Research Research Report Report Number Number 33 33
Health Health Review Review Committee Committee
Includes Includes the the Conunentary Conunentary by by the the Institute~s Institute~s
University University of of Massachusetts Massachusetts Medical Medical School, School, Worcester, Worcester, MA MA
Community Community Medicine, Medicine, Environmental Environmental Health Health Sciences Sciences Program, Program,
Air Air Resources Resources Board, Board, Sacramento, Sacramento, and and Department Department CA; CA; of of and and Family Family
of of Environmental Environmental Toxicology; Toxicology; University University of of California, California, Davis, Davis, CA; CA; California California
Medicine, Medicine, Center Center for for Occupational Occupational and and Environmental Environmental Health, Health, and and Department Department
Division Division of of Occupational Occupational and and Environmental Environmental Medicine, Medicine, Department Department of of Internal Internal
S. S. J. J. Katharine Katharine Hammond, Hammond, Thomas Thomas Smith, Smith, Susan Susan R. R. Woskie Woskie
J. J. Marc Marc B. B. Schenker, Schenker, Steven Steven Samuels, Samuels, Norman Norman Y. Y. Kado, Kado,
in in Railroad Railroad Workers Workers
Markers Markers of of Exposure Exposure to to Exhaust Exhaust Diesel ------rr..tHEALTH EFFECTS INSTITUTE I.J:T
The Health Effects Institute (HEI) is a nonprofit corporation founded in 1980 to assure that objective, credible, high-quality scientific studies are conducted on the potential human health effects of motor vehicle emissions. Funded equally by the U.S. Environmental Protection Agency (EPA) and 28 automotive manufacturers or marketers in the United States, HEI is independently governed. Its research projects are selected, conducted, and evaluated according to a careful public process, including a rigorous peer review process, to assure both credibility and high scientific standards. HEI makes no recommen dations on regulatory and social policy. Its goal, as stated by former EPA Administrator William D. Ruckelshaus, is "simply to gain acceptance by all parties of the data that may be necessary for future regulations:'
The Board of Directors Archibald Cox Chairman Donald Kennedy Carl M. Loeb University Professor (Emeritus), Harvard Law School President, Stanford University William 0. Baker Walter A. Rosenblith Chairman (Emeritus), Bell Laboratories Institute Professor (Emeritus), Massachusetts Institute of Technology
Health Research Committee Richard Remington Chairman Robert F. Sawyer University of Iowa Foundation Distinguished Professor of Preventive Class of 1935 Professor of Energy, University of California at Berkeley Medicine and Environmental Health, University of Iowa John W. Tukey Joseph D. Brain Senior Research Statistician and Donner Professor of Science Emeritus, Cecil K. and Philip Drinker Professor of Environmental Physiology, Princeton University Harvard University School of Public Health Mark J. Utell Leon Gordis Professor of Medicine and Toxicology, University of Rochester School of Professor and Chairman, Department of Epidemiology, Johns Hopkins Medicine University, School of Hygiene and Public Health Gerald N. Wogan Curtis C. Harris Professor of Toxicology, Massachusetts Institute of Technology Chief, Laboratory of Human Carcinogenesis, National Cancer Institute Roger 0. McClellan President, Chemical Industry Institute of Toxicology
Health Review Committee Arthur Upton Chairman Robert M. Senior Professor and Chairman, Institute of Environmental Medicine, New York Professor of Medicine and Director, Respiratory and Critical Care University Division, The Jewish Hospital at Washington University Medical Center Bemard Goldstein James H. Ware Professor and Chairman, Department of Environmental and Community Dean of Academic Affairs and Professor of Biostatistics, Harvard Medicine, University of Medicine and Dentistry of New Jersey, Robert University School of Public Health Wood Johnson Medical Center Mary C. Williams Gareth M. Green Professor of Medicine (Cell Biology), Boston University School Associate Dean for Education, Harvard School of Public Health of Medicine Millicent W. P. Higgins W. Kent Anger Special Consultant to the Committee Associate Director for Epidemiology and Biometry, National Heart, Lung Associate Director for Occupational and Environmental Toxicology, and Blood Institute The Oregon Health Sciences University Herbert Rosenkranz Chairman, Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh
Officers and Staff Andrew Sivak President and Treasurer Martha E. Richmond Consulting Staff Scientist Richard M. Cooper Corporate Secretary Ann Y. Watson Consulting Staff Scientist Debra N. Johnson Controller Judith Zalon Lynch Director of Administration and Finance L. Virgi Hepner Publications Manager Kathleen M. Nauss Director for Scientific Review and Evaluation Gail V. Allosso Assistant to the Director of Administration and Finance Jane Warren Director of Research Robin A. Cuozzo Accounting Assistant William F. Busby, Jr. Senior Staff Scientist Jean C. Murphy Research Assistant Brenda E. Barry Staff Scientist Mary-Ellen Patten Administrative Assistant Aaron F. Cohen Staff Scientist Kate Rose Publications Assistant Maria G. Costantini Staff Scientist Hannah J. Protzman Secretary Bemard Jacobson Staff Scientist Joyce L. Speers Secretary Debra A. Kaden Staff Scientist Carolyn N. White Secretary Alison M. Dorries Consulting Staff Scientist Patricia White Receptionist
Copyright © 1990 by Health Effects Institute. § The paper in this publication meets the minimum requirements of the Printed at Capital City Press, Montpelier, VT. ANSI Standard Z39.4B-19B4 (Permanence of Paper) effective with Report Library of Congress Catalogue No. for the HE! Research Report Series: Number 21, December 19BB, and with Report Numbers 25 and 26 excepted. WA 754 R432. Reports 1 through 20, 25, and 26 are printed on acid-free coated paper.
42 42 ...... Survey Survey Health Health A. A. 18 18 ...... Squares Squares Least Least Generalized Generalized
42 42 ...... Appendices Appendices 17 17 ...... Strains Strains Bacterial Bacterial of of Sensitivity Sensitivity
Differential Differential for for 37 37 Accounting Accounting Form: Form: Model Model
......
• • References • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 0 0
17 17 ...... Diagnostics Diagnostics Model Model and and 37 37 Transformations Transformations ...... Acknowledgments Acknowledgments
16 16 Models Models Regression Regression for for 36 36 Factors Factors Predictive Predictive Other Other ...... Findings Findings the the of of Implications Implications
16 16 ...... 36 36 Models Models Phenanthrene Phenanthrene ...... Power . Study Study
16 16 ...... Effects Effects 35 35 Exposure Exposure for for Model Model Interaction Interaction ...... Compounds Compounds
Occurring Occurring Naturally Naturally of of Activity Activity Antimutagenic Antimutagenic 15 15 ...... Construction Construction Index Index
35 35 ...... Factors Factors Dietary Dietary 14 14 ...... Exposure Exposure Exhaust Exhaust Diesel Diesel of of Indices Indices
34 34 ...... Exhaust Exhaust Diesel Diesel 14 14 ...... Analyses Analyses Smoking Smoking Preliminary Preliminary
34 34 ...... Mutagenicity. Mutagenicity. Urinary Urinary 14 14 ...... Strategy Strategy General General
34 34 ...... Exposure Exposure Phenanthrene Phenanthrene 14 14 ...... Methods Methods Statistical Statistical
33 33 ...... Exposure Exposure Smoke Smoke 14 14 ...... Management Management Data Data
Tobacco Tobacco
Environmental Environmental and and Particle Particle Respirable Respirable
13 13 ...... Chemistry Chemistry Clinical Clinical
33 33 ......
Exposure Exposure Exhaust Exhaust Diesel Diesel
12 12 ...... Urine Urine of of Testing Testing Mutagenicity Mutagenicity
33 33
......
Discussion Discussion
11 11 ...... Methods Methods Extraction Extraction and and Collection Collection Urine Urine
33 33
...... Analyses Analyses Other Other
11 11 ...... Extracts Extracts Diesel Diesel
29 29
...... Exposure Exposure Exhaust Exhaust Diesel Diesel
11 11 ...... Method Method Assay Assay Microsuspension Microsuspension
29 29
...... Mutagenicity Mutagenicity Urinary Urinary
11 11
......
Chemicals Chemicals
of of Predictors Predictors Exhaust Exhaust Non-Diesel Non-Diesel Other Other
11 11
......
Mutagenicity Mutagenicity
28 28 ...... Smoking Smoking Cigarette Cigarette Passive Passive of of Markers Markers
10 10
......
Strategy Strategy
Sampling Sampling
27 27 ...... Smoking Smoking Cigarette Cigarette Active Active of of Markers Markers
...... 9 9 ...... Phenanthrene Phenanthrene
27 27 4 ...... 4 Aim Aim Specific Specific
...... 9 9 ......
Nicotine Nicotine
26 26 ...... Exposure Exposure Phenanthrene Phenanthrene
...... 8 8 ...... Particles Particles
Respirable Respirable
24 24 ...... Exposure Exposure Particles Particles Respirable Respirable
...... 7 7 ...... Control Control Quality Quality
24 24 ...... Smoking Smoking Cigarette Cigarette
...... 7 7 ......
Approach Approach
General General
23 23 ...... Population Population Study Study
...... 7 7 ......
Hygiene Hygiene Industrial Industrial
23 23 3 ...... 3 Aim Aim Specific Specific
6 6
......
• • Epidemiology • • • • 0 0
23 23 ...... Samplers. Samplers. Personal Personal
6 6 Methods...... Methods......
from from Particles Particles Respirable Respirable of of Mutagenicity Mutagenicity
5 5
......
