United StatM
             Environmental Protection
             Agency
Health Effects
Research Laboratory
Research Triangle Park.
North Carolina 27711
                                EPA/600/1-90/001
                                Fbo'ruary 1990
             Research and Development
             Indoor  Air  -  Health
            LJ1
Neurotoxic Effects  of
Controlled Exposure  to  a
Complex  Mixture of Volatile
Organic  Compounds

                         38101
  EJBD
k ARCHIVE
  EPA
.  600-
> 1-
  90-
s, 001

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DISCLAIMER:  This manuscript has been reviewed by the Health Effects Laboratory,
U.S.   Environmental  Protection  Agency, and approved for publication as an EPA
document.  Approval does not signify that the contents necessarily  reflect  the
views and policies of the Agency.  Mention of trade names or commercial products
does not constitute endorsement or recommendation for use.

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                       FINAL REPORT (Deliverable 2520)


            NEUROTOXIC EFFECTS OF CONTROLLED EXPOSURE TO A COMPLEX

                    MIXTURE OF VOLATILE ORGANIC COMPOUNDS


                   Principal Investigator:   David A. Otto

                                                     2
                       Co-Investigators:   Lars Molhave
                                              3
                              H.Kenneth Hudnell

                              George Goldstein

                                 John O'Neil1

                                                   4
                         Statistician:   Dennis House

                      Technical Support:   Gregory Rose
                                Jon Berntsen

                                Wayne Counts
                               Shirley Fowler1

                                 Scott Meade


           Human Studies Division, Health Effects Research Laboratory,
                 U.S. Environmental Protection Agency, RTF, NC
             2
              Institute of Environmental and Occupational Medicine,
                    University of Aarhus, Aarhus, Denmark

          Neurotoxicology Division, Health Effects Research Laboratory,
                 U.S. Environmental Protection Agency, RTF, NC

       Research and Regulatory Support Division, Health Effects Research
           Laboratory, U.S. Environmental Protection Agency, RTF, NC

            NSI Environmental Services, Inc., Research Triangle Park,
                                  NC  27711

                 CE Environmental, Inc., Chapel Hill, NC  27514

     Sources of Support:  EPA Contract 68-02-4179  (Combustion Engineering)
                         EPA Contract 68-02-3800  (Subject Recruitment)




 Repository Material                        u  H    „    us EPA
            /„     ,,                          Headquarters and Chemical Libraries
Pern lanent Collection                          EPA west a* Room 3340
                                                          Mailcode 3404T
                                                     1301 Constitution Ave NW
                                                      Washington DC 20004
                                                          202-566-0556

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                               PREFACE

      In  October  1986  Congress  passed  the  Superfund   Amendments  and
Reauthorization Act (SARA, PL 99-499) that Includes Title IV - The Radon Gas
and Indoor Air Quality Research Act.  The Act directs that  EPA undertake a
comprehensive Indoor air research program.

      Research program requirements under Superfund Title IV  are specific.
They  Include  Identifying, characterizing, and  monitoring  (measuring)  the
sources and levels of Indoor air pollution; developing Instruments for Indoor
air quality data collection; and studying high-risk  building types.   The
statute also requires research directed at Identifying effects of Indoor air
pollution  on  human health.    In  the area of mitigation and control  the
following are required:  development of measures to prevent  or abate Indoor
air pollution; demonstration of methods  to reduce or eliminate  Indoor air
pollution; development of methods to assess the  potential for  contamination
of new construction from soil gas,  and examination of design  measures for
preventing  Indoor  air pollution.   EPA's  indoor  air research program  is
designed to be responsive In every way to the legislation.

      In  responding  to  the  requirements  of  Title  IV  of  the  Superfund
Amendments, EPA-ORD has organized the Indoor Air Research Program around the
following categories of research:  A)  Sources of  Indoor Air  Pollution;  B)
Building Diagnosis and Measurement Methods; C) Health Effects; D) Exposure
and Risk (Health Impact)  Assessment; and E) Building Systems and Indoor Air
Quality Control Options.

      EPA  is  directed  to   undertake  this  comprehensive   research  and
development effort not only  through in-house work but also  in  coordination
with other Federal  agencies,  state and local governments,  and private sector
organizations having an interest in indoor air pollution.

The ultimate goal of SARA Title IV is the dissemination of information to the
public.  This activity includes the  publication  of  scientific  and technical
reports in the areas described above.  To support these research  activities
and other  interests  as well, EPA publishes its results  in  the  INDOOR AIR
report series.   This  series  consists  of five subject categories:  Sources,
Measurement, Health, Assessment,  and Control.  Each report is  printed In  a
limited quantity. Copies may be ordered while supplies last  from:

      U.S. Environmental  Protection Agency
      Center for Environmental Research Information
      26 West Martin Luther King Drive
      Cincinnati, OH 45268

When EPA supplies are depleted, copies may be ordered  from:

      National Technical  Information Service
      U.S. Department of Commerce
      5285 Port Royal Road
      Springfield, VA 22161

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TABLE OF CONTENTS
A.  EXECUTIVE SUMMARY	J
B.  BACKGROUND AND RATIONALE	*
C.  METHODS	;
      C.I   Subject Selection and Characterizaton	/
      C.2   Physical Facilities	8
      C.3   VOC Generation and Monitoring	9
      C.4   Equipment Readiness  and Pilot  Study	9
      C.5   Experimental Design	1°
      C.6   Behavioral Battery	11
      C.7   Subjective Reactions	11
      C.8   Confirmatory and Exploratory  Hypotheses	12
      C.9   Data  Analysis  Plan	13
      C.10 Risks and  Safeguards	1*
 D  RESULTS	15
      D.I   Comfort Meter  (Potentiometer)	15
      D.2   Symptom Questionnaire	15
      D.3   Auditory  Digit Span (ADS)	16
      D.4   NES Main  Test  Results	17
               D.4a  Finger Tapping 	17
               D.4b  Visual Digit Span (VDS)	17
               D.4c  Continuous Performance Test (CPT)	17
               D.4d  Symbol Digit Substitution (SDS)	18
               D.4e  Serial Digit Learning (SDL)	18
               D.4f  Pattern Memory (PM)	18
               D.4g  Switching Attention (SWATT)	18
               D.4h  Mood Scales	19
       D.5   NES Secondary Test Results	19
               D.5a  Associate Learning/Recall	19
               D.5b  Pattern Comparison	19
               D.5c  Simple Reaction Time  (SRT)	20
               D.5d  Grammatical Reasoning	20
               D.5e  Horizonal Arithmetic	20
       D.6   Water Consumption	20
       D.7   Learning/Practice Effects	20
  E.  DISCUSSION	22
       E.I   Confirmatory  Hypotheses	^
       E.2   Exploratory Hypotheses	22
                E.2a   Sensory Irritation	22
                E.2b   Controlled  Climate Variables	23
                E.2c   Motor Performance		23
                E.2d   Mood  Scale Alterations	24
                E.2e   Cognitive  Performance	24
        E.3   Effects  of  Practice and  Learning	25
  F.  REFERENCES	28

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LIST OF TABLES

1.   Symptoms of Sick Building Syndrome	30
2.   Subject Selection Criteria	30
3.   VOC Mixture	31
     Experimental Design	31
5.   Measurement Variables	32
6.   Comfort Ratings (Potentiometer Readings)	34
7.   Symptom Questionnaire	35
8.   Symptom Response Changes Over Time	37
9.   Yes-No Question Responses	38
10.  Auditory Digit Span (without 12 subjects)	39
11.  Auditory Digit Span (All subjects)	40
12.  Finger Tapping	41
13.  Visual Digit Span	41
14.  Continuous Performance Task	42
15.  Symbol Digit Substitution	42
16.  Serial Digit Learning	43
17.  Pattern Memory	43
18.  Switching Attention Test	44
19.  Mood Scale	45
20.  Associative Learning	46
21.  Pattern Comparison	46
22.  Simple Reaction Time	47
23.  Grammatical Reasoning	47
24.  Horizonal Arithmetic	48
25.  Serial Order Effects:  Auditory Digit Span	49
26.  Serial Order Effects:  Visual Digit Span	50
27.  Serial Order Effects:  Pattern Memory	50
28.  Summary of Serial Order Effects	51
LIST OF FIGURES

1.   Schematic of Test Protocol	53
2.   Comfort Rating Scale Results	54
3.   Headache, Odor and Air Quality Responses	55
4.   Odor, Eye and Throat Irritation	56
5.   Control Question Responses	57
6.   Yes-No Question Responses	58
7.   Auditory Digit Span	59
8.   Visual Digit Span	60
9.   Continuous Performance Task	61
10.  Switching Attention Test		62
11.  Mood Scale Difference Scores	63
12.  Serial Order Effects:  Auditory Digit Span	64
13.  Serial Order Effects:  Visual Digit Span	65
14.  Serial Order Effects:  Confusion Scale	66
15.  Serial Order Effects:  Pattern Memory		67

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APPENDICES

A.  VOC Generation and Monitoring Methods	.68
B.  Instructions for Administering and Scoring the
    Auditory Digit Span Test	79
C.  Description of Neurobehavioral Evaluation System
    (NES) Tests	84
D.  Symptom Questionnaire	91
E.  Linear Analysis Rating Scale (Potentiometer)	93

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A.  EXECUTIVE SUMMARY
     Growing public awareness of the potential health hazards of chemicals found
in  the  indoor  environment  has  given rise to the definition of a new problem
designated as sick (or tight) building syndrome (SBS).  SBS is characterized  by
irritation  of  the  eyes,  nose  and  throat.  Preliminary evidence from Danish
studies (Molhave et al, 1986;  Kjaergaard et al, 1989) also indicates effects on
cognitive  functions  such  as memory.  Many of the chemicals found indoors that
have been linked with SBS are volatile organic compounds  (VOCs).   Congress  has
mandated  EPA to characterize the sources, real-world exposure levels and health
risks of indoor pollutants.  Little information is currently  available  on  the
health  effects of exposure to VOC mixtures found in new  or renovated buildings.
EPA has established an intramural research program  to  address  this  question.
The current report details the results of the initial study in this program.

     Objectives of this study were to confirm and extend  the pioneering work  of
Molhave  et  al  (1986) and to identify sensitive measures for use in subsequent
studies  of  sick-building  syndrome.   Molhave  et  al.   demonstrated  sensory
irritant  and  neurobehavioral effects of controlled human exposure to a complex
VOC mixture at concentrations of 5 and 25  mg/m3,  representing  the  usual  and
maximal  levels  found  in  new  homes.  The design of  the Danish experiment was
problematic, however.  A  repeated-measures design was  used  in  which  subjects
completed  clean air and  exposure conditions in the morning and afternoon of the
same day.  Three types of confounding may have occurred in this design—practice
effects,  VOC  exposure   carry-over  effects  from  morning  to  afternoon,  and
time-of-day  differences.   The  present  study  was  designed  to  minimize  or
eliminate  these  problems  by  scheduling  a  separate training day to minimize
practice effects, testing all subjects at the same time of day (afternoon),  and
separating clean air and  exposure runs by a minimum interval of one week.

     Other important differences between studies included the subject population

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and  tests  used.   The Danish study included both males and females aged 18-64,
all with documented histories of chemical sensitivity.  The  present  study  was
limited to young adult males aged 18-39 with no history of chemical sensitivity.
Only two neurobehavioral performance tests were used in the Danish study,  while
fourteen  tests  were used in the present study to fully characterize the nature
of the possible neurobehavioral effects of VOC exposure.

     Results of this study confirm the adverse subjective reactions of  subjects
to a 25 mg/m3 concentration of volatile organic compounds reported by Molhave et
al.  Ratings of general discomfort (defined as irritation of the eyes, nose  and
throat), symptom questionnaire responses on odor intensity, air quality, eye and
throat irritation, headache and drowsiness, and mood scale measures  of  fatigue
and  confusion  all  differed  in  predicted  directions  between  clean air and
exposure  conditions.   However,  no  convincing  evidence  was  found  of   any
neurobehavioral  impairment  associated  with exposure to the VOC mixture.  That
is, we were unable to confirm the Molhave et al report that  VOCs  impair  short
term memory as measured by auditory digit span.  Nor were other tests of memory,
sensorimotor performance,  or  more  complex  cognitive  functions  affected  by
exposure  to  this  concentration of VOCs.  In summary, clear adverse subjective
reactions to VOC exposure, but no functional (neurobehavioral) impairments, were
found.

     Confirmation  of  sensory  irritation  in  a  normal,  healthy  adult  male
population  is  quite  important.   This  finding  demonstrates  that subjective
reactions to  VOCs  at  levels  found  in  new  buildings  are  not  limited  to
"complainers"  or  chemically sensitive subgroups in the general population.  In
fact, subjects in this study probably represent the subgroup least likely to  be
affected  by VOC exposure.  Sensory irritation, headache and drowsiness are also
of  considerable  importance  in  the  community   and   workplace.    Perceived
dissatisfaction  with  air quality in the home, office, market place, or factory
can lead to reduced productivity, decreased quality of  life  among  home-owners
and  workers,  and  could  result in mass exodus from buildings with air quality
problems.  This  is an important factor in the sick-building  syndrome.   Further
consideration  should  be  given  to  the  relevance  of subjective reactions in

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evaluating the adverse health effects of indoor pollutants.

     Subjects in this study and previous Danish studies found the odor  of  VOCs
to  be  very  strong  and  unpleasant.   The unmistakable odor of VOCs confounds
efforts to conduct a double-blind experiment.  Subjective reactions to  the  VOC
mixture  can  be  attributed, at least in part, to the unpleasant odor.  If this
were the only objectionable characteristic of VOCs, we would  probably  conclude
that  VOCs  at  concentrations found in the indoor environment do not constitute
any significant health risk.  The time course of responses to odor and  symptoms
of  irritation  differ,  however, suggesting that odor accounts for only part of
the subjective reaction to VOCs.   Further  study  is  needed  to  differentiate
olfactory and trigeminal components of the VOC response.

     Failure to find any functional consequences of exposure to the VOC  mixture
however,  raises  important  questions  in  assessing  health risks and planning
subsequent indoor air studies.  For instance, we cannot conclude that a 25 mg/m3
concentration  of  VOCs poses no neurotoxic risk to susceptible subgroups of the
general population, or even  to  the general population.   Further  study  with  a
chemically sensitive group of subjects is needed and the relationship of age and
gender to VOC response needs to be explored.   It  is  also  possible  that  the
neurobehavioral   tests  used  were not sensitive (or difficult) enough to detect
effects.  If subjects find  that VOCs  irritate  the eyes  and  throat  and  induce
headaches,  one would expect performance  to be impaired, at least indirectly, by
distraction or impaired attention.

     Another possibility   to  consider  is  physiological  endpoints.   If  VOCs
irritate  the  eyes,  nose   and   throat,  we   should examine secretions of these
tissues for evidence of inflammation.  Kjaergaard et al (1989) recently reported
evidence  of  inflammatory   response  in  eye  fluids  following  VOC  exposure.
However, no evidence of inflammatory  response  in  nasal  secretions  was  found,
consistent  with  symptom   response   in   the   present  study.   Further study of
inflammatory effects of VOC  exposure  in eye  fluid and other mucosal tissues  are
needed.

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B.  BACKGROUND AND RATIONALE.
     "Sick Building Syndrome" costs industry  and  the  government  considerable
time  and  dollars  in  lost  vorktime,  worker  dissatisfaction,  and temporary
abandonment of facilities (Melius et al, 1984).  Beyond the economic  impact  is
the  question  of  possible  adverse  health  effects  of exposure to indoor air
pollutants found in new and renovated  homes  and  commercial  buildings.   Most
people  spend  the  majority  of  their  lives  indoors (NRC, 1981).  Therefore,
agencies and institutions responsible for protecting human health therefore need
to  carefully study the potential adverse effects of chemicals commonly found in
indoor environments.

     Molhave and his colleagues  (1986)  have  studied  the  health  effects  of
controlled  exposure  to  mixtures  of  volatile organic compounds found in new-
construction Danish homes.  Subjects were healthy adults selected by means of  a
questionnaire  survey  to  identify  individuals who have experienced discomfort
associated  with  indoor  air  quality.   Subjects   were   exposed   to   three
concentrations  (0,  5, 25 mg/m3) of a mixture containing 22 VOCs in the climate
chamber at the Institute of Hygiene, Environmental  and  Occupational  Medicine,
University  of  Aarhus.   The  composition  of  this  mixture  was based on VOCs
commonly found in newly constructed Danish homes (Molhave and Holler, 1979)  and
is  similar  to the levels and types of VOCs found in U.S.  homes (White, 1987).
Several  measures  of  performance  and  sensory   irritation   were   obtained.
Subjective  ratings  of  sensory irritation and air quality varied significantly
with VOC exposure.  Results also indicated impaired memory as  measured  by  the
Digit  Span  Test.   The  implications of these results are that exposure to low
level mixtures of VOCs commonly found in  new  or  renovated  buildings,  levels
hitherto  considered to be harmless to human health, may impair-the performance,
productivity, comfort and well-being of workers and home-owners.

     These findings  provide  preliminary  evidence  of  neurotoxic  effects  of

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exposure to complex low-level mixtures of VOCs, although further study is needed
to determine the precise nature of the  observed  effects.   For  instance,  are
these  findings limited to a particular subset of the population who dislike the
odor of VOCs?  Is  the  response  psychosomatic  or  does  low-level  mixed  VOC
exposure  produce  measurable  decrements  in objective health endpoints?  Which
health endpoints are most  sensitive  to  VOC  exposure—e.g.,  sensory,  memory
pulmonary, or immunological measures?  Are there other vulnerable subsets of the
general population such as young children, elderly, asthmatics or  persons  with
allergies?   Are  VOC exposure effects due to single constituents of the mixture
or the combination of several constituents?  Systematic study of these questions
under  controlled laboratory conditions is necessary to clarify the human health
risks of exposure to indoor air pollutants.  Very little evidence  is  currently
available  concerning   the  neurotoxic  effects  of  low-level human exposure to
complex VOC mixtures.   Controlled exposure studies  are  also  necessary  before
undertaking related field studies.

