee's findings and recommendations on the
scientific basis for a revised national ambient air quality standard for carbon
monoxide can be found in Appendix C.

Chairman, Clean Air Scientific Advisory Committee:

Dr. Morton Lippman                 i
Institute of Environmental Medicine
New York University
New York, NY  10016

Staff Director, Science Advisory Board:

Dr. Terry F. Yosie                 !
U.S. Environmental Protection Agency
Science Advisory Board
401 M Street, S.W.
Washington, DC  20460

Members and Consultants:

Dr. Stephen M.  Ayres
Department of Internal  Medicine
St. Louis University Medical Center
1325 S. Grand Blvd.
St. Louis, MO  63104               ;

Dr. Edward Crandall                i
Division of Pulmonary Disease
Department of Medicine
University of California
Los Angeles, CA  90024

Dr. Robert Dorfman                 !
Department of Economics
Harvard University
325 Littauer
Cambridge, MA  02138

Dr. Ronald J. Hall
Ministry of the Environment
Box 39                             j
Dorset, Ontario  TOA1EO

Dr. Ian T. Higgins
Department of Epidemiology         i
University of Michigan             ;
109 Observatory Street
Ann Arbor, MI  48109
                                       vl

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              CLEAN AIR SCIENTIFIC ADVISORY COMMITTEE (continued)
Dr. Warren B. Johnson, Jr.
Director, Atmospheric Sciences Center
Advanced Development Division
SRI International
333 Ravenswood Avenue
Menlo Park, CA  94025

Dr. Lewis H. Kuller
Professor and Chairperson
Department of Epidemiology
Graduate School of Public Health
University of Pittsburg
130 Desoto Street
Pittsburgh, PA  15260

Dr. Timothy Larsen
Department of Civil Engineering
Mail Stop FC-05
University of Washington
Seattle, WA  98195

Dr. Victor Laties
Environmental Health Sciences Center
School of Medicine, Box RBB
University of Rochester
Rochester, NY  14612

Dr. Lawrence Longo
Division of Perinatal Biology
Loma Linda University
School of Medicine
Loma Linda, CA  92350

Mr. Donald Pack
1826 Opalocka Drive
McLean, VA  22102

Dr. John Seinfeld
Department of Chemical Engineering
206-41
California Institute of Technology
Pasadena, CA  91125

Mr. Bill Stewart
Executive Director
Texas Air Control Board
6330 Highway 290 East
Austin, TX  78723
                                      VI

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               CLEAN AIR SCIENTIFIC ADVISORY COMMITEE (continued)
Dr. Michael Treshow
Department of Biology
University of Utah
Salt Lake City, UT  84112

Dr. James Ware
Department of Biostatisties
Harvard School of Public Health
677 Huntington Avenue
Boston, MA  02115
                                     VI I I

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                               TABLE OF CONTENTS

                                                                      Page

INTRODUCTION	    1

MECHANISMS OF ACTION	•;	    2

HEALTH EFFECTS OF LOW LEVEL CO EXPOSURES 	    6

  CARDIOVASCULAR EFFECTS 	    7
  NEUROBEHAVIORAL EFFECTS 	    12
  EFFECTS OF CO EXPOSURE ON FIBRINOLYSIS 	    15
  PERINATAL CO EFFECTS 	    17

POPULATIONS AT RISK	    19

SUMMARY AND CONCLUSIONS 	    23

REFERENCES 	    27

APPENDIX A	    A-l

APPENDIX B 	    B-l

APPENDIX C 	    C-l
                                       IX

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                                  INTRODUCTION
     On April 30, 1971, the Environmental Protection Agency promulgated (36 FR
8186) national  ambient air quality standards  (NAAQS) for carbon monoxide (CO)
under section  109 of  the  Clean Air Act.  Identical primary  and  secondary
standards were  set  at  levels of 9 ppm,  8-hour  average,  and 35 ppm, 1-hour
average, neither to be exceeded more than once per year.   The scientific bases
for these standards  are  contained in the Air Quality  Criteria Document for
Carbon  Monoxide (U.S.  Department of Health,  Education, and  Welfare,  March,
1970, AP-62).   The  1971  standards were primarily based on work by Beard and
Wertheim (1967)  suggesting that low-level  CO exposures resulting  in carboxy-
hemoglobin (COHb) levels of  2 to 3 percent are associated with impairment of
ability to discriminate time intervals, a central nervous  system  (CNS) effect.
The revised Air Quality Criteria Document for Carbon Monoxide (U.S.  EPA, 1979)
indicated that this study is no longer considered to provide credible evidence
for such CNS effects occurring at 2-3% COHB and, therefore, does not represent
a sound  scientific  basis  for the standard as discussed in an August 18, 1980
EPA proposal notice  (45 FR 55066).  However, medical evidence published since
1970 indicated  at the  time of proposal that aggravation of angina and other
cardiovascular diseases may occur at COHb levels as low as 2.7 to  2.9 percent.
Assessment of this  and  other medical  evidence  led EPA to propose, on August
18, 1980, retention of the 8-hour primary standard level  of 9 ppm  and revision
of the 1-hour standard level  from 35 ppm to 25 ppm (45  FR  55066).
     The 1980 proposal was  based in part on several  health studies  conducted
by Dr.  Wilbert Aronow (Aronow et al. ,  1972;  Aronow and  Isbell, 1973;  Aronow et
al., 1974; Aronow et al.,  1974; Aronow and Cassidy, 1975;  Aronow et  al., 1977;
Aronow,   1978).    Based  on  evaluation  of these studies  in  1979 by  EPA staff,
their expert  consultants,  and the Agency's  Science  Advisory  Board,  it was
concluded that  these studies  demonstrate  human health effects  of carbon
monoxide that should be  considered  by the Agency in reconfirming  existing or
proposing new NAAQS  for  CO.   The Aronow studies were an important element in
identifying the  blood carboxyhemoglobin (COHb)  levels that represent a health
concern  for  sensitive  individuals.   This  "critical" range was defined as
2.7-3.0 percent.  An additional study by Aronow (1981)  later reported findings

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suggesting that aggravation of angina symptoms, i.e., small but statistically
significant decreases  (~10%)  in time to onset of exercise-induced angina, may
occur in angina patients at COHb levels as low as 2.0 percent.
     Since the CO  standard was  proposed by EPA  in  1980,  news media reports
appearing in  early 1983  indicated that the Food and Drug Administration (FDA)
raised questions regarding the technical  adequacy of several studies conducted
by Dr. Aronow on  experimental drugs, leading to FDA rejection of use of the
drug study data.   While  there was no specific direct  evidence that similar
problems might exist  for the CO studies conducted by Dr. Aronow, EPA judged
that an independent assessment of these studies was advisable prior to a f,inal
NAAQS decision on  CO.   An expert committee was empaneled by EPA and met with
Dr. Aronow to discuss  his studies and to examine limited available data and
records from  his  CO studies.   In their report, the committee (chaired by Dr.
Stephen M. Horvath,  Director  of the Institute of Environmental Stress, Uni-
versity of California-Santa,Barbara) concluded that EPA should not rely on Dr.
Aronow1s data due  to  concerns regarding problems associated with the studies
which substantially limit the validity and usefulness of those study results
(Horvath et a!.,  1983).   Dr.  Aronow submitted a detailed  reply to EPA that
disputed, but did  not  effectively refute, the major points  raised by the com-
mittee report (Aronow, 1983).
     The main purpose  of the  present addendum  is to reevaluate the scientific
data base concerning health effects associated with exposure to CO at ambient
or near  ambient exposure  levels,  in  light of the diminished  value  of the
Aronow studies  and taking  into  account any new findings that have become
available beyond those reviewed  in the revised Air Quality Criteria Document
for CO (U.S.   EPA,   1979).   This addendum is, accordingly, organized to provide:
(1) a  concise summary  of key  health effects information discussed in the 1979
document as pertinent  to characterization of health effects associated with
relatively low level  CO  exposures;  and (2) an overview of the limited new
evidence bearing on the  subject which has become available in the past several
years.
                             MECHANISMS OF ACTION
     The  1979  Criteria Document discussed extensive evidence indicating that
the binding  of CO  to  hemoglobin,  producing COHb  and  decreased oxygen  carrying

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capacity, results In decreased oxygen transport and uptake in most body tissues.
The resulting hypoxic state (impairing normal biochemical-physiological cellular
processes as a  function of increasing external CO exposure and consequently
increasing blood  COHb  concentrations),  it was concluded, probably represents
the main mechanism  of  action  underlying the  induction of  toxic effects by low
level CO exposures.
     Several important relationships between COHb levels and other physio-
logical parameters  discussed  in the 1979 Criteria Document have continued to
be the  subject  of evaluation  since  then.  Of much  importance  is the relation-
ship between external  CO exposure levels and consequent  increases  in blood
COHb levels.   Many factors,  discussed  in the  1979  Criteria  Document, can
affect  the  rate at  which COHb increases above pre-existing endogenous levels
of COHb in response to  inhalation of exogenous CO.   These include, for example,
the pattern of external CO exposure, as in the case of acute short-term exposures
                                                i
to high CO concentrations versus longer term exposure to  relatively low levels
of CO.   CO exposure-COHb  concentration  relationships  have been modeled by
Coburn  (Coburn  et al. , 1965), taking into account several pertinent  factors
(see Chapter 9  of the  1979 EPA  CO  Criteria  Document for discussion  of the
Coburn  model equations).   COHb levels predicted by  the  Coburn equations, as
depicted  in Figure  1,  are widely accepted as  the currently  best available
modeled estimates of  COHb levels likely to  result from varying CO concentra-
tions,  exposure  durations  and exercise levels.  It should be noted that some
questions have been raised regarding the specific mathematical approach employed
by Coburn in solving his equations to predict COHb concentration as a function
of time, considering appropriate physiological parameters (Venkatram and Louch,
1979; Ott and Mage, 1980; Marcus, 1980; Goldsmith, 1981;  Joumard et al., 1981).
However, the proposed  alternative approaches yield very similar estimated COHb
levels  to those projected by  Coburn's. approach.  In addition, actual blood COHb
concentrations  observed in response to  particular  external CO exposure situa-
tions have been consistent with those predicted by Coburn (Peterson and Stewart,
1975),  although further experimental verification would be useful to demonstrate
that the  Coburn equation accurately predicts uptake  and  excretion of  CO under
widely  varying  conditions.
     The  exact  mechanisms responsible for the  hypoxia  induced by CO  are  not
known.   The most widely accepted mechanism of  CO  toxicity has been  attributed

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X
O
O
                                                        20 LPM 50 PPM CO •
                            50 LPM 50 PPM CO
                                      4

                                   HOURS
     Figure 1. Blood COHb concentrations predicted by Coburn equations to
     occur as a function of exposure duration and ambient carbon monoxide
     concentrations under resting (10 LPM), light exercise {20 LPM), or heavy
     exercise (50 LPM) conditions. LPM  = liters per minute ventilation rate.
    Source:  U.S. EPA (1979).