Aims Aims
• • Specific • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 0 0 21 21 Extracts Extracts Diesel Diesel of of Relationships Relationships Dose-Response Dose-Response
. . 5 5 . . Excretion Excretion Mutagen Mutagen
of of
Kinetics Kinetics the the and and Exposure Exposure 21 21 ...... Extracts Extracts
...... 4 4 ...... Assay Assay Mutagenicity Mutagenicity Diesel Diesel for for Sensitivity Sensitivity Assay Assay Microsuspension Microsuspension
...... 3 3 ...... Contaminants Contaminants Air Air for for Markers Markers of of Use Use 21 21 2 ...... 2 Aim Aim Specific Specific
...... 3 3 ...... Exhaust Exhaust Diesel Diesel of of Composition Composition Complex Complex 19 19 ...... Smoke Smoke Tobacco Tobacco
Environmental Environmental for for Marker Marker a a as as Nicotine Nicotine ...... 2 2 ...... Exposure Exposure of of Variability Variability
18 18 . . . . Exhaust Exhaust Diesel Diesel for for Marker Marker a a as as Phenanthrene Phenanthrene ...... 2 2 ...... Assessment. Assessment. Exposure Exposure
18 18 ...... 1 ...... 1 Aim Aim Specific Specific ...... 1 1 ...... Introduction Introduction
18 18 ...... Results Results ...... 1 1 ...... Abstract Abstract
J. J. Woskie Woskie R. R. Susan Susan Smith, Smith, Thomas Thomas
Hammond, Hammond, J. J. Katharine Katharine S. S. Kado, Kado, Y. Y. Norman Norman Samuels, Samuels, Steven Steven Schenker, Schenker, B. B. Marc Marc REPORT REPORT INVESTIGATORS' INVESTIGATORS'
Workers Workers Railroad Railroad in in Exhaust Exhaust Diesel Diesel to to Exposure Exposure of of Markers Markers
33 33 Number Number Report Report Research Research
CONTENTS CONTENTS OF OF TABLE TABLE
ii ii
Phenanthrene Phenanthrene ...... 58 58 References References ...... 61 61
Total Total Respirable Respirable Particle Particle Concentration Concentration ...... 57 57 Conclusions Conclusions ...... 61 61
Environmental Environmental Monitoring Monitoring ...... 57 57 Implications Implications for for Future Future Research Research ...... 60 60
Technical Technical Evaluation Evaluation ...... 57 57 Other Other Factors Factors ...... 60 60
Study Study Design Design ...... 57 57 Diesel Diesel Exhaust Exhaust Exposure Exposure ...... 59 59
Goals Goals and and Objectives Objectives ...... 56 56 Cigarette Cigarette Smoke Smoke Exposure Exposure ...... 59 59
Justification Justification for for the the Study Study ...... 56 56 Urine Urine Monitoring Monitoring ...... 59 59
Scientific Scientific Background Background ...... 53 53 Personal Personal Samples Samples ...... 58 58
Regulatory Regulatory Background Background ...... 53 53 Mutagenicity Mutagenicity of of Respirable Respirable Particles Particles from from
Introduction Introduction ...... 53 53 Cigarette Cigarette Smoke Smoke ...... 58 58
HEALTH HEALTH REVIEW REVIEW COMMITTEE'S COMMITTEE'S Health Health COMMENTARY COMMENTARY Effects Effects Institute Institute
Regression Regression Variables. Variables...... 46 46
D. D. Urinary Urinary Mutagenicity Mutagenicity Correlation Correlation with with Multivariate Multivariate
Abbreviations Abbreviations ......
51 51 per per Milliliter Milliliter Equivalent Equivalent of of Urine Urine ...... 45 45
About About the the Authors Authors ...... 51 51 Methods Methods for for Computing Computing the the Number Number of of Revertants Revertants
C. C. Comparison Comparison of of Point-Deletion Point-Deletion and and Tangent-Slope Tangent-Slope F. F. Phenanthrene Phenanthrene as as a a Marker Marker for for Diesel Diesel Exhaust Exhaust ...... 49 49
Thiocyanate Thiocyanate Concentration Concentration ...... 43 43 Controls Controls ...... 48 48
B. B. Effects Effects of of Storage Storage on on Urinary Urinary Mutagenicity Mutagenicity and and Statistical Statistical E. E. Descriptions Descriptions of of Mutagenic Mutagenic Activity Activity
1 1
of of risk risk
increased increased
the the about about
concerns concerns
raised raised
States States United United 95616. 95616. CA CA Davis, Davis, California, California, of of University University Medicine, Medicine,
Environmental Environmental
and and Occupational Occupational of of the the in in Division Division use use engine engine Professor, Professor, diesel diesel in in Associate Associate and and increase increase projected projected the the and and haust haust
Chief Chief Division Division Schenker, Schenker, B. B. Marc Marc Dr. Dr. to to
addressed addressed
be be
may may
Correspondence Correspondence
ex diesel diesel of of mutagenicity mutagenicity known known the the 1970s, 1970s, late late the the In In 1 1
INTRODUCTION INTRODUCTION job job by by exposure exposure exhaust exhaust diesel diesel known known with with correlated correlated
and and samples, samples, personal personal of of subset subset a a in in measured measured was was marker, marker,
potential potential a a as as concentration, concentration, Phenanthrene Phenanthrene grouping. grouping. job job
by by
exposure exposure exhaust exhaust diesel diesel of of assessments assessments priori priori a a with with exposure. exposure. exhaust exhaust diesel diesel with with associated associated risks risks cer cer
correlated correlated
smoke smoke tobacco tobacco environmental environmental for for can adjusted adjusted the the was was about about conclusions conclusions draw draw to to which which on on basis basis no no vides vides
that that
concentration concentration particle particle respirable respirable but but location, location, pro work work the the study study Our Our sensitivity. sensitivity. greater greater or or levels levels exposure exposure higher higher
in in
existed existed smoke tobacco tobacco environmental environmental by by require require would would contamination contamination studies studies epidemiologic epidemiologic for for mutagenicity mutagenicity
when when exposure exposure exhaust exhaust diesel diesel of of marker marker good good urinary urinary a a of of not not use use was was The The urine. urine. the the in in undetectable undetectable were were exposure exposure
concentration concentration particle particle Respirable Respirable samples). samples). of of 306 306 levels levels (n (n these these = = day day with with associated associated mutagens mutagens the the study, study, this this in in
single single a a for for
data data usable usable provided provided subjects subjects four four additional additional smoke smoke an an tobacco tobacco environmental environmental or or exhaust exhaust diesel diesel to to exposure exposure
protocols; protocols;
two-day two-day 151 151 completed completed subjects subjects of of Eighty-seven Eighty-seven levels levels the the at at that, that, conclude conclude We We mutagens. mutagens. urinary urinary of of els els
lev low low very very observe observe to to study study the the of of power power components. components. statistical statistical limited limited within-person within-person and and between-
the the and and field; field; the the into into in in them them variation variation study study to to residual residual ability ability divided divided that that limited limited the the analysis analysis least-squares least-squares
and and excretion, excretion, and and generalized generalized a a metabolism, metabolism, including including absorption, absorption, exhaust exhaust mutagenicity, mutagenicity, diesel diesel urinary urinary of of minants minants
of of kinetics kinetics unknown unknown deter the the independent independent concentrations; concentrations; analyze analyze particle particle to to used used respirable respirable were were analyses analyses sion sion
than than accurate accurate more more markers markers of of regres use use the the Multiple Multiple work. work. despite despite and and estimates estimates home home at at exposures exposures and and habits habits
exposure exposure exhaust exhaust diesel diesel inexact inexact assay; assay; personal personal on on day day mutagenicity mutagenicity second second urine urine the the of of end end the the at at questionnaire questionnaire
the the of of sensitivity sensitivity a a the the completed completed exhaust; exhaust; diesel diesel subjects subjects to to and and exposures exposures recorded, recorded, place place was was shift shift study study the the
work lower-than-expected lower-than-expected on on the the smoked smoked include include These These cigarettes cigarettes of of number number mutagenicity. mutagenicity. The The enzyme). enzyme). S9 S9 without without
urinary urinary and and or or exposure exposure with with exhaust exhaust TA98 TA98 diesel diesel strain strain between between Salmonella Salmonella assay; assay; association association preincubation preincubation cro cro
an an of of absence absence the the to to (mi assay assay contributed contributed have have may may microsuspension microsuspension factors factors sensitive sensitive a a Several Several using using mutagenicity, mutagenicity,
for for
and and cotinine) cotinine) (nicotine, (nicotine, smoking smoking cigarette cigarette of of markers markers enzyme). enzyme). S9 S9 by by tivation tivation
for for
analyzed analyzed were were and and shifts shifts work work study study ac the the of of end end metabolic metabolic the the with with TA98 TA98 strain strain tester tester Salmonella Salmonella the the (using (using
at at
collected collected were were samples samples Urine Urine smoking. smoking. cigarette cigarette mutagenicity mutagenicity passive passive urinary urinary lower lower with with associated associated dependently dependently
and and
active active for for concentrations concentrations particle particle in respirable respirable was was the the fish) fish) adjust adjust and and sprouts, sprouts, Brussels Brussels (cabbage, (cabbage, foods foods certain certain
to to
measured measured was was samples samples the the in in Nicotine Nicotine of of shifts. shifts. ingestion ingestion work work tive tive Self-reported Self-reported mutagenicity. mutagenicity. urinary urinary and and smoke smoke
consecu two two over over taken taken samples samples tobacco tobacco air air personal personal environmental environmental using using or or by by haust haust exhaust exhaust diesel diesel of of markers markers tween tween
ex
diesel diesel to to workers workers railroad railroad individual individual of of be found found exposure exposure be be the the could could association association no no but but urine, urine, the the in in present present
measured measured We We workplace. workplace. the the in in exposure exposure were were exhaust exhaust diesel diesel mutagens mutagens of of of of concentrations concentrations detectable detectable nonsmokers, nonsmokers,
marker marker biological biological a a as as mutagenicity mutagenicity urinary urinary of of Among Among usefulness usefulness mutagenicity. mutagenicity. urinary urinary and and exposure exposure exhaust exhaust diesel diesel
the the
evaluate evaluate to to undertaken undertaken was was study study This This marker. marker. between between standard standard present present was was association association no no for, for, controlled controlled was was
a a of of absence absence the the by by limited limited been been has has also also smoking smoking exposure exposure cigarette cigarette After After exhaust exhaust day. day. study study the the on on smoked smoked cigarettes cigarettes
diesel diesel of of Measurement Measurement received. received. dose dose of of internal internal the the of of number number the the mates mates and and mutagenicity mutagenicity urinary urinary between between served served
esti accurate accurate be be not not may may exposure exposure of of ob measures measures was was accurate accurate relationship relationship dose-response dose-response a a smokers, smokers, Among Among
even even and and exposure, exposure, of of estimates estimates crude crude used used have have studies studies ical ical
studies. studies. epidemiological epidemiological in in investigation investigation
epidemiolog many many However, However,
exposure. exposure.
exhaust exhaust
diesel diesel
from from
further further for for useful useful be be may may technique technique this this and and samples, samples, filter filter
cancer cancer lung lung of of risk risk the the in in increase increase
small small
a a
demonstrated demonstrated
personal personal few few a a in in particles particles respirable respirable of of extracts extracts from from
sured sured
have have studies studies epidemiological epidemiological Recent Recent
carcinogen. carcinogen.
man man
mea also also was was activity activity Mutagenic Mutagenic exhaust. exhaust. diesel diesel for for marker marker
hu potential potential a a and and mutagen mutagen
known known a a is is
exhaust exhaust Diesel Diesel
a a as as promising promising is is phenanthrene phenanthrene that that suggested suggested locations locations
work work exhaust-exposed exhaust-exposed diesel diesel from from samples samples area area in in particles particles
ABSTRACT ABSTRACT
respirable respirable to to phenanthrene phenanthrene of of ratio ratio constant constant A A grouping. grouping.