     The  type of symptoms affected by VOC exposure  in  the Molhave  et  al  study
has  been  identified   as   the Sick Building Syndrome  (SBS) by WHO (1982).  This
syndrome  is characterized by subjective and physiological  symptoms  of  sensory
irritation of mucosal  tissue and other symptoms  as  shown  in Table 1.

     These symptoms are acute,  nonspecific,   and   cannot  be  ascribed  to  any
specific  cheraical(s)   found   in   the problem  environment.  In the Molhave et al
study,  individual VOCs  in  the  complex mixture  were present  at  concentrations
well  below  levels   that   would be expected  to  produce irritant effects for any
single  chemical in  the   mixture.   Molhave   (1986)  has   hypothecized   that  the
summation of   effects   of   exposure  to  the mixture, rather than any single con-
stituent  chemical,  may  be  the  cause   of   the   effects  observed  in  the  study.
Molhave  (1986) has  further hypothecized  that the  observed effects are mediated
by trigeminal nerve function which is generally   considered   to  be  the  neuro-
physiological substrate of  the chemical  sense.
 the
The objective of the present study was to confirm the principal findings of
 Aarhus   study—i.e.,  increased  perception  of  sensory  irritation  and

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discomfort associated with exposure to a  low-level  complex  mixture  of  VOCs.
Molhave  et al also reported an effect on digit span performance suggestive of a
VOC-related impairment of short-term memory.  The functional significance of the
digit  span finding, however, requires clarification.  The observed effect could
be a primary neurotoxic effect of VOC exposure or a secondary effect of  sensory
irritation  (e.g.,  distraction).   Digit  span  performance  also reflects both
attentional and memory processes.  The  present  study  included  a  battery  of
neurobehavioral  tests designed to clarify the nature and validity of the Danish
findings.  Response to low-level VOC exposure  was  studied  in  normal  healthy
adults.

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C.  METHODS
C.I Subject Selection and Characterization.
     Normal  healthy  adult  volunteers  were  recruited  by   the  UNC   Subject
Recruitment  Office  (operated  under  EPA Contract 68-02-3800).  Advertisements
posted on campus  and  published  in  local  newspapers  were  used   to  recruit
subjects.   Selection  criteria  based  on  those  used  in the Danish study are
specified in Table 2.  In brief, healthy, non- smoking Caucasion males  aged  18
to  40  with  no  history  of  asthma, allergic rhinitis, cigarette use or other
medical conditions were referred for physical examination and  allergy  testing.
It  should  be noted that these requirements were more stringent than the Danish
study which included smokers and persons up to 64 years  of  age.   Smokers  and
persons  over 40 years were excluded from the present study in order  to minimize
the variability in performance and sensory irritation that might  be  associated
with these variables.

     Exposure history was assessed by a questionnaire adapted  from Molhave et al
(1986).   Exposure  history  was  used  to  exclude individuals with  significant
occupational or recreational  exposures  that  could  influence  neurobehavioral
measures  and   to  exclude  individuals  with histories of chemical sensitivity.
Volunteers who met selection criteria were referred for physical examination and
allergy skin testing.  A standard scratch test for 18 common allergens including
airborne pollens, cat and dog danders, molds and house  dust   was  administered.
Subjects  with  positive  skin  allergy  tests were not included in the study in
accordance with the selection criteria of Molhave et al (1986).

     Payment for participation  conformed to guidelines established  by  the  EPA
Clinical  Research  Branch.  A  base  rate of $7 per hour and bonus for completion
of the study was paid to subjects.   $30  was  paid  to  potential  subjects  who
completed the physical examination.  Subjects who qualified for participation in
th* study earned $84 more, depending on the number of hours devoted to  training

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and  running  in  the  control  and exposure sessions.  Subjects also received a
bonus of $30 for completion of the experiment.  Subjects completing  all  phases
of the study earned a maximum of $144.

     Payments were made at the end of the  experiment  or  upon  termination  of
participation.   Although a subject could elect to discontinue the experiment at
any time due to discomfort, concern or other reasons, if the  subject  chose  to
leave for non-medical reasons, only fees earned to that point were paid.  On the
other hand,  if  the  investigators  terminated  an  individual  on  medical  or
technical  grounds  once  an  exposure  session  had  commenced,  the subject(s)
received full payment for that session.   Subjects  commuting  from  Durham  and
Raleigh  were  paid an additional $6 and $11, respectively, and all parking fees
were paid.

     To conform to the Federal Privacy Act, each subject was assigned  a  number
at  the time of physical examination.  Subject privacy was maintained thereafter
by exclusive use of the subject number for subject identification.

     Seventy-six subjects completed the VOC study.  Data from ten subjects  vere
discarded  for  technical  reasons  including the addition of the auditory digit
span test after seven subjects had been run and the  accidental  termination  of
exposure during one session with three subjects.  Therefore, data from sixty-six
subjects vere used in statistical analyses.  The mean age of subjects  was  25.0
yrs.  (s.d.  5.6).

C.2 Physical Facilities.

     The study vas carried out in the  exposure  facilities  of  the  EPA  Human
Studies  Division  in  Chapel  Hill  (Strong,  1978).  The environment including
temperature, humidity, light levels and pollutant concentration vas continuously
controlled  and  monitored  by  computer.   Personnel  operating  this  facility
included  EPA  psychologists  and  technicians  and  a  team  of  technical  and
engineering personnel from CE, Inc.  vho operate and maintain the facility under
contract  to EPA.  These personnel vere highly trained individuals vho  routinely
                                       8

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administer  other  similar  protocols.   A  licensed  physician  was immediately
available to the operating team during all experiments.

C.3 VOC Generation and Monitoring.

     The complex VOC mixture described by Molhave et al  (1986) was used   in   the
present  study with the substitution of one compound (1,1-dichloroethane  for  1,2
dichlorethane).  The Molhave mixture of 22 volatile organic compounds  is shown
in Table 3.  The compounds were selected on the basis of previous Danish  surveys
(Molhave and Holler, 1979) including the 10 most commonly  found and  the 10 found
in  greatest concentration in indoor nonindustrial environments, but ex-  eluding
known carcinogens.  That is, this mixture is characteristic of the   chem- icals
and concentrations to which residents in new Danish and  American homes are often
exposed.  Any compounds classified  by the International  Agency for   Research  on
Cancer   (IARC  monographs)  as Group 1 (chemicals for which there is "sufficient
evidence from epidemiological studies to support a causal  associa- tion   between
the  exposure  and cancer") or Group 2 (chemicals that are probably  carcinogenic
to humans—i.e.,  "exposures for which there  [is] at least  limited   evidence  of
carcinogenic!ty  to humans" or "sufficient evidence in animals") were excluded.

     The concentration  of  individual constituents of  the mixture shown in Table
3  is  specified  in ug/m3,  each of which  is below  the  threshold limit value (TLV)
for occupational exposure.  TLV  is  the concentration  of  a   chemical  to which
workers may  be  continuously exposed for  8 hours without  adverse health effect as
defined by OSHA. A   system   for  generating   and  monitoring the   VOC   mixture
equivalent   to  that  used  in  the  Danish study was  constructed  by CE,  Inc.  A des-
scription  is contained  in Appendix  A.  Subjects  were  exposed   to   a  25 mg/m3
concentration  of VOCs  measured   as  a toluene  equivalent concentration with  a
flame  ionization detector (FID).  The  duration of exposure was 2.75  hours.

C.4 Equipment Readiness and  Pilot  Study.

     Prior  to initiating the  main study,  a  pilot  study   was   conducted   to   test
equipment,  to train  the operating staff,  and  to  check-out  the protocol to insure

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that the testing schedule ran smoothly.  Prior to exposing subjects to  the  VOC
mixture,  extensive  testing  of  the  gas  generating and monitoring system was
conducted by CE to ensure that the system operated reliably and to  ensure  that
gas  concentrations  did  not  vary by more than +3.0% once the target level (25
mg/m3) was reached.  When  CE  completed  testing  of  the  gas  generating  and
monitoring  equipment, two groups of three subjects (total N=6) were run through
the complete protocol in order to work out any scheduling problems and to  train
the  operating  staff.   Both  groups  were  run  under  control (clean air) and
exposure conditions  as  specified  for  the  main  protocol  in  the  following
sections.

C.5 Experimental Design.

     Molhave et al (1986) used a repeated measures design  in  which  each  sub-
ject  completed  both  clean air control and VOC exposure (either 5 or 25 mg/m3)
conditions on the same day.  Half the subjects received clean air in  the  morn-
ing  and  half  in the afternoon in a counterbalanced design.  There are several
problems with this design including possible confounding from carry-over effects
(i.e.,   carry-over  effects  on  afternoon  control  performance  from  morning
exposure), from practice or fatigue effects (session 1 to session 2),  and  from
time-of-day effects  (morning vs.  afternoon).  Carry-over effects were prevented
by separating control and exposure sessions by a week or more.  This  separation
also  reduced,  but  did not eliminate practice effects.  It also permitted each
subject  to complete  exposure and control sessions  at  the  same  time  of  day,
eliminating  time-of-day  confounding.   A  disadvantage  was that subjects were
required to return for an extra day,  introducing  the  possibility  of  subject
drop-out as well as  other possible confounding factors such as different weather
conditions or subject mood and preparedness.

     To  simplify the design further, only one exposure level—25 mg/m3—was run.
Control  and  exposure  sessions  were  administered  in, a  double-blind manner
counter-balanced across subjects similar  to  the  design  shown  in  Table  AA.
Although   the   chamber  could  accommodate  four  subjects  per  run,  subject
availability limited testing on many days to less than four as  shown  on  Table
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4B.

C.6 Behavioral Battery

     A battery of behavioral tests was used to assess which behavioral  processes
are affected by VOC exposure.  The battery included  the digit span  test  reported
by Molhave et al  (1986)  to  be  impaired  by  VOC  exposure.   This   test  was
administered  in  the  same  manner as Molhave et al—i.e., the paper-and-pencil
form derived from tfechsler  (1955)  with  auditory   stimuli  presented   by   tape
recorder.   Instructions  for  the  administration   and scoring of  this  test are
contained in Appendix B.  Since performance of  the  digit  span  test   includes
components  of attention as well as memory, other behavioral tests  were  included
to evaluate which processes are affected by VOC exposure.  A main set   of  eight
computerized  behavioral  tests developed by Baker and Letz (1985)  and  described
in Appendix C was used.  The tests included motor speed, memory,  attention  and
mood.   These  tests were administered twice during  each testing session—at the
beginning and end.  A secondary set  of  five  other sensorimotor,  memory  and
cognitive  tests  were  administered  once  midway through each testing session.
These  tests are also described in Appendix C.

C.7  Subjective Reactions

     A 25-item questionnaire related  to  sensory  irritation,  discomfort,  and
indoor air quality  was used  to obtain subjective ratings as described by Molhave
et al  (1986).  This questionnaire is presented in Appendix D.   The feeling  of
irritation  in   the eyes,   nose  and  throat  was also measured at 15-30 minute
intervals using   a  linear   potentiometer  described in  Appendix   E.   In  the
questionnaire subjects were  asked to indicate  the degree of perceived discomfort
by adjusting  the  position of a vertical marker on a  video screen by means  of  a
joystick.   The   range   of   response varied between  two well-defined extremes  of
items  such as dryness of the nose or skin, irritation of the   throat,   and   room
noise,   temperature or  humidity.   The  linear  potentiometer is  a device  that
reduces the  questionnaire   to  a  single  scale  representing  the collective
irritation of eyes, nose and throat.  The potentiometer consisted of an aluminum
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box with a sliding lever which  electronically  and  continuously  recorded  the
comfort rating of a particular subject during the experiment.

C.8 Confirmatory and Exploratory Hypotheses.

     Confirmatory and exploratory  hypotheses  specified  a  priori  are  stated
below.   Measurement  variables  from  individual  tests  and questionnaires are
listed in Table 5.

      a.  Confirmatory Hypotheses.

          (1)  Exposure to complex VOC mixtures will result in higher ratings
               of sensory irritation as measured by the linear analog rating
               scale (potentiometer).
          (2)  Exposure to complex VOC mixtures will result in impaired
               cognitive performance relative to clean air as follows:
               i.  Short-term memory as measured by span length (sum of forward
                   and backward span lengths) in the auditory digit span test
                 will be
                   reduced;
              ii.  Coding time (msec) for symbol-digit substitution items will
                   be longer;
             iii.  Selective attention, as measured by mean time/item in the
                   switching attention test (incompatible trials) will be
                   longer.

      b.  Exploratory Hypotheses.

          Exposure to complex VOC mixtures will:

          (3)  Increase sensory irritation as indexed by questionnaire ratings;
          (4)  Not influence ratings of controlled climate variables—i.e., air
               temperature, humidity, light level and sound level;
          (5)  Not affect motor performance on the finger tapping test;
          (6)  Alter selected dimensions of mood—i.e., tension, confusion and
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               fatigue—as indexed by pre-post mood scale difference scores;
          (7)  Impair other measures of cognitive performance as indexed by
               continuous performance, serial digit learning and pattern memory
               tests.
C.9 Data Analysis Plan.

     The main analyses of the experiment were done to determine  if  there  were
any VOC effects.  Secondary analyses of most variables were done to determine if
learning or practice effects were present and, if so, to determine the nature of
those  effects.   In the main or confirmatory analysis were:  (1) auditory digit
span (forward plus backward), (2) potentiometer  (comfort  meter)  measurements,
(3)  symbol-digit  substitution (mean response time) and (4) switching attention
test (reaction time in alternating condition).  All other variables and analyses
are considered as secondary or exploratory in nature.

     Variables measured only once during each testing session were analyzed by a
paired  t-test on the  two measurements for each subject.  Variables with pre and
post measurements were analyzed by a paired t-test on the  air  and  VOC  change
(post-pre)  for each subject.  This analysis is equivalent to a two-way analysis
of variance with subjects as one factor  and  treatment  changes  as  the  other
factor.   The  two  response profiles of each of the 25 questions in the comfort
questionnaire and the  response profiles of the comfort meter  measurements  were
tested  for  parallelism  using  multivariate  analysis  of variance of repeated
measures methods.  If  the parallelism hypothesis for a  question  was  rejected,
the  three  additional  hypotheses were tested in order to explain the nature of
the lack of parallelism.  The first hypothesis compared the mid-exposure changes
on air and VOC, the second hypothesis compared the post-exposure changes and the
third hypothesis compared the changes from  mid  to  post-exposure.   The  first
three  questions  required   either a yes or no response and were analyzed in the
above manner by coding "1" for yes and "0" for no.  In other words, the  profile
analyzed was the proportion  of "yes" responses at each measurement point.
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     Pretest questionnaire responses were analyzed by the nonparametric  McNemar
test for the significance of changes.  This test was used in order to answer the
question of whether or not potential covariates vere changing from one  exposure
to the other.

     All  variables  except  questionnaire  and  comfort  meter  responses  vere
analyzed  to  determine if learning or practice effects vere present and, if so,
as far as possible, to determine the nature of  those  effects.   For  variables
vith  pre-  and post-measurements during each exposure, the exposure vas ignored
and the actual order of the four measurements vas analyzed by fitting  a  first,
second  and  third  degree polynomial to the means.  For variables vith only one
measurement during each  exposure,  the  exposure  vas  ignored  and  the  first
measurement  vas  compared  to  the second to determine if learning effects vere
present.

C.10 Risks and Safeguards.

     The concentrations of all volatile organic compounds  used  in  this  study
vere  belov  the  TLVs for occupational exposure by orders of 10 to 1000 (ACGIH,
1982).  Although mucosal irritation and discomfort vere reported by subjects  in
the  study  conducted by Molhave et al (1986), these effects vere gone vithin 24
hours   as   indicated   by   follow-up   questionnaires   (Molhave,    personal
communication).   Reported  effects  on digit span performance vere marginal and
transient as veil.  There is no evidence of  any  permanent  adverse  effect  of
acute  VOC  exposure  at  these  levels.  There is no known risk involved in the
performance of the computerized behavioral battery or discomfort rating  instru-
ments.

     In the unlikely event of medical emergency, a  licensed  physician  vas  be
present  in  the EPA facility at all times during the experiment.  Comprehensive
emergency procedures used as general practice in the operation of  the  facility
could  have  been  activated  vithin  seconds of the occurrence of an emergency.
Emergency equipment vas kept at the site of the exposure chambers at all  times.
Furthermore,   subjects   vere  kept  under  constant  surveillance  by  trained
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biomedical personnel via closed circuit TV monitoring, direct  observation,  and
voice (intercom) communication.
D.  RESULTS

D.I Comfort Meter (Potentiometer)

     Comfort  meter  response  profiles  for  exposure  and   clean   air   were
significantly  different  (p  <  .001)  as shown in figure 2.  That  is, subjects
expressed much more discomfort  (irritation  of  mucous  membranes)  during  VOC
exposure  than  clean air.  An abrupt divergence of comfort levels occurred when
VOC exposure commenced at the 75 min.  point in testing.  The maximum difference
in   comfort  levels  occurred  about  75  min.   after  exposure  onset.   Mean
potentiometer readings (and standard errors) at each sampling point  are shown in
Table 6.

D.2 Symptom Questionnaire

     A 25-item symptom questionnaire was administered three  times   during  each
testing  session—during  pre-exposure baseline, immediately after VOC build-up,
and at the end of the 2.75 hr.  exposure period.  Mean response  levels  at  the
three  measurement  points  for  continuous  items  1-22  are  shown in Table 7.
Significant main effects  of VOC exposure (25 mg/m3) compared to clean  air  were
found  for  six items including headache, odor level, eye and throat irritation,
drowsiness and air quality.   Figures 3 and 4 illustrate  these  findings.   Five
environmental  variables  (light and noise levels,  temperature, humidity and air
movement) were held constant  during  testing.  Responses to control variables did
not vary with exposure (Figure 5).