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to the preferential binding of CO to hemoglobin which produces hypoxia by re-
ducing 02 transport by red blood cells to  the  tissues  and impedes the dis-
sociation of 02 from hemoglobin in the capillaries.  However, other mechanisms
have been postulated for  reducing oxygen transport.  It  is possible for CO to
bind to  intracellular  hemoproteins  such as myoglobin and cytochrome oxidase,
which depends on the relationship of oxygen tension (P02) and CO tension (PCO)
to CO binding  constants (Coburn,  1979).  The affinity of cytochrome oxidase
for CO is similar  to that for oxygen compared to myoglobin (30-50x) and hemo-
globin (220x) which would make it less likely to be responsible for impairment
of facilitated  diffusion  of  oxygen to the  mitochondria.  However,  if steep
oxygen tension  gradients exist between the extracellular  and intracellular
environment, then the P02 surrounding the mitochondria!  terminal oxidase would
be low enough  to have  increased binding with CO.   This hypothesis was tested
by Coburn (1979) in studies on isolated vascular smooth  muscle.  He concluded
that significant CO binding  to cytochrome  oxidase was  unlikely to be an iji
vivo mechanism  of  CO  toxicity in that particular tissue.  Myoglobin was also
unlikely because it  is absent or present  in only  small  quantities.   It is
possible that  CO binds to hemoproteins other than hemoglobin, myoglobin, or
cytochrome  oxidase.  Cytochrome P-450,  tryptophan  deoxygenase, and tryptophan
catalase all have high  enough binding affinities for CO in specific tissues to
be considered as possible candidates (Coburn, 1979).
     the binding of  CO to myoglobin in heart and skeletal  muscle may be high
enough to  reduce  intracellular oxygen  transport  in those tissues (Coburn,
1979; Agostoni  et al., 1980).   Using  a  computer simulation  of a three-
compartment  model  (arterial  blood, venous  capillary blood,  and tissue myo-
globin), Agostoni  et  al.  (1980) predicted  that conditions would be favorable
for formation  of carboxymyoglobin at COHb  levels  of  5-10%,  particularly in
areas where  the P02 was physiologically low (e.g., in subendocardium) and when
conditions  of  hypoxia, ischemia,  or increased metabolic  demands  were present.
This could  provide theoretical support for  experimental evidence of myocardial
ischemia, such  as  electrocardiographic irregularities and decrements in work
capacity discussed later.  However, it is  not known whether binding of CO to
myoglobin could cause  health effects (e.g., decreases in maximal oxygen con-
sumption during exercise) occurring at COHb levels as low as 4-5%.  Additional
research is  needed before this possibility  can be more definitively evaluated.

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     Whatever  the specific biochemical-molecular mechanisms involved  in  the
 induction  of CO toxicity in specific  tissues or organ systems, it is thought
 that  COHb  concentrations  in  the blood  represent  a meaningful and  useful
 physiological  marker by which to gauge the internal CO dose present at a given
 time due to  the combined cumulative contributions of:  (1) baseline endogenous
 production of  CO  by internal body processes; and (2) added CO body burden(s)
 resulting from inhalation exposure to  exogenous sources of CO.
                  HEALTH EFFECTS OF LOW-LEVEL CO EXPOSURES

     In  evaluating  CO-induced health effects in humans,  a crucial question
that must be addressed concerns blood COHb levels demonstrated to be associated
with effects  on specific organ systems. The COHb levels of concern vary with
the  patterns  and concentrations  of external CO exposure  pertinent to the
objectives of any particular health risk evaluation.  For example, as discussed
in the 1979 CO Criteria Document (U.S. EPA, 1979),  some occupations inherently
involve  exposure  to  high levels of CO, at times including frequent exposures
to CO levels well in excess of 100-200 ppm under certain workplace conditions.
On the other  hand, it  is relatively unusual  for members of the general public
to encounter such high CO exposure levels.
     As examples of possible extreme CO exposure situations encountered by the
general  public, the  1979 Criteria  Document noted that the following scenarios
may result in exposure to unusually high ambient levels of CO: (1)  On a large
city freeway where traffic has come to a halt, the  ambient CO level may exceed
44 ppm;  (2)  Inside  a closed automobile where cigarettes are being smoked, CO
concentrations may exceed  87  ppm;  (3)  In enclosed, unventilated garages, CO
levels in excess  of  100 ppm have  been found; and (4)  In a heavily traveled
vehicular tunnel, a 1-hour maximum of 218 ppm CO was recorded.  Under relatively
mild exercise conditions likely  to occur in such exposure situations for any
sustained period of time, the Coburn equations predict COHb blood concentrations
of <10 percent,  assuming exposure durations of less than 8 hrs.   More often,
the  general  (non-smoking)  population is exposed to substantially lower CO
levels (<20-50 ppm)  sustained  over 1  to 8 hr.  periods,  potentially resulting
in maximum COHb levels of 6-7 percent but much more  often in COHb levels below
2-3 percent.  The present  evaluation  is,  therefore, focused mainly on health
effects  observed  at  blood  COHb  levels  below 10 percent.  The  latter COHb
                                     6

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levels are most  pertinent  for present objectives, i.e.,  the  development of
criteria for ambient air quality standards.
     Since the writing of the 1979 Criteria Document, several  new studies have
been published that contribute to the CO health effects data base.  These new
studies, including both human and animal  toxicology data,  are  concisely reviewed
below within a context of integrating the new information  with evidence previ-
ously reviewed in the 1979 Criteria Document.

1.   Cardiovascular Effects
     The most extensive studies on the cardiovascular effects of CO have been
those involving maximum aerobic capacity (V0_   ).  Previous data reviewed in
                                            ^max
the revised CO Criteria Document (U. S. EPA, 1979) demonstrated statistically
significant decreases in V02    when COHb levels ranged from 7-20% under con-
ditions of short-term maximal exercise (Ekblom and Huot, 1972; Pirnay et al.,
1971; Vogel and Gleser,  1972).   In another study (Horvath et al.,  1975),  the
critical level  at which COHb  marginally  influenced (P <  0.10) VtL     was
approximately 4.3%.   In this study, work time to exhaustion was also reduced by
4.9 and 7% when COHb levels had attained 3.3 and 4.3%,  respectively.   Reductions
in V0_    following exhaustive treadmill  exercise have since been  confirmed at
5% COHb. In a double blind experiment (Stewart et al. ,  1978; Klein et al.,  1980),
6 physically fit male fire fighters were randomly exposed to either CO or fil-
tered air twice a week for 3 weeks and exercised to exhaustion.   On exposure
days, the subjects breathed a bolus of 20,000 ppm CO for 47 seconds followed by
30 ppm CO for 4 hours.   This resulted in a sustained elevation of  COHb at 5.0-
5.5% saturation for 4 hours.  Similar decrements in maximal exhaustion times were
noted following the acute exposure and at the end of 4 hours.   No  adaptation to
this hypoxic stress was observed after 4 hours of exposure or over the 3 weeks
of testing.   Significant decreases in total  exercise time (3.8%)  and V0_    (3%)
were also previously reported by Weiser et al.  (1978)  in a project designed to
determine the effect of 5% COHb exposure on healthy young men residing in Denver
at an altitude of 1610m.   Blood COHb concentrations were quickly increased to
5.1% from a  resting level  of 1.0% by adding a bolus of 100% CO to a closed-
circuit system from which the subjects rebreathed.  The decrement in V0_    was
consistent with those reported above and the authors concluded that changes in

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exercise  performance  following CO exposure at this altitude were similar to,
but not greater than, changes  occurring at sea level.
     The  effects of lower CO exposure levels have also been investigated under
conditions  of short-term maximum exercise duration (Drinkwater et a!., 1974;
Raven  et  a!., 1974a,b).   In this; series of studies, a walking test with pro-
gressively  increasing grade was used on subjects continuously breathing 50 ppm
CO at  either  of two ambient temperatures, 25°C or 35°C, with a relative humidity
of 20%.   The two populations  consisted of young  (23+  years) and middle-aged
(48+ years) subjects, both smokers and nonsmokers.  During the duration of the
test,  COHB  levels in nonsmokers increased from 0.6-0.9% to 2.3-2.7%, while levels
in smokers  rose from 2.6-3.2%  to 4.1-4.5%.  Control studies conducted on these
subjects while they breathed filtered air indicated that COHb decreased in both
smokers and nonsmokers in the  absence of experimental CO exposures.   These stud-
ies did not find any reduction in;maximum aerobic capacity.  In fact, the only
statistically significant effect related to CO was a small decrease (<5%) in ab-
solute exercise time consistently observed in the nonsmoking subjects but not in
the smokers (Drinkwater et a!., 1974; Raven et al., 1974a).  These observations
extend those found earlier by  Ekblom and Huot (1972), who reported a large de-
crease (38%) in work time at 7% COHb.
     The revised CO Criteria Document (U.  S.  EPA,  1979) also noted that oxygen
uptake during  short exposures  and submaximal work was  apparently not affected
even at COHb concentrations of 15-20%.   Recently,  DeLucia et al.  (1983) reported
that COHb levels of 7.3% in nonsmokers and 9.3% in smokers did not  induce any
effects involving subjective symptoms,  pulmonary function, exercise metabolism,
or blood  parameters  in  healthy subjects.   In a controlled experimental study
designed to test for potential  syhergism between 03 and CO, 24 male and female
volunteers were evaluated while performing moderate aerobic exercise at 50% of
V0_    . After 100 ppm CO exposure for approximately 1  hr., COHb levels  in non-
smokers rose  from  1.0-2.1%  to  6.0-9.6% and COHb levels in smokers  rose from
1.9-5.1% to 6.6-11.8%.   DeLucia et al.  (1983) attributed the lack of CO effects
as possibly being due to large cardiovascular reserves  found in healthy subjects.
     It should be  noted  that  healthy young subjects were used in most  of the
above  studies  evaluating the  effects of CO on work capacity.   A recent study
by Calverley  et  al.  (1981)  demonstrated a decrease in  walking distance in 15
                                     8

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patients with  severe  chronic bronchitis and emphysema at a mean COHb concen-
tration of  12.3%.   They evaluated 11 men and 4 women with  severe reversible
airway obstruction  [FEV-j Q  = 0.56 ± 0.2  (SD) liters;  FVC = 1.54 ± 0.4 (SD)
liters] who were  hypoxic [Pa02 = 5.2 ± 4.9 (SD)  mm Hg]. All patients were
medically stable  at the time  of  the study and smokers were asked  to stop
smoking for 12 hours before each session.  Each subject walked while breathing
air and oxygen before and after exposure to 200 ppm CO in  air, which raised
their COHb concentrations from 1.1-5.4% to 9.6-14.9%.   There was a significant
reduction in walking  distance  when the  patients breathed  either air or oxygen
after exposure  to  CO.   A significant increase in walking distance  when the
patients breathed  oxygen  after exercise was abolished by CO exposure.  There
was no relationship to normal smoking habits of the patients.   It is therefore
quite possible  that individuals  with hypoxia due to bronchitis or emphysema
are more susceptible  to CO during submaximal work loads  typical of everyday
exercise.
     Other cardiovascular effects of CO are thought to be of greater concern,
i.e., those affecting individuals suffering from chronic angina.   However,  the
precise COHb levels at which such cardiovascular effects occur in angina patients
are much less  well  defined than COHb concentrations associated with various
health effects  discussed above.  Angina pectoris is a symptom of pressure and
pain in the chest  produced during mild exercise or excitement because of in-
sufficient oxygen  supply to the heart muscle.  Angina patients exposed to low
levels of CO while resting have been reported to exhibit statistically sig-
nificantly reduced time to onset of exercise-induced angina at mean  COHb levels
of 2.9 (range,  1.3-3.8 percent) and 4.5 percent (range,  2.8-5.4 percent) and to
experience significantly increased duration of angina attacks during exercise at
a mean COHb level  of 4.5 percent (Anderson et al.,  1973).   Certain questions have
been raised regarding  the  design and conduct of the study, the small number
(N=10) of subjects studied, and the absence of credible independent  confirmation
of its findings.  A reevaluation of the Anderson et al.  (1973)  study,  addressing
major points of concern, has been conducted recently (see Appendix A), and  found
that the study, in fact, provides reasonably good evidence for the hastening of
angina occurring in angina patients at COHb levels  of 2.9 to 4.5 percent.   The
Aronow et al.  (1973)  and Aronow (1981)  studies were  previously accepted as
demonstrating decreased time to onset of angina in  exercising patients at COHb