Woskie Woskie J. J. R. R. Susan Susan Smith, Smith, Thomas Thomas
Hammond, Hammond,
Katharine Katharine S. S. Kado, Kado, J. J. Y. Y. Norman Norman Samuels, Samuels, Steven Steven Schenker B. B. Marc Marc , ,
1
Workers Workers Railroad Railroad in in Exhaust Exhaust Diesel Diesel to to Exposure Exposure of of Markers Markers
REPORT REPORT INVESTIGATORS' INVESTIGATORS'
2 2
An An important important consideration consideration in in studies studies of of biological biological gens gens can can be be compared. compared.
exposures exposures against against which which the the variability variability of of urinary urinary muta species. species.
ception ception (Woskie (Woskie et et al. al. 1988a,b). 1988a,b). Thus, Thus, there there is is a a range range of of exhaust, exhaust, because because is is not not specific specific for for it it a a single single chemical chemical
strated strated clearly clearly that that exposure exposure to to diesel diesel exhaust exhaust is is not not an an ex it it is is particularly particularly useful useful for for complex complex mixtures, mixtures, such such as as diesel diesel
taminants. taminants. Our Our previous previous work work with with railroads railroads has has demon cause cause of of its its low low cost cost and and the the ease ease of of sample sample collection, collection, and and
seen seen in in both both the the composition composition and and the the concentration concentration of of con mutagenicity mutagenicity is is convenient convenient for for occupational occupational studies studies be
tions, tions, jobs, jobs, work work areas, areas, individuals, individuals, and and time. time. Variability Variability is is minki minki et et al. al. 1977; 1977; Everson Everson 1986). 1986). Measurement Measurement of of urinary urinary
substantially substantially variable variable across across companies, companies, industrial industrial opera posures posures to to other other known known or or suspected suspected carcinogens carcinogens (Hem
Occupational Occupational exposure exposure to to air air contaminants contaminants generally generally is is sequently sequently it it has has been been associated associated with with occupational occupational ex
among among cigarette cigarette smokers smokers by by Yamasaki Yamasaki and and Ames Ames (1977); (1977); sub
Variability Variability of of Exposure Exposure
urine urine as as a a result result of of exogenous exogenous exposure exposure was was first first observed observed
ity ity and and chromosomal chromosomal changes. changes. Mutagenic Mutagenic activity activity in in the the
differences differences within within and and between between groups. groups.
by by the the exposure. exposure. Examples Examples of of the the latter latter include include mutagenic
variability variability of of urinary urinary mutagenicity mutagenicity relative relative to to exposure exposure
specific specific chemicals chemicals or or their their metabolites, metabolites, or or of of effects effects caused caused
group group men men with with similar similar job job descriptions descriptions and and examine examine the the
vironmental vironmental exposures exposures in in biological biological specimens specimens may may be be of of
daily daily personal personal exposure exposure over over several several months, months, but but we we could could
posure posure sampling. sampling. Measurements Measurements of of occupational occupational or or en
requirement, requirement, it it was was not not practical practical to to measure measure each each subject's subject's
validate validate exposure exposure measurements measurements made made by by personal personal ex
lecting lecting urine urine samples samples at at the the end end of of each each day. day. For For the the second second
biological biological specimens. specimens. Biological Biological measurements measurements may may also also
posure posure of of each each subject subject on on two two consecutive consecutive days days and and by by col
agents agents may may be be estimated estimated better better by by measurements measurements of of dose dose in in
the the first first requirement requirement was was fulfilled fulfilled by by measuring measuring the the ex
Individual Individual differences differences in in reactions reactions to to environmental environmental
exposure exposure over over the the previous previous several several months. months. For For our our study, study,
individual. individual.
ately ately prior prior to to urine urine sampling; sampling; and and (2) (2) estimation estimation of of average average
not not be be accurate accurate
estimates estimates ofthe ofthe
internal internal dose dose received received
by by
an an
components: components: (1) (1) determination determination of of daily daily exposure exposure immedi
these these factors, factors, even even accurate accurate measurements measurements of of exposure exposure
may may
Ideally, Ideally, an an exposure exposure assessment assessment strategy strategy should should have have two two
ing, ing, nonoccupational nonoccupational exposures, exposures, and and drug drug use. use. Because Because of of
over over time. time.
posures. posures. These These include include age, age, gender, gender, genotype, genotype, diet, diet, smok
with with location location or or work work activity activity and and are are generally generally not not stable stable
that that cause cause individual individual differences differences in in reactions reactions to to similar similar ex
sorption, sorption, and and ingestion. ingestion. Air Air concentrations concentrations vary vary widely widely
Estimates Estimates of of dose dose may may also also be be affected affected by by many many factors factors
possible possible routes routes of of entry entry into into the the body: body: inhalation, inhalation, skin skin ab
pothesis pothesis of of no no association. association.
problem problem is is further further complicated complicated by by the the need need to to evaluate evaluate three three
sensitivity, sensitivity, and and
may may thus thus
bias bias the the results results
toward toward the the
null null hy
ject: ject: composition, composition, concentration, concentration, and and time time profile. profile. The The
measures measures in in epidemiological epidemiological
studies studies
may may
reduce reduce
the the study study
dimensions dimensions of of exposure exposure must must be be determined determined for for each each sub
exposure exposure measurements. measurements.
The The use use
of of inaccurate inaccurate
exposure exposure
because because exposure exposure conditions conditions are are highly highly variable. variable. Three Three
sification sification or or work work
area, area,
but but
these these are are
not not always always
based based
on on
posures posures for for studies studies of of human human health health effects effects is is very very difficult difficult
dustry dustry as as indicators indicators of of exposure. exposure. Some Some studies studies
use use job clas job
Characterization Characterization of of environmental environmental or or occupational occupational ex
studies studies use use indices indices as as crude crude as as employment employment in in a a certain certain in
EXPOSURE EXPOSURE
ASSESSMENT ASSESSMENT
dose dose (Axelson (Axelson 1985). 1985). Many Many occupational occupational epidemiology epidemiology
been been limited limited or or inaccurate inaccurate determination determination of of exposure exposure or or
the the early early studies studies of of diesel diesel exhaust-exposed exhaust-exposed workers, workers, has has determinants determinants of of urinary urinary mutagenicity. mutagenicity.
spiratory spiratory disease disease due due to to environmental environmental agents, agents, including including use use respirable respirable particles, particles, and and to to evaluate evaluate the the independent independent
A A major major weakness weakness in in the the epidemiological epidemiological studies studies of of re adjust adjust the the measurements measurements of of diesel diesel exhaust exhaust exposure exposure that that
been been reviewed reviewed (Ishinishi (Ishinishi et et al. al. 1986; 1986; McClellan McClellan 1987). 1987). have have an an independent independent determination determination of of cigarette cigarette smoke smoke to to
1985), 1985), and and the the health health effects effects of of diesel diesel exhaust exhaust exposure exposure have have ble ble particle particle concentrations. concentrations. is is important, important, therefore, therefore, It It to to
Hall Hall and and Wynder Wynder 1984; 1984; Schenker Schenker mental) mental) et et al. al. 1984; 1984; smoking smoking Wong Wong may may et et al. al. also also contribute contribute to to measured measured respira
of of long-term long-term exposure exposure to to diesel diesel exhaust exhaust (Howe (Howe et et al. al. 1983; 1983; dently. dently. Cigarette Cigarette smoke smoke from from active active and and passive passive (environ
epidemiological epidemiological studies studies were were begun begun to to evaluate evaluate the the effects effects study study to to measure measure the the dose dose of of cigarette cigarette smoke smoke indepen
(Schenker (Schenker and and Speizer Speizer 1979; 1979; Schenker Schenker 1980). 1980). Several Several new new (Benowitz (Benowitz et et al. al. 1986). 1986). These These methods methods were were used used in in this this
inadequate inadequate duration duration of of exposure, exposure, or or limited limited follow-up follow-up as as cotinine cotinine (a (a metabolite metabolite of of nicotine) nicotine) in in the the blood blood or or urine urine
but but the the studies studies were were often often limited limited studies studies by by small small have have sample sample used used more more size, size, specific specific biochemical biochemical markers markers such such
haust haust exposure exposure had had previously previously shown shown conflicting conflicting results, results, traditionally traditionally has has been been done done by by questionnaire, questionnaire, but but newer newer
of of lung lung cancer cancer among among workers workers with with occupational occupational diesel diesel ex urine. urine. Control Control for for smoking smoking in in epidemiological epidemiological studies studies
National National Research Research Council Council 1981). 1981). Epidemiological Epidemiological studies studies rette rette smoking, smoking, a a known known cause cause of of mutagen mutagen excretion excretion in in the the
cancer cancer from from diesel diesel exhaust exhaust exposure exposure (Pepelko (Pepelko et et al. al. 1979; 1979; markers markers for for diesel diesel exhaust exhaust is is control control for for the the effects effects of of ciga
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
3 3
reference. reference. your your for for report report
this this of of end end the the at at A A appears appears interest. interest. of of abbreviations abbreviations of of agent agent list list A A the the
with with proportionately proportionately vary vary to to shown shown 2 2
been been has has concentration concentration whose whose indicator indicator or or compound compound
single single a a is is marker marker A A "markers:' "markers:' measure measure and and identify identify to to is is and and collect collect to to developed developed were were Methods Methods exhaust. exhaust. diesel diesel for for
exposure exposure
quantify quantify to to how how of of problem problem the the to to solution solution One One marker marker possible possible another another as as therefore, therefore, selected, selected, was was threne threne
Phenan 1984). 1984). Hites Hites and and Behymer Behymer 1983; 1983; Hites Hites and and (Jensen (Jensen
Contaminants Contaminants Air Air for for Markers Markers of of Use Use
sources sources combustion combustion other other or or engines engines spark-type spark-type do do than than
phenanthrene phenanthrene more more produce produce to to seem seem engines engines
diesel diesel
that that
directly. directly.
exhaust exhaust diesel diesel measure measure to to possible possible not not is is sult, sult, it it
literature literature the the in in reports reports other other with with
consistent consistent
was was
finding finding
are
As As
air. air.
the the in in is is else else what what and and analyzed, analyzed, is is it it emission emission
This This 1985). 1985). al. al. et et (Hammond (Hammond
concentration concentration
highest highest the the
in in
after after long long
how how
conditions, conditions,
operating operating and and type type engine engine the the
present present (PAH
hydrocarbon hydrocarbon ) ) aromatic aromatic
polycyclic polycyclic
the the
be be to to
2
composition, composition, fuel fuel the the on on depending depending varies varies but but fixed, fixed, not not
phenanthrene phenanthrene found found we we engines, engines, diesel diesel
railroad railroad
from from haust haust
is is
therefore, therefore,
exhaust, exhaust,
diesel diesel of of composition composition The The oxidation). oxidation).
ex of of samples samples previous previous our our In In
sought. sought.
was was
marker marker
specific, specific,
by by
example, example,
(for (for
formed formed be be may may compounds compounds additional additional and and
more more another, another, so so area, area, an an in in particles particles of of
sources sources
important important
contaminants contaminants air air
other other
with with interacts interacts it it atmosphere, atmosphere, the the in in
other other be be may may still still there there made, made, is is
correction correction
this this
after after Even Even
mixing mixing
turbulent turbulent and and
diffusion diffusion by by diluted diluted is is mixture mixture this this
marker. marker. a a as as used used are are particles particles
respirable respirable
the the
before before
tracted tracted
As As
sources. sources.