     Different patterns of change over time in response levels were  apparent for
different  symptoms.   Further analysis of temporal patterns (Table  8) indicated
that the perceived odor level increased dramatically after VOC  exposure  onset,
but  then  decreased  significantly  with continued exposure at a constant level
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(Figure 3).  A similar (but inverse)  pattern  vas  observed  for  air  quality.
Headache,  eye  and  throat  irritation  also  increased significantly after VOC
onset, but remained elevated for  the  duration  of  VOC  exposure  (Figure  4).
Increased  drowsiness, relative to clean air, was significant only at the end of
exposure.

     Three other questions solicited binary (yes-no)  responses  concerning  air
quality as follows:

      (1)  Do you feel now that staying in the chamber is comfortable?

      (2)  Would you be satisfied with this air quality in your own home?

      (3)  Would you ventilate your home more?

VOC exposure in each case elicited significant change in  perceived  comfort  as
shown  in  Figure  6  and Table 9.  Sixty-four percent more of the subjects, for
instance, indicated that they would  not  be  satisfied  with  the  air  quality
perceived  after VOC exposure build-up (mid) than at the same point during clean
air.  A small (14%) but significant reduction in dissatisfaction occurred by the
end of exposure.

D.3 Auditory Digit Span (ADS)

     Six scores were derived from the ADS test including the  standard  measures
of  the  longest spans recalled in forward and backward conditions and the total
(forward + backward) span.  Three variant  measures,  designated  as  "weighted"
scores, were also obtained as described in the methods section.

     A problem was encountered in the administration of ADS to some subjects who
failed  to  reverse  numbers  in  the backward condition.  That is, numbers were
written down from right-to-left on the answer sheets, enabling these subjects to
complete the backward condition without mentally reversing numbers.  The problem
was observed in 12 subjects, despite explicit instructions and training  to  the
contrary.   Therefore,  ADS  results  were analyzed both with and without the 12
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aberrant subjects.  Hean pre- and post-exposure scores are shown  in  Tables  10
(without  12  subjects)  and  11  (all  subjects).   VOC exposure did not affect
auditory digit span in the predicted manner.   One  measure  (standard  backward
span   length)  varied  significantly  with  VOC  exposure,  but  in  the  wrong
direction—i.e., that is, auditory digit span (backwards)  was  slightly  longer
during exposure than clean air.  ADS results are shown in Figure 7.

D.4 NES Main Test Results
     D.4a Finger Tapping

     As predicted, finger tapping speed was not altered by  VOC  exposure.   The
mean   numbers  of  finger  taps  obtained  with  preferred,  non-preferred  and
alternating hands are shown in Table 12.
     D.4b Visual Digit Span (VDS)

     Three scores were derived from the VDS test  including   the  longest  spans
recalled  in  forward and backward directions and the  total  (forward + backward)
span.  These scores are comparable to the  standard  scores   obtained  form   the
auditory digit span test.  Mean pre- and post-exposure scores are shown in Table
13 and Figure 8.  Forward span length increased slightly but  significantly  more
under clean air than VOC exposure.  Backward and total span  lengths did not vary
with exposure.
     D.4c Continuous Performance Test  (CPT)

     Three CPT measures  were  analyzed—mean  reaction   time   (RT),  errors  of
omission,  and  false positives.  The  primary measure  (RT) did  not vary with VOC
exposure, nor did omission errors.  A  marginal trend (p =  .061)  toward increased
false  positives  was observed during  VOC exposure.  Mean  scores are provided in
Table 14 and illustrated in Figure  9.
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     D.4d Symbol Digit Substitution (SDS)

     Two scores were obtained from the SDS test—the mean  number  of  incorrect
responses per trial, and the mean time to complete a trial (requiring nine digit
substitutions).  Hean scores and results are shown in Table 15.   Neither  score
was affected by VOC exposure.
     D.4e Serial Digit Learning (SDL)

     Two measures were analyzed from the SDL test, a performance score described
in  the  methods  section,  and  a variant of the performance score in which the
total number of trials to reach criterion was added to  the  performance  score.
Unlike  related  auditory  and  visual digit span tests, smaller scores indicate
better performance in the SDL test.  Mean scores and  results  are  detailed  in
Table 16.

     SDL performance was not affected by VOC exposure.
     D.4f Pattern Memory (PM)

     Three measures were analyzed including the mean  response  latency  of  all
trials,  mean  response  latency for correct trials only, and the mean number of
correct trials.  Results and means are shown in Table 17.  VOC exposure did  not
affect pattern memory.
     D.4g Switching Attention (SWATT)

     Seven response-time measures were derived from  the  SWATT  test  including
mean  response  times  for  three  basic  conditions—side,  direction  and  the
alternation of side and direction (switching).  The switching condition was then
broken  down  into  four  subsets of trials—side compatible, side incompatible,
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direction compatible, and direction incompatible.  Mean response times for  each
of  these  measures are presented in Table 18.  Response times improved over the
testing session (pre - post difference) for all measures during clean air.   The
pre  -  post  difference  was less or in the opposite direction for six of seven
measures  during  VOC  exposure,  although  none   of   the   differences   were
statistically  significant.   Response times also increased markedly relative  to
the complexity of  the  type  of  trial—the  side  direction  is  easiest,  the
direction condition is more difficult, and the alternating (switching) direction
condition is most difficult as shown in Figure 10.
     D.4h Mood Scales

     Mean scores obtained for the five mood scales  during  clean  air  and  VOC
exposure are shown in Table 19.  Post - pre change scores differed significantly
during VOC exposure on two scales (confusion and fatigue)  as  predicted.   That
is,  increases  in  fatigue  and  confusion over the 4-hour testing session were
greater during VOC exposure than during clean air as shown in  Figure  11.

D.5 NES Secondary Test Results
     D.5a Associate  Learning/Recall

     Two measures were obtained  for   this   test—the   total   number   of   correct
 pairings  over   3  trials   and   the   number of  correct  pairings  made during  the
 subsequent  recall trial.  Means  and  results are  shown  in  Table   20.   Associate
 learning and  recall  were not affected by VOC exposure.
      D.5b  Pattern Comparison

      Three measures  were  analyzed  including the number of correct   trials,   mean
 latency  for   all trials,  and  mean latency for correct trials only.   Mean  scores
 and results are  presented in Table 21.   VOC exposure did not  affect  any  pattern
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comparison measures.
     D.5c Simple Reaction Time (SRT)

     Mean reaction-times were examined for preferred  and  non-preferred  hands.
Means and results are shown in Table 22.  SRT was not altered by VOC exposure.
     D.5d Grammatical Reasoning

     Three measures were obtained from this test  including  the  mean  response
time for individual trials, the total number of errors, and a composite score of
the total number of correct responses minus the total number of errors.  None of
these measures vas affected by VOC exposure as shown in Table 23.
     D.5e Horizontal Arithmetic

     Three measures were obtained including the number  correct,  mean  response
time  for  all  trials  and  the  mean  response  time  for correct trials only.
Slightly more problems were answered correctly during VOC exposure  (p  =  .031)
than during clean air as shown in Table 24.

D.6 Water Consumption

     Subjects consumed slightly more water during VOC exposure  (2.60  cm)  than
during clean air (2.36 cm), but this difference was not significant.

D.7 Learning/Practice Effects

     First, Second and third degree polynomials were fitted to the  four  serial
order  means  of  measures  from each of the NES main tests to determine whether
learning or practice effects occurred  beyond  testing.   A  significant  linear
contrast  implies  that  there  are  learning or practice effects across the two
training  sessions  (or  four  serial  test  administrations).   A   significant
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quadratic  effect implies that the learning or practice effect is approaching or
has reached asymptote.  A significant cubic effect  within  the  present  design
suggests  that  learning  or  practice effects occur within each testing session
(pre-post comparison).  The  absence  of  linear,  quadratic  or  cubic  effects
implies  no  learning  or  practice  effects.   Most measures showed significant
serial order effects within or across testing sessions.  The auditory digit span
test  illustrates  significant  linear trends for all measures (Figure 12, Table
25).  The total span length measure on the Visual Digit Span  Test  (Figure  13,
Table  26) demonstrates a clear quadratic pattern.  An example of a cubic effect
in the absence of linear or quadratic trends  occurred  on  the  Confusion  Mood
Scale  (Figure  14).   Response latencies on the pattern memory test exhibited a
complex serial order pattern in  which  all  three  contrasts  were  significant
(Table  27, Figure 15).  Table 28 summarizes the results of polynomial contrasts
for all computerized NES tests used in this study.
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E.  DISCUSSION

     E.I Confirmatory Hypotheses.

     Four  confirmatory  hypothese  were  specified,  only  one  of  which   was
confirmed.   As  hypothesized,  subjects  expressed  considerably  more  general
discomfort on the linear analog rating scale of eyes, nose and throat irritation
during  VOC  exposure  than clean air.  This finding is consistent with previous
Danish reports (Molhave et al., 1986;  Kjaergaard et al, 1989).

     Other hypotheses that VOC exposure would impair  cognitive  performance  of
short-term  memory  on  the  auditory digit span test, coding time on the symbol
digit substitution test, and selective attention on the switching attention test
were  not  confirmed.   Contrary  to  Molhave  et al's (1986) report, short-term
memory was not affected by VOC exposure.

     E.2 Exploratory Hypotheses

     Five exploratory hypotheses were specified, four of which were confirmed.

     E.2a Sensory Irritation.

     Symptoms of sick building syndrome, listed in Table 1,  include  irritation
of the eyes, nose and throat;  mental fatigue, and headache.  Subjects expressed
more irritation of eyes and throat (but not nose) during exposure to  VOCs  than
clean  air.   Subjects  also  reported  more  headache and drowsiness during VOC
exposure, consistent with Danish studies (Molhave et al,  1986;   Kjaergaard  et
al, 1989).  Symptom questionnaire results provide a more detailed picture of the
irritant effects of VOC exposure than the general measure of discomfort provided
by  the  linear  analog  rating  scale.   Although  the potentiometer rating was
defined as an index of "eye, nose and throat irritation", responses to  specific
symptom  questions  suggest  that  VOCs  irritated  the  nose less than eyes and
throat.  This finding  has  implications  for  future  VOC  studies—i.e.,  that
measures  of  inflammatory process in eye fluids and throat secretions may yield
more useful information than in nose secretions.  Failure to find a  significant
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increase  in  nose irritation is not in accord with the SBS profile, although it
is consistent with Kjaergaard et al's (1989) report that VOC  exposure  did  not
produce any signs of inflammation in nasal secretions.

     The most dramatic reactions to VOC exposure in the  present  study  and  in
previous  Danish  studies  were  obtained  on  ratings of odor intensity and air
quality.  Subjects exposed to the 25 mg/m3  VOC  mixture  found  the  odor  very
strong  and  unpleasant.   Subjects  similarly  reported  that  VOCs degrade air
quality  very  much.   Molhave  et  al  (1988),  moreover,  found  a  systematic
dose-dependent  relationship  of perceived odor intensity and air quality at VOC
exposure levels of 3, 8, and 25mg/m3.

     The strong and unmistakable odor of the VOC mixture  poses  a  methodologic
challenge—i.e.,  VOCs  and  clean  air  cannot be administered to subjects in a
double-blind manner.  Subjects know unequivocally when  exposure  begins.   This
problem was reduced by using a counterbalanced design.  The rapid habituation of
olfactory sensation also helps to mitigate this problem.  Results of the present
study, for instance, indicate that perceived odor intensity decreased at the end
of the testing period, while eye, throat, headache, fatigue and confusion scores
either increased or remained elevated during the 2.75 hour exposure period.  The
irritant effects of VOC exposure thought to be mediated by the trigeminal  nerve
(Molhave,  1986),  can  thus be distinguished from olfactory response over time.
Further study is needed to characterize in detail the time course  of  olfactory
and trigeminal response to VOCs.

     E.2b Controlled Climate Variables

     It was further hypothesized that ratings of  five  climate  variables  (air
temperature,  humidity,  light  and noise levels, and air flow), which were held
constant throughout testing, would not vary with VOC exposure.  This  hypothesis
was confirmed, providing construct validity to the symptom questionnaire.

     E.2c Motor Performance.

     The finger tapping test provides a simple, straightforward measure of motor
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performance.  Since motor dysfunction has not been associated with VOC exposure,
we predicted no effects on finger tapping.  This hypothesis was confirmed.

     E.2d Mood Scale Alterations.

     Pre-post differences on three mood scales—tension, confusion and fatigue—
associated  with  VOC exposure were hypothesized.  This hypothesis was confirmed
for confusion and fatigue, but not tension.  These findings are consistent  with
SBS  syndrome  as veil as the drowsiness item on the symptom questionnaire.  The
lack of effect on the tension and anger scales is of some interest, particularly
in  view  of  the perceived increase in discomfort and sensory irritation.  Many
constituents of the mixture are organic solvents with known  anesthetic  or  CNS
depressant  properties—e.g.,  n-butylacetate, 1-1-dichloroethane, n-hexane, and
p-xylene (Anger and Johnson, 1985).  The anesthetic or narcotic effects  of  the
solvents  may account for the absence of tension or anger in subjects during VOC
exposure.

     E.2e Cognitive Performance.

     The final exploratory hypothesis concerning the impairment  of  performance
on  three different tests of cognitive performance (continuous performance test,
serial digit learning and pattern memory) was not confirmed.  The results of all
memory,  sensoriraotor  and  other types of cognitive tests were very consistent.
None of these tests showed any convincing evidence that  VOC  exposure  impaired
performance,  despite  marked  subjective reactions on the symptom questionnaire
and comfort rating scale.  Nor is this finding particularly surprising  in  view
of  the  low  aggregate  exposure  level  (7  ppm toluene equivalent) of the VOC
mixture.  Neurobehavioral effects  have  seldom  been  observed  below  300  ppm
toluene  (Johnson  et  al, 1987).  Concentrations of individual chemicals in the
mixture were below TLVs by factors of 10 to 100 or more.

     Molhave et al (1986) reported a small reduction in digit  span  during  VOC
exposure.   The  consistent  negative  results on fourteen neurobehavioral tests
argue rather persuasively against any functional  impairment  in  healthy  young
adult  males  exposed  to  this  concentration  (25  mg/m3)  of volatile organic
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compounds.  Exposure and clean air sessions were run  in  the  morning  and  and
afternoon  of the same day in the original Danish study, giving rise  to possible
confounding from exposure carry-over effects, practice or fatigue  effects,  and
time-of-day  effects.   Modifications  in  the  experimental design,  to mitigate
these confounding factors render the present  results  more  reliable than  the
original study.

     On the other hand, several important differences  in  the  subject  samples
used  in  the  two  studies could possibly account for the discrepant digit span
findings.  (1) Molhave et al carefully selected  chemically  sensitive  subjects
with  documented  histories  of  indoor  climate problems.  Chemically sensitive
subjects were excluded from the present study.  (2) The range  of  ages  in  the
Danish  study (18-60 years) was much wider than the present study  (19-39 years).
Older subjects could be more sensitive than younger subjects  to   VOC exposure.
(3)  Finally,  the  Danish  study  included  equal numbers of males and females.
Maizlich et al (1987) have shown that females perform more poorly  than males  on
many NES tests.  Whether cognitive performance is more susceptible to impairment
by VOC exposure in women then men,  in  older  than  younger  subjects,  and  in
chemically  sensitive  compared  to  normal  healthy subjects are  questions that
require further study.

     E.3 Effects £f Practice and Learning

     The question of practice or learning effects is  an  important   concern  in
repeated-measures  studies  employing neurobehavioral tests.  Such effects could
exaggerate or obscure effects of chemical exposure if not controlled  properly in
the  experimental  design.   It is also of considerable interest to the research
community to characterize the effects of practice and learning in  a widely  used
neurotoxicity  testing  system.  For the present discussion the terms "practice"
and  "learning"  will  be  used  synonymously.   Although  the  study was   not
specifically  designed  to  examine  practice  effects,  the  study   provided  a
convenient and extensive dataset to evaluate this question.

     Nine tests were administered  twice on each of two  test days,  yielding  four
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replications.   Test days were separated by at least one week.  Practice effects
were evaluated by testing for linear, quadratic and cubic  trends  in  the  data
arrayed in the serial order of presentation.  As noted previously, a significant
linear  trend  implies  extended  practice  effects   across   the   four   test
administrations, quadratic effects imply that practice effects approach or reach
asymptote levels, and cubic effects imply transient practice effects  (within  a
testing session).

     Results of polynomial tests for serial  order  effects  are  summarized  in
Table 28 including data on nine tests repeated four times.  Evidence of extended
practice effects (significant linear trends) were found on five  tests—auditory
and  visual  digit  span, continuous performance, symbol digit substitution, and
switching attention tests.  Results of  three  secondary  NES  tests  (associate
learning,  pattern  comparison,  and  grammatical reasoning tests), administered
once during each testing session, also suggest extended practice effects.   Only
one  of  five mood scales (tension) exhibited a significant linear trend.  Tests
which do not appear to have extended practice effects  include  finger  tapping,
serial  digit  learning, other mood scales, simple reaction time, and horizontal
addition.  Finger tapping and two mood scales (fatigue and confusion),  however,
showed  significant  cubic  trends  which  imply  within-session effects, but no
between-session effect.  Mood scales, in  particular,  are  designed  to  assess
short-term   subjective   reactions  to  experimental  variables.   Fatigue  and
confusion scales, if valid, should vary over a demanding task and then return to
previous  values  after  resting or at the beginning of another testing session.
Significant quadratic effects were observed in two tests—visual digit span  and
pattern  memory—and  marginal quadratic trends were found for some variables in
serial digit learning, switching attention, and the tension  scale.   All  three
trends occurred in the pattern memory test.