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levels of 2.0-3.0 percent.  However, these Aronow findings are now most appro-
priately interpreted as providing, at most, suggestive evidence for such effects
occurring at COHb levels below 3 percent.  More conclusive statements regarding
this issue will not be possible until the results of independent studies attempt-
ing to replicate such findings become available.
     Another cardiovascular effect  of possible concern is that of  increased
blood flow  that occurs  as a compensatory  response to  CO exposures  (Ayres et
a!., 1969;  Ayres et  a!.,  1970; 1979).  This response might result in coronary
damage or other vascular effects due to added  stress  on  the cardiovascular
system.   However, inconclusive results have been obtained in community epidemi-
ology studies  examining  the  relationship between CO exposure, mortality from
myocardial  infarction  (heart attack), sudden death due  to  arteriosclerotic
heart disease,  and cardiorespiratory complaints (Goldsmith  and Landau, 1968;
Kurt, et a!.,  1978;  Kurt, et al. ,  1979).  Hence, the  possibility of such an
                                  F
association remains  in question,  and further research  is also needed in order
to clarify this issue.
     The results of one controlled human exposure study reported by Davies and
Smith (1980) are suggestive of possible  effects on cardiac function at low to
moderate CO exposure in healthy individuals.   In a series  of replicated experi-
ments, six matched groups of young human subjects lived in a closed-environment
exposure chamber for 18 days.   They were exposed continuously to 0,  15, or 50 ppm
of CO in air during the middle 8 days.  Standard 12-lead electrocardiograms were
recorded from each subject during the control,  exposure,  and recovery periods.
Although statistical  evaluation of the data was not reported, unequivocal  P-wave
changes were observed during the CO exposure period in 3 of  15 subjects at 15 ppm
CO (2.4% COHb) and 6 of 15 at 50 ppm (7.1% COHb) compared to none of 14 at 0 ppm
(0.5% COHb).  The changes were evenly distributed among nonsmoking subjects and
subjects who had stopped smoking 3 days before the start of  CO exposure.   In addi-
tion, one subject,  later identified as having evidence of myocardial ischemia,
showed marked S-T changes at 15 ppm.  In a separate pilot study by these inves-
tigators with both smokers and nonsmokers at 75 ppm CO (10.9% COHb in nonsmokers;
14.9% COHb in smokers)  significant EKG changes were demonstrated in 7 of 10 sub-
jects.   In  most cases,  the CO-induced changes remained 4 days after exposure
ceased.   The authors concluded that  P-wave abnormalities demonstrated in this
study were due to interference of normal  atria! pacemaking or conducting tissue
activity by CO.  In addition, they speculate that CO has a specific toxic effect,

                                     10

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  on the myocardium rather than (or in addition to) a generalized decrease in 0-
  transport to the tissue.
       Most animal studies  on  the cardiovascular effects of CO have been con-
  ducted at exposure concentrations  resulting in rather .high (>15-20 percent)
  COHb levels.  Only two  studies are relevant for present discussion.  Becker
  and Haak (1979) exposed 11 adult mongrel dogs to increasing concentrations of
  CO 1 hr. after coronary artery  ligation.  These sequential exposures produced
  step-wise increases in the COHb level  from 4.9% to 17.0%.   Myocardial  ischemia,
  as indicated by the amount of S-T segment elevation in epicardial electrocar-
  diograms, increased  significantly  at  the  lowest COHb level  and increased
  further with increasing CO exposure.   These changes occurred in the absence of
  altered heart, rate, blood pressure,  left atria!  pressure,  cardiac output,  or
  blood flow  to ischemic myocardium.  Flow to  non-ischemic myocardium increased
  with CO exposure at  a rate approximately double the increase in COHb.  They
  concluded that low level  exposure to CO can  significantly augment ischemia in
  acute myocardial infarction,  apparently through a reduction in oxygen  supplied
  to the  ischemic tissue.   They suggested, however, that the hypoxia induced by
  CO was  more severe  than could be accounted  for  by a reduction in tissue 0?
'  delivery alone.
       Foster (1981) investigated  the  arrhythmogenic effects of CO during the
  initial minutes of acute  myocardial  ischemia in 8 mongrel dogs.  Since each
  dog served  as its  own control, brief  occlusions of the coronary artery were
  performed sequentially both before and after the administration of 100 ppm CO
  which  raised  COHb  levels  to  10.4%.    Bipolar  epicardial  electrograms  were
  recorded in each experiment from the  ischemic and non-ischemic myocardial
  zones.  An  additional 6  dogs  were  used to  confirm the  reproducibility of
  ischemic conduction slowing during successive occlusions  in the absence of CO.
  There was no  significant  increase  in  ischemic myocardial conduction slowing
  after  CO.   This lack of  arrhythmogenic  effect of CO was  supported by the
  absence of  increased  incidence of  spontaneous ventricular tachycardia during
  ischemia after CO  administration.   The author concludes that clinically en-
  countered COHb  levels may not have sufficient arrhythmogenic effect to be of
  significant health importance during the initial  minutes  of myocardial  ischemia.
                                       11

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2.  Neurobehavloral Effects
    •—'	       '                            i

     The 1979 CO Criteria Document; noted that statistically significant effects
on central nervous system (CNS) functions have been most clearly shown to occur
at COHb levels of 5-17 percent.  This is indicated by studies which demonstra-
ted decrements  in vigilance,  visual perception,  manual  dexterity, learning
ability,  and performance  of complex sensorimotor  tasks such  as  driving
(Bender et al., 1971; Schutte, 1973; O'Donnell et al., 1971; McFarland et al. ,
1944;  McFarland,  1973;  Putz et al. , 1976;  Salvatore,  1974; Wright et al.,
1973;  Rockwell  and Weir,  1975;  Rjummo and  Sarlanis,  1974).   An evaluative
review by  Laties  and Merigan  (1979) substantially  agreed with  these  conclu-
sions.   Also, as reviewed in the 1979 Criteria Document, some studies (Horvath
et al., 1971;  Fodor  and Winneke,  1972; Groll-Knapp  et al.,  1972; Putz et al.,
1976)  have reported significant decrements in vigilance performance (defined as
the ability to detect small changes in one's external  environment occurring at
unpredictable times)  to be  associated with  COHb  levels  in the 3.0-7.6% range;
and one study by Beard and Grandstaff (1975) reported that vigilance effects may
occur at  levels   as  low as 1.8 percent COHb. The lowest COHb levels at which
vigilance  decrements  occur,  however,  are a matter of considerable dispute in  ,
view of numerous  other  studies not finding such effects at COHb levels below
5.0 percent  (Haider  et al.,   1975* Winneke,  1974;  Winneke  et  al. ,  1976;
Christensen et al., 1977;  Benignus; and Otto, 1977).
     Since the writing of the  1979; Criteria Document,  several new studies have
been published which contribute to*the data base of CO-related neurobehavioral
effects.   In the  following,  each  of the  several  neurobehavioral  endpoints are
re-evaluated by integrating new findings.
                                  I                             -
Vigilance—Since the  1979 document was written, several new pieces of research
on CO and vigilance have appeared.  In an evaluative review of the CO-vigilance
literature, Benignus  et al.  (1983) concluded that all of the studies (except
one) cited in  the 1979 document had serious  credibility  flaws due  to  (a) non-
replication of the work,  (b) gross statistical abuse, (c) non-concentration-
related effects or (d) combinations of the above.
     The single study from the 1979 document  which had  no  serious flaws was
that of Putz et al.   (1976),  which  demonstrated  vigilance decrements at 5%
                                     12

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COHb.  Since that  time  the same group of  researchers  have replicated these
results on  independent  subjects  (Putz et al., 1979) and have reported their
earlier results  in  peer-reviewed literature (Putz, 1979).   The fact that the
experimental design and details  of the tasks were  not the same in the two
experiments implies that the effects were robust and thus lends further cre-
dibility to these  results.   The  credibility of these findings would be appre-
ciably increased if an  independent group of researchers also were to  success-
fully replicate the study.
     Three other studies on CO and vigilance have appeared since 1979 (Benignus
et al., 1983,  Davies  et al., 1981,  Roche  et  al.,  1981),   all of which used
COHb levels of 5-7%.   None of the studies found significant effects on vigi-
lance.   It  is  noteworthy that all of them used vigilance paradigms different
from those of  Putz et al (1976).  Quite possibly the parameters and conditions
under which vigilance was studied are so sensitive that unless the proper con-
ditions exist, the effects of such COHb levels will not be detected.
     It appears safe to  conclude that, at least under some conditions, reliable
but small decrements in  vigilance occur at about 5% COHb.   The fact that 5% and
higher levels  of COHb were not observed to produce vigilance decrements in many
studies is probably a reflection of  (a) low experimental test sensitivity in the
those  cases or (b) the  small effects of CO at these levels coupled with what
is probably a  rather low-slope  concentration-effects  curve  in  the  region of
COHb less than 20%.  It must be  emphasized that these explanations for the many
no-effect studies  are conjectural.
                                                         *
Sensory and Time Discrimination—Benignus  et  al.   (1983)  concluded that  no
highly  reliable  evidence for time discrimination  decrements  exists.   The  ele-
gant concentration-related decrements in dark adaption beginning  at  5%  COHb
demonstrated by  McFarland  et al. (1944) remain to be replicated.  The importance
of replication should not  be overlooked since the decrements were (a) dose-related
and  (b) showed a decrement  at the  lowest non-zero COHb  level.
     Davies et al. (1981) tested  visual sensitivity in dark-adapted  subjects
who  had been  exposed continuously to 50 ppm CO for a total of 5 days  (COHb  of
7%  by  the end of each day).  They reported no effects  of  CO.   Luria and  McKay
(1979)  reported  that untrained  observers showed no  decrement in a night vision
test,  eye movement, or  visual evoked potential at COHb  levels of 9  percent.  The
                                      13

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fact that  McFarland et al. (1944)  used  bolus  exposure methods, whereas the
above investigators used continuous low  level exposure, is perhaps significant.
McFarland et al. (1944) also used highly trained observers, which had the effect
of reducing variance and thus increasing the sensitivity of their study.  Cer-
tainly the McFarland et al. (1944) study cannot be said to have been invalidated
by newer data.
     Some support for visual sensitivity decrements due to CO has been provided
by a study measuring the electroretinogram in  anesthetized cats (Ingenito and
Durlacher, 1979).  When cats were exposed to 1000 ppm CO the electroretinogram
was decreased in amplitude by 30 minutes after the start of exposure, at which
time the mean  COHb was 7.5%.   Further decreases were  concentration-related.

Complex Sensorimotor Performance and Driving—The  conclusion  of the  1979
document, that driving-like tasks are impaired at COHb levels of 5% or greater,
has since been substantially strengthened.   Three articles have been published
(Putz et al.,  1976,  1979;  Putz, 1979), using variations of the same task and
experimental  design but in two independent groups of subjects.   In all cases, it
was reported that 5% COHb produced decrements in compensatory tracking, a hand-
eye-coordination task.   In all cases, however, the decrements occurred only during
high task difficulty.   Other tasks such as reciprocal tapping and digit manipula-
tion were  not  affected by COHb levels of  up to 5% (Mihevic et al. , 1982).