other other and and additives additives fuel fuel from from material material organic organic
sub be be should should particles particles cigarette cigarette of of
amount amount
the the of of
mate mate
in other other
and and
combustion; combustion; during during formed formed are are that that dioxide) dioxide)
esti an an areas, areas, work work some some in in particles particles
respirable respirable
of of
source source
nitrogen nitrogen
and and
oxide oxide
(nitric (nitric oxides oxides nitrogen nitrogen fuel; fuel; the the in in fur fur
common common a a is is smoke smoke cigarette cigarette
Because Because area. area.
an an
in in cles cles
sul
from from
aerosol aerosol
acid acid
sulfuric sulfuric and and dioxide dioxide sulfur sulfur present: present:
parti respirable respirable of of source source
predominant, predominant, the the
least least at at or or
only, only,
also also
is is
material material
Inorganic Inorganic condense. condense. may may materials materials volatile volatile
the the is is exhaust exhaust if if exhaust exhaust diesel diesel
of of
marker marker a a be be
can can
air air
the the
the the cool, cool,
emissions emissions
as as but but vapors, vapors, lose lose rapidly rapidly particles particles
in in particles particles respirable respirable of of mass mass the the
submicron-sized, submicron-sized,
are are cles cles
hot hot
the the
emission, emission,
after after
Immediately Immediately 1985). 1985). al. al. et et (Hammond (Hammond
parti exhaust exhaust diesel diesel Because Because proposed. proposed. were were
markers markers eral eral
compounds compounds aromatic aromatic polycyclic polycyclic including including products, products, sis sis
sev and and smoke, smoke, diesel diesel with with used used was was approach approach
similar similar A A
pyroly of of variety variety wide wide a a and and compounds, compounds, oxygenated oxygenated bons, bons,
particles. particles. respirable respirable and and nicotine nicotine tween tween
hydrocar fuel fuel unburned unburned compounds: compounds: organic organic of of
hundreds hundreds
be relationship relationship the the estimate estimate to to used used were were subgroups subgroups free free
and and particles particles soot soot carbonaceous carbonaceous are are constituents constituents
main main
The The
diesel
and and studies studies Chamber Chamber smoke. smoke. tobacco tobacco vironmental vironmental
interact. interact. continuously continuously particles particles and and vapors vapors the the and and
aerosol, aerosol,
en to to
exposure exposure for for marker marker a a as as nicotine nicotine use use to to and and nicotine nicotine
and and vapors, vapors, gases, gases, of of mixture mixture complex complex a a is is exhaust exhaust
Diesel Diesel
analyze analyze and and collect collect to to method method a a developed developed we we study, study, this this of of
Exhaust Exhaust Diesel Diesel of of Composition Composition part part As As Complex Complex smoke. smoke. cigarette cigarette airborne airborne for for marker marker a a as as used used be be can can
nicotine nicotine airborne airborne nicotine, nicotine, of of fraction fraction constant constant a a contains contains
generally generally smoke smoke sidestream sidestream Because Because 1984). 1984). al. al. et et (Rickert (Rickert sampling. sampling. urine urine preceding preceding
ventilation ventilation of of degree degree of of regardless regardless cigarettes, cigarettes, of of brands brands ent ent shift shift work work the the on on measured measured be be should should individual individual an an for for
differ among among found found is is smoke smoke sidestream sidestream of of levels levels nicotine nicotine exposure exposure mutagenicity, mutagenicity, urine urine and and exposure exposure exhaust exhaust diesel diesel
the the in in variability variability Little Little 1987). 1987). al. al. et et (Hammond (Hammond nicotine nicotine between between relation relation the the estimate estimate to to and and variable, variable, highly highly is is
of of amount amount
constant constant
roughly roughly a a emits emits cigarette cigarette the the of of end end ing ing setting setting this this in in exposure exposure personal personal Thus, Thus, !J,g/m 905 905 and and
35 35 . . 3
burn the the that that shown shown have have Experiments Experiments smoke. smoke. tobacco tobacco tal tal between between interval interval the the in in were were electricians electricians for for concentrations concentrations
environmen of of component component major major the the is is air, air, the the into into directly directly particle particle respirable respirable the the of of percent percent 95 95 example, example, For For 1988b). 1988b). al. al.
cigarette cigarette the the from from emitted emitted is is which which smoke, smoke, Sidestream Sidestream et et (Woskie (Woskie categories categories job job within within exposures exposures variable variable highly highly
the the confirmed confirmed workers workers railroad railroad among among work work 1983). 1983). earlier earlier Our Our Chortyk Chortyk and and (Schlotzhauer (Schlotzhauer ways ways other other in in rette rette
ciga the the of of design design the the changing changing and and ventilation, ventilation, creasing creasing posures. posures.
in smoke, smoke, the the filtering filtering by by achieved achieved is is smoke smoke ex mainstream mainstream characterize characterize to to strategies strategies sampling sampling statistical statistical use use to to
in in nicotine nicotine Reducing Reducing 1983). 1983). Chortyk Chortyk and and (Schlotzhauer (Schlotzhauer necessary necessary is is it it Thus, Thus, processes. processes. transport transport the the or or emissions emissions
matter matter particulate particulate total total to to nicotine nicotine of of ratio ratio the the in in ations ations either either affect affect may may behavior behavior individual's individual's exposed exposed the the setting, setting,
vari small small only only but but tar, tar, and and nicotine nicotine of of yields yields smoke smoke occupational occupational stream stream an an in in and and air); air); uncontaminated uncontaminated with with example, example,
main in in increases increases large large produce produce may may duration duration and and (for (for volume volume materials materials other other with with them them dilute dilute to to mixing, mixing, turbulent turbulent as as
puff puff in in Increases Increases lungs. lungs. the the into into and and cigarette cigarette the the such such through through processes, processes, transport transport random random permits permits individual individual posed posed
drawn drawn is is that that smoke smoke the the smoke, smoke, mainstream mainstream in in 1985) 1985) port port ex the the to to source source the the from from contaminants contaminants of of passage passage time; time;
Re Commission Commission Trade Trade (Federal (Federal nicotine nicotine of of levels levels various various over over composition composition and and strength strength in in vary vary emissions emissions source source
contain contain cigarettes cigarettes of of brands brands Different Different many: many: compounds. compounds. of of are are sands sands variability variability extreme extreme this this for for reasons reasons The The minutes. minutes.
thou of of mixture mixture complex complex very very a a is is which which smoke, smoke, even even cigarette cigarette or or hours, hours, days, days, over over magnitude magnitude of of orders orders by by vary vary
for for marker marker a a as as nicotine nicotine airborne airborne of of use use the the is is example example to to good good found found been been have have contaminants contaminants air air of of Concentrations Concentrations
al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers
analyze phenanthrene on personal samplers as part of this methane was the most efficient method, the volumes of research. urine they extracted, which had relatively high concentra tions of diesel-associated mutagens, were small. Liquid ex traction becomes impractical for concentrating larger volumes of urine as well as for processing larger numbers MUTAGENICITY ASSAY of urine samples, and the use of solid adsorbents, such as Investigators have estimated exposure to complex mix XAD-2, becomes more appealing. Since XAD has been the tures by determining mutagenic activity in physiologic adsorbent used in studies measuring mutagenic activity in fluids (Legator et al. 1976; Yamasaki and Ames 1977; Falck the urine of cigarette smokers, and the data base for using et al. 1980; Dolara et al. 1981; Baker et al. 1982; Sorsa et al. the resin method of extraction is larger than for other 1985). Urine has been the most widely used physiologic methods, we considered its use in the current study. One fluid because it is convenient to collect, is noninvasively ac factor that makes the XAD method of extraction favorable is quired, and is one of the major routes of excretion for muta that it traps mutagens in urine, but differentially does not gens. There have been several short-term tests used for this trap histidine, which can cause the histidine prototrophs of purpose (Vainio 1985), but the most widely used and vali Salmonella to revert. If present, these "pseudorevertants" dated is the Salmonella microsome assay (Ames et al. can increase the background mutagenic activity. 1975). To detect the low levels of mutagenic activity in the urine In studies in which the Salmonella test was used for de of nonsmokers, an assay with greater sensitivity than the tecting mutagenic activity in physiologic fluids (mostly standard plate incorporation test of the Salmonella micro urine), samples collected from control individuals who some assay would be helpful. Numerous modifications of were not exposed to high concentrations of complex muta the Ames test procedure have been reported to increase its genic mixtures generally had low or undetectable muta sensitivity to selected compounds (Gletten et al. 1975; genic activity (Yamasaki and Ames 1977; Falck et al. 1980; Green et al. 1977; Malling 1977; Yahagi et al. 1977; Andrews Aeschbacher and Chappuis 1981; Kriebel et al.. 1985). et al. 1978; Mitchell1978). For example, Falck and cowork Numerous studies have been conducted on the mutagenic ers (1980) have used a fluctuation test to detect low levels of ity of cigarette smokers' urine, but very few on nonsmokers' mutagen in the urine of rubber workers, but these results are or passive smokers' urine (Bos et al. 1983; Sorsa et al. 1985). not directly comparable with the existing data base of muta Studies of urinary mutagenicity from passive smoking have gens tested in the standard plate incorporation test. often not accounted for such important covariates as urine We have previously reported a simple (microsuspension) dilution (Bos et al. 1983). Methods of detecting mutagenic modification of the Salmonella assay that is more sensitive activity in human physiological fluids have been reviewed than the standard plate incorporation test, as judged by ab by Everson (1986) and Venitt (1988). solute amounts of mutagen added for a specific mutagenic With respect to diesel particulate matter, Belisario and response (Kado et al. 1983, 1985). The modification consists coworkers (1984) reported on the bioavailability of the of adding 10 times more bacteria (approximately 109 per in mutagens adsorbed to diesel particles. They administered cubation tube) and 5 to 10 times less metabolic enzyme, diesel particles to rats by intraperitoneal injection and compared to the plate incorporation method. The total mix found mutagenic activity in the rats' urine 24 hours later. ture volume is approximately 0.2 mL, and the mixture is in The dose of particulate matter required to double the muta cubated for 90 minutes before it is poured, according to the genic response (using Salmonella strain TA98 with meta standard protocol. Results from this procedure (expressed bolic activation by S9) over that of control rats was approxi as revertants per plate) can be compared to the data base ex mately 100 to ZOO mg/kg of body weight. Urine was tested isting for mutagens and complex mixtures compiled from directly, without a concentration step. The authors also the standard plate assays. This modification has been vali reported that passing urine through XAD-2 resin resulted dated with a number of standard mutagens and with ciga in about 60 percent recovery of the mutagens added. Higher rette smokers' urine, air particulate matter from community recoveries, of 70 to 90 percent of mutagenic activity, were air, and diesel exhaust. The range for increased sensitivity, obtained by passing urine in succession through XAD-2 based on absolute amounts of mutagen required for a spe and XAD-7 resins. cific mutagenic response, has been about 10 to 50 times that Belisario and colleagues (1985) further evaluated these of the standard plate incorporation assay of Ames and as procedures for extracting mutagenic metabolites from the sociates (1975). This is based on the slopes of the linear por urine of rats administered diesel particulate matter. Al tion ofthe dose-response curve for each compound or com though they reported that direct extraction with dichloro- plex mixture. The amount of metabolic; activation with S9
4
5 5
uri- using using for for methods methods laboratory laboratory validate validate and and refine refine To To 2. 2. in in excretion excretion mutagen mutagen of of kinetics kinetics the the on on reported reported have have We We
smoke. smoke. tobacco tobacco environmental environmental notably notably particles, particles, spirable spirable 1984). 1984). Hayatsu Hayatsu and and
re
of of sources sources other other from from exposure exposure measure measure to to refined refined Kobayashi Kobayashi 1982; 1982; al. al. et et (Recio (Recio others others by by confirmed confirmed been been has has
be be
to to also also were were Methods Methods concentration. concentration. phenanthrene phenanthrene observation observation This This hours. hours. of of period period a a in in detoxified detoxified or or excreted excreted
as as
such such exposure, exposure, diesel diesel of of markers markers specific specific more more were were mutagens mutagens that that suggesting suggesting evening, evening, previous previous the the lected lected
to to compared compared be be to to was was particles, particles, respirable respirable to to posure posure col urine urine than than activity activity mutagenic mutagenic less less had had smokers smokers arette arette
ex personal personal exposure, exposure, exhaust exhaust diesel diesel for for marker marker dard dard cig from from morning morning the the in in collected collected urine urine that that reported reported (1977) (1977)
stan The The studies. studies. epidemiological epidemiological in in exposure exposure haust haust Ames Ames and and Yamasaki Yamasaki mixtures. mixtures. complex complex to to exposure exposure ing ing
ex
diesel diesel estimating estimating for for methods methods validate validate and and refine refine To To 1. 1. regard especially especially humans, humans, in in excretion excretion mutagen mutagen of of kinetics kinetics
the the concerning concerning available available is is follows: follows: as as were were information information little little project project this this Very Very of of cult. cult. aims aims specific specific The The
diffi also also was was collection collection urine urine and and characterization characterization posure posure
ex for for period period time time of of AIMS AIMS choice choice the the SPECIFIC SPECIFIC Thus, Thus, urine. urine. the the in in ance ance
appear their their and and mutagens mutagens the the of of deposition deposition the the between between
be be would would relationship relationship kinetic kinetic the the what what unclear unclear was was it it tain, tain,
uncer are are mutagens mutagens the the of of many many of of identities identities the the and and plex, plex, workers. workers.