     The implication of  the serial order analysis is that practice  or  learning
effects  occur in the performance of most neurobehavioral tests commonly used in
neurotoxicity testing of humans.  It should be noted  that  these  effects  were
observed  despite  an  initial training session.  Data from the training session
were not included in the present analysis because the length and  complexity  of
                                       26

-------
some  tests  varied  on  training  and testing days.  However, a single training
session does not appear to be adequate to eliminate practice/learning effects on
most  NES  tests.   Further parametric studies need to be done to systematically
evaluate  the  practice/learning  characteristics  of  each  NES   test.    Task
complexity,  length  of  task  and  interest  trial  interval  must be carefully
assessed in order to determine optimal parameters for the use of  NES  tests  in
repeated-measures studies in the laboratory as well as in the field.
                                       27

-------
14.  REFERENCES


American Conference of Government Industrial Hygienists.  Threshold Limit Values
for  Chemical  Substances and Physical Agents in the Workroom Environ- ment with
Intended Changes for 1982.  ACGIH Publ.  Office, Cincinnati, 1982.

Bach B, Molhave L, Pedersen OF.  Human reactions during controlled exposures  to
low  concentrations  of  organic  gases  and  vapors  known as normal indoor air
pollutants:  Performance tests.  In Berglund et al  (eds)  Indoor  Air,  Vol  3,
Sensory  and  Hyperreactivity  Reactions to Sick Buildings.  Swedish Council for
Building Research, Stockholm, 1984, pp397-401.

Hathaway SR, McKinley JC.  The Minnesota Multiphasic Personality Inventory.  The
Psychological Corporation, 304 East 45th St, New York, NY 10017, 1966.

Hollingshead AB.  Two-factor index of social position.   Unpublished  ms.,  Yale
University,  1957  (Available  from the author, 1965 Yale Station, New Haven, CT
06520).

IARC Monographs on the Evaluation of  the  Carcinogenic  Risk  of  Chemicals  to
Humans, Volumes 1-41, Lyon, 1971-1986.

Johnson BL (ed).  Prevention  of  Neurotoxic  Illness  in  Working  Populations.
Wiley, New York, 1987.

Kjaergaard S, Molhave L, Pedersen O.F.  Human reactions to a mixture  of  indoor
air volatile organic compounds.  Environment Int., 1989 (in press).

Letz R, Baker EL.  Computer-administered neurobehavioral testing:   On  defining
its  limitatins.   In:   Environmental Health, Doc 3, World Health Organization,
Copenhagen, 1985:158-162.

Maizlich N, Schenker  M,  Weisskopf  C,  Seiber  J,  Samuels  S.   A  behavioral
evaluation  of  pest  control workers with short-term, low-level exposure to the
organophosphate Diazinon.  Am J Indus Med, 1987, 12:153-172.

Melius J, Wallingford K, Carpenter J, Keenlyside R.  Indoor  air  quality:   the
NIOSH experience.  Am Conf Indus Hyg, Rep 10, 1984:  3-7.

Molhave L.  Indoor air quality in relation to sensory irritation due to volatile
organic compounds.  ASHRAE Transactions, Vol 92(1), paper 2954, Dec 1986.

Molhave L, Bach B, Pedersen OF.  Human reactions during controlled exposures  to
low  concentrations  of  organic  gases  and  vapors  known as normal indoor air
pollutants.  In Berglund et al (eds) Indoor Air, Vol 3, Sensory  and  Hyperreac-
tivity  Reactions  to  Sick  Buildings.   Swedish Council for Building Research,
Stockholm, 1984, pp431-436.
                                       28

-------
Molhave L, Bach B, Pedersen  OF.   Human  reactions  to  low  concentrations  of
volatile organic compounds.  Environment Int., 1986, 12:  in press.

Molhave L, Jensen JG, Larsen S.  Acute afnd  subacute  subjective  reactions  to
volatile  organic  air  pollutants.  Unpublished Report, Inst.  Environ.  Occup.
Med., Univ.  of Aarhus, Denmark, Dec.  1988.

Molhave L, Holler J.   The  atmospheric  environment  in  modern  Danish  dwell-
ings-Measurements  in  39m  flats.   in  Indoor Climate, P Fanger and 0 Valbjorn
(eds), SBI, Horsholm, Denmark, 1979, pp!71-186.

National Research Council, Committee on Indoor Pollutants.   Indoor  Pollutants.
National Academy Press, Washington, DC, 1981.

Strong AA.  Description of the CLEANS Human Exposure System.   EPA-600/1-78-064,
Health  Effects  Research  Laboratory,  U.S.   Environmental  Protection Agency,
Research Triangle Park, NC, Nov 1978.

Wechsler D.  Wechsler Adult Intelligence Scale Manual.   Psychological  Corpora-
tion, New York, 1955.

White JR.  A Review  of  Indoor  Air  Contaminants  and  Recommendations  for  a
"Typical"  Indoor  Air  Mixture.   Internal  Report,  Air and Energy Engineering
Laboratory, U.S.  Environmental Protection Agency,  Research Triangle, 1987.

World Health Organization.  Indoor Air  Quality  Research.   EURO  Reports  103,
Stockholm, 1986.
                                        29

-------
TABLE 1.  SYMPTOMS OF SICK BUILDING SYNDROME (WHO, 1982)
          Irritation of eye, nose and throat
          Dry mucous membrane and skin
          Erythema
          Mental fatigue, headache
          Airvay infections, cough
          Hoarseness of voice, wheezing
          Nonspecific hyperreactivity reactions
          Nausea, dizziness
        TABLE 2.  SUBJECT SELECTION CRITERIA
              Age: 18-40 years
              Race: Caucasion
              Gender:  Male
              Non smokers
              No pre-existing diseases or
              abnormal conditions such as:
                 *chronic airway disease
                 *cardiac disease
                 *Asthma
                 *Chronic bronchitis
                 *tuberculosis
                 *Psychiatric disorder
                 *Neurological disorder
                 *Skin disease
                 *Chronic eye disease
                 *0besity
                 *Cancer or other severe disease
                 *Alcohol or Drug Addiction
                 *IgE Atopy (negative skin test
                  to common agents)
              Native language must be English
              Subjects must be able to perform
                 tasks without difficulty
              Normal psychological profile
               as determined by MMPI
                              30

-------
         TABLE 3.   MOLHAVE et al (1986) VOC MIXTURE
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
Compound
n-Hexane
n-Nonane
n-Decane
n-Undecane
1-Octene
1-Decene
Cyclohexane
3-Xylene
Ethylbenzene
1,2, 4-Trimethylbenzene
n-Propylbenzene
a-Pinene
n-Pentanal
n-Hexanal
Iso-propanol
n-Butanol
2-Butanone (MEK)
3-Methyl-3-Butanone
4-Methyl-2-Pentanone
n-Butylacetate
Ethoxyethylacetate
1,1-Dichloroe thane
Ratio
1
1
1
0.1
0.01
1
0.1
10
1
0.1
0.1
1
0.1
1
0.1
1
0.1
0.1
0.1
10
1
1
cone. (ug/m3)
825
825
825
75
8
825
75
8250
825
75
75
825
75
825
75
825
75
75
75
8250
825
825
*Based on a mixture with a total concentration of 25 mg/m3



              TABLE 4.   EXPERIMENTAL DESIGN



 A.  TREATMENT           SESSION 1              SESSION 2




         A                   0                     25



         B                  25                      0





 B.
No. of Subjects
in Chamber
1
2
3
4
Frequency of Occurrence
during Testing
20
23
15
10
                                31

-------
                       TABLE 5.  MEASUREMENT VARIABLES
     A.  MAIN NES TESTS
     TEST
          MEASURE
C/E*
Finger Tapping
Visual Digit Span
Continuous
Performance
Test

Symbol Digit
Substitution

Serial Digit
Learning

Pattern Memory
Switching Attention
Mood Scales
flaps for preferred, non preferred,
 and alternating condition

Total span length (forward+backward)
Forward span length
Backward span length

Mean Reaction Time
tMisses
#False Alarms

Mean msec/digit
terrors
^trials attempted + score

Mean msec/trial
#correct

Mean msec/side condition
Mean msec/direction condition
Mean msec/switching condition

Tension
Depression
Anger
Fatigue
Confusion
 E
 E
 E

 E
 E
 E

 C
 E
 E
 E

 E
 E
 C

 E
 E
 E
 E
 E
                                       32

-------
TABLE 5 (cont.)

     B.  SECONDARY NES TESTS
     TEST
      MEASURE
C/E
Associate Learning

Associate Recall

Pattern Comparison



Simple Reaction Time




Grammatical Reasoning


Horizontal Arithmetic
# Correct (trials 1-3)

# pairs correctly recalled

mean latency of trials 2-25,
excluding incorrect  trials
# trials correct

mean RT for preferred
hand (P2-P5) and
non preferred hand (N6-N9),
excluding block 1

mean response time
# errors

# correct
mean response time
E

E
E
E
E
E

E
E
      C.   OTHER TESTS
 Auditory  Digit  Span
 Comfort  Scale
 (Potentiometer)
 Symptom Questionaire
total span length (Fwd + Bwd)
forward span length
backward span length

difference profile
between VOC exposure
and clean air

difference between VOC
exposure and clean air
for individual  items
 C
 E
 E
                                                                    E
 *C=Conformity,  E=Exploratory  Measures
                                        33

-------
                TABLE 6

      MEAN POTENTIOMETER READING
         BY EXPOSURE AND TIME

EXPOSURE








AIR
















VOC








TIME
(MINUTES)
0
15
30
45
60
75
90
105
120
135
150
165
180
195
210
225
240
0
15
30
45
60
75
90
105
120
135
150
165
180
195
210
225
240

MEAN*
.048
.059
.234
.235
.235
.310
.321
.361
.362
.438
.461
.531
.467
.476
.586
.572
.570
.103
.106
.373
.382
.380
.570
.921
1.084
1.136
1.277
1.303
1.131
1.166
1.178
1.223
1.222
1.223

S.E.
.017
.020
.045
.045
.045
.056
.055
.056
.056
.069
.070
.081
.077
.078
.097
.093
.094
.033
.033
.055
.054
.054
.074
.096
.100
.106
.117
.123
.115
.116
.118
.123
.131
.131
* N=63
Ho: F(16,47) = 3.98 (P<.001)

-------
                              TABLE 7

                           QUESTIONNAIRE
QUESTION
1. BODY/SKIN
IRRITATION
2. NOISE LEVEL
3. EYES DRY / WATERY
4 . HEADACHE
5. FACIAL SKIN
TEMPERATURE
6. SLUGGISHNESS
7. GDOR LEVEL
8. EYE IRRITATION
9. BODY TEMPERATURE
10. -I5HT INTENSITY
11. ZOUGHING
TIME
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
: POST
ft
MEAN(tt)
1.77
3.35
5. 18
11.48
12.91
13.80
12.71
12.23
11.30
2.00
2.12
3.41
14.00
12.30
11.33
6.33
8.27
9.83
8.73
6.53
0.68
2.56
5.21
7.59
14.03
11.05
10.05
17.11
17.35
18.08
1.38
3.94
1.05
tIR
S.E.
0.51
0.73
0.80
0.76
0.82
0.89
0.36
0.45
0.56
0.61
0.53
0.76
0.37
0.55
0.63
0.85
0.95
1.09
0.95
0.89
0.91
0.57
0.87
0.97
0.46
0.56
0.66.
0.44
0.48
3. 55
3.41
0.32
0.37
V
MEAN ( * )
1.73
5.26
6.39
10.88
13.45
14.09
13.30
12.39
11.24
1.64
5.05
6.97
14.53
12.82
13.21
5.70
8.70
11.71
B. 96
21.68
17.77
3.70
9.68
10.71
13.86
11.58
11.55
17.44
17.68
18.42
1.1*
2.06
1.82
OC
S.E.
0.48
0.90
0.92
0.75
0.81
0.88
0.34
0.59
0.60
0.48
0.82
1.04
0.39
0.63
0.60
0.84
0.94
1.10
0.99
0.80
1.09
0.76
1.04
1.13
0.49
0.62
0.64
0.50
a. 52
: 0.53
: 13.41
: 3.57
: 0.55
F«
(P)
1.71
( .190)
1.37
( .261)
0.47
( .626)
9.52
( < . 001 )
2.05
(.138)
2.42
C . 098 )
77.90
«.001)
4.13
(.021)
1.84
( .167)
0.00
( . 999 )
: 2.31
( . 108 ) ;
* N
«
•66
& 64 Degre<
              es of Freedom
                                    35

-------
                             TABLE 7
                             (Can t)
                         QUESTIONNAIRE
QUESTION
12. FACE IRRITATION
13. HUMIDITY
14. AIR MOVEMENT
15. NOSE IRRITATION
16. TEMPERATURE
17. TIRED SLEEPY
IB. SKIN. BODY
no IST /DRY
19. THROAT IRRITATION
20. AIR QUALITY
21. ^ACIAL SKIN
MOIST/DRY
22. CONCENTRATION
TIME
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
f.
MEAN(*») !
1.70
3.35
4.55
11.62
11.38
11.44
17.86
19.79
20.09
4.33
5.26
6.85
10.82
8.61
7.70
6.97
8.79
10.45
12. 88
12.03
11.76
2.48
3.92
4.45
11.00
11.59
11.52
12.65
12.23
11.86
2.42
2.55
3.20
IIR
S.E.
0.46
0.76
0.84
0.56
0.61
0.65
0.67
0.59
0.62
0.80
0.89
1.03
0.42
0.47
0.53
0.89
1.02
1.02
0.37
13.60
13.56
0.60
0.79
0.92
0.74
0.80
0.87
0.51
0.56
0.59
0.59
0.55
0.66
V,
MEAN ( * )
2.36
4.33
5.17
12.24
12.50
12.24
17.88
19.41
20.23
5.12
7.77
9.41
10.47
8.36
7.95
5.98
8.08
12.15
13.02
12.00
12.24
1.71
5.65
6.18
10.74
19.98
18.36
12.45
12.17
11.53
2.26
3.42
4.03
'DC
S.E.
0.56
0.79
0.86
0.54
0.62
0.60
0.65
0.62
0.57
0.85
0.97
1.11
0.35
0.46
0.44
0.90
0.91
1.05
0.42
0.42
0.47
0.46
0.94
1.00
0.77
0.70
0.80
0.49
0.52
(3.56
0.56
0.69
0.75
F*
(P)
0.08
( . 924 )
0.24
( .784)
0.50
( .612)
1.16
( .319)
0.60
( .550)
4.38
( .017)
3.36
i .696)
4.41
( .016)
32.86
( <.001)
0.08
( . 922 5
1.75
( .183)
« 2 4 e>4 Degrees  of
                                     36

-------
                             TABLE 8
                       ANALYSIS OF QUESTIONS
                      4,  7,  8, 17, 19, AND 20
QUESTION
4 . HEADACHE
7. ODOR LEVEL
8. EYE IRRITATION
17. TIRED / SLEEPY
19. THROAT
IRRITATION
20. AIR QUALITY
CONTRAST
(*)
1
2
1
2
3
1
2
3
1
2
3
1
2
3
1
2
3
CONTRAST
MEAN
3.29
3.92
0.63
15.02
10.96
-4.06
3.33
1.98
-1.35
0.28
2.69
2.41
2.50
2.50
0.00
8.65
7.10
-1.55
F(*«)
16.04
15.99
0.70
153.16
79.04
12.79
8.24
2.34
1.01
0.05
4.65
7.80
6.73
6.84
0.00
66.47
39.92
5.30
P
<.001
<.001
0.406
<.001
<.001
<.001
0.006
0.131
0.318
0.815
0.035
0.007
0.012
0.011
0.999
<.001
<.001
0.024
*1:  Change to mid on VOC - Change to mid on AIR
 2:  Change to post on VOC - Change to post on AIR
 3:  Contrast 2 - Contrast 1
** 1 and 65 Degrees of Freedom
                             37

-------
                      TABLE 9

                   QUESTIONNAIRE
                 (YES-NO QUESTIONS)
QUESTION
23
24
25
TIME
PRE
MID
POST
PRE
MID
POST
PRE
MID
POST
(•
PROP.
"YES"
0.95
0.92
0.80
0.79
0.73
0.74
0.35
0.3B
0.36
>IR
5.E.
0.03
0.03
0.05
0.05
0.06
0.05
0.06
0.06
0.06
V
PROP.
"YES"
0.97
0.62
0.59
0.79
0.09
0.23
0.33
0.83
0.73
/DC
S.E.
0.02
0.06
0.06
0.05
0.04
0.05
0.06
0.05
0.06
F*
(P)
15.19
«.001)
44.68
«.001)
22.35
( <.001 )
# N=66
t 2 & 64 Degrees of Freedom
                    TABLE 9A

                  QUESTIONNAIRE
                 (YES-NO QUESTIONS)
: QUEST I ON
i
: 23
1
: 24
1
25
CONTRAST
(*)
1
2
3
1
2
3
1
2
3
CONTRAST
MEAN
-0.32
-0.23
0.09
-0.64
-0.51
0.13
0.47
0.39
-0.08
F(«>
26.62
12.60
1.51
90.14
50.63
4.84
42.18
28.59
2.03
P
<.001
<.001
0.223
<.001
<.001
0.031
<.001
< .001
0.159
 *1: Change  to mid  on  VOC  -  Change  to  mid  on AIR
 2: Change  to post on VOC - Change to cost on AIR
 3: Contrast 2  - Contrast 1
 ** 1 and 65 Degrees of  Freedom
                             38

-------
                       TABLE 10
                  AUDITORY DIGIT SPAN
                 (WITHOUT 12 SUBJECTS)
   TEST
 FORWARD
(STANDARD)
 BACKWARD
(STANDARD)
  TOTAL
(STANDARD)
 FORWARD
(WEIGHTED)
 BACKWARD
 (WEIGHTED)
  TOTAL
 (WEIGHTED)
           TRT


           AIR

           VOC


           AIR

           VOC

           5 S5SS^SS
           AIR

           VOC


           AIR

           VOC
            AIR

            VOC


            AIR

            VOC

TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST

MEAN*

7.85
8.09
7.89
8.09
7.43
7.24
7.22
7.54
15.28
15.33
15.11
15.63
60.61
62.74
60.96
61.91
53.43
54.43
53.81
58.59
114.04
117.17
114.80
120.50

S.E.