Sleep and Activity—Although there  has been one new publication in this area
(Groll-Knapp et al., 1982), the conclusions remain the same.   Marginal increases
in deep  sleep and  concomitant  decreases  in  rapid-eye-movement sleep were
reported at 8% COHb due to exposure to 100 ppm CO for 8 hrs.  during sleep.   As
before,  the changes did not reach statistical  significance when appropriate
corrections were made (Benignus and Muller, 1982).

Central  Nervous System Electrical  Activity—No significant changes have occurred
in this area  of research since 1979.  Marginal, nonsignificant effects have
been reported  by Benignus et al.  (1983) in the electroencephalogram  alpha
frequency band  and  by  Groll-Knapp et al.  (1982) on evoked potentials.  Both
groups  of investigators produced COHb levels of 5-6 percent.
                                     14

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3.  Effects of CO Exposure on Fibrinolysis

     An area of  growing interest,  which was not extensively discussed in the
1979 Criteria Document, concerns possible effects of CO on  fibrinolysis.  The
fibrinolytic system  is  an integral  part of homeostatic mechanisms and it has
been suggested that  derangement  of that system may contribute to the patho-
genesis of  thrombosis.  The  plasminogen activator may be  released  from the
blood vessel wall  by a variety of stimuli,  including anoxia, electric shock,
and either  local  or  systemic administration of vasoactive substances such as
epinephrine, acetylcholine,  serotonin,  or histamine.  The finding that anoxia
can release plasminogen activator has  suggested  to investigators that the
presence of COHb with its associated anoxia  could  also  be associated with
increased plasminogen activator.
     Resting fibrinolytic activity is often low and its measurement may yield
conflicting results  that  do  not  readily permit intra-individual  comparisons.
The relationship  of  tissue activator to vascular activator  suggests that they
are components of two separate fibrinolytic mechanisms:  (1)  the vascular acti-
vator being part of a humoral system whose main role is maintenance of vascular
patency and  (2)  the  basic activator concerned with tissue  repair and wound
healing.

Animal  Studies—Fibrinolytic activity was studied in 12  rabbits  (6 control
rabbits) continuously exposed for 8 weeks to an ambient concentration of 50 ppm
CO (Kalmaz  et al.,  1977).   The COHb levels reached  30.9% by the end of the
exposure period.  Significant  increases in  whole blood clotting time, serum
fibrin/fibrinogen degradation products•,  and acceleration  of whole blood clot
lysis occurred.   Euglobulin lysis time was significantly accelerated by the end
of the first week of exposure.   Although these fibrinolytic activity changes are
of interest, their relationship to CO exposure is far from clear.
     In another study, Kalmaz et al.  (1978)  continuously exposed rabbits to 50
ppm for 8 weeks or intermittently to 300 ppm for 4 weeks.   Acceleration of the
whole blood clot lysis and euglobulin lysis times was observed in all  CO-exposed
groups.   Microscopic examination of large vessels showed endothelial damage —
a possible source for a plasminogen activator  release.   A more recent study by
Kalmaz et al.  (1980) involved exposing rabbits to ambient air,  50 ppm CO for
                                     15

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24 hours/day continuously for 8 weeks, and 300 ppm CO for 8 hours/day, 5 days/
week  for 4 weeks.  They found  a  consistent change in circulating  platelet
quantity for all CO-exposed rabbits.  However, their conclusion that prolonged
exposure to low levels of CO may influence changes in circulating platelet counts
and/or congenital  platelet  function disorders in man has not been confirmed.
     In a recent iji vitro study by Hartiala et al .  (1982) a 5-minute exposure to
100% CO  was  found to have neither  an effect  on  the decrease  in prostacyclin
(PGIg) production  nor a direct effect on ADP-induced aggregability of human
platelet-rich plasma.  This led the authors to conclude that CO is not respon-
sible  for  the temporary increase :in platelet aggregability after cigarette
smoking.
Human Studies — Workers chronically  exposed  to high levels of carbon monoxide
(approximately 100 ppm with  occasional  levels of 200-400 ppm) were  studied by
El-Attar and Sairo (1968).  They found an accelerated clot lysis time suggestive
of an enhancement of blood fibrinolytic activity.   Levels of COHb present were
only crudely determined  and  were not related to any specific individual.  CO
poisoning of 21 workers  was  suggested by the  presence of subjective  symptoms,
Headache, blurred vision,  etc.   Twenty-eight workers, presumably some of the
first group of 21, were restudied after a one day exposure to ambient CO (levels
not given).   This group also exhibited accelerated lysis times but to a lesser
degree.  In 15 control  subjects no fibrinolytic activity could be  detected.
This study,  although suggestive, jwas  too  poorly controlled to be  of real
value.   A similar suggestive study was made by Alexieva et al .  (1975) on 100
workers in a coke-chemical plant, i Some increase in fibrinogen was observed in
these  chronically  exposed individuals.  Panchenko  et al.  (1977) have also
suggested that a  relationship between blood coagulation and CO existed.   The
data presented are far from conclusive.
     The possibility  that exposure to  other substances in  addition to  CO
resulted in alterations  of fibrinolytic activity was evaluated by Janzon and
Nilsson (1975).   Smokers and nonsmbkers were studied by techniques superior to
those used by the  above investigators.   Smokers and  nonsmokers had  the  same
fibrinolytic activity when smokers were studied after 12 hours abstention from
smoking.  Smoking 6 cigarettes during 3 hours was associated with an increased
                                     16

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fibrinolytic activity in blood.  They believed that this increase was probably
due to the  combined effects of nicotine and carbon monoxide.   Mansouri  and
Perry (1982) studied the alteration of platelet aggregation by cigarette smoke
and carbon  monoxide.   Inhalation  by healthy adults of CO sufficient to raise
the COHb  level  to  between 4.5% and 11% was found to be responsible for inhi-
bition of platelet  aggregation, which returned to normal  after five hours.
However,  there  was  no  consistent  correlation reported  between COHb  levels  and
alterations in platelet aggregation.
     Brinkhouse (1977) exposed 23 men (15 nonsmokers) for 4 hours to either 0,
50, or 100 ppm CO.   COHb levels on the days of carbon monoxide exposure reached
2.17% (50 ppm) and 4.15% (100 ppm).  Platelet count,  prothrombin time, partial
thromboplastin time, thrombin time, fibrin split products, factor VIII, and pla-
telet aggregation were  determined before and after the exposure.  Coagulation
parameters were not significantly affected by the CO exposures.  A review by
Haft (1979) discusses the role of platelets in the etiology and natural' history
of coronary artery disease.   It is clear thaf smoking increases the activity of
platelets and cigarette smokers have  shortened platelet survival time.  Endo-
thelial injury  may  be  facilitated by  serum CO, by levels  of circulating cate-
cholamines, and other factors.
     In conclusion, the effects on fibrinolytic activity of exposure to carbon
monoxide are  far from  clear.   The studies on acute exposure to  CO do  not  spe-
cifically implicate this pollutant in the observed alterations in fibrinolytic
activity, and the  studies  on chronic exposure are too poorly controlled to
confirm any definite effects on the blood coagulation system.

4.  Suggestive Evidence for Perinatal CO Effects

     The  Criteria  Document  (U.S.  EPA, 1979) provides  discussion  of results
from certain animal toxicology studies which point toward the possibility that
CO exerts perinatal effects on the fetus or newborn.   With long-term exposures
of pregnant animals to CO, fetal COHb levels have been shown to be higher than
maternal  COHb levels,  and fetal elimination of CO was  slower than maternal  CO
elimination.  The ability of CO to decrease the oxygen transport capacity of
maternal   and  fetal  hemoglobin  may result in interference  in  fetal tissue
oxygenation during important developmental stages.   Whereas normal  adults have
                                     17

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reserve capacity and compensatory responses which enable them to handle moder-
ately high COHb  levels without irreversible consequences, the fetus may under
normal situations be  operating  close to critical levels  in  terms  of tissue
oxygen supply.  Thus, even moderate CO exposures may have a deleterious effect
on fetal development  (Longo,  1977) but this,  too, remains to be demonstrated
along with pertinent  dose-response relationships.   In several  animal  studies
in which pregnant females  were  exposed to CO, deleterious effects were gen-
erally reported in the offspring (e.g., reduced birthweight,  increased newborn
mortality,  and lower  behavioral  activity levels) even when no effects on the
mothers were  detected.   In human studies, similar effects have been reported
in children of mothers who smoked cigarettes during pregnancy,  suggesting that
expectant mothers and their  unborn  children  may also represent population
groups at special risk  for CO effects, but this remains to be more clearly
defined along with any pertinent dose-response relationships.
                                   i
     Sudden  infant  death  syndrome'(SIDS)  is  characterized  by  the sudden,
normally unexplained  death of an  infant.   Research has suggested numerous
possible etiological  factors  (e.g.^  disease,  temperature, maternal smoking,
and pollutant levels),  but numerous questions remain regarding which are the
most significant factors involved.  [Seasonal incidence of SIDS has been studied
in several  epidemiological studies  (Peterson,  1966;  Bergman et al. ,  1972;
Bonser et al., 1978).  A pattern which appears to be consistent across countries
in the  northern  hemisphere is increasing  incidence in October, peaking in
December and  January, remaining  high  until May,  and then declining sharply in
June and remaining low until October.
     CO has  been  hypothesized to be associated  with  seasonal  variations  in
SIDS incidence rates, based on certain  epidemiologic  data.   Hoppenbrouwers et
al.  (1981)  have  reported that increased  seasonal  incidence  of SIDS in Los
Angeles County during winter  may be at  least partially explained  by higher
levels of CO, sulfur dioxide  (SO-), nitrogen dioxide (N02)  and hydrocarbons
(HC).  They  suggest that higher ambient  levels  of  these pollutants may be
implicated in chronic hypoxia, which  often precedes death from  SIDS.   Related
to this hypothesis,  they found that  SIDS  cases in Los Angeles were correlated
to daily mean levels of the  above pollutants and peaks in these pollutant
levels preceded seasonal increases in SIDS by seven weeks.  The authors further
report that  the  lifespans  of  infants dying from SIDS were longer (1) if born
in low rather than high pollution areas and (2) if born in months of low versus
high pollution.
                                     18

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Finally, a direct  proportionality  was reported between exposure to pollution
for infants from conception  to two months of age and bimonthly rate of SIDS,
     In an editorial letter assessing the results of the Hoppenbrouwers et al.
study, Goldstein (1982) commented on the presence of indoor sources of CO, NOp
and lead.  She  further pointed out that  during  colder months infants spend
most of  their time indoors where concentrations of these pollutants can  far
exceed ambient  levels.  Thus,  she  suggests that  the relationship between  SIDS
and ambient pollution  levels may be only  coincidental  and the evidence for  it
is at  best suggestive  and in  need of further confirmation before any causal
relationships are inferred.
     Maternal  smoking has been related to SIDS in several studies (Bergman and
Wiesner, 1976; Lewak et al.,  1979;  Peterson,  1981).   Because CO is only one of
numerous pollutants  found in  cigarette smoke,  however,  it is difficult  to
infer  a  causal  relationship  between CO and SIDS.  Other factors associated
with  SIDS  include  passive smoking,  younger maternal  age,  short intervals
between pregnancies, gestational age  of less than 40  weeks, birth weight of
less than 3000 g.,  lower socioeconomic status, and male sex.   Thus, the number
of potentially confounding factors  makes finding an association between CO and
SIDS extremely difficult.