com very very is is exhaust exhaust diesel diesel of of unexposed unexposed composition composition among among than than chemical chemical the the workers workers exhaust-exposed exhaust-exposed diesel diesel
Because Because kinetics. kinetics. elimination elimination and and among among mutagenicity mutagenicity biotransformation, biotransformation, urinary urinary tion, tion, of of level level higher higher a a be be would would
absorp material's material's the the there there depends on on depends that that because because time time of of detectable detectable period period a a be be over over would would theoretically theoretically but but posure, posure,
excreted excreted be be to to likely likely are are lungs lungs ex daily daily workers' workers' to to these these in in mutagenicity mutagenicity deposited deposited total total of of relation relation apparent apparent the the
compounds compounds Mutagenic Mutagenic shops. shops. reduce reduce repair repair would would engine engine in in This This urine. urine. the the electricians electricians in in mutagens mutagens diesel diesel clearing clearing
and and machinists machinists example, example, for for slow of of present, present, are are buildup buildup a a be be exhaust exhaust may may diesel diesel there there exposure exposure of of months months during during
of of amounts amounts significant significant where where then then areas areas in in half-lives, half-lives, work work elimination elimination personnel personnel long long with with materials materials of of fraction fraction
railroad railroad Some Some mutagens. mutagens. of of excretion excretion significant significant a a is is urinary urinary in in ifthere ifthere and and increases increases rates, rates, elimination elimination different different with with
proportionate proportionate in in result result to to presumed presumed chemicals chemicals is is of of mutagens, mutagens, elimination elimination contains contains represents represents activity activity total total the the ever, ever,
which which exhaust, exhaust, diesel diesel of of How inhalation inhalation exposure. exposure. study, study, of of this this period period In In ied. ied. eight-hour eight-hour an an after after immediately immediately
stud being being effect effect the the to to relevant relevant is is highest highest be be that that to to day day each each assumed assumed on on be be posure posure could could activity activity mutagenic mutagenic urinary urinary
ex personal personal subject's subject's each each determine determine mutagens, mutagens, to to is is smoke smoke study study cigarette cigarette to to miological miological similar similar pattern pattern a a in in eliminated eliminated
epide an an in in assessment assessment were were exposure exposure for for exhaust exhaust approach approach diesel diesel ideal ideal from from The The If If mutagens mutagens 1989). 1989). al. al. et et Claxton Claxton
1986; 1986; Lazardis Lazardis and and (Lofroth (Lofroth S9 S9
with with than than S9 S9 without without tivity tivity
EXCRETION EXCRETION
MUTAGEN MUTAGEN
ac less less
much much have have
smoke smoke
tobacco tobacco environmental environmental of of tracts tracts
OF OF
KINETICS KINETICS THE THE
AND AND
EXPOSURE EXPOSURE
Ex 1983). 1983). Schuetzle Schuetzle 1981; 1981; (Claxton (Claxton S9) S9) (TA98 (TA98 S9 S9 by by + + tion tion
activa metabolic metabolic with with activity activity the the matches matches almost almost which which
S9), S9), - engines. engines. (TA98 (TA98 diesel diesel S9 S9 adding adding heavy-duty heavy-duty from from from from activation activation metabolic metabolic without without
collected collected particles particles from from extracts extracts activity activity and and facilities facilities mutagenic mutagenic railroad railroad the the at at considerable considerable have have particles particles diesel diesel from from
collected collected matter matter particulate particulate Extracts Extracts airborne airborne from from particles. particles. of of extracts extracts types types exhaust exhaust two two the the of of activity activity mutagenic mutagenic
diesel diesel using using assay, assay, the the the the by by of of exemplified exemplified are are characteristics characteristics differences differences mutagenic mutagenic the the of of chemical chemical The The mutagens. mutagens.
some some examined examined also also We We smoke. smoke. associated associated cigarette cigarette the the and and of of exhaust exhaust elimination, elimination, diesel diesel the the as as well well as as absorption, absorption,
to to exposed exposed individuals individuals the the among among in in differences differences particularly particularly in in result result humans, humans, in in might might which which smoke, smoke, cigarette cigarette of of
mutagens mutagens of of doses doses absorbed absorbed that that from from monitoring monitoring for for different different is is assay assay particles particles pension pension diesel diesel of of makeup makeup chemical chemical
microsus Salmonella Salmonella the the and and used used we we physical physical the the investigation, investigation, However, However, this this In In elimination. elimination. of of pattern pattern similar similar
a a follow follow would would exhaust exhaust diesel diesel from from mutagens mutagens that that study study section). section). Mutagenicity Mutagenicity the the in in
this this for for assumed assumed we we processes, processes, combustion combustion from from rived rived below below (described (described engines engines diesel diesel heavy-duty heavy-duty and and light- from from
de are are exhaust exhaust diesel diesel and and smoke smoke cigarette cigarette both both Because Because extracts extracts diesel diesel using using investigated investigated were were parameters parameters These These
mL). mL). (0.2 (0.2 used used mixture mixture incubation incubation of of volume volume hours. hours. small small the the and and
seven seven approximately approximately mixtures), mixtures), is is complex complex half-life half-life and and the the and and compounds compounds of of 0.1/hour, 0.1/hour, variety variety mately mately a a for for minutes minutes
approxi is is 90 90 constant constant rate rate approximately approximately is is calculated calculated The The (optimum (optimum kinetics. kinetics. preincubation preincubation order order of of use use gets," gets,"
first "tar more more following following body body provide provide to to the the used used from from bacteria bacteria of of eliminated eliminated are are and and concentration concentration hours) hours) five five higher higher
to to and and three three (in (in number number rapidly rapidly larger larger the the absorbed absorbed are are include include assay, assay, assay assay microsuspension microsuspension microsuspension microsuspension
Salmonella Salmonella the the the the of of by by detected detected sensitivity sensitivity as as in in mutagens, mutagens, increase increase the the the the for for that that gest gest reasons reasons Major Major
sug cigarette cigarette single single a a smoking smoking after after patterns patterns excretion excretion gen gen test. test. incorporation incorporation plate plate the the for for than than
muta
The The 1985). 1985). al. al. et et (Kado (Kado smokers smokers cigarette cigarette of of urine urine the the less less times times 10 10 to to 5 5 about about is is response response specific specific a a for for required required
al. al. et et Schenker Schenker B. B. M. M.
6 6
~ ~ Numerator Numerator number number ~ ~ of of subjects. subjects. b b Denominator Denominator number number of of samples. samples.
was was 87. 87.
Some Some subjects subjects provided provided samples samples a a on on more more than than one one visit. visit. Those Those individuals individuals are are counted counted separately separately for for each each visit visit in in this this table. table. The The total total number number of of subjects subjects
Total Total 19/37 18/35 18/35 19/37 29/60 29/60 8/16 8/16 57/158 57/158 131/306 131/306
Apr Apr 6 6 '85 '85 1/2 1/2 0/0 0/0 olo olo 9/20 9/20 17/56 17/56 27/78 27/78
Feb'85 Feb'85 5 5 10/18 10/18 7/13 7/13 4/8 4/8 5/10 5/10 3/6 3/6
29/55 29/55
Jan Jan '85 '85 o!o o!o 4 4 1/4 1/4 1/2 1/2 0/0 0/0 18/58 18/58 20/64 20/64
o!o o!o Dec Dec '84 '84 3 3 3/6 3/6 4/8 4/8 2/4 2/4 19/38 19/38 28/56 28/56
olo olo 2 2 Nov Nov '84 '84 olo olo 11/22 11/22 4/8 4/8 o!o o!o 15/30 15/30
Jun Jun '84 '84 1 1 4/7 4/7 0/0 0/0 7/14 7/14 1/2 1/2 o/o o/o 12/23 12/23
Visit Visit Date Date Braker Braker Carman Carman Clerk Clerk Engineer Engineer Shop Shop Worker Worker Totalb Totalb
Job Job Groupb Groupb
Table Table Number Number of of Study Study 1. 1. Subjects Subjects and and Samples Samples by by Visit Visit and and Job Job Groupa Groupa
these these factors. factors. (sometimes (sometimes done done on on preceding preceding day). day).
nonoccupational nonoccupational exposures, exposures, and and any any interactions interactions among among • • Subjects Subjects recruited recruited and and informed informed consent consent forms forms signed signed
including including diesel diesel exhaust exhaust exposure, exposure, cigarette cigarette smoking, smoking, Day Day 1 1
study study samples samples (specific (specific aim aim 3) 3) for for specific specific determinants, determinants,
sections. sections.