0.14
0.13
0.15
0.14
0.11
0.15
0.13
0.11
0.22
0.22
0.24
0.21
1.99
1.71
1.95
2.04
1.61
1.73
1.74
1.69
3.31
2.89
3.32
3.14
CHANGE
(POST
-PRE)

0.24

0.20

-0.19

0.31

0.06

0.52

2.13

0.94

1.00

4.78

3.13

5.70

P-value
**


0.869



0.028



0.132



0.638



3.096



0.462

* N=54 for all means
** Ho: Change on VOC -
                        Change on AIR = 0
                            39

-------
     TABLE 11
AUDITORY DIBIT SPAN
  (ALL SUBJECTS)

TEST


FORWARD
( STANDARD )


BACKWARD
(STANDARD)


TOTAL
( STANDARD )


FORWARD
(WEIGHTED)


BACKWARD
(WEIGHTED)


TOTAL
(WEIGHTED)


TRT

AIR

VOC

AIR

voc

AIR

VOC

AIR

VOC

AIR

VOC

AIR

voc


TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST

MEAN*

7.80
8.11
7.86
8.03
7.17
7.17
7.14
7.38
14.97
15.27
15.00
15.41
59.95
62.47
60.62
61.58
50.95
50.20
52.52
54.47
110.91
115.67
113.15
119.05

S.E.

0.13
0.12
0.13
0.13
0.15
0.13
0.13
0.14
0.24
0.20
0.22
0.22
1.72
1.54
1.76
1.90
1.82
1.68
1.58
1.65
3.20
2.69
3.02
3.00
CHANGE
(POST
-PRE)

0.30

0.17

0.00

0.24

0.30

0.41

2.52

0.95

2.24

4.95

4.76

5.89

P-value
**


0.503



0.259



0.721



0.481



0.208



0.715

* N=66 for all means
*» Ho: Change on VOC -
     Change on AIR = 0
          40

-------
                        TABLE 12
                      'INGER TAPPING
                                           CHANGE
   NUMBER
    TAPS
  'REFERRE
   HAND)
;   NUMBER
ITAPS (NON-:
I PREFERRED
   HAND)

   NUMBER
:    TAPE
:  (ALTER-
:  NAT ING)
TRT

, AIR
1
: voc
;
: AIR
!
: VOC

TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POET
MEAN*

116.6
113.3
114. 1
113.5
106.3
102.7
104.7
102.1
5 . E •

1.9
1 .9
1.8
1.9
1.7
1.4
1.5
1 e
(POST
-PRE )

-3.3

-0.6

-3.6

-2.6
P— vai ue
x*


0.213



0.396

AIR

VOC
PRE
POET
PRE
POST
101.0
 99.9
102.4
 98. 9
* . c
                        1.9
        -1 .1

        -3.5
 * N=66 for all means
 *» Ho: Change on VOC - Change on AIR = 0
                          TABLE  13
                     VISUAL  DIGIT SPAN

TEST

LONGEST
FORWARD
SPAN
LENGTH
g=^^»^sg?~ •— -•
LONGEST
BACKWARD
SPAN
: LENGTH

TOTAL

TRT

AIR

VOC
B*^^B«v^e^*e
^V^^^E^Bfi
AIR

voc

AIR


TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST

MEAN*

7.68
8.20
7.97
8.06
7.53
7.62
7.59
7.83
15.21
15.62

S.E.

0.17
0.16
0.17
0.16
0.21
0.20
0.21
0.21
0.34
0.32
CHANGE
(POST
-PRE)

0.52

0.09

0.0"

• 0.24

0 . 61

P-value
XX


0.030



0.512



             VOC
      PRE
      POS"
        15.56
        15.8*
        E.34
                0.41e
 « N=do *or all means
 ** no: Change on VOC  -  Change on .'SIR = C

-------
                       TABLE 14
              CONTINUOUS PERFORMANCE TASK

TEST


MEAN
REACTION
TIME

TOTAL
OMISSION
ERRORS
============

TOTAL
FALSE
POSITIVES

TRT

AIR

VOC

AIR

VOC

=======
AIR

VOC


TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST
EtEssrsass
PRE
POST
PRE
POST

MEAN*

376.3
380.4
375.5
383.7
0.26
0.48
0.26
0.35
==========
0.91
0.88
0.74
1.20

5 . E .

4.23
4.08
4.14
4.37
0.11
0.14
0.08
0.09
========
0.14
0.19
0.14
0.15
CHANGE
(POST
-PRE)

4.1

8.2

0.22

0.09
==========

-0.03

0.46

P- value
**


0.156



0.406

=========


0.061

* N=66 for all means
»> Ho: Change on VOC - Change on AIR = 0
                         TABLE  15
                SYMBOL-DIGIT  SUBSTITUTION
TEST


MEAN
NUMBER
CORRECT
as • SB • ' ' I sssssssa
MEAN
TIME
;
TRT

AIR

VOC

AIR

VOC

TIME

ESSSSSSS&
PRE
POST
PRE
POST
PRE
POST
PRE
POST
MEAN*

.028
.027
.034
.029
15.05
14.61
14.94
14.50
S.E.

'35^5 XS SS ^E ^5 S
.008
.009
.007
.009
0.27
0.27
0.24
0.25
CHANGE
(POST
-PRE)

-0.001

-0.005

-0.44
"
-0.44
                                                  P-value
                                                    **
                                                   0.812
                                                   0.954
 * N=66 for all  means
 ** Ho: Change on  VOC  -  Change on AIR = 0

                             42

-------
10
       TABLE 16
SERIAL DIGIT LEARNING

TEST


SCORE


SCORE
-»• NUMBER
OF
TRIALS

TRT

AIR

VOC

AIR

VOC


TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST

MEAN*

2.55
2.48
2.70
2.89
6.45
6.30
6.80
7.06

S.E.

0.2B
0.34
0.27
0.34
0.44
0.53
0.45
0.52
CHANGE
(POST
-PRE)

-0.07

0.19

-0.15

0.26
==========

P-value
X*


0.54B



0.539

     *  N=66  for  all  means
     ** Ho:  Change on VOC - Change on AIR = 0
                            TABLE 17
                         PATTERN MEMORY

TEST


LATENCY
(ALL
TRIALS)
LATENCY
( CORRECT
TRIALS
ONLY)

NUMBER
CORRECT


TRT

AIR

VOC

AIR

VOC

AIR

VOC


TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST

MEAN*

5.21
4.29
5.08
4.31
4. 98
4.02
4. 82
4.08
23.3
22.7
23.2
22.7

S.E.

0.1B
0.14
0.16
0.14
0.17
0.13
0.15
0.14
0.20
0.23
0.17
0.24
CHANGE
(POST
-PRE)

-0.92

-0.77

-0.96

-0.74

-0.6

-0. 5

P-value
**


0.359



0.157



0.B56

     * N=66 for all means
     ** Ho: Change on VOC
        Change on AIR = 0
             43

-------
11
        TABLE 18
SWITCHING ATTENTION TEST

TEST


SIDE
CONDITION


DIRECTION
CONDITION


SWITCHING
CONDITION


SWITCHING-
SIDE
COMPAT .

SWITCHING-
SIDE
INCOMPAT.

SWITCHING
DIRECTION
COMPAT .

SWITCHING
DIRECTION
INCOMPAT.

TRT

.AIR

VOC

AIR

VOC

AIR

VOC

AIR

VOC

AIR

VOC

AIR

VOC

AIR

VOC


TIME

PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST

MEAN*

319.5
315.7
320.3
321.1
432.2
428. 0
430.9
419.9
489.3
474.9
485.9
483.4
426.4
416.4
410.6
414.6
444.1
427.5
446.0
438.7
550.4
537.0
551.5
553.7
536.1
518.8
535.6
526.8

S.E.

6.2
7.1
6.6
6.7
6.9
8.4
7.7
7.7
13. B
13.8
16.6
15.4
14.7
14.6
14.5
15.0
15.4
17.0
22.1
18.8
14.0
13.6
17.6
16.9
13.8
13.2
15.6
14.4
CHANGE
(POST
-PRE)

-3.B

0.8

-4.2

-11.0

-14.4

-2.5

-10.0

4.0

-16.6

-7.3

-13.4

2.2

-17.3

-8.8

P-value



0.611



0.473



0.341



0.337



0.561



0.321



0.532

        * N=66 for all means
        ** Ho: Change on VOC - Change  on  AIR  =  0

-------
12
                                TABLE 19
                               MOOD SCALE
============
TEST

TENSION
SCORE


DEPRESSION
SCORE


ANGER
SCORE


FATIGUE
SCORE


CONFUSION
SCORE

TRT
AIR

VOC

AIR

VOC

AIR

VOC

AIR

VOC

AIR

VOC

TIME
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
PRE
POST
MEAN*
1.97
1.89
1.94
1.98
1.57
1.58
1.50
1.57
1.18
1.20
1.23
1.22
2.68
2.75
2.58
2.82
1.94
1.97
1.93
2.10
5 .E .
0.07
0.06
0.08
0.08
0.05
0.06
0.05
0.05
0.04
0.05
0.06
0.04
0.09
0.09
0.09
0.08
0.06
0.06
0.07
0.08
CHANGE
(POST
-PRE)

-0.08

0.04

0.01

0.07

0.02

-0.01

0.07

0.24

0.03

0.17
P-vai ue


0 . 1 53



0.197



0.722



0.045



0.026

* N=66 for all means
*» Ho: Change on VOC - Change on
                                         AIR
0

-------
13
      TABLE 20
ASSOCIATIVE LEARNING
TEST
TOTAL
CORRECT
RECALL
TRT
AIR
VOC
AIR
VOC
MEAN*
21.15
22.06
7.94
8.02
S.E.
0.62
0.55
0.17
0.21
DIFF-
ERENCE:
VOC-AIR
0.91
0.0B
V ^HB ^ ^ ^^ •«
P-value
X*
0.132
0.747
         * N=66 for all means
         «» Ho: VOC - AIR  =  0
                             TABLE  21
                       PATTERN  COMPARISON
TEST
NUMBER
TRIALS
CORRECT
LATENCY
(ALL
TRIALS)
LATENCY
( CORRECT
: ONLY )
TRT
AIR
VOC
AIR
VOC
AIR
VOC
MEAN*
24.80
24.85
3.07
3.07
3.07
3.07
S.E.
0.06
0.05
0.09
0.08
0.09
0.08
DIFF- ;
ERENCE: IP-value
VOC-AIR I »*
0.05 ! 0.581
t
0*00 : 0.968
0.00 : 0.969
          * N=fa6  for  all  means
          ** Ho:  VOC  -  AIR  =  0
                                    46

-------
14
      TABLE 22
SIMPLE REACTION TIME
TEST
REACTION
TIME
(PREFERRED
HAND)
REACTION
TIME (NON-
PREFERRED
HAND)
TRT
AIR
VOC


AIR
VDC

MEANX
244.3
246.4


253.7
254.3

S . E .
3.38
3.41


3.91
3.54

                                          :  DIFF-
                                           ERENCE:
                                           VOC-AIR
                                               2.1
                              p-value
                                x»
                               0.455
                                               0.6    0.862
         * N=66  for  all  means
         *« Ho:  VOC  -  AIR = 0
                             TABLE 23
                       GRAMMATICAL REASONING
TEST
MEAN
RESPONSE
TIME
SUM
OF
ERRORS
SCORE
TRT
AIR
VOC
AIR
VOC
AIR
VOC
MEAN*
3396
3375
6.7
6.2
: 54.5
: 54 . o
: DIFF-
S.E. 1ERENCE:
! VOC-AIR
96 I
101 : . -21
0.9 :
0.9 : -0.5
1 t ;
: l.o. 0.4
._______.
P-value
IX
0.737
0.4B2
: 0.694
          * N=66 for all means
          xx HO: VOC - AIR  =  0
                                      47

-------
15
                           TABLE 24
                      HORIZONTAL ADDITION
           TEST
 NUMBER
 CORRECT

 RESPONSE
TIME (ALL
 TRIALS)
=======:===
 RESPONSE
TIME (COR-
RECT ONLY)
            TRT
                    AIR
                    voc
                    AIR
                    VOC
                    AIR
                    voc
EAN*
58.5
56.9
3.26
3.25
3.25
3.24
S.E.
0.18
0.11
0.09
0.0B
0.09
0.08
DIFF-
ERENCE:
VOC-AIR
0.4
-0.01
-0.01
P— va lue
**
0.031
0.766
0.806
        *  N=66 for all means
        ** Ho: VOC - AIR = 0

-------
16
                               TABLE 25
                         AUDITORY DIGIT SPAN
                            (ALL SUBJECTS)
TEST


FORWARD
( STANDARD )


BACKWARD
( STANDARD )


TOTAL
( STANDARD )


FORWARD
(WEIGHTED)


BACKWARD
(WEIGHTED)


TOTAL
(WEIGHTED)

ORDER

1
2
3
4
1
1
^
3
4
i
2
3
4
1
2
3
4
1
2
3
4
1
2
3
4
MEAN*

7.74
8.06
7.92
8.08
7.03
7.11
7.27
7.44
14.77
15.17
15.20
15.52
58.76
61.64
61.82
62.41
50.73
54.32
52.74
56.35
109.50
115.95
114.56
118.76
S.E. i CONTRAST
i
0.13 !
0.13 ILINEAR
0.13 ! QUADRAT 1C
0.12 JCUBIC
0.15 !
0.14 iLINEAR
0.13 ! QUADRAT 1C
0.13 ICUBIC
0.24 !
0.22 ILINEAR
0.21 ! QUADRAT 1C
0.19 ICUBIC
1.68 !
1.82 ILINEAR
1.77 I QUADRAT 1C
1.64 1 CUBIC
1.80 !
1.64 ILINEAR
1.60 1 QUADRAT 1C
1.72 ICUBIC
3.22 ;
2.94 ILINEAR
2.96 [QUADRATIC
2.75 ICUBIC
P-value


0.042
0.379
0.081

0.002
0.644
0.836

<.001
0.788
0.301

0.022
0.291
0.521

0.002
0.994
0.036

<.001
0.465
0.053
          N=66 for all means
                                49

-------
17
     TABLE 26
VISUAL DIGIT SPAN
TEST
LONGEST
FORWARD
SPAN
LENGTH
LONGEST
BACKWARD
SPAN
LENGTH

TOTAL


1 ORDER
: 1
> 4U
• 3
: 4
: 1
: 2.
• 3
: 4

1 <^
: 3
: 4
MEAN*
7.55
8.00
B.li
6.26
7.26
7.71
7.B6
7.74
i4.ee
15.71
15.97
16.00
5 .£ .
0.17
0.15
0.17
0.17
0.21
0.20
0.20
0.21
0.34
0.31
0.32
0.33
; CONTRAST
! LINEAR
! QUADRAT I C
! CUBIC
J
I LI NEAR
! QUADRAT 1C
i CUBIC

I LINEAR
! QUADRAT 1C
.'CUBIC
P-vaiue
<.001
0.129
0.377

0.005
0.024
0.957

<.001
0.007
0.556
         N=66 for all  means
                            TABLE 27
                          PATTERN MEMORY
TEST
LATENCY
(ALL
TRIALS)
LATENCY
( CORRECT
TRIALS
ONLY)

NUMBER
CORRECT

ORDER
1
2
3
4
1
2
3
4
i
_j"
T
', **
MEAN*
5.73
4. SB
4.56
4.02
5.47
4.25
4.33
3.84
23.1
22.2
23.4
23.1
S.E.
0.17
0.16
0.14
0.12
0.16
0.14
0.14
0.12
0.20
0.23
0.17
: 0.23
CONTRAST
LINEAR
QUADRATIC
CUBIC

LINEAR
QUADRATIC
CUBIC

LINEAR
QUADRAT I C
CUBIC
P— value :
<.00l :
<.001 :
<.00l :

<.001 :
<.001 :
<.00i ;

0.10B •
0.053
X .001
               for ail means
            50

-------
              TABLE 28.   SERIAL ORDER/PRACTICE EFFECTS (p-Values)




A.  NES MAIN TESTS

1.


2.


3.


4.

5.



7.






8.