                             POPULATIONS AT RISK

     The 1979 Criteria Document  directed attention toward identification of
sensitive population groups  at special  risk for CO-induced  health effects.
One key  concept in defining  special risk groups for  CO effects is the idea
that any preexisting or concomitant physiological  or  pathological condition
which  interferes or interacts with  oxygen absorption into blood or its transport
to  and perfusion of body tissues  can logically be expected  to exacerbate
CO-induced health  effects  associated  with the hypoxic effects  of CO.  Thus,
certain  large segments of the general population can  be reasonably hypothe-
sized  as likely to be at greater risk for experiencing CO-induced health effects
than healthy, non-smoking adults.
     These probable risk groups include:  (1) fetuses and young infants;  (2) the
elderly, especially those with reduced cardiopulmonary functions attributable to
a variety of factors associated with typical  aging processes; (3) other,  younger
                                     19

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individuals with  overt,  severe cardiac damage or acutely severe respiratory
diseases, e.g., pneumonia; (4) individuals with chronic bronchitis or emphysema;
(5) individuals with symptoms (e.g.; angina) indicative of chronic cardiovascular
disease; (6) individuals with hematological diseases, (e.g., anemia) that affect
oxygen-carrying capacity or transport in the blood;  and (7)  persons with geneti-
cally unusual  hemoglobin  forms associated  with decreased oxygen capacity.  In
addition to the above, one might  reasonably expect that individuals  under the
influence of certain drugs, used for recreational or medicinal purposes, may be
at greater risk for CO-induced effects due to interactive effects between CO and
certain pharmacological agents.  Lastly, under high altitude conditions (where
reduced levels of atmospheric oxygen exist), increased vulnerability to CO health
effects of both the above sensitive!population groups and otherwise, non-sensitive
healthy individuals might be expected.
     As  noted  in the  1979  Criteria  Document,  relatively  little  concrete
experimental or observational  evidence currently exists by which most groups
(or conditions) listed above have been clearly demonstrated to be associated
                                    I
with  increased risk  for CO-induced  health  effects.   Nor  have  clear-cut
quantitative lowest-observed-effect levels or dose-response relationships been
delineated for the  occurrence of ct) effects among the above "at risk" groups
                                    !                             i
or in conjunction with special interacting circumstances  (i.e., drug usage or
high altitude  residence)  that might exacerbate CO effects.  Only very limited
discussion, therefore,  can be provided  here  regarding CO risk factors  and
sensitive groups likely at special risk for CO .effects.
     In regard to the first group, fetuses and young infants,  certain evidence
was alluded to earlier from animaljtoxicology studies indicating that higher
levels of COHb and increased CO excretion time occur among fetuses in compari-
son to  their mothers  exposed to CO; and  some  deficits  in postnatal  growth and
development were noted in the offspring of dams exposed to CO during pregnancy.
Also, analogous perinatal effects  were noted  in  human  infants  born to mothers
who smoked during pregnancy, and associations between CO exposure and SIDS have
been hypothesized.  However, insufficient evidence exists at this time by which
                                    \
to estimate CO exposure levels at which any CO effects on human fetuses or new-
born infants may  occur or whether the latter  types of  effects  seen in smoking
mothers are due  specifically to  CO versus  other components of tobacco smoke
either singly or in combination.
                                     20

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     Little  specific  evidence directly demonstrates  the  increased vulnera-
bility of  the  elderly for CO health effects  in  precise quantitative terms.
Given the  increased  vulnerability  of  the aged to many different  kinds of
stress,  including  thermal  stress  (hot  or cold) which taxes decreased reserve
capacities to  maintain adequate  cardiovascular  delivery  of oxygen to  body
tissues, it  must  be expected that CO  exposure would  render the elderly more
vulnerable to  the  effects of other types of  cardiovascular stresses.   Also,
conversely, it is reasonable to hypothesize that interactive effects involving
other stress factors  might lead to exacerbation of CO-induced health effects
or  their  occurrence at lower external CO  exposure  levels than in younger>
healthy adults.   Similarly, younger individuals with overt,  severe cardiac
damage or  insufficiency or severe acute respiratory diseases,  e.g., pneumonia,
can be  expected  to be more vulnerable to CO  (i.e., either CO exacerbation of
other disease  effects or, conversely,  increased susceptibility to CO health
effects) due to  reduced reserve capacities to cope with  stress generally or
increased  sensitivity of  already  compromised organs  or tissues, e.g.,  heart
muscle, to the hypoxia induced by CO.
     Turning to  individuals with  chronic bronchitis and emphysema, again,
reduced reserve  capacities for dealing with  cardiovascular stresses and al-
ready reduced  oxygenation of blood should exacerbate or  hasten the onset of
health  effects associated with  CO-induced  hypoxia.   Analogously, angina
patients or others with obstructed coronary arteries  but not manifesting overt
symptoms such  as angina,  should be at  greater risk for CO health effects.   Both
the Anderson et  al.  (1973) and several Aronow publications  on  angina patients
reported findings  previously accepted  as  demonstrating the  increased vulnera-
bility of  angina patients  to CO in terms of hastening of the onset of exercise-
induced angina.   These papers,  furthermore,  appeared  to confidently establish
2-3% blood COHb as the range of COHb values associated with the onset of statis-
tically significant effects  indicative of CO  exacerbation of angina and, prob-
ably, associated  hypoxic  effects  on cardiac  muscle.   It is  now clear that the
Anderson findings  (as of yet not  independently confirmed) can be appropriately
interpreted  as providing  reasonably good evidence for exacerbation of  angina
symptoms occurring at approximately 2.9 to 4.5 percent COHb; and the possibility
of  such effects occurring  at lower COHb levels cannot be ruled out at this time.
                                     21

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     In  regard  to individuals  with|anemia being  at  special  risk for  CO  health
effects,  the 1979 Criteria Document noted that  CO  poisoning is similar to
anemia,  wherein the oxygen capacity of  the blood is decreased  because the
affinity of hemoglobin (Hb) for binding oxygen is reduced.  The 02 dissociation
curve  for  anemics is  similar to that for normals except  that  it is shifted to
the right.  However, when curves from individuals with 50% reductions in Hb con-
tent are compared with dissociation curves for blood with 50% COHb content, there
are striking  differences.   Consequently, care must be taken  to avoid overly
simplistic  extrapolations  regarding  the  likely impact  of  particular  CO
exposures  on  anemia patients  due to additional  (CO-induced) reductions of 02
carrying capacity beyond  the  reductions already evident  in their blood.  The
specific CO  exposure  levels and associated blood COHb concentrations at which
anemia patients may be at increased risk for specific CO  health effects remain
to be  clearly delineated, but little doubt  exists  regarding  the likelihood
that the effective CO exposure (and COHb)  levels are distinctly lower than for
healthy, non-anemic individuals.
     Another logically-hypothesized "at risk" group for CO effects are individuals
with unusual  hemoglobin types  that;result in chronic elevations  of COHb blood
levels even  in  the absence of external CO exposure.  Normal adult hemoglobin
has a  relative  affinity  or equilibrium constant (M) for CO of  about 200 for
most animal  species,  but  has  been -reported  to be as high as 240 to 250 in
humans (Roughton, 1970).   There are1 approximately 350 human hemoglobin variants,
including those found in the fetus, sickle cell anemics,  and other individuals
with hemoglobinopathy (hemoglobin disorders).  Approximately one fourth of the
hemoglobin variants now known are considered unstable,  i.e., they denature and
precipitate when  red blood cells  or  hemolyzates are exposed to heat,  red
oxdyes, or isopropanol (Bunn et al.,, 1977).  One of these variants,  hemoglobin
Zurich (HbZ)  has  been found to have an affinity  for CO which  is  approximately
65 times that of normal  hemoglobin (Zinkham et al., 1980; Giacometti et al.,
1980;  Zinkham et  al., 1983).  This !results in chronic elevation  of endogenous
COHb levels ranging from 3.9 to 6.7% in nonsmoking HbZ individuals.
     As  for  possible  drug-induced enhancement of CO effects, whereas  there
exists little specific data directly  supporting the idea,  it is logical  to
suspect that  individuals  who use certain  drugs would be  at  increased risk for
experiencing health effects associated  with CO exposure.  For example,  drugs
                                     22

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with primary or secondary CNS depressant effects should be expected to exacer-
bate neurobehavioral  effects  of CO;  and  drugs which  have  primary  or secondary
cardiac  stimulant  effects might worsen CO-related  cardiac effects.  Any  vaso-
constrictive drugs  would be expected to  reduce  0«  delivery to  various  organs,
thus also  exacerbating  CO effects  on exercise, cardiac  or  neurobehavioral
function.  Further  speculations about drug-CO interactions are possible, but
it should be emphasized  that these are predictions based on theoretical grounds
that rely  heavily  on  our  current  understanding  of  hypoxia as the likely
main mechanism  underlying the induction  of  CO-induced  health effects.  Unfor-
tunately, few data currently  exist by which to judge  the likely  validity of
such speculations.
     Hypothesized enhancement  or exacerbation of CO  health effects under high
altitude conditions,  similarly,  rests heavily on  hypoxia  as the  key mechan-
ism of  CO  toxicity.  The interactive effects of high altitude hypoxia and CO
exposure, however,  appear  to be  more complex  than might  be simplistically
expected (Collier and Goldsmith, 1983).   For example, the 1979 Criteria Document
notes that adaptation  to high altitudes occurs and alludes to the fact that,
analogously, adaptation  to CO may alter the position  of  the 0? dissociation
curve as a function of extensive prior CO exposure.  Thus, individuals living
in high altitude situations with frequently elevated ambient air CO levels may
adapt to both the  high altitude and  CO-induced hypoxia, whereas other  indivi-
duals newly entering high  altitude and elevated ambient  CO conditions might
experience CO effects  at concentrations  below those effective at lower alti-
tudes.

                            SUMMARY AND CONCLUSIONS
     As was stated  at  the outset, the main  purpose of  the  present addendum is
to reevaluate the  scientific  data base concerning health  effects associated
with exposure  to CO  at  ambient or near-ambient exposure  levels.  The re-
evaluation includes both (1)  summarization of information contained  in  the
revised EPA Air Quality  Criteria Document for CO (U.S. EPA, 1979) and  (2) new
information and studies  that  have become available  beyond that  reviewed in
the 1979 document.   The  most  important points of information reviewed and key
conclusions derived from this  evaluation of the CO health effects data base  are
summarized below.
                                     23

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Mechanisms of Action—The binding of CO to hemoglobin, producing COHb and de-
creasing the oxygen-carrying capacity of blood, appears to be the main mechanism
of action underlying the induction of toxic effects of low-level CO exposures.
The precise mechanisms  by  which toxic effects are induced via COHb formation
are not yet fully understood, but likely include the induction of a hypoxic state
in many tissues of diverse organ systems.   Alternative mechanisms of CO-induced
toxicity (besides COHb)  have been hypothesized, but none  have yet been  demon-
strated to operate at relatively low (near-ambient) CO exposure levels.   Blood
COHb levels, then, are currently accepted as representing a useful  physiological
marker by which to  estimate  internal CO burdens due to the combined contribu-
tion of  (1)  endogenously derived CO and (2) exogenously derived CO resulting
from exposure  to  external  sources 'of CO.  COHb levels likely to result from
particular patterns  (concentrations, durations, etc.) of  external CO exposure
can be reasonably well estimated from equations developed by Coburn.