4. 4. To To analyze analyze the the concentration concentration of of urinary urinary mutagens mutagens from from
and and analyses analyses of of specific specific samples samples are are covered covered in in subsequent subsequent
or or
urinary urinary mutagens mutagens
were were
also also to to be be
evaluated. evaluated.
presented presented below below and and in in Figure Figure Details Details about about the the 1. 1. handling handling
mutagenicity. mutagenicity.
Nondiesel Nondiesel
sources sources
of of respirable respirable
particles particles
day day sampling sampling period. period. An An overview overview of of the the data data collection collection is is
at at the the
end end of of
each each
work work
shift shift for for measurement measurement
of of urinary urinary
sources sources of of data data were were collected collected from from each each subject subject over over a a two
secutive secutive
work work
shifts. shifts. Urine Urine
samples samples
were were to to be be
collected collected
After After obtaining obtaining informed informed consent, consent, several several separate separate
personal personal samples samples of of airborne airborne contaminants contaminants on on two two con
work work locations locations that that were were geographically geographically close close to to
each each
other. other.
haust haust was was to to be be characterized characterized by by questionnaire questionnaire and and by by
out out the the entire entire work work shift, shift, and and this this could could be be done done
only only in in
unexposed unexposed workers. workers. Occupational Occupational exposure exposure to to diesel diesel ex
sonal sonal sampling sampling pumps pumps on on each each subject subject frequently frequently
through
jects jects recruited recruited from from among among diesel diesel exhaust-exposed exhaust-exposed and and
reasons. reasons. For For example, example, it it was was necessary necessary to to monitor monitor
the the per
3. 3. To To conduct conduct a a field field survey survey of of railroad railroad workers, workers, with with sub
separate separate visits, visits, but but this this was was not not always always possible possible for for logistic logistic
carcinogens. carcinogens.
attempt attempt was was made made to to distribute distribute job job groupings groupings across across the the
particles, particles, which which
contain contain
known known
mutagens mutagens
and and potential potential women women were were employed employed in in the the job job categories categories surveyed. surveyed. An An
nary nary
mutagenicity mutagenicity
to to
assess assess exposure exposure
to to diesel diesel
exhaust exhaust category category and and work work location. location. All All subjects subjects were were male; male; no no
Subjects Subjects for for each each visit visit were were selected selected on on the the basis basis of of job job
hygienist, hygienist, and and at at least least two two assistants. assistants.
Figure Figure 1. 1. Outline Outline of of study study design. design. for for each each visit visit consisted consisted of of an an epidemiologist, epidemiologist, an an industrial industrial
highest highest in in the the winter winter (Woskie (Woskie et et al. al. 1988b). 1988b). The The study study team team
(revertants/1-lmol (revertants/1-lmol of of creatinine) creatinine)
Mutagemcity Mutagemcity (±S9) (±S9)
diesel diesel exhaust exhaust exposures exposures among among railroad railroad workers workers were were
Nicotine, Nicotine, cotinine cotinine
Thiocyanate Thiocyanate
winter winter months months because because our our previous previous work work had had shown shown that that Creatinine Creatinine
Unnalysis Unnalysis
April April 1985 1985 (Table (Table 1). 1). Most Most sampling sampling was was done done during during the the
TOXICOLOGY: TOXICOLOGY: URINE URINE SAMPLING SAMPLING
Six Six field field visits visits were were completed completed between between June June 1984 1984 and and
work work activities, activities, and and cigarette cigarette smoking smoking (ASP, (ASP, ETS, ETS, (Garshick (Garshick phenanthrene) phenanthrene) et et al. al. 1987). 1987).
Diesel Diesel
exhaust exhaust
mar'Kers mar'Kers
(amb1ent (amb1ent n1cotine) n1cotine)
on on all all current current employees, employees, including including job job categories, categories, usual usual
urinary urinary mutagens mutagens
Environmental Environmental tobacco tobacco smoke smoke (ETS) (ETS)
Respirable Respirable particles particles
(ASP) (ASP)
ure ure 1). 1). Previous Previous surveys surveys of of this this railroad railroad had had provided provided data data
field field visits visits to to a a single single railroad railroad facility facility in in New New England England (Fig
All All data data collection collection was was performed performed through through a a series series of of
Other Other exposures exposures
Select Select
study study populat10n populat10n
MedicatiOns MedicatiOns
EPIDEMIOLOGY EPIDEMIOLOGY
Survey Survey railroad railroad workers workers D1et D1et
Tobacco Tobacco smoke smoke exposure exposure
/t /t
I I
POPULATION POPULATION SELECTION SELECTION
JObJhome JObJhome
Diesel Diesel exhaust exhaust exposure exposure history, history,
Health Health history history
METHODS METHODS
EPIDEMIOLOGY: EPIDEMIOLOGY: QUESTIONNAIRE QUESTIONNAIRE
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
7 7
sam- of of
contamination contamination
detect detect
to to obtained obtained were were used) used) not not personal personal measure measure to to developed developed method method the the of of validation validation the the
but but
trains, trains,
sampling sampling into into
assembled assembled media media (sample (sample blanks blanks of of Details Details field. field. the in in and and chamber, chamber, exposure exposure vironmental vironmental
Field Field day. day.
each each of of
end end the the
at at
rechecked rechecked was was calibration calibration the the en an an in in laboratory, laboratory, the the in in conducted conducted were were studies studies tion tion
and and
use, use,
each each before before
calibrated calibrated
was was equipment equipment The The Field. Field. Valida them. them. analyze analyze accurately accurately and and collectors, collectors, the the from from
them them desorb desorb efficiently, efficiently, materials materials measures. measures. these these collect collect control control to to quality quality methods methods specific specific
had had method method each each developing developing on on addition, addition, In In focused focused condition. condition. assessment assessment operating operating exposure exposure good good in in project, project, the the
of of phase phase equipment equipment early early the the maintain maintain in in to to used used were were contracts contracts Therefore, Therefore, respectively. Service Service sponse. sponse. phenanthrene, phenanthrene,
re and and nicotine nicotine instrumental instrumental the the set set to to mixtures, mixtures, used used were were contaminant contaminant these these standards standards for for chosen chosen Calibration Calibration ers ers
drift. drift. mark instrument instrument or or two two the the measure measure contamination contamination to to or or detect detect to to exhaust, exhaust, diesel diesel to to concurrently concurrently or or smoke smoke co co
tobac analyzed analyzed blanks were were blanks environmental environmental to to laboratory laboratory exposure exposure and and personal personal Reagent Reagent measure measure to to Laboratory. Laboratory. able able
avail were were methods methods suitable suitable no no began, began, project project
this this
When When
Control Control Quality Quality
hours. hours. 10 10 to to 7 7 from from
duration duration in in ranging ranging thus thus possible, possible, whenever whenever collected collected were were
phenanthrene. phenanthrene. move move
samples samples Full-shift Full-shift
period. period.
sampling sampling
the the
during during
present present
re not not does does nicotine nicotine for for filter filter treated treated the the example, example, for for tors; tors;
concentration concentration air air the the of of average average
time-weighted time-weighted a a
represents represents
collec lower lower for for destined destined materials materials the the of of any any remove remove not not do do
sample sample the the used, used, was was air air of of rate rate
flow flow
constant constant a a
Since Since
devices devices upstream upstream the the that that demonstrated demonstrated have have and and materials, materials,
zone. zone. breathing breathing subject's subject's the the near near
location location
a a from from
drawn drawn
designated designated the the of of collectors collectors efficient efficient be be to to devices devices these these of of
was was Air Air sampling. sampling. personal personal by by
measured measured
were were
taminants taminants
each each shown shown have have tests tests Laboratory Laboratory 2). 2). (Figure (Figure vapors vapors ganic ganic
con air air other other and and exhaust exhaust
diesel diesel to to
exposures exposures
Personal Personal
or other other and and phenanthrene phenanthrene capture capture to to foam foam polyurethane polyurethane
of of plugs plugs
2.4-cm-long 2.4-cm-long three three of of series series a a and and vapor; vapor; nicotine nicotine Approach Approach General General
capture capture to to filter filter bisulfate-treated bisulfate-treated a a criteria); criteria); [1968] [1968] giene giene
HYGIENE HYGIENE
INDUSTRIAL INDUSTRIAL
Hy Industrial Industrial Governmental Governmental of of Council Council American American the the on on
based based 3.5 3.5 at at ).lm, ).lm, cutoff cutoff percent percent (50 (50 preseparator preseparator cyclone cyclone lon lon
inhalation. inhalation. smoke smoke of of depth depth the the ny 10-mm 10-mm a a through through pass pass that that particles particles respirable respirable the the lect lect
and and
smoked smoked
cigarette cigarette of of
brand brand the the included included questions questions ing ing col to to filter filter high-efficiency high-efficiency a a series: series: in in arranged arranged collectors collectors
smok
The The
A). A).