TEST
Finger
Tapping

Visual
Digit
Span
Continuous
Performance
Test
Symbol Digit
Substitution
Serial Digit
Learning
Pattern
Memory
Switching
Attention





Mood
Scales



MEASURE
Preferred Hand
Nonpreferred
Alternating
Fvd Span
Bvd Span
Total
React ion- time
Omissions
False Positives
Correct
Response Time
Score
Score + tt trials
Latency (all trls)
Latency (corr. trls)
Side
Direction
Switching
Sw. Side Compat.
Sw. Side Incomp.
Sw. Dir. Compat.
Sw. Dir. Incomp.
Tension
Depression
Anger
Fatigue
Confusion
LINEAR
NS
NS
NS
<.001
0.005
<.001
0.024
0.087
0.012
0.090
<.001
NS
NS
<.001
<.001
(0.085)
NS
<.001
<.001
0.004
<-001
<.001
0.002
NS
NS
NS
NS
QUADRATIC
NS
NS
NS
NS
0.024
0.007
NS
NS
NS
NS
NS
(0.102)
NS
<.001
<.001
NS
NS
NS
(0.065)
NS
NS
(0.072)
(0.089)
NS
NS
NS
NS
CUBIC
0.014
<.001
0.009
NS
NS
NS
0.003
NS
NS
NS
NS
NS
NS
<.001
<-001
NS
NS
NS
(0.097)
NS
NS
NS
NS
(0.062)
NS
0.004
0.02
                                        51

-------
Table 28 (cont.)
B.  AUDITORY DIGIT SPAN TEST
Forward (Std)
Backward (Std)
Total (Std)
Forward (Wtd)
Backward (Wtd)
Total (Wtd)
0.042
0.002
<.001
0.022
0.002
<.001
NS
NS
NS
NS
NS
NS
(0.081)
NS
NS
NS
0.036
(0.053)
C.  NES SECONDARY TESTS               DIFFERENCE (ORDER 2 - ORDER 1)
1.  Associate         # Correct                  0.022
    Learning          Recall                       NS

2.  Pattern           # Correct                 (0.095)
    Comparison        Latency (All)              <.001
                      Latency (Corr. trls)       <.001

3.  Simple            Preferred Hand               NS
    Reaction-Time     Nonpreferred                 NS

4.  Syntactic         Mean Response Time         <.001
    Reasoning         f Errors                   <.001
                      Score                      0.001

5.  Horizontal        # Correct                    NS
    Arithmetic        Response Time (All)          NS
                      Response Time (Corr.)        NS
                                       52

-------
                            FIGURE  1
                    SCHEMATIC OF TEST PROTOCOL
 COMFORT RATING  SCALE
SYMPTOM QUESTIONNAIRE
      NES MAIN BATTERY
    AUDITORY DIGIT SPAN
      NES FILLER TESTS
                       PRE-EXPOSURE
            BUILDUP
CONSTANT EXPOSURE
     I  I    I  I   I     I     I  I    I   I
I	
                                                I	I
                      0
         60         120         180

          Time Expired (Minutes)
                 240

-------
                            FIGURE  2
                   COMFORT RATING SCALE RESULTS
       1.4 r	
Ln
      AIR
                                                              VOC
60
90     120   150

 TIME (minutes)
                                             180
210
240

-------
                             FIGURE  3
                    HKADACHE, ODOR & AIR QUALITY
Ln
       24
       20
    ^D
    ID
     II 16
       12
        8
    O
    o
    CO
    <
    LJ
        0 '—
AIR      VOC
  HEADACHE
                              AIR      VOC
                                  ODOR
AIR      VOC
AIR  QUALITY

-------
                           FIGURE 4
                  ODOR, EYE & THROAT IRRITATION
cr»
            AIR      VOC

               THROAT
   VOC
EYE
AIR     VOC

   ODOR

-------
                            FIGURE 5
                    CONTROL QUESTION RESPONSES
Ln
        0
             AIR      VOC
                 NOISE
AIR      VOC
   LIGHT
 AIR      VOC
TEMPERATURE

-------
                 FIGURE  6
     YES-NO  QUESTION RESPONSES
            "DO YOU FEEL NOW THAT STAYING
to          IN THE CHAMBER IS COMFORTABLE?"
| ,i .	.	y

I ,f                                               !"^~
&J t
£UJ-

O Mr-
O  r
O 014.

§  '
t &3»-
O
g  -
fe oL
K   PRE                    MID                    POST
                      EXPOSURE PERIOD
           "WOULD YOU BE SATISFIED WITH THIS
«          AIR QUALITY IN YOUR OWN HOME?"
 «f -^	_        	.  JS.
   f   -^—                 .	 • ,
a.  :		 ~             	   _
i   PRE                    MID                    POST
                      EXPOSURE PERIOD
        "WOULD YOU VENTILATE YOUR HOME MORE?"
to
W 1,	. «
1  !
< OJf
r
                       *.__	                    I VDC
    PRE                    MID                   POST
                      EXPOSURE^gERIOD

-------
                             FIGURE  7
                         AUDITORY DIGIT SPAN
Ln
vo
                                                                  AIR

                                                                  VOC
              PRE      POST
                FORWARD
PRE      POST
  BACKWARD
PRE      POST
    TOTAL

-------
              FIGURE  8
            VISUAL DIGIT SPAN
PRE     POST
  FORWARD
PRE      POST
  BACKWARD
PRE      POST
    TOTAL

-------
              FIGURE 9
  CONTINUOUS PERFORMANCE TASK
400
             MEAN REACTION TIME
                             voc
I o.u
            TOTAL OMISSION ERRORS
                             VOC
             TOTAL FALSE POSITIVES

-------
                                FIGURE  10

                        SWITCHING ATTENTION TEST
UJ
   600
   550
   500
   450
UJ
en
z

CL  400
cn
UJ
CE
   350
   300
                      -G


                      -A
       PRE
POST
	L_

PRE
-•

 . J_-_

 POST
                               600
                               550
                               500
                                                                u
                                                                Q)
                                  UJ


                               450 £

                                  UJ
                                  en


                               400 a.
                                  in
                                  UJ
                                  tr
                                                             350
                                                             300
                                                                          SIDE
                                                                        DIRECTION
                                          SNITCHING
                                    SHITCHING-SIDE

                                     COMPATIBLE
                                    SHITCHING-SIDE

                                    INCOMPATIBLE
                                  SWITCHING-DIRECTION

                                     COMPATIBLE
                                       SWITCHING-DIRECTION

                                         INCOMPATIBLE
                  AIR
                VOC

-------
                              FIGURE  11
                      MOOD SCALE DIFFERENCE SCORES
ON
U>


LiJ
O
^
X
O
bJ
OH
Q-
Z>
H-
CO
O
Q.


0.30
0.25


0.20


0.15

0.10
0.05
0.00


-0.05
r\ 1 n




-


-

-
-

.



                  AIR


                  VOC
             TENSION DEPRESSION   ANGER
             (p=.153)     (p=-197)     (p=.722)
EATIGUE  CONEUSION
(p=.045)     (p=.026)

-------
12
    16.5
16
    15.5
             TOTAL



            FORWARD



            BACKWARD
     15
 (O
 CO
                               FIGURE 12

                          SERIAL ORDER EFFECTS

                           AUDITORY DIGIT SPAN
 b:
 o
 Q
     7.5
    6.5
                              -j-
                                              _i_
                               2               3

                                    ORDER

-------
13
    16.5
     16 -
    15.5
     15
 CO

 co
 2 H.5X
 '*^
 fc

 O

 o
     7.5
     6,5
                             FIGURE 13


                         SERIAL ORDER EFFECTS


                           VISUAL DIGIT SPAN
                                   ORDER
                                  65

-------
UJ
UJ
2.06


2.04


2.02


2.00
CL  1.98
(/I
LU
^  1.96
1.94


1.92

1.90
          CONFUSION
                           FIGURE 14
                       SERIAL ORDER EFFECTS
                         CONFUSION SCALE
CONTRAST
p-VALUE
LINEAR
QUADRATIC
CUBIC
 0.639
 0.761
 0.027
                                         	L-
                                           3
                                                         4
                            SERIAL ORDER

-------
                          FIGURE  15
                      SERIAL ORDER EFFECTS
                        PATTERN MEMORY
   6.00
Ld
O
CL
CO
   5.50 -
   5.00
   4.50
   4.00
   3.50
                                          CONTRAST
                                          LINEAR
                                          QUADRATIC
                                          CUBIC
p-VALUE
< 0.001
< 0.001
< 0.001
                             SERIAL ORDER

-------
                                   APPENDIX A
        DESIGN AND PERFORMANCE OF A SYSTEM TO CONTROL VOC CONCENTRATIONS
           WITHIN AN ENVIRONMENTAL LABORATORY USED FOR HUMAN EXPOSURE
                                  Jon Berntsen
                            C-E Environmental, Inc.
                             Chapel Hill, NC  27514
                                 INTRODUCTION

     The Environmental Protection  Agency's  Clinical  Environmental  Laboratory
(CEL)  on the campus of the University of North Carolina at Chapel Hill, is used
to provide human exposure data which can withstand vigorous technical and  legal
                                                      3         3
challenge.   This  facility  consists  of  of two 84 m  (3000 ft ) environmental
laboratories which are used to expose human volunteer subjects  to  constant  or
varying  concentrations  of  03>  N02,  S02, CO and water soluble aerosols.  The
pollutant delivery system and the aerosol delivery  system  are  under  computer
control.  The computers are used to monitor the concentration of the pollutants,
control the pollutant delivery systems, validate the data as it is taken,  alarm
the  operator  of  unsafe  conditions  and record the data for later use.  These
chambers have been in continuous use since 1977 exposing subjects to  pollutants
and  measuring  responses  with  a  computerized  Physiological Data Acquisition
System.

     The EPA's interest in  indoor  air  pollution  and  the  effects  of  these
pollutants  on  humans has expanded the requirements of the facility.  To enable
the exposure of  humans  to  carefully  controlled  concentrations  of  Volatile
Organic  Compounds  (VOCs),  the pollutant delivery system was modified to allow
injection and monitoring of the VOC compounds  in  one  of  the  chambers.   The
following  sections  discuss  the design and operation of the VOC generation and
monitoring system.
                                       68

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                                SYSTEM DESIGN

Existing System Constraints

     The VOC monitoring and delivery  system was designed  to be  compatible  with
the  existing  pollutant  control  system  and  chambers.   In contrast to other
chambers, these chambers recirculate  air  with  a   flow  of  3.8  mVsec  (8000
ftVmin)  through  the  main loop.  Fresh  (make-up)  air is provided at a rate of
0.56 m3/sec (1200 ftVmin) to replace the same  amount  of  recirculating  air
exhausted  through  a  purge damper,  leaks or the chamber door when opened.  The
total volume  of  the  system,  chamber  and  associated  duct  work,  has  been
calculated  to  be 175 m3 (6180 ft3)  with  a time constant of about 5 minutes for
exchanging 67K of the chamber air.  For  the purpose  of controlling  a  pollutant
gas concentration, the chambers behave as  a one pass chamber with a total volume
of 175 m3 and a flow rate of 0.56 m3/sec except  for the amount  of  pollutant
needed to bring the concentration to  the desired value.

     Due to space constraints, the VOC generator was connected  to  the  aerosol
generator.  The aerosol generator uses .38 mVsec (800 ftVmin) for sweep air to
dry and remove the aerosol particles  from  the generator.  This  air  also  moves
aerosols  from the generator into the main air duct  where the aerosols are mixed
with chamber air.  This sweep air and the  aerosol delivery ducts are used by the
VOC generator.

     The chamber design is shown in Figure 1.  This   figure  shows  recirculated
air  through  the main chamber air loop, the fan and bypass to force air through
the VOC generator, the location of  the  makeup  air injection  point  and  the
location of the exhaust air outlet.

VOC Generator Design Considerations

     The VOC generator was designed to meet a variety of  stringent requirements.
                                        69

-------
The generator had to deliver precisely controlled amounts of a vaporized mixture
of highly flammable VOC's to the  chamber.   The  amount  of  vapor  had  to  be
automatically  controlled  by an existing computer and associated system.  Since
the mixture contains compounds with boiling points between 69 °C and 181 °C, the
generator   must   vaporize  all  constituents  without  changing  the  relative
concentrations  of  any  components.   The  generator  must  also  vaporize  the
components  with  a minimum of chemical reactions so that only the components in
the mixture are in the chamber without generating impurities.   Since  VOCs  are
flammable, the generator must minimize the danger of explosion or fire.

     The other major factor  is  the  chamber  design.   Since  the  chamber  is
multipass with large internal volume and a relatively low air exchange rate, the
generator uses an on-off type of control instead of proportional control.  Since
the  design  of  the  existing  system  has  a  30 second control cycle, the VOC
generator also incorporates this 30 second controlling  period.   Thus  the  VOC
concentration  can  be  controlled  at  a  steady  concentration  by turning the
solenoid valve on at the start of the 30 second period and off sometime  in  the
middle of the period.  Since the computer reports 2 minute averages, the effects
of this on and off control are further minimized.

VOC Generator Design

     The VOC generator (Figure 2) is a modification  of  a  design  by  Molhave,
which  was  scaled  up  and  changed  to  meet increased flow requirements.  The
mixture of hydrocarbons is placed in an Erlemeyer flask.  The top of  the  flask
has  been  modified  to  accept  a  screw type fitting.  This fitting allows the
connection of a N» pressure line and a mixture delivery line.  A Fairchild Model
81  precision pressure regulator is used to control the pressure over the liquid
mixture.  This pressure was held at 3 psig for these tests but can be varied  to
control the liquid feed rate.  A dip tube which almost touches the bottom of the
flask removes liquid from the flask.  The flask is placed on a magnetic  stirrer
and agitated throughout the run to keep the mixture homogeneous.

     The mixture delivery tube was connected through a T-connection to  a  Nupro
                                       70

-------
SS-SS2-KZ  fine  metering  valve.  The metering valve vas fitted with Kalrez and
Teflon seals.  The other end of the T-connection was connected to a tube several
feet  long.   This  tube was uncapped after the mixture was pressurized to allow
the mixture to fill the delivery tube.  This tube was securely capped after  the
delivery tube was filled.  The outlet of the metering valve was connected to the
inlet of a Sierra Model 840VO-020 solenoid valve.  The valve  has  a  0.02  inch
orifice  with  Kalrez  seals.  The outlet of the solenoid valve was connected to
the (evaporator) through  a  sprayer.   This  sprayer  was  constructed  of  two
concentric  tubes.   The  inner  capillary tube allowed the mixture to enter the
evaporation container.  About 0.05 liters/sec (0.1 CFM) of nitrogen  was  passed
through  the  outer  tube.  The flow of nitrogen helped spray the mixture on the
bottom of the evaporation container  and  also  forced  the  vapor  out  of  the
container.

     The evaporator is made of glass and during operation is placed in  a  large
brass block (15 cm in diameter and 15 cm high).  The evaporator is about 8 cm in
diameter and 10 cm high.  A fritted glass disk separates the evaporator into two.
sections.   The  fritted  glass  keeps  any  liquid  from moving from the bottom
section to the top section.  An arm on one side of  the  bottom  section  allows
access  for  the  sprayer.   The  angle of this arm allows the VOC mixture to be
sprayed on the bottom of the container.  Another arm, sealed at the  bottom  and
almost  touching  the  bottom  of  the container, contains a thermocouple (Omega
Model C03-E) probe.  The large brass block contains six 250 V cartridge  heaters
(McMaster-Carr  P/N  3618K615)  which were controlled by an Omega Model CN5001E1
proportional controller.  For this mixture of compounds, the block was heated to
250  °C.   After  the VOC mixture has evaporated, the nitrogen forces it through
the fritted glass into the top section.  Here the vapor is diluted with  another
0.5  liters/sec  (1  CFM)  of  nitrogen.   The flow of nitrogen was measured and
controlled with an Omega Model FM064-63ST flowmeter.

     The concentrated VOC vapor is then diluted with about .02 m3/s (40 CFM)  of
air  from  the  chamber.   This  air  cools  the  mixture  and  dilutes  the VOC
concentration below the lower explosive limit of the compounds.   A  vertical  4
inch glass pipe 5 feet long allows any liquid to condense and run down the glass
                                        71

-------
into a trap.  After passing through this condenser,  the  VOC  mixture  is   then
diluted  with  about  .36  m3/s  (760 CFM) of the chamber air.  This air is  then
center injected into the main air duct where it is  further  diluted  and  mixed
with main recirculated air and passed through the chamber.

     The use of nitrogen to dilute the VOC vapor and to  force  it  out  of   the
evaporator  reduces  the explosion and fire risk, prevents the oxidation of  VOCs
and minimizes the time for reactions caused by the heat  required  to  evaporate
the compounds.

VOC Monitoring and Control System

     The VOC concentration in the chambers is monitored by  two  GowMac  23-500R
total hydrocarbon analyzers.  These analyzers use a flame ionization detector  to
monitor the hydrocarbon concentration in the chamber.

     The analyzers sample from a 3 inch glass sample manifold used by all of the
gaseous  pollutant  analyzers  in  the  facility.   This manifold draws a sample
through an inverted glass funnel.  The opening of this funnel is 3  meters   from
the  floor  and  in  the  center of the chamber ceiling.  The manifold exits the
chamber near the ceiling of the chamber and passes over  the  back  top  of   the
instrument  racks  containing  the gas analyzers.  Chamber air is pushed through
the sample manifold by the 0.1 inch of water positive pressure  in  the  chamber
and two sample manifold exhaust blowers.  The flow though the sample manifold  is
0.2 m3/s.

     Each analyzer has a tap consisting of a I/A inch teflon line inserted   into
the  glass  pipe  so  the  opening  is in the middle of the glass pipe.  For the
hydrocarbon analyzers a Mace Model 800-2334-2-0 solenoid valve is used to  allow
the analyzers to sample from the sample manifold or a calibration manifold.  The
sample is pushed into the analyzer with a Thomas  Model  107CA14TFEL  pump   with
teflon coated piston, valves and cylinder.

     A PDF 11/35 computer measures the output of each analyzer once per  second.
These  samples  are  used  to  generate  2 minute averages which are reported  to
                                       72

-------
investigators as the chamber concentration.  For control purposes, the  computer
samples  the  concentration  in  the chamber every 30 seconds and determines the
percent on time for the VOC valve accordingly.  This value  is  converted  to  a
voltage and sent to the VOC control card.  Depending on the state of the system,
this card uses a voltage from a potentiometer in the manual mode or the  voltage
from the computer in the automatic mode, to control the duty cycle of the valve.
This allows either automatic  or  manual  control  of  the  generator.   In  the
automatic  mode, another signal from the computer forces the card to synchronize
with the computer.