CO Exposure Levels—Evaluation  of human CO exposure situations  indicates that
occupational exposures  in  some workplace situations can regularly exceed 100
ppm CO, often  leading to COHb levels of 10 percent or more.  In contrast,  such
high exposure  levels are much less commonly encountered by the non-occupationally
exposed general public.  More  frequently,  exposures to less  than 25-50  ppm  CO
for any extended period of time occur among the general population and, at the
low exercise levels usually  engaged in under such circumstances, resulting COHb
levels most typically remain below 2-3 percent among non-smokers.  Those levels
can be compared to the physiologic norm for non-smokers, which is estimated to
be in  the  range of 0.3 to 0.7  percent COHb.  Baseline COHb concentrations in
smokers, however, often greatly exceed  3  percent,  reflecting absorption of  CO
from inhaled smoke.

Health Effects of Low Level  CO  Exposures—Four types of health effects  reported
or hypothesized to be associated with CO exposures (especially those producing
COHb levels below 10 percent) were evaluated:  (1) cardiovascular effects; (2)
neurobehavioral effects;  (3) fibrinolysis effects;  and (4) perinatal  effects.
In  regard to  cardiovascular effects,  decreased oxygen uptake  capacity and
resultant decreased work capacity under maximal exercise  conditions  have  been
clearly  shown  to  occur in healthy young adults starting  at  5.0 percent COHb;
                                      24

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and several studies observed small decreases in work capacity at COHb levels as
low as 2.3 to 4.3%.  These cardiovascular effects may have health implications
for the general population in terms of potential curtailment of certain physi-
cally demanding occupational or recreational activities under circumstances of
sufficiently high  CO  exposure.   However,  of greater concern at more typical
ambient CO exposure levels are certain cardiovascular effects (i.e., aggravation
of angina symptoms during exercise) likely to occur in a smaller,  but sizeable,
segment of the general population.  This group, chronic angina patients, is pre-
sently viewed as the most sensitive risk group for CO exposure effects, based on
evidence for aggravation of angina occurring in patients at COHb levels of 2.9
to 4.5 percent  COHb.   Such aggravation of  angina  is thought to represent an
adverse health effect for several reasons articulated in the 1980 proposal  pre-
amble (45  FR  55066),  and the Clean Air Scientific Advisory Committee (CASAC)
concurred with  EPA's  judgment on  this matter (see  Appendix B).  Dose-response
relationships for  cardiovascular  effects  in coronary artery disease patients
remain to be more conclusively defined, and the possibility cannot be ruled out
at this time that  such effects may occur at levels below 2.9 percent COHb (as
hinted at by the results of the now-questioned Aronow studies).
     No reliable evidence demonstrating decrements in neurobehavioral  function
in healthy young  adults  has been reported  at  COHb levels below 5%.  Much of
the research at 5% COHb did not  show any effect even when behaviors similar
to those  affected  in  other  studies were involved.  However, if any  CO  effects
on neurobehavioral functions in fact occur below 5% COHb,  then none of the sig-
nificant-effects studies would have found such decrements,  because none of them
used COHb levels below 5%.  Other workers who failed to find CO decrements at 5%
or higher  COHb  levels may have employed tests  not sufficiently sensitive to
reliably detect small effects of CO.   From the empirical evidence, then, it can
be said that  the  COHb levels in the 5% range do produce decrements in neuro-
behavioral function.  However, it cannot be said confidently that COHb  levels
lower than 5%  would be without effect.   One important point made in the 1979
document should be reiterated here.   Only young, healthy adults have been stu-
died using demonstrably sensitive tests and COHb levels of 5% or greater.  The
question of groups at special  risk for CNS effects, therefore, has not been
explored.   Of special note are those individuals who are taking drugs which have
primary or secondary  depressant  effects which  would be  expected to  exacerbate
CO-related neurobehavioral decrements.   Other groups at possibly increased risk
                                     25

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for CO-induced  neurobehavioral  effects  are the aged and  ill but these groups,
also, have not been evaluated for such risk.
     In contrast to the data available which demonstrates associations between
cardiovascular and  neurobehavioral  effects  and relatively low-level CO expo-
sures, much  less clear  evidence exists  that  other types  of health effects are
associated with  low-level  CO exposures.  For example, only  relatively weak
evidence points towards possible CO effects on fibrinolytic activity and, then,
generally only  at rather high CO exposure levels.  Similarly, whereas certain
data also suggest that  perinatal effects (e.g. reduced  birth weight, slowed
postnatal development,  Sudden  Infant Death Syndrome) are associated with CO
exposure, insufficient  evidence presently  exists  by which to either confirm
such associations qualitatively or to establish any pertinent exposure-effect
relationships.

Population Groups at Risk for Ambient CO Exposure Effects—Angina patients or
others with  obstructed  coronary arteries, but not yet manifesting overt  symp-
tomatology of coronary  artery disease,  appear to be  best established as a
sensitive group within  the general population that is at increased risk for
experiencing  health  effects  (i.e.  exacerbation of cardiovascular symptoms) of
concern at ambient or near-ambient'CO exposure levels.   Several other probable
risk groups  were  identified, i.e.:  (1) fetuses and young  infants; (2) the
elderly  (especially those with  compromised  cardiopulmonary  functions);  (3)
younger individuals with severe  cardiac or acutely severe respiratory diseases;
(4)  individuals  with chronic bronchitis or  emphysema;  (5)  individuals with
hematological diseases  (e.g.  anemia) that affect oxygen carrying capacity or
transport in the blood; (6)  individuals with genetically unusual forms of hemo-
globin associated with  reduced  oxygen carrying capacity; and (7) individuals
using medicinal or recreational  drugs having CNS depressant properties.   However,
little empirical evidence currently is available by which to specify particular
COHb levels at which such individuals are likely to experience specific health
effects associated with ambient  or near-ambient CO exposures.   Nor does unambi-
guous evidence yet  exist which  clearly  establishes that  healthy non-sensitive
individuals or those in the  above probable risk categories are affected at lower
CO exposure levels under high altitude conditions than CO exposure concentrations
effective at  lower altitudes.
                                     26

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                                      34

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                 APPENDIX A

   EPA HEALTH EFFECTS RESEARCH LABORATORY
REEVALUATION OF ANDERSON ET AL. (1973) STUDY
                     A-l

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 XDW%
QSp
                UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
                          HEALTH ERFECTS RESEARCH LABORATORY
                                RESEARCH TRIANGLE PARK
                                NORTH CAROLINA 27711

   DATE:  February 1, 1984

SUBJECT:  Evaluation of Anderson et al.. (1973)  Study of Carbon Monoxide  (CO)
          Effects on Angina Patients.

   FROM:  John J. O'Neil, Ph.D., Chief
          Clinical Research Branch, ITD/HERL

     TO:  Lester D. Grant, Ph.D.
          Director, ECAO (MD-52)

          THRU:  F. Gordon Hueter, Ph.D., Director  f
                 Health Effects Research Laboratory, RTP  (MD-58)
        This memo replaces one dated August 19, 1983.   Changes have been
   made to clarify seme points.  The discussion of carboxyhemoglobin (COHb)
   analysis (section la) and Table 1 have been notably expanded.  An analysis
   of possible reasons for the differences between the measured values of
   COHbreported by Anderson et al. and estimates of COHb calculated using
   the Coburn-Forster Kane Equation are also offered.   The conclusions at
   the memo remain the same.         i

        Thank you for the opportunity to carment on the paper by Einar Anderson
   and his co-workers which is entitled "Effect of Low-Level Carbon Monoxide
   Exposure on Onset and Duration of Angina Pectoris."  I have re-read it,
   have discussed it with several colleagues, especially Dr. Vernon Benignus,
   and would offer the following observations by way of review.

        This is a good paper.  The work appears to be carefully done, the
   data seem reasonable, and the paper is well written.  Critical review
   reveals some flaws in the design  and conduct of the research, but this is
   typical of most scientific work and, therefore, significance of these
   conroents may be interpreted differently by different readers.

   DESIGN                           !

        1)   ST segment depression reported in  this paper is not useful
   and does  not constitute  any physiological support to the observations
   regarding the onset and  duration  of angina.  The authors recognize this
   shortcoming.  The important data  in the paper  are the measurement of time
   to onset  of pain  and the measurement of the duration of pain.

         2)   The paper would have been considerably strengthened by more
    careful selection of the subject  population.   For example,  5 subjects
   were smokers  and  5 were  non-smokers and one subject was  taking  digitalis.
    In my opinion,  the study would  have been strengthened considerably had
    the subject population been more  homogeneous.
                                        A-2

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     3)  The number of subjects (n) is very small.  Oily ten subjects
were studied and of these there are three for whom there are missing data
points.  I have been told anticdotally that this study was intended as
a pilot study.  It is unfortunate that it was not possible to design
and complete the proper follow-up study.  Given the situation, this is
valuable data to have published and should be used to develop our under-
standing of the response of angina patients to carbon monoxide (CO).

METHODS AND RESULTS

     1)  It is disconcerting that the measured COHb levels appear to be lower
than we would predict on the basis of our knowledge of CO-COHb kinetics.  Why
is this measured value lower than we would expect?

          a)  Measurement of GOHb.  The measurements were done in
     triplicate using the Buchwald analysis.  As best as I can determine
     there is no evidence which causes me to doubt this analysis.
     However, this technique is less precise than other methods that are
     used today.  That is, more scatter occurs in the data for repeated
     measurements at a given concentration.   At low COHb concentrations
     this scatter might have a disproportionately large effect which, in
    .turn,  might lead to an over-estimation of the "zero" COHb sample analy-
     sis.   If this occurred, estimations of COHb concentration calculated
     using the Coburn-Forster-Kane equation would be over-estimates because
     this initial measurement is also used as the initial concentration in the
     Coburn-Forster-Kane equation.

          b)  It is also possible that the exposure mask used in this study
     was loose fitting and leaked.  This would reduce the exposure level
     and the final COHb levels.

          c)  Though not reported in the paper, the subjects were apparently
     allowed to rest ad libitum during the study.   This amounted to approxi-
     mately 10-15 minutes out of the hour. -This "rest" period would have
     reduced the exposure time and the final COHb levels.

           Using a minute ventilation of 5 1/min,  a blood volume of 5500 ml
     and a  hemoglobin concentration of 15 g/lOOml,  Dr.  Benignus calculated a
     predicted COHb with the Cobum-Forster-Kane Equation.   These data are
     presented in Table 1.  He concludes that there is an apparent and unex-
     plained inconsistency between the predicted value for COHb and that
     actually measured.
                                      A-3

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    Table 1.   Concentrations of Carboxyhemoglobin Reported by Anderson
       et al.  and Estimated Using the Coburn-Forster-Kane Equation
Exposure Concentration (ppm)
Initial measurement (% COHb)
Final measurement (% COHb)
Coburn-Forster-Kane Calculated Value
(% COHb) 1
Difference (% COHb)
50
1.4
2.9
3.3
+0.4
100
1.6
4.5
5.8
+1.3
          d)   The Cobum-Forster-Kane equation was derived to deal with endo-
     genously produced CO.   Its application to situations involving exogenous
     CO, though widely done, may be inappropriate.

          e)   There exist,  therefore, at least three possible explanations
     for the differences in the reported COHb measurements and the estimates
     derived using the Coburn-Forster-Kane equation.

              1) The final measurement of COHb in the study by Anderson
          et al. underestimates the true COHb concentration.