Appendix Appendix
(see (see habits habits smoking smoking on on questions questions three three through through air air drew drew pump pump sampling sampling The The plugs. plugs. foam foam
specific specific
and and
diet, diet,
week, week,
preceding preceding the the during during mutagens mutagens polyurethane polyurethane cylindrical cylindrical of of series series a a and and filter filter a a both both ploys ploys
nonoccupational nonoccupational
potential potential
to to exposure exposure of or or of use use subject, subject, em that that PAHs PAHs for for sampling sampling personal personal of of method method a a develop develop to to
the the
around around
cigarettes cigarettes
smoking smoking were were who who workers workers of of number number chose chose we we Therefore, Therefore, sampling. sampling. personal personal for for not not but but pling, pling,
days, days,
study study
the the on on
exhaust exhaust diesel diesel to to exposure exposure of of perception perception sam high-volume high-volume for for used used been been has has foam foam Polyurethane Polyurethane
the the
on on
questions questions specific specific
included included questionnaire questionnaire The The sampling. sampling. air air during during pressure pressure back back little little very very troduces troduces
in days. days. two two foam foam preceding preceding polyurethane polyurethane over over of of diet diet and and structure structure exposures, exposures, open-cell open-cell place place the the and and
nonwork cost, cost, and and smoking, smoking, time time analysis analysis exposure, exposure, diesel diesel reduces reduces and and history history substantially substantially medical medical which which cally, cally,
on on subjects subjects ultrasoni by by extracted extracted completed completed be be can can foam foam questionnaire questionnaire survey survey Polyurethane Polyurethane Health Health • • collected. collected.
are are
samples samples of of
hundreds hundreds which which in in studies studies epidemiological epidemiological subjects. subjects. from from collected collected urine urine Spot Spot • •
for for inappropriate inappropriate is is resin resin with with sampling sampling therefore, therefore, sample; sample;
completed. completed. log log hygiene hygiene trial trial
each each for for expensive expensive and and
time-consuming time-consuming
is is
which which
extracted, extracted,
indus and and shift shift of of end end at at collected collected samples samples and and Pumps Pumps • •
Soxhlet
be be
must must
resin resin XAD-2 XAD-2
Second, Second,
day. day.
the the throughout throughout
begun. begun. log log
hygiene hygiene
Industrial Industrial
shift. shift.
of of ning ning
rate rate
flow flow
constant constant
maintain maintain to to
able able not not are are
pumps pumps pling pling
begin at at
attached attached
filters filters and and pumps pumps
sampling sampling
Personal Personal
• •
sam personal personal many many and and substantially, substantially, train train sampling sampling the the
2 2
Day Day
of of drop drop pressure pressure the the increase increase they they First, First, disadvantages. disadvantages. two two
completed. completed.
collection collection
specimen specimen
biological biological on on sheet sheet Data Data • • have have they they but but sampling, sampling, personal personal for for used used been been also also have have
resin resin XAD-2 XAD-2 subjects. subjects. of of all all tubes tubes from from Small Small collected collected 1984). 1984). urine urine Bidleman Bidleman Spot Spot • • and and (You (You foam foam
polyurethane polyurethane
or or resin resin XAD-2 XAD-2 as as such such sorbents, sorbents, downstream downstream sampling. sampling. personal personal the the during during
use use
sampling sampling
area area for for schemes schemes collection collection high-volume high-volume quality quality sample sample and and location, location, work work smoked, smoked, cigarettes cigarettes
many many
therefore, therefore,
sampling; sampling; during during filter filter the the from from volatilize volatilize activities, activities, work work on on completed completed log log hygiene hygiene industrial industrial and and
compounds compounds
low-vapor-pressure low-vapor-pressure These These matter. matter. particulate particulate shift shift of of end end at at collected collected samples samples and and pumps pumps Sampling Sampling • •
airborne airborne collect collect to to used used filters filters by by efficiently efficiently collected collected not not is is
tions. tions.
PAHs, PAHs, weight weight molecular molecular
lower lower
many many like like
Phenanthrene, Phenanthrene,
condi field field and and temperature, temperature, area, area, work work activity, activity, work work
1987). 1987).
al. al. et et (Hammond (Hammond published published been been have have smoke smoke daily daily of of begun begun log log hygiene hygiene Industrial Industrial shift. shift. of of beginning beginning
tobacco tobacco
environmental environmental for for
marker marker a a as as nicotine nicotine to to at at exposure exposure attached attached cassettes cassettes filter filter and and pumps pumps sampling sampling Personal Personal • •
al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers
.. pling. All weighing was done in a room with controlled temperature (70° ± 5°F) and relative humidity (50 ± 10 D Cyclone percent). Quality control was maintained by periodically weighing ~Cassette a control filter, and checking zero and a 10-mg tare weight. The control filter monitored effects caused by humidity ~Filter Respirable particles changes, and the zero and the 10-mg weight monitored the J balance's zero and calibration, which may drift. The zero Stainless-steel filter was checked after sets of 10 filters. If the zero changed by more than ± 5 J.Lg, the balance was recalibrated and the C J Teflon 0-ring previous 10 filters were reweighed. The control filter and tare weights were weighed at the beginning and end of a set ~ Sodium bisulfate-treated filter J Nicotine (>90%) of 20 filters. If the control filter weight changed by more than ± 7 J.Lg, the balance was recalibrated and the previous ~ Supportpad 20 filters were reweighed. If the tare weight changed by more than ± 3 percent, the balance was recalibrated. ~Cassette The air flow, drawn by a small pump through each per sonal sampling train, was calibrated ( ± 5 percent) in the field with a rotameter that had been calibrated with a bub Polyurethane foam ble flow meter (primary standard) before and after each use. Volatile Collected air volumes that required corrections due to Polyurethane foam organic differences (of more than 10 percent) in barometric pres
compounds sure or in temperature were corrected during data entry and analysis. Polyurethane foam Timed samples were collected as breathing zone personal +- Pump samples. Each sample was collected over a single work shift (7 to 10 hours). The following information was collected on Figure 2. Schematic of sampling train for personal samples. each sample: 1. Identifier number ples. All equipment was checked for constant flow and for 2. Interstate Commerce Commission (ICC) job code battery capability before each field trip and was serviced as 3. Sample type needed. 4. Pump number Respirable Particles 5. Flow rate 6. Sample number Respirable particles were collected on a 37-mm Teflon 7. Start and stop times coated fiber filter (Emfab TX40 HI20-WW, Pallflex Corp., Putnam, CT). Each filter was placed in a polystyrene cas 8. Worksite (facility type) sette (Millipore Corp., Bedford, MA) on a stainless-steel 9. Work location (specific) backup screen (MSA Corp., Pittsburg, PA) that had been 10. Shift precleaned by soaking in 1:1 (v/v) nitric acid:water for one 11. Date hour, then rinsed with deionized distilled water, methanol, 12. Meteorologic data (outdoor temperature, barometric and dichloromethane. A Teflon washer (37 mm outer di pressure) ameter x 32 mm inner diameter x 0.5 mm thick) was used 13. Indoor temperature and humidity (measured) to separate the stainless-steel backup screen from the treated filter used to collect nicotine. The washer was 14. Location of sample (indoor or outdoor) precleaned by ultrasonication with detergent and water, 15. Number of cigarettes smoked while subject wore sam then rinsed with deionized distilled water, dichloro pler (subject counted cigarettes in pack before and after methane, and methanol. The mass of particles collected on shift) the first filter in the sampling train was determined After sampling, filters were removed from cassettes and gravimetrically by weighing the filter before and after sam- placed in tin storage canisters (Freund Co., Chicago, IL)
8
9 9
rna- of of fraction fraction The The extracted. extracted. repeatedly repeatedly and and phenanthrene phenanthrene Chro- with with packed packed column column stainless-steel stainless-steel Va-in Va-in by by (6-ft (6-ft
of of 0.2 0.2 or or 50 50 ).lg ).lg with with ).lg ).lg spiked spiked were were Plugs Plugs efficiency. efficiency. tion tion analysis analysis chromatographic chromatographic gas gas for for removed removed immediately immediately
collec the the evaluate evaluate to to and and procedure procedure extraction extraction optimal optimal the the was was heptane heptane ofthe ofthe aliquot aliquot An An seconds). seconds). 30 30 for for heptane heptane the the
determine determine to to used used were were experiments experiments field field and and Laboratory Laboratory through through bubbled bubbled was was ammonia ammonia (gaseous (gaseous heptane heptane moniated moniated
am of of 11L 11L 250 250 with with extraction extraction liquid-liquid liquid-liquid by by tracted tracted filter. filter. glass-fiber glass-fiber
ex was was base base free free The The again. again. vortexed vortexed Teflon-coated Teflon-coated was was 25.4-cm 25.4-cm mixture mixture x x the the 20.4- a a and and with with (Model2000) (Model2000) Sampler Sampler
base, base, free free the the form form to to added added was was High-Volume High-Volume hydroxide hydroxide Metals Metals sodium sodium N N 10 10 General General a a of of using using series, series, in in diameter) diameter) in in
mL mL 2 2 then then ethanol; ethanol; of of J.lL J.lL 100 100 mm mm with with 76 76 x x water water of of long long em em mL mL 2 2 (7.2 (7.2 in in plugs plugs vortexed vortexed three three with with shop shop repair repair railroad railroad
were were filters filters Sample Sample the the from from chromatography. chromatography. gas gas collected collected and and was was desorption desorption samples samples high-volume high-volume of of set set small small a a
aqueous aqueous by by determined determined plugs, plugs, was was samples samples 37-mm-diameter 37-mm-diameter these these the the in in with with Nicotine Nicotine sampling sampling to to addition addition In In
months. months. six six 20°C. 20°C. - least least at at at at for for freezer freezer the the in in conditions conditions stored stored these these and and under under lab lab the the to to returned returned
stable stable were were samples samples were were the the they they that that until until ice ice confirmed confirmed dry dry on on nicotine nicotine of of stored stored amounts amounts were were extenders extenders the the
known known with with spiked spiked sampling, sampling, filters filters after after with with Tests Tests Immediately Immediately collection. collection. bags. bags. sample sample plastic plastic Ziploc Ziploc in in stored stored
of of months months six six to to four four within within were were analyzed analyzed plugs plugs were were foam foam samples samples These These polyurethane polyurethane containing containing extenders extenders the the
analysis. analysis. to to prior prior laboratory, laboratory, the the in in sampling, sampling, 20°C 20°C after after - at at and and Before Before freezers freezers 2). 2). in in (Figure (Figure sampling sampling personal personal
and and field, field, for for the the in in cassette cassette ice ice dry dry filter filter on on the the of of stored stored base base were were the the to to filters filters treated treated connected connected The The was was extender extender
The The diameter). diameter). in in mm mm 37 37 x x long long em em (7.5 (7.5 extender extender cassette cassette filter. filter. primary primary the the
a a
within within series series in in plugs plugs three three from from assembled assembled was was lector lector on on collected collected nicotine nicotine of of amount amount the the of of percent percent 1 1 than than less less
col sample sample The The hood. hood. a a in in air-dried air-dried were were plugs plugs the the lution, lution, contained contained filters filters backup backup the the of of All All hours. hours. seven seven to to one one for for
so excess excess out out squeezing squeezing After After solution. solution. fresh fresh with with repeated repeated unit unit collection collection the the through through drawn drawn was was Air Air present. present. smokers smokers
was was process process the the and and forceps, forceps, with with out out squeezed squeezed was was liquid liquid with with rooms rooms in in found found smoke smoke tobacco tobacco environmental environmental the the is, is,
excess excess the the solution, solution, the the from from removed removed were were plugs plugs minutes; minutes; that that smoke, smoke, mainstream mainstream exhaled exhaled and and sidestream sidestream of of mixture mixture
15 15 for for ultrasonication ultrasonication by by dichloromethane dichloromethane percent percent 5 5 ing ing aged aged an an in in resulting resulting chamber, chamber, the the in in cigarettes cigarettes smoking smoking
contain heptane heptane with with extracted extracted were were plugs plugs foam foam urethane urethane actively actively people people four four with with conducted conducted were were experiments experiments
poly 20 20 about about of of sets sets analysis: analysis: for for used used was was that that procedure procedure These These 1987). 1987). al. al. et et (Hammond (Hammond chamber chamber environmental environmental m3 m3
same same the the to to according according it it extracting extracting by by use use before before cleaned cleaned 34- a a in in smoke smoke tobacco tobacco environmental environmental ambient ambient of of trations trations
was was foam foam polyurethane polyurethane The The OH). OH). (Spencerville, (Spencerville, Products Products concen high high to to exposed exposed was was cassette cassette collection collection the the backup, backup,
Foam Foam Flexible Flexible from from purchased purchased were were foam foam polyurethane polyurethane the the to to lost lost vapor vapor nicotine nicotine of of amount amount the the measure measure To To backup. backup.