System Calibration The VOC concentration in  the  Molhave  study  was  25  mg/m3
toluene   equivalent.    Molhave  calibrated  FID  analyzers  with  toluene  and
controlled the VOC concentration  with  instrument  output  at  the  same  level
obtained with the toluene calibration standard.  The 25 mg/m3 was converted to 7
ppm toluene equivalent for these studies.  The  VOC  analyzers  were  calibrated
with toluene and the concentrations reported in terms of toluene.  A standard of
toluene diluted with nitrogen was diluted to  the  various  concentrations  with
ultra  high  purity zero air with the dilution calibration system which has been
used for 12 years.  This system is  automatically  controlled  by  the  computer
which  after  the  calibration has been specified by the operator, automatically
calibrates the analyzers.  The standard  calibration  consisted  of  three  span
points  (5,  7  and 9 ppm) and a zero point.  The zero was taken first, then the
highest point was taken twice with the first one thrown out.

System Operation The following procedure was  used  to  run  a  session  of  the
protocol.

      1. The instruments were calibrated the morning of the protocol  using  the
         method given above.

      2. A new solution was mixed  up.   Several  safeguards  were  followed  to
         assure  that the mixture was made correctly.  The operator used a check
         sheet to remove a bottle of  each  of  the  chemicals  from  a  storage
         cabinet.   Each  chemical  was pipetted using its own premarked pipette
                                        73

-------
         (both with chemical name and the volumne) from the bottle to the flask.
         As this was done, the chemical was checked off on the check sheet.  The
         bottle was then placed back in storage.

      3. After the mixture was prepared, it vas connected to the system.

      A. The  generation  system  vas  turned  on  and  heated  to  the   proper
         temperature during instrument calibration and chemical mixing.

      5. The  results  of  the  calibration  were  then  checked  and  the   new
         calibration constants loaded into the computer.

      6. The computer vas instructed to monitor the  VOC  concentration  in  the
         chamber.

      7. Just before VOC generation  was  initiated,  the  ambient  chamber  VOC
         concentration   was  measured  and  the  exposure  level  (7  ppm)  was
         incremented by the ambient VOC level.  For example, if the ambient  VOC
         level was 0.37 ppm, the exposure level vas set at 7.37 ppm.

      8. Seventy-five  minutes  after  commencement  of  testing,  the  operator
         initiated  exposure  by  activating  the  automatic  control of the VOC
         concentration in the chamber.
                              SYSTEM PERFORMANCE

     This system has been used in two different studies for a total of 110 hours
of  VOC  control during a protocol.  The two studies used a mixture very similar
to the tventy-tvo compounds Molhave used.  Figure 3 shovs  a  plot  of  the  VOC
concentration  in  the chamber over the period of the protocol.  For this study,
the subjects were in the chamber while the VOC control was established  and  the
system equilibrated to the required valve.

-------
                     MAKEUP  AIR
COMPUTER
SVI
CTRL
   FID
ANALYZER
                                                         PURGE
                r
                             MIXER
         A
                   CHAMBER
                                          t
   voc
GENERATOR
                               FAN
                            Figure 1

                     Overall Chamber Design

-------
  Spil
  NITROGEN
    o\
VOC
MIXTURE
                             Figure 2

                       VOC Generator Design
                      METERING   SOLENOID
                        VALVE     VALVE
NITROGEN 2 l/m
                                SPRAYER

                                GLASS
                                EVAPORATOR
                                BRASS BLOCK
                                                                             DAMPER
                                         HEATERS

-------
                                  H«  IN OHMM* ft

                                  I'll/il  IHUlll
I .
4 .
3 -
i .
I  .
I . —
 lit   HI   9M    911   IN    (41
 I     I     I     I      I     I     I
t»M   Itl*  1IM   »•<•  UN   lilt   I1M
1141   II
  	 »CtU»l
  	  MIIWD
  TIM
                                       Figure 3

                           Typical  Plot of VOC  Control

-------
REFERENCES

Molhave L, Bach B and Pedersen 0.   Klimakammerundersogelse  af  geneforekomster
hos  personer,  der  udsaettes for organiske gasser og dampe fra byggematerialer
[Human reactions to volatile  organis  gases].   Project  Report  ROO-52(22411),
Institute of Hygiene, Universitetsparken, Aarhus, Denmark, 1985.

Strong AA;  "Description of the  CLEANS  Human  Exposure  System".   U.S.   EPA.
EPA-600/1-78-064, 1978.

Crider VL, Landis DA, Weaver FH.  In "Generation of Aerosolos and Facilities for
Exposure Experiments".  Villeke K, (ed.) Ann Arbor, Michigan, 1980:493-515.

Glover DE, Berntsen JH, Crider, VL, Strong AA.  "Design  and  Performance  of  a
System  to  Control  Concentrations  of  Common  Gaseous  Air  Pollutants within
Environmental  Laboratories  Used  for  Human  Exposure".   J.   Environ.   Sci.
Health, A16(5), 1981, pp501-522.
                                       78

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                                   APPENDIX B

              AUDITORY DIGIT SPAN TEST DESCRIPTION AND INSTRUCTIONS

                               H. Kenneth Hudnell

          Neurotoxicology Division, Health Effects Research Laboratory
                 U.S. Environmental Protection Agency, RTF, NC

     The  following   tape-recorded  instructions   were   given   to   subjects.
"Attention   Subjects:   you  will now be given another version of the digit span
test.  Unlike  the  Digit Span Test you took earlier on the  computer,  this  test
will   be  administered by a cassette recording played over your speakers.  Please
write  your  responses  on the answer sheet attached  to the clipboard.  Please pick
up  the clipboard at  this time  and assume a comfortable writing position.

     The  answer sheet should say "auditory digit span -  pretest"  at  the  top.
The test  will  have two parts,  Digit Forward and Digit Backward.  Please find the
section labeled Digit Forward.   As you can see on  the answer sheet, blank spaces
are  provided  for  you to write down each of the digits in the string.  The first
string will  contain 3 digits.  Please listen carefully as the digits are  called
out.   Be sure  not  to repeat the digits verbally, but you may rehearse them in
your mind.   After  the string has been called out,  there will be a  pause  during
which  you   are  to   write  down the digits as you remember them and in the same
order  as  they were presented.  Please do not begin writing until all  digits  in
the  string  have  been  called  out.   After the pause, a different string of 3
digits will  be presented and then followed by a pause during which you can write
your  answers.   This process  will be repeated until 2 trials with strings of up
to 9 digits have been presented.

     After the digit  forward test has been completed there  will  be  a  1  min.
pause.    Then,  the  digit backwards test will begin.  This test is exactly like
the digit forward  test except  that you are to  write  down  the  digits  in  the
reverse order of their presentation.  That is, the last digit you hear should be
                                       79

-------
the first digit you write on the answer line.  The  digit  backwards  test  will

begin with a 2 digit string and end with an 8 digit string.  Please remember, do

not repeat the digits out loud and do not begin to write your answers until  the

pause  begins  after  all the digits in the string have been called out.  Please

prepare to begin the digit forward test.  Write the digits in the same order  as

they are presented.  The test will begin now.

         Trial 1 of 3 digits (say 1-4-7            and  pause 10 sec.)
         Trial 2 of 3 digits (say 3-7-2            and  pause 10 sec.)
           "   1 "  4   "    ( » 2-9-5-6           and    "   12 sec.)
           "   2 "  4   "    < " 2-6-5-4           and    "   12  "  )
           "   1 "  5   "    ( " 2-8-7-4-5         and    "   14  "  )
           "   2 "  5   "    ( " 2-9-6-7-5         and    "   14  ")
           "   1 "  6   "    ( n 1-6-9-2-4-1       and    M   16  "  )
           "   2 "  6   "    ( " 4-7-6-1-2-8       and    "   16  "  )
           M   1 "  7   "    ( " 5-7-1-6-2-4-9     and    M   18  "  )
           "   2 "  7   "    ( " 1-5-2-8-9-4-6     and    "   18  "  )
           "   1 "  8   "    ( " 7-3-9-8-4-5-2-7   and    "   20  "  )
           "   2 "  8   "    < " 8-7-4-2-5-4-3-1   and    "   20  "  )
          •"   1 "  9   "    ( " 3-7-4-9-7-3-8-4-2 and    "   22  "  )
           "   2 "  9   "    ( " 1-3-2-5-2-7-8-4-8 and    "   22  ")

The digit forward test is now completed.  We  will  pause  for  1  min.   before

beginning  the digit backwards test,  (pause 1 min.) Please prepare to begin the

digit backwards test.  Please write the digits in the  order  opposite  that  in
which they are presented.  The test will begin now".
         Trial 1 of 2 digits (say 3-5            and pause 10   sec.)
         Trial 2 of 2 digits (say 4-2            and pause 10   sec.)
           "   1 "  3   "    ( " 2-9-2           and   "    12   "  )
           "   2 "  3   "    ( " 3-2-5           and   "    12   "  )
           "   1 "  4   "    ( " 2-7-9-4         and   "    14   "  )
           "   2 "  4   "    ( " 2-8-9-4         and   "    14   "  )
           "   1 "  5   "    ( " 3-6-7-5-1       and   "    16    " )
           "   2 "  5   "    ( " 2-6-9-7-3       and   "    16    " )
           M   1 "  6   "    ( " 1-4-8-3-9-3     and   "    18   "  )
           "   2 "  6   "    ( " 7-3-6-2-4-1     and   "    18   "  )
           "   1 "  7   "   . ( " 4-1-6-7-9-5-1   and   "    20    " )
           "   2 "  7   "    ( " 2-9-6-7-2-1-4   and   "    20   "  )
           "   1 "  8   "    ( " 1-4-2-6-7-5-6-9 and   "    22   "  )
           M   2 "  8   "    ( " 1-6-1-3-9-8-3-7 and   "    22   "  )

Similar instructions were given for the post test.  Digits presented in the post
test were:
                                       80

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              FORWARD
               2-8-6
               3-2-6
              1-5-7-3
              9-3-5-1
             6-8-1-9-2
             8-2-5-7-3
            2-8-6-1-3-7
            2-4-8-7-5-1
           9-5-1-3-1-7-2
           6-3-5-7-2-8-5
          1_5_9_5_7_3_9_2
          1-6-9-3-4-8-6-2
         2-8-1-5-2-6-1-7-3
         4-1-4-8-2-6-9-4-1

Digit Sequences used on day 2 vere as follows:

         FORWARD PRE-TEST

              6-1-3
             3-5-1-7
             8-9-4-2
            5-9-8-1-6
            2-8-9-4-6
           3-1-4-2-4-7
           3-7-9-2-6-1
          2-9-3-7-9-4-6
          8-5-1-7-2-4-3
         7-1-4-6-8-3-2-9
         9-4-7-3-2-9-5-6
        1-6-2-5-1-8-2-7-3
        7-3-9-3-8-4-5-7-2
                                                  BACKWARD
                                                     TT~
                                                     2-8
                                                    5-2-7
                                                    1-3-8
                                                   3-7-9-5
                                                   6-1-4-7
                                                  2-8-9-5-6
                                                  2-9-5-2-1
                                                 1-3-4-8-2-1
                                                 5-9-3-8-7-2
                                                4-5-7-4-1-6-7
                                                3_9_5_9_1_5_7
                                               1-7-4-3-2-9-1-6
                                               9-4-8-2-7-1-3-2
                                              BACKWARD PRE-TEST
                                                     T-3
                                                     2-6
                                                    1-4-2
                                                    1-8-7
                                                   5-2-4-1
                                                   2-4-6-5
                                                  3-1-5-4-6
                                                  6-3-5-1-7
                                                 2-9-1-3-8-4
                                                 3-6-7-5-2-8
                                                7-3-7-1-8-1-4
                                                6-1-3-7-9-8-2
                                               2-9-4-7-5-1-4-9
                                               4-2-4-1-7-3-6-8
POST TEST-FORWARD
       7-4-1
       2-8-9
      1-6-8-7
      5-2-7-1
     2-7-3-1-4
     l_5_8-3-9
    9-3-4-7-6-1
    4-6-2-8-1-3
   3-7-8-9-1-3-5
   3-2-4-5-7-9-6
  2-7-8-6-9-3-4-8
  9-4-3-5-8-2-1-5
 8-3-7-4-6-9-2-3-1
 5-2-3-1-4-9-2-6-8
                                                    POST TEST-BACKWARD
                                                            539
                                                            3-1
                                                           2-9-5
                                                           2-6-9
                                                          1-4-5-8
                                                          3-2-8-6
                                                         7-3-2-4-1
                                                         5-6-1-4-3
                                                        7-3-5-1-2-6
                                                        8-5-2-7-3-1
                                                       3-2-3-5-4-2-1
                                                       4-3-6-5-7-8-2
                                                      1-5-3-9-4-8-2-6
                                                      1-6-9-3-8-2-7-1
                               81

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                             Digit Span Answer Sheet
Subject ID Number :



Date              :
Session # (Circle one): 147



Day   (Circle one)     :  0 1 2
A.  Digit Forward



    3.  (1) 	



        (2)	






    4.  (1)	



        (2)	





    5.  (1)	



        (2)	





    6.  (1)	



        (2)	






    7.  (1) 	



        (2)	





    8.  (1) 	



        (2)	





    9-  (1)	



        (2)
B.  Digit Backward



    2.  (1) 	



        (2)	





    3.  (1) 	



        (2)	






    4.  (1) 	



        (2)	





    5.  (1)	




        (2)	





    6.  (1) 	




        (2)	





    7.  (1)	



        (2)	





    8-  (1)	



        (2)
                                       82

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               INSTRUCTIONS FOR SCORING AUDITORY DIGIT SPAN DATA


     ADS data will be scored in two ways, the standard method (std) and the weigh
ted
method (wtd).

Procedure.

     Both Methods.  Start with the 3 digit forward string, trial 1.  Use  the
answer key to see if all 3 digits are correctly recorded on the answer sheet.
If so, put a checkmark at the right end of the string.  If no, put an X at the
right end.  Repeat this process for each string in the digit forward test.
Then score the digit backward test.

     Std  Method.  At the bottom of the answer sheet under the forward responses,
write the number of the string that corresponds to the longest string in  which
at least 1 of the 2 trials was answered correctly for that string and all
previous strings.  For example, if at least 1 trial is correctly answered on
strings 3, 4, 5 and 6 but neither trial is answered correctly on the string of
7 digit, the score is "std = 6".  Repeat this procedure for the digit backwards
test.  At the center, record the total score.  For example, if the forward
score is 6 and the backwards score is 5, write "Tstd = 11".

     Wtd  Method.  For the digit forward test, multiply each string number (3-9)
by the number of trials answered correctly (0,1 or 2).  Add all products
together.  For example, if the scores are:

string           # trials correct          the products are
   3                     2                    3 x 2 = 6
   4                     2                    4x2 = 8
   5                     2                    5 x 2 = 10
   6                     1                    6x1 = 6
   7                     0                    7x0 = 0
   8                     1                    8x1=8
   9                     0                    9x0 = 0

     The product sum = 38.  Write "Wtd = 31" under the forward responses.  Repeat
this procedure for the backwards test.  Record the total Wtd score at the
bottom of the answer sheet in the center.  For example, if the forward score is
38 and the backwards score is 26, write "Twtd = 64".

     Note.   For the backwards test, remember that the answer must be in the
reverse order of presentation to be correct.
                                       83

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                                   APPENDIX C

                NEUROTOXICITY TESTING OF VOC MIXTURES IN HUMANS
     In order to characterize the neurobehavioral effects of VOC exposure, a
microprocessor-based test battery was used.  The neurobehavioral evaluation
system (NES) developed by Baker et al (1985) for neurotoxicity field testing
contains a menu-driven battery of tests selected on the basis of proven
sensitivity to environmental toxicity and WHO recommendations (Johnson et al.,
1987).  A variety of sensorimotor and cognitive functions including reaction
time, motor coordination, attention, memory and spatial integration were
assessed.  The NES battery has been used previously in the study of nitrous
dioxide (Greenberg et al, 1985), styrene, alcohol, is currently being used by
CDC in the AGENT ORANGE project, and in NHANES III (National Health and
Nutrition Survey).  Specific tests to be used in the VOC study included finger
tapping, continuous performance, digit span, symbol-digit substitution, visual
memory and switching attention. Individual tests are described below.

                          NES TESTS USED IN VOC STUDY
  (1)  Finger tapping (mean no.)                         3 min.
  (2)  Digit span (no. correct fwd, bvd)                 7 min.
  (3)  Continuous Performance Test (msec, errors)        7 min.
  (A)  Symbol-digit substitution (sec/digit, errors)     5 min.
  (5)  Serial digit learning (no. correct, no.  trials)   7 min.
  (6)  Pattern memory (no. correct, msec)                5 min.
  (7)  Switching attention ((msec/no, correct)           7 min.
  (8)  Mood scales (5 scale avs.)                        5 min.
                                       84

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                                              Total time:     46 min.

                        Descriptions of Individual Tests


     (1)  Finger-Tapping.  This test evaluates motor response speed.  The
subject is instructed to press a button as many times as possible within a
1-second interval.  In the first trial, the subject presses one button with the
left hand,  The second trial tests the right hand, and in the third trial, the
subject alternately taps two buttons with the preferred hand.  This test
provided a straightforward measure of VOC effect on motor response.  It was a
negative control measure since VOCs were not expected to affect motor response
per se.

     (2)  Digit Span.  The auditory version of this widely used clinical test
has been adapted from the Vechsler Adult Intelligence Scale (Wechsler, 1955).
Subjects must enter into the computer progressively longer series of digits
which are presented visually at a rate of one per sec by the computer. New
digit sequences are created at each span length.  After incorrectly responding
to two trials at a span length, the task changes such that the digits must be
entered in reverse order.  This test has been used as a measure of short-term
memory and attention in studies of solvent and lead toxicity (Hanninen, 1979).
Molhave et al (1986)  reported that digit span performance was impaired by
exposure to the complex VOC mixture, a finding that the present study attempted
to replicate.