              2) The initial measurement of COHb in the study by Anderson
          et al. overestimates the true COHb concentration and when
          "this number is used in the Cobum-Forster-Kane equation, it
          results in an overestimation of the final COHb concentration.

              3) The Coburn-Forster-Kane equation over-estimates COHb
          concentrations in the range of interest for this study.

     2)  There is considerable variability in the data reported.  The following
table gives the mean and range of the time to onset of pain for each day of
the study.
                     Table 2.  Time to Onset of Pain
                               Mean (Range)
           Mon (Air)

           Air (Control)

           50 ppm

           100 ppm

           Friday (Air)
294   (215-480)

325   (220-435)

264.5 (65-390)

263.5 (85-425)

300   (185-485)
                                    A-4

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The exposures on Monday and Friday were included to demonstrate the com-
parison of two air exposures separatead by the study itself.  The air
control and the exposures to 50 or 100 ppm CO were randomized on Tuesday
Vfednesday> and Thursday. The data for Monday and Friday are both considerably
closer to the data for 50 and 100 ppm CO exposure than to that for the air
control.  This is, I believe, associated with the small number of subjects
used in the study.  Dr.  Benignus and his colleagues analyzed the data for
the different air days and did not show any significant differences
between these days.

STATISTICAL ANALYSIS

     Dr. Benignus has developed an analysis of the paper by Anderson et
al. and I have excerpted part of his critique here.

             Critique of the Study by Anderson et al. (1973)


                            Vernon A. Benignus


          The statistical tests used in the study were anticonserv-
     ative (Benignus and Muller, 1982) and thus would have tended
     toward showing a significant effect even if none were present
     in the population.

          The data of Anderson et al. were reanalized using one
     multivariate analyses of variance for each of the two measures
     (time to onset of angina and duration of angina).  Multivariate
     tests were used because the data for 0, 50 and 100 ppm exposure
     were collected from the scjme subjects.  Since two separate
     significance tests were to be run (one for time to onset and
     one for duration) Bonferroni corrections were used to keep
     experimentwise a = .05.  Each test would then be evaluated
     at a/2 = .025.

                      Table A.   Overall Test Results
Variable
Time to Onset
Duration
F
7.72
3.10
df
2,8
2,7
P<
.014
.11
     Table A shows the results of the overall tests.   This table
     agrees with Anderson et al.   that time to onset of angina
     would have been significantly affected by CO exposure but
     does not agree with Anderson et al.  that duration of Angina
     is affected by CO exposure.   Stepdown tests of the time to
     onset data revealed that the 50 ppm exposure showed a CO

                                   A-5

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          effect, p<.017 and the 100 ppm exposure showed a CD
          effect p<.002.
                  Table B.  Means of Time to Onset Data

             CO Level	Mean time to onset of Angina
             0 ppm

            50 ppm

           100 ppm
310

265

264
     Table B shows the mean times to onset for the 0, 50, and
     100 ppm exposure days.  While both the 50 and 100 ppm
     exposures produced shorter ^.imes to onset, they did not
     differ among themselves.  (The fact that the p value for
     the 100 ppm day was lower than for the 50 ppm day was due
     to the fact that the data on -the 100 ppm day were more
     closely correlated with the 0 ppm values.  Thus Table B
     shows a puzzling non-dose-related finding.)

         Inspection of Table 1 in Anderson et al. (1973),
     reveals that all COHb values were related to the exposure
     level so that the non-dose-related findings in time to
     onset remain unexplained.  To be sure, the variability
     was high on all days and with the small number of subjects,
     such non dose related findings can occur even if the effects
     are dose related in the population.  This is especially
     true since the COHb values were both close to the lower
     limits for CO effects and the general dose-effects curve
     can be plausibly argued to have a rather low slope.

         The tests of significance employed in this critique are
     quite conservative.  Thus, the fact that significant results
     would have been reported even if appropriate and conservative
     tests had been done, lends (credibility to the study.  On the
     other hand, the non dose related findings are mildly disturbing.
     Considering (1) the small number of subjects, (2) the high
     variability of the data, (3) 'the fact that both doses were
     near the no-effects level and (4) the low p values on the
     significance tests, the data are strongly suggestive of an
     effect but are sorely in need of replication and extension.
     More subjects should be run and a wider dose range
     should be studied to be able adequately to quantify a
     dose-effects curve.

Conclusions

     Several aspects of the study were of above average quality.  The study
was conducted in a double-blind fashion; very commendable, especially by
comparison to the extant literature.  Only a small number of variables
                                   A-6

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were studied, thus minimizing the chances of finding sane spurious variable
which accidentally covaried with CD exposure; another positive quality.    v

     When compared to an ideal study the study by Anderson et al. has
several flaws in its design and execution and the results have inconsistencies.
However, when compared to the extant literature, the design and execution
of this study is commendable.  None of the inconsistencies are of a major
nature and several plausible explanations exist for them.  The results of
this study suggest that angina is exacerbated by small increases in COHb.

     This study is sorely in need of replication and extension.  More subjects
should be run and a wider dose range should be studied to be able to adequately
quantify the dose response relationships.  Even if no inconsistencies were
present in this study, it would be rash to rely entirely upon one study
with 10 subjects.  It would be equally irresponsible to disregard these
findings.  The greatest imprudence would be to fail to do the follow-up
studies suggested by these results.
                                   A-7

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            APPENDIX B

 CASAC LETTER (AUGUST 31, 1982) TO
EPA ADMINISTRATOR CONCERNING ISSUES
INVOLVED IN SETTING OF NAAQS FOR CO
                B-l

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            UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
                        WASHINGTON, D.C. 20460
                          \August 31, 1982
                                                          OFFICE OF
                                                       THE ADMINISTRATOR
Mrs. Anne M. Gorsuch      \
Administrator
U.S. Environmental Protection Agency
Washington, D.C.  20460

Dear Mrs. Gorsuch:        !

     The Clean Air Scientific Advisory Committee (CASAC) met
on July 6 to provide its advice on several issues related to
the ambient air quality standard for carbon monoxide.  The
Committee had previously ajavised the Administrator of the
scientific adequacy of the criteria document and staff paper
in a closure memorandum dated October 9, 1979.

     At its most recent meeting the Committee provided advice
to the Agency on .four issues.  These included:  1) setting a
revised eight-hour carbon monoxide standard that includes five
allowable exceedances; 2) the role and significance of the 1981
study published by Dr. Wilbert Aronow; 3) sensitivity analysis
and exposure analysis predictions of carboxyhemoglobin (COHb)
levels and ambient CO concentrations under alternative air
quality standards;. 4) range of scientifically acceptable
alternative standards for CO.

     I would like to briefly summarize for you the Committee's
views on each of these issues.

     1.  Development of a Multiple Exceedance 8-Hour Standard.

     The CASAC reached a consensus that a multiple exceedance
standard has both scientific as well as administrative merit.
From a scientific point of view this approach recognizes the
stochastic or random-like (character of meteorological events;
administratively, it reduces the element of chance in determining
compliance with the standard.  In recommending that you adopt
a multiple exceedance standard, the Committee notes that an
increase in the number of allowable exceedances will, in effect
relax the existing standard if the standard level 'remains
unchanged.  In order to provide protection to the public health
with an adequate margin of safety you should consider the impact
of a multiple exceedance standard upon ambient CO concentrations
and levels of blood COHb.

                              B-2

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      2.   Role  of  the  1981 Aronow  Study.

      CASAC  reached  no overall consensus on the significance
 which the Agency  ought to attribute to the Aronow study.  The
 study reported a  10 percent  reduction in the time to onset of
 angina during  treadmill exercise  at blood carboxyhemoglobin
 levels of 2 percent.   CASAC  discussed the fact that the response
 observed  at 2.0%  COHb was more subtle than that observed at
 higher levels  (2.7  -  2.9% COHb) and speculated that even more
 subtle responses  might be found at COHb levels below 2.0%.  The
 Committee concluded that there may be no physiological response
 threshold for  carbon  monoxide.  One CASAC consultant, while
 noting that the study data are solid and irrefutable, concluded
 that  activity  and exposure patterns of angina patients are far
 different from the  general population.  He also observed that
 there is  no reason  to believe that changes in the time of
 onset of  angina during treadmill  exercise are a valid biologic
 endpoint  for the  determination of an adverse health effect.
 Another Committee consultant, however, concluded that shortening
 of exercise time  prior to the onset of an angina attack clearly
 is an adverse health  effect.

      While  reaching no consensus  on the role of this study, the
 Committee's earlier position as stated in the October 9, 1979
 closure memorandum  —• that.the critical effects level for COHb
 occurs between  2.7% — 3.O% and that the onset of angina represents
 an adverse  effect —  remains as the CASAC consensus on this
 issue.

      3.   Scientific and Technical Adequacy of Sensitivity and
          Exposure Analyses.

      The  sensitivity  and exposure analyses were prepared by the
 Agency to compare the  relationship between ambient CO concentrations
 and various levels  of blood COHb.  In addition, the analyses
 estimated the number  and distribution of individuals who were
 projected to experience various COHb levels under alternative
 CO standards.

      CASAC has reviewed the exposure and sensitivity analyses
 and has concluded that both are scientifically acceptable given
 the current state-of-the-art of the scientific community's
 ability to model physiological and other parameters related to
 this pollutant.   Specifically, the Committee would draw to your
 attention two of  its conclusions on these analyses:  1) the
Agency's use of the Haldane constant with a value set at 218 is
 a reasonable selection among a variety of physiological parameters
                              B-3

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discussed in the sensitivity analysis; and 2) the draft preamble
states that an Agency objective is to keep 99% of the population
below a COHb level of 2.5%.  Since there may be no threshold
concentration level for carbon monoxide below which no adverse
effects will be experienced by anyone, and since one hundred
percent protection is not feasible, a social policy choice must
be made to limit societal risk from this pollutant.  From a
scientific standpoint the 99% objective is within the realm of
reason, but there may be other than scientific factors you wish
to consider in reaching a decision on this particular issue.

     4.  Scientifically Acceptable Range for the 8-Hour CO
         Standard.          ;

     In commenting upon the staff's proposals for a revised
8-hour CO standard set at 9 parts per million (p.p.m.) with
five exceedances, or 12 p.p.'m. with one exceedance, the Committee
made the following consensus observations:
          o  a standard set at 12 p.p.m.
     is not scientifically acceptable
with 5 exceedances
          o  a standard established at 12 p.p.m. with 1
     would provide a very small margin of safety
               exceedance
          o  the scientific evidence alone cannot identify an
     exact level at which to set a standard for carbon monoxide.
     Given the need to protect sensitive members of the population
     from this pollutant, the Committee advises you to choose a
     standard level and a corresponding number of exceedances that
     will limit COHb below tihe critical effects level of 2.7 - 3.01,
     with an adequate margin of safety.

     The Committee appreciates the opportunity to advise you on
the carbon monoxide standard and hopes that its comments will
be useful as you finalize tlie standard.  We urge you to proceed
expeditiously in this matter because the criteria document and
staff paper, reviewed by CASAC more than three years ago, will
be increasingly subject to challenge because of any newly published
literature on this pollutant.  In addition, both the private
sector and individual citizens need to know the standard level
for the next five years for [planning purposes and for reassurance
that public health is being adequately protected.