of of sheets sheets One-inch-thick One-inch-thick foam. foam. open-cell open-cell polyurethane polyurethane a a as as collector collector nicotine nicotine primary primary the the after after filter filter treated treated
of of diameter) diameter) in in mm mm 37 37 x x thick thick em em (2.4 (2.4 plugs plugs cylindrical cylindrical second second a a placing placing by by checked checked was was vapor vapor nicotine nicotine of of Loss Loss
small, small, on on adsorption adsorption by by collected collected was was Phenanthrene Phenanthrene
2). 2). ure ure
(Fig particles particles airborne airborne the the collected collected
that that filter filter untreated untreated the the Phenanthrene Phenanthrene
from from downstream downstream and and washer washer Teflon Teflon the the under under pad, pad, port port
sup cellulose cellulose a a on on cassette cassette 37-mm 37-mm a a in in mounted mounted was was filter filter linearity. linearity.
coated coated
The The
filters. filters. the the on on bisulfate bisulfate sodium sodium of of mg mg 10 10 to to 7 7 left left and and quantification quantification of of limits limits chromatographic chromatographic gas gas on on based based
treatment treatment This This dry. dry. air air to to allowed allowed then then and and
glass, glass, watch watch a a nicotine, nicotine, 0.2 0.2 of of ).lgim
concentration concentration average average an an is is hours hours 3 3
in in
grade) grade) (reagent (reagent bisulfate bisulfate sodium sodium aqueous aqueous percent percent 4 4 with with eight eight for for Lpm Lpm 1.7 1.7 at at sampling sampling field field for for detection detection of of limit limit
soaked soaked were were Corp.) Corp.) Pallflex Pallflex HI20-WW, HI20-WW, TX40 TX40 (Emfab (Emfab filters filters The The contamination. contamination. smoke smoke cigarette cigarette limited limited with with areas areas
glass-fiber glass-fiber Teflon-coated Teflon-coated salt. salt. nonvolatile nonvolatile a a form form to to nicotine nicotine in in collected collected be be would would that that material material of of amounts expected expected of of
the the
with with binds binds
that that bisulfate, bisulfate, sodium sodium acid, acid, an an with with filter filter a a end end lower lower the the represents represents which which spikes, spikes, 0.5-1-!g 0.5-1-!g for for percent percent 2 2
treating treating by by collected collected was was it it so so alkaline, alkaline, is is vapor vapor Nicotine Nicotine 98 98 of of ± ± recovery recovery average average an an to to led led ethanol ethanol of of amount amount small small
a a of of Addition Addition variable. variable. highly highly was was but but percent, percent, 75 75 of of age age
Nicotine Nicotine
aver an an recovered recovered medium medium extraction extraction water water pure pure A A ciency. ciency.
effi extraction extraction overall overall the the determine determine to to used used was was filters filters
room. room.
clean clean onto onto spiked spiked nicotine nicotine of of amounts amounts known known of of
Recovery Recovery
weighing weighing humidity-controlled humidity-controlled the the in in conditioning conditioning of of hours hours
complete. complete. virtually virtually was was heptane heptane the the into into
nicotine nicotine
of of
tion tion
to 24 24 to 18 18 after after reweighed reweighed were were filters filters The The laboratory. laboratory. the the to to
extrac liquid-liquid liquid-liquid the the that that determined determined tests tests Laboratory Laboratory
return return on on 20°C) 20°C) (- freezer freezer a a to to transferred transferred and and trip trip field field the the
detector). detector). nitrogen-selective nitrogen-selective 170°C; 170°C; during during ice ice dry dry with with stored stored were were filters filters The The above. above. described described
at at
separation separation
isothermal isothermal hydroxide; hydroxide; potassium potassium percent percent 3 3 washers, washers, Teflon Teflon the the as as procedure procedure same same the the by by precleaned precleaned
contained contained that that L L Apiezon Apiezon
percent percent 10 10 with with coated coated W W mosorb mosorb were were liners liners and and canisters canisters storage storage The The liners. liners. Teflon Teflon with with
al. al. et et Schenker Schenker B. B. M. M.
10 10
mL!minute. mL!minute. Aliphatic Aliphatic hydrocarbons hydrocarbons eluted eluted first, first, between between 2 2 air air contaminants contaminants emitted emitted indoors indoors accumulate accumulate more more and and ex-
methyl methyl ether ether in in heptane heptane as as the the mobile mobile phase, phase, operated operated at at 2 2 are are closed closed when when the the outdoor outdoor temperature temperature drops. drops. As As a a result, result,
umn umn (Perkin (Perkin Elmer) Elmer) with with a a solution solution of of 3 3 percent percent tert-butyl tert-butyl cause cause doors doors and and windows windows that that provide provide ventilation ventilation indoors indoors
phase phase liquid liquid chromatography chromatography using using an an amino-cyano amino-cyano col door door temperature temperature was was an an important important factor factor in in exposure, exposure, be
fluorimeter. fluorimeter. Class Class fractionation fractionation was was performed performed by by normal shops, shops, had had the the highest highest exposures exposures to to diesel diesel exhaust. exhaust. Out
254 254 nm, nm, a a Waters Waters Data Data Module, Module, and and a a McPherson McPherson spectra chanics chanics and and electricians, electricians, who who work work in in large large open open repair repair
programmer, programmer, M440 M440 ultraviolet ultraviolet light light detector detector monitored monitored at at smoke smoke exposure exposure but but no no exposure exposure to to diesel diesel exhaust. exhaust. Me
tor, tor, M6000 M6000 and and M45 M45 solvent solvent delivery delivery systems, systems, M660 M660 solvent solvent offices, offices, had had the the highest highest level level of of environmental environmental tobacco tobacco
uid uid chromatograph chromatograph equipped equipped with with a a WISP WISP automated automated injec air air contaminants contaminants (Woskie (Woskie et et al. al. 1988a). 1988a). Clerks, Clerks, who who work work in in
matography matography was was performed performed on on a a Waters Waters (Milford, (Milford, MA) MA) liq minants minants of of their their level level of of exposure exposure and and the the composition composition of of
step step followed followed by by an an analytical analytical separation. separation. All All liquid liquid chro ture ture during during the the exposure exposure measurements measurements were were major major deter
chromatographic chromatographic analysis, analysis, including including a a class class fractionation fractionation workers' workers' job job titles, titles, work work locations, locations, and and the the outdoor outdoor tempera
The The extract extract solutions solutions were were analyzed analyzed by by a a two-stage two-stage liquid liquid Our Our previous previous work work in in railroads railroads showed showed that that railroad railroad
gen gen to to 500 500 !lL. !lL.
Sampling Sampling Strategy Strategy
were were combined combined and and concentrated concentrated under under a a stream stream of of nitro
dichloromethane dichloromethane in in heptane; heptane; these these extraction extraction solutions solutions
for for breakthrough. breakthrough. by by ultrasonication ultrasonication twice twice in in fresh fresh solutions solutions of of 5 5 percent percent
in in the the cassette cassette extenders extenders was was analyzed analyzed separately separately to to check check polyurethane polyurethane foam foam plugs plugs from from each each sample sample were were extracted extracted
field, field, each each of of the the three three polyurethane polyurethane foam foam plugs plugs in in series series if if desired, desired, they they could could be be analyzed analyzed separately. separately. The The three three
through. through. For For selected selected personal personal samples samples collected collected in in the the three three plugs plugs normally normally would would be be analyzed analyzed together together although, although,
urethane urethane foam foam plug plug in in series series to to check check for for possible possible break containing containing three three plugs plugs was was chosen chosen for for field field work, work, and and all all
were were expected expected to to be be highest, highest, with with an an extra, extra, fourth, fourth, poly amounts amounts were were on on the the third third plug. plug. Therefore, Therefore, a a sampling sampling train train
railroad railroad repair repair shops, shops, where where diesel diesel exhaust exhaust concentrations concentrations the the phenanthrene phenanthrene was was found found on on the the first first plug, plug, and and trace trace
plugs. plugs. Spiked Spiked filters filters and and sampling sampling trains trains were were tested tested in in the the tioned tioned across across three three or or four four plugs plugs run run in in series. series. Virtually Virtually all all
did did not not affect affect PAHs PAHs collected collected on on the the polyurethane polyurethane foam foam shops shops to to determine determine how how airborne airborne phenanthrene phenanthrene is is parti
was was tested tested at at the the same same time time to to insure insure that that the the treated treated filters filters Collectors Collectors also also were were placed placed in in diesel diesel locomotive locomotive repair repair
smoke. smoke. A A second second set set of of samplers samplers without without the the treated treated filter filter
a a flame flame ionization ionization detector. detector.
volunteer volunteer smokers smokers were were generating generating environmental environmental tobacco tobacco
235°C 235°C final final temperature, temperature, at at a a rate rate of of 4°C 4°C per per minute, minute, using using
ter ter and and then then sampling sampling in in the the environmental environmental chamber chamber while while
with with a a temperature temperature program program of of 75°C 75°C initial initial temperature temperature to to
known known amounts amounts of of the the standard standard PAH PAH solutions solutions to to a a blank blank fil
fused fused silica silica column, column, operated operated at at 30 30 mL mL of of nitrogen/minute, nitrogen/minute,
urethane urethane foam foam plugs plugs (Figure (Figure 2) 2) -was -was tested tested by by adding adding
then then determined determined by by gas gas chromatography chromatography on on a a 15-m 15-m DB-5 DB-5
The The entire entire sampling sampling train-filter, train-filter, treated treated filter, filter, and and poly
The The amount amount of of PAHs PAHs on on each each of of these these collection collection devices devices was was
at at 253 253 nm nm and and emission emission wavelength wavelength at at 372 372 nm. nm. thane thane foam foam sections sections was was extracted extracted and and analyzed analyzed separately. separately.
fluorimeter, fluorimeter, which which was was set set to to monitor monitor excitation excitation wavelength wavelength and and analyzed analyzed separately, separately, and and each each of of the the three three polyure
acetonitrile. acetonitrile. Phenanthrene Phenanthrene was was measured measured with with the the spectra Front Front and and back back sections sections of of the the XAD-2 XAD-2 resin resin were were extracted extracted
Hesperion, Hesperion, CA) CA) with with an an aqueous aqueous solution solution of of 62 62 percent percent in in a a solution solution of of 5 5 percent percent dichloromethane dichloromethane in in heptane). heptane).
(no (no end end capping) capping) (Vydac, (Vydac, 201TP54; 201