     (3)  Continuous Performance Test (CPT).  This test measures sustained
visual attention by having the subject press a button upon seeing a large
letter "S" when it is projected on a video display.  Letters flash briefly (for
about 50 msec) on the screen at a rate of one per second for five minutes.
Recording and storage of individual response latencies permits the computation
mean (and standard deviation) of reaction times.  Omission and commission
errors are also recorded.  Molhave et al (1986) failed to observe any effect of
VOC exposure on "graphic" CPT performance, but the present test is very
                                       85

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different and has proven to be sensitive to toxicant exposure in previous
studies (Baker et al, 1988; Erstin et al, 1988; Mahoney et al, 1988).

     (4)  Symbol-Digit (SD) Substitution Test.  This test is similar to the
Digit-Symbol Substitution Test from the WAIS.  SD evaluates speed and coding
ability and has been found useful in previous studies of lead, carbon disulfide
and solvent mixtures (Anger and Johnson, 1985).  In this adaptation, nine
symbols and digits are paired at the top of the screen and the subject has to
press the digit keys corresponding to a re-ordered test set of the nine
symbols.  The time required to complete each SD set and the number of digits
incorrectly matched are recorded.  Five sets of  nine SD pairs are presented in
succession. The pairing of symbols vith digits is varied between  sets to avoid
learning.

     (5)  Serial Digit Learning (SDL).  This is a test of short-term memory and
attention in which subjects are presented a series of 10 digits on the display
screen at a rate of one per second.  Upon completion they are asked to enter
that series into the computer.  The series is presented until two successive
trials are performed correctly, or at most 8 times.  The number of correct
digits entered on each trial is recorded.  The series is changed for each test
session to avoid learning effects.  This test is a variant of the digit-span
test found to be sensitive to solvent and lead exposure (Anger and Johnson,
1985) and is similar to Supraspan in the neurotoxicity test battery developed
by Eckerman et al (1985).  Based on the digit span findings of Molhave et al
(1986), we predicted that SDL performance would be impaired by VOC exposure.

     (6)  Pattern Memory (PM) Test.  This test of short term visual memory
involves the brief presentation of a block-like pattern followed by three
similar figures, one of which is identical with the original pattern.  The
degree of correspondence of the two incorrect patterns to the correct one
varies.  The number of correct responses and the response time are recorded.
Vhile digit span and serial digit learning  assessed verbal memory, PH examined
if VOC exposure impaired visual memory.
                                       86

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     (7)  Switching Attention (SVAT).  This test evaluated the effect of VOC
exposure on selective attention with little memory demands.  During  two initial
sets the subject is trained to respond differentially to target stimuli on the
left or right side of the screen, and then to stimuli in the center  of the
screen  pointing to the left or right.  The target stimuli are large rectangles
with inset arrows pointing left or right which may appear on the right or left
side of the screen.  In the third set of trials, these two response  conditions
are mixed.  When the word on the screen is "SIDE", the subject must  press the
left button if the target stimulus is on the left and vice versa.  When the
word on the screen is "DIRECTION", then the subject must press the right button
if the arrow points to the right and vice versa.  The word on the screen is
randomly assigned each trial,   the numbers of incorrect responses and mean
react ion-times are recorded.  This test was chosen to clarify whether the
Molhave et al digit span finding was due to memory or attentional impairment.

     (8)  Mood Scales.  This test consists of a series of adjectives or phrases
with which subjects rate themselves on their feelings.  Similar instruments
have been used to evaluate the efficacy of psychotherapeutic drugs and to
classify individuals with various neurobehavioral disorders (McNair  et al,
1971) The present test contains 25 items and yields a five-dimensional mood
profile (tension, depression,  anger, fatigue, confusion) by combining ratings
on individual items.  Prior studies have shown that this approach is sensitive
in the evaluation of central nervous system effects  of occupational lead
exposure (Baker et al, 1984).   Molhave et al (1986) reported that subjective
ratings of sensory irritation (comfort) and air quality were altered by VOC
exposure.  The mood scale was chosen to clarify these findings by assessing
which dimension(s) of mood were affected by VOC exposure.

     (9)  Associate Learning/Recall.  This test is similar to the paired
associate learning test in the Wechsler Memory Scale (Wechsler, 1945).  Three
letter names (e.g. Jan, Pat, Tom) are paired with occupations (e.g.  florist,
lawyer, plumber).  Nine pairs were presented on the video monitor for two sees.
each with one sec. intervals between pairs.  After all pairs were presented,
each name was displayed and subjects were required to select the appropriate
                                       87

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alternative from a list of occupations.  Three trials were presented with the
same pairings, but in different order.  The score consisted of the total number
of correct pairings in trials 1-3.  A single recall trial vith the same
pairings vas presented about 20 mins. later.  This test provided an alternative
type of memory challenge for testing VOC exposure effects.

     (10)  Pattern Comparison.  This test is similar to the Pattern Memory Test
described above and utilizes the same 10x10 character arrays.  Three patterns
are presented simultaneously on the video screen.  Two patterns are identical
and the third is slightly different.  The task is to identify the odd array. 25
trials were presented.  Two measures were recorded—the mean latency of correct
trials 2-25 and the number of correct trials.  Schmedtje et al (1988) found
that performance on this test improved after administration of
dextroamphetamine and worsened after scopalomine.

     (11)  Simple Reaction Time (SRT).  SRT is the oldest tool in the
psychologist's armamentarium.  This visual task requires subjects to
button-press as fast as possible when a large square is displayed on the
screen.  Nine blocks (five with the preferred hand and four with the
nonpreferred hand) containing 12 trials each and variable intertrial intervals
of 2-4 sees were used.  Mean reaction times, excluding block one, were
calculated for each hand.  SRT tests have been used extensively in occupational
field studies including workers exposed to solvents (Anger and Johnson, 1985).

     (12)  Grammatical Reasoning.  This test was adapted from Baddeley (1968).
Each trial consists of a logical sequence such as "A FOLLOWS B: AB" to which
the subject must respond "true" or "false".  Sequences vary along four
dimensions: active-passive, positive-negative,  true-false, and whether the
first letter  is A or B.  Two blocks of 32 trials each were presented.  Mean
response  time for all correct trials and the number of correct trials were
recorded.  This was the most difficult NES test used.  Little evidence of the
utility of this test in neurotoxicity testing is available.

     (13)  Horizontal Addition Test.  Subjects were presented with 60 simple
                                       88

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addition problems in which three single digits were presented in a row.

Subjects were instructed to sum the digits as rapidly and accurately as

possible.  Responses had to be entered on the keyboard.  Mean response times

and number of correct trials were recorded.  Subjects made very few errors.

This test served primarily as a filler.
                                   REFERENCES

Anger WK, Johnson BL.  Chemicals affecting behavior.  In: O'Donoghue JL (ed)
     Neurotoxicity of Industrial and Commercial Chemicals.  CRC Press, Baton
     Rouge, 1985.

Baddeley, AD.  A 3 minute reasoning test based on grammatical transformation.
     Psychonom Sci 1968, 10:   341-342.

Baker E, Letz R, Fidler A, etal.  A computer-based neurobehavioral evaluation
     system for occupational and environmental epidemiology: Methodology and
     validation studies.  Neurobehav Toxicol Teratol 1985, 7:369-377.

Baker E, Letz R, Eisen EA et al.  Neurobehavioral effects of solvents in
     construction painters.  J Occup Med, 1988, 30:116-123.

Eckerman D, Carroll J, Force D, et al.  An approach to brief field   testing
     for neurotoxicity.  ibid:387-393.

Estrin WJ, Moore P, Letz R, Wasch HH.  The P300 event-related potential in
     potential in experimental nitrous oxide exposure.  Clin Pharmacol Ther,
     1988, 43:86-90.

Greenberg B, Moore P, Letz R, Baker E.  Computerized assessment of human
     neurotoxicity: Senitivity  to nitrous oxide exposure.  Clin Pharmacol Ther-
     apeutics, 1985, 38:656-660.

Hanninen H.  Psychological test methods:  Sensitivity  to  long term chemical
     exposure at work.  Neurobehav Toxicol Teratol, 1979, l(supp  1):157-161.

Mahoney P, Moore PA, Baker EL,  Letz R.  Experimental nitrous oxide exposure as
     a model system for evaluating neurobehavioral  tests.  Toxicol.,  1988,
     49:449-457.

McNair DM, Lorr M, Drpopleman LF.  BITS Manual—Profile of Mood States.  San
     Diego: Educational and Testing Service, 1975.

Molhave L, Bach B, Pedersen OF.  Human reactions  to low concentrations of
     volatile organic compounds.  Environ Internal, 1986, 12:167-175.
                                        89

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Schmedtje, J.F., Oman C., Letz, R., Baker, E.L.  Effects on scopalomine and
     dextroamphetamine on human performance.  Aviat Space Environ Med, 1988,
     59:407-410.

Vechsler D.  A standardized memory scale for clinical use.  J Psychol 1945,
     19:87-95.

Vechsler D.  Vechsler Adult Intelligence Scale Manual.  Psychological Corp-
     oration, Nev York, 1955.
                                       90

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                                   APPENDIX D

                             SYMPTOM QUESTIONNAIRE

SUBJECT ID:	


DO YOU FEEL NOW THAT STAYING IN THE CHAMBER IS COMFORTABLE?    (Y/N)

WOULD YOU BE SATISFIED WITH THIS AIR QUALITY IN YOUR OWN HOME? (Y/N)

WOULD YOU VENTILATE YOUR HOME MORE?  (Y/N)
(1)   DOES THE SKIN OF YOUR BODY FEEL IRRITATED OR ITCHY?
  NOT AT ALL |	1	1  VERY MUCH

(2)   HOW IS THE NOISE LEVEL IN THE ROOM?
  VERY QUIET |	1	1  VERY NOISY

(3)   DO YOUR EYES FEEL DRY OR WATERY?
  VERY DRY |	1	1  VERY WATERY

(4)   DO YOU HAVE A HEADACHE RIGHT NOW?
  NOT AT ALL |	1	1  VERY STRONG

(5)   DOES THE SKIN ON YOUR FACE FEEL WARM OR COLD?
  VERY COLD |	1	1  VERY WARM

(6)   DO YOU FEEL SLUGGISH?
  NOT AT ALL |	1	1  VERY MUCH

(7)   HOW IS THE ODOR LEVEL IN THE ROOM?
  NO ODOR |	1	1  VERY STRONG

(8)   DO YOU FEEL ANY IRRITATION IN YOUR EYES?
  NOT AT ALL |	1	1  VERY STRONG

(9)   DOES THE SKIN OF YOUR BODY FEEL WARM OR COLD?
  VERY COLD |	1	1  VERY WARM

(10)  HOW IS THE LIGHT LEVEL IN THE ROOM?
  VERY DARK |	1	1  VERY BRIGHT

(11)  HAVE YOU BEEN COUGHING?
  NOT AT ALL |	|	1  A LOT

(12)  DOES YOUR FACE FEEL IRRITATED OR ITCHY?
  NOT AT ALL |	1	1  VERY MUCH
                                       91

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(13)  HOW IS THE AIR HUMIDITY IN THE ROOM?
  VERY DRY |	1	1  VERY HUMID

(14)  HOW IS THE MOVEMENT OF AIR IN THE ROOM?
  VERY STILL |	1	1  VERY DRAFTY

(15)  DOES YOUR NOSE FEEL DRY OR IRRITATED?
  NOT AT ALL |	1	1  VERY MUCH

(16)  HOW IS THE AIR TEMPERATURE IN THE ROOM?
  VERY COLD |	1	1  VERY HOT

(17)  ARE YOU TIRED OR SLEEPY?
  NOT AT ALL |	1	1  VERY MUCH

(18)  DOES THE SKIN OF YOUR BODY FEEL MOIST OR DRY?
  NOT AT ALL |	1	1  VERY MUCH

(19)  DOES YOUR THROAT FEEL IRRITATED?
  NOT AT ALL |	1	1  VERY MUCH

(20)  HOW IS THE QUALITY OF AIR IN THE ROOM?
  VERY GOOD |	1	1  VERY BAD

(21)  DOES THE SKIN ON YOUR FACE FEEL DRY OR MOIST?
  VERY DRY |	1	1  VERY MOIST

(22)  HOW HARD DID YOU HAVE TO CONCENTRATE ON THESE QUESTIONS?
  VERY LITTLE |	1	1  VERY MUCH
                                       92

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                     APPENDIX E
  THE USE OF LINEAR POTENTIOMETERS FOR SUBJECTIVE
         ASSESSMENTS OF INDOOR AIR QUALITY
                 12              3
     Lars Molhave ,  David Otto ,  Gregory Rose
Institute of Environmental and Occupational Medicine
                University of Aarhus
                  Aarhus, Denmark

        2
         Human Effects Research Laboratory
       U.S. Environmenatal Protection Agency
            Chapel Hill, NC  27599-7315
              3
               NSI Environmental, Inc.
         Research Triangle Park, NC  27711
                         93

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Introduction

     The principle of potentiometer assessment is called modified magnitude
estimation and is based on a linear analog rating scale (Bond and Lader, 1974).
Subjects may continuously indicate individual or collective ratings of any
specified indoor climate factor presented in the form of questions with defined
continuous answers between two well defined extremes.

Measuring Principles

     Each subject has his own rating box (Fig. 1) with a sliding pointer that can
 be
positioned along a 60 mm track.  The device can be used in two modes. Ratings
can be signalled by external prompt (e.g., a visual prompt on the subject's
video screen or verbal instruction from the investigator) or ratings can be
initated spontaneously by the subject whenever a change in indoor air quality
is perceived.  In this study comfort ratings were signalled every 15-30 minutes
by video prompts or verbal instruction.  The extremes were defined as "no
irritation" and "unacceptably strong irritation".  Prior to commencement of
testing, the potentiometer pointer was positioned at the "no irritation"
extreme.
  Instructions to the Subject should include:

  a)  Definition of the endpoints and rating scale.

  b)  The nature of the indoor climate factor (s) to be rated.

  c)  That the measurement scale must be established by the subject
      himself.

  d)  That only the pointer should move and that nobody else should see
      the setting of the pointer.
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  e)  That the scale includes a neutral range (e.g., at the "no
      irritation" end).

  f)  That the initial position of the pointer has no particular
      relationship to the present IAQ.

  g)  Operation of the box and pointer must be demonstrated and explained
      to subjects prior to its use.

  h)  The time intervals at which ratings are to be made should be
      specified.

  i)  If external timing is used, the subjects should be instructed to go
      to the box and make a simulated rating even if they don't feel any
      change.  They may delay their rating if they are occupied with
      other tests when the sound signal appears.

  j)  The pointer must be set in a neutral position (i.e., no irritation)
      at the start of the tests.

  k)  Subjects should not move the pointer back to neutral and then to
      a new position when a rating is required.  That is, if a change in
      rating is desired, the pointer should be moved from its previous
      position directly to the new position.

The Equipment

     The rating  box  is  shown  in  Figure  1.   The  box  contained  a  linear
potentiometer   with   sliding   pointer  which  could  be  positioned  along  a
one-dimensional 60 mm axis.  A 5 volt power source in the breakout box  supplied
the voltage across the resistive arm to give a O to 5 volt output range.  Due to
a non-linear response in the 4 to 5 volt range,  the  slide  was  limited  to  a
maximum  output  of 4 volts.  Thus 0 volts corresponded to "no irritation" and 4
volts corresponded to "unacceptabley  strong  irritation".   Each  of  the  four
potentiometer  outputs  was directed to an input channel of an analog-to-digital
                                       95

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realtime module mounted in  the  backplane  of  the  Compaq  Plus  microcomputer
control  station.   A  BASIC  algorithm  automatically  sampled each of the four
channels simultanelusly every 15 minutes throughout the test day.  Sampling  was
initiated  manually  two  minutes  after testing commenced to permit subjects to
complete the initial comfort rating.  Subjects vere prompted on  video  monitors
while  testing  and  by  operators over loudspeakers during break periods to set
their comfort level prior  to  the  15  minute  auto  sampling.   The  algorithm
converted  the  sampled  digital units back to voltage units to store and report
irritation in a range of 0.0 to 4.0.
                                       96

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  4   VOLTS  0
RATING BOX 1
RATING BOX 2
RATING BOX 3
RATING BOX 4
                 i
BREAKOUT BOX

 (WITH 5 VOLT

POWER SUPPLY)

    FIGURE  1
COMPAQ PLUS
   WITH
    A/D
 CONVERTER

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     The calibration of the potentiometers with the analog-to-digital  converter

was  checked  prior  to each test day and stored by the computer in 0, 25%, 50%,

75%, 100% increments of the slide of the  potentiometer  which  corresponded   to

measures of 0.0, 1.0, 2.0, 3.0, and 4.0.


                          Instructions to the Subjects

     "An aluminum Comfort Meter is located to the right of your  computer.   The
lid should be open to show a sliding brass knob with a pointer that can be moved
from "No irritation" to "Strong irritation".  You will use this comfort meter  to
rate  the  degree  of  comfort or irritation that you feel in your eyes, nose  or
throat during the experiment.  There are no scale markings on the  box,  so  you
will have to establish your own scale when you start".

     "Remember, we want to monitor the level of comfort or irritation  that  you
feel  in  your  eyes,  nose  and  throat  during  the  experiment.   You will  be
instructed at regular intervals, both by computer prompts and over the intercom,
to  indicate  your comfort level.  Do not interrupt an ongoing test to reset the
comfort meter unless instructed to do so.  During rest  periods  between  tests,
however,  you  are  encouraged  to indicate any change in comfort level that you
feel.  Please close the lid after each rating in order to  prevent  others  from
seeing your comfort rating".

REFERENCE

Bond A., and Lader H.  The use of analoque scales in rating subjective feelings.
     Br.  J.  Med.  Psycholo.  1974, 47:211-218.
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