                              Sincerely yours,
cc:  Dr. John W. Hernandez
     Kathleen Bennett
     Dr. Terry F. Yosie
                              Sheldon K. Friedlander
                              Chairman, Clean Air Scientific
                                Advisory Committee
                               B-4

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                       APPENDIX C

     CASAC LETTER (MAY 7, 1984) TO EPA ADMINISTRATOR
     CONCERNING FINDINGS AND RECOMMENDATIONS ON THE
SCIENTIFIC BASIS FOR A REVISED NAAQS FOR CARBON MONOXIDE
                           C-l

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                UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
                              WASHINGTON, D.C.  20460
                                   May 17, 1984
                                                                       OFFICE OF
                                                                   THE ADMINISTRATOR
Honorable William D. Ruckelshaus
Administrator
U.S. Environmental Protection Agency
401 M Street, S.W.
Washington, D.C.  20460

Dear Mr. Ruckelshaus:

     The Clean Mr Scientific Advisory Committee  (CASAC) has completed its
review of two documents related to thb development of revised primary
National Ambient Air Quality Standards (NAAOS) for Carbon Monoxide (CO).
Ihe documents were the Revised Evaluation of Health Effects Associated with_ ,
Carbon Monoxide Exposure;  An Addendum to the 1979 Air Quality Criteria
Document for Carbon Monoxide written by the staff of the Office of Research
and Development (ORD), and a staff pajpef entitled Review of the NAAQS for
Carbon Monoxide; 1983 Reassessment of Scientific and Technical Information
prepared by the Office of Air Quality! Planning and Standards (OAQPS).  The
Committee unanimously concluded that both documents represent a scientifically
balanced and defensible summary of the current basis of our knowledge of
the health effects literature for this pollutant.

     As you know, the latest CASAC review of the CO documents took place in
an atmosphere of great scientific uncertainty and controversy due to the
fact that a group of scientists condupting a revi-ew of the protocols for a major
series of peer reviewed studies, carried' out by Dr. Wilbert Aronowr had "shortly
before concluded that adequate standardized procedures for scientific
research were not utilized in those studies.  Confronted with this situation,
Agency staff in both ORD and OAQPS moved quickly and resolutely to analyze
the remaining scientific basis for the Clean Air Act requirement to finalize
a revised CO standard.  Hie CASAC concludes that, even without the use of
the Aronow studies to determine a critical effects level from CO exposures,
there remains a sufficient and scientifically adequate basis on which to
finalize the CO standard.

     As a result of its review of the information contained in these docu-
ments, the CASAC recommends that you consider choosing the 8-hour and
1-hour carbon monoxide standards to maintain approximately current levels
of protection.  A more extended analysis of the factors that led to this
reconmendation is contained in the enclosed report.
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     Thank you for the opportunity 'to present the Committee's views on  this
important public health issue.
                                           Sincerely,
                                           Morton Lippnanry1; Chairman
                                           Clean Air Scientific
                                             Advisory Canmittee
Enclosure

cc:  Mr. Alvin Aim
     Mr. Joseph Cannon
     Dr. Bernard Goldstein
     Dr. Tterry Yosie
                                        C-3

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       CASAC Findings and Recommendations on the Scientific Basis for
                    a Revised NAAQS for Carbon Monoxide

Addendum to the CO Air Quality Criteria Document


     1.  A key issue in the evaluation of public health risks from carbon

monoxide (CO) exposures concerns the relation between CO in air and its

displacement of oxygen .in blood hemoglobin.  The index for this displacement,

known as carboxyhemoglobin (COHb), is expressed as a percentage of the

blood hemoglobin.  There is a scientific consensus that relatively low

levels of COHb are associated with critical (i.e., health impairing) health

effects.  The discussion of the scientific evidence thus centers on what

percentage of, COHb causes a critical effect.

     On October 9, 1979, CASAC submitted a report to the Administrator

concluding that the critical COHb level occurred within a range of 2.7—3.0%.

The Cornittee reached this finding following an extensive review of the

scientific literature, including a series of studies performed by Dr. Wilbert

Aronow.  CASAC expressed some reservations about one of these studies

(Aronow, 1978 which reported effects at levels  [1.8%] well below the  2.7-

3.0% range) in view of the fact that some confounding factors in the study

protocols were not appropriately accounted for.  The Committee further

recommended that "given the uncertainties stemming from the methodological

approach,  [the Agency].. .should utilize the  [1978 Aronow] study for margin

of safety considerations rather than using it for the determination of a

threshold value"   (CASAC report, October 9, 1979, p.5).  On August 31,

1982 CASAC sent a  follow-up report on several issues related to the NAAQS

for carbon monoxide.   In that report the Committee reaffirmed its prior

findings on the critical COHb effects level.  It should be noted that

CASAC1 s 1982 recommendations were reached after the Committee members
                                     i

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 had an opportunity to review an additional (1981)  study by Dr.  Aronow which



 concluded  that a 10%  reduction in the time to onset of an angina attack



 occurred during treadmill  exercise with 2% CDHb.



     A review of the most  recent update of this scientific literature in



 the August 1983 draft  EPA  Addendum to the  00 Air Quality Criteria Document



 persuades  CASAC that  there is no significant reason to substantively alter



 its previous  findings.  An elaboration of  CASAC's  current reasoning  on



 several issues will clarify the Committee's position,   These  include:



           A,   The  role of  the Aronow  studies



     A key question raised about Aronow's  work was" whether or not the



 procedures used insured that the studies were double blind.   A double



 blind  protocol is  one  in which  neither the subjects nor the laboratory



 technicians conducting the experiments  and collecting  the  data are aware of



 key parameters  of  the study (exposure conditions,  timing,  etc.) and  the



 results of the  responses by the  experimental  grsap and the-control group.



 It  is  apparent  that such double  blind procedures were  not  applied in Aronow's



work because technicians who were directly  involved with the subjects  knew



some of the important parameters of the  study.  The lack of quality  assurance



checks represents another  issue of concern.   In these  respects, the  results



of Aronow's work do not meet a reasonable standard  of  scientific quality



for a study of the kinds of responses of interest, and  therefore, they should



not be used by the Agency  in defining the critical COHb level.



          B.  Ihe role of the Anderson study



     The 1973 study by Anderson et al. reported that angina patients exposed



to low CO levels while at rest experienced a statistically significant reduction



in time to onset of exercise induced angina at average COHb levels of 2.9% and



4.5%.  Ihe study further concluded that there was  a significantly lengthened



                                         C-5

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angina attack during exercise at an average COHb level of 4.5%.  The 1983
CO criteria document addendum noted concerns expressed by some parties
about the study due to the small number of subjects studied, apparent
inconsistencies between predicted and observed COHb levels, the possibility
that the protocols were not .truly double blind, and the lack of subsequent
confirmatory findings.
     CASAC reached several conclusions concerning this study.  It was trou-
bled that so few patients were included in the study design and that there was
uncertainty about the exposures to which the patients were subjected. The
Ccnroittee agreed that it is important to replicate such a study, but the notion
that a study has no validity until it's been replicated is flawed.  Based
                                     I
upon its current knowledge of how the study was conducted, CASAC presumes
that double blind protocols were, in fact, observed and that discrepancies
between observed and predicted COHb levels are not as great or as serious
as originally suggested.  In summary; while CASAC treats the Anderson et
al. study with caution, it can find no substantive reason at this tine to
dispute the reported values, and it reccmnends that the Agency not disregard
its findings.                        j
          C.  Additional studies     [
     CASAC wishes to point cut two sets of additional studies which lend
support to concerns about low level CO exposures.  In 1974, both Raven et al.
and Drinkwater et al. reported  statistically significant decreases (less
than 5%) in exercise time for work capacity in healthy, nonsmoking young
and middle aged ran at approximately 2.3 - 2.8% COHb.  Also, a 1980 controlled
human exposure study by Davies & Smith, observed changes in electrocardiogram
(EKG) measurements in a small number of healthy nonsmoking young men at
2.4% COHb.  Such CO induced changes are a cause for public health concern
and should be factored into the Agency's thinking for setting  a standard
with an adequate margin of safety.
                                     ;     c-e

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          D.  Use  of the Coburn-Fcster-Kane (CFK)  equation
     The CFK model is  the most important  available tool  for analyzing  a
 number  of physiologically important variables  (blood volume and endogenous
 CO production rate, for example)  in order to project a relationship
 between ambient CO exposures and  resulting COHb  levels.  Wiile this
 model,  like any model, is subject to the  need  for  additional evaluation
 of COHb in different population groups, it is  reasonable to conclude that
 the CFK equation accurately predicts CO uptake under differing exposure
 conditions.
          E.  Summary of cardiovascular effects
     The Committee unanimously agrees that:  1)  the key  mechanism of CO toxicity"
 is the  decreased oxygen carrying  capacity resulting frcm the greater af-
 finity  of blood hemoglobin for carbon monoxide than for  oxygen; 2) reduction
 in time to the onset of an angina attack  is  a medically  significant event
 and should be considered an adverse health effect; and 3) following a
 review  of the peer reviewed scientific literature  (not including the Aronow
 studies), the critical effects level for  NAAOS setting purposes is
 approximately 3% COHb (not including a margin  of safety).

     2.  A second  important public health issue  in setting a NAAQS for  carbon
monoxide concerns CD-induced central nervous system effects.  Decreased
vigilance or sensory-motor function is a health effect which the standard
ought to protect against.  CASAC's position  is that such behavioral effects
are observed between 5-8% COHb.
     3.  The Committee was asked  to address the  issue of the role of CO in
Sudden  Infant Death Syndrome (SIDS).  A review of the current scientific
                                       C-7

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literature leads to the conclusion that there is not a sufficient scientific
basis to establish a connection between a CO exposure level and SIES.

OftOPS Staff Paper Review of the NAA05 For Carbon Monoxide

     Based upon the addendum to the revised Air Quality Criteria Document
for Carbon Monoxide, QAOPS developed a staff paper analyzing alternative
ranges of concentration levels for a final promulgated standard.  The current suite
of primary standards is set at 9 parts per million (ppm) for the 8-hour averaging
time and 35 ppm for the 1-hour average.
     CASAC was asked to advise the Agency on several issues associated with
the proposed ranges.  The following discussion responds to the Agency request.
     1.  CASAC reaffirms the judgment it reached in its October 1979 report
         that reduction in the time to onset of angina aggravation represents
         an adverse health effect.
     2.  The Committee concurs with the Agency that R-hour and 1-hour
         standards are the appropriate averaging times, but it recommends
         that there be additional discussion and more explicit comparison
         in the regulatory package concerning the relationship between the
         two averaging times, particularly in terms of what attainment of
         the 8-hour standard portends for the health protection provided by
         the 1-hour standard.
     3.  The factors identified by OAOPS for margin of safety
         consideration are appropriate.  Underlying CASAC's view of
         the margin of safety, however, is its traditional belief that
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    where the scientific data, as  in this case, are subject to large



    uncertainties, it is desirable for the Administrator to consider a



    greater margin of safety than  the  numerical values of OOHb generated



    by the Coburn equation might otherwise suggest.



4.  The QAQPS staff recommends that the Administrator retain or



    select an 8-hour primary standard  in the range of 9 to 12 ppm.



    With regard to the 1-hour primary  standard, the staff recomnends



    that a selection be made within the range of  25 to 35 ppm.  CASAC



    concurs that the proposed ranges for both the 8-hour and 1-hour



    primary standards are scientifically defensible.  Given the uncer-



    tainties within the scientific data base and  Discussion of margin



    of safety issues, the Committee reconmenrts that you consider



    choosing standard limits that  maintain approximately current



    levels of protection.
                                 C-9
                                          •&U. S. GOVERNMENT PRINTING OFFICE: 1984/759-102/10639.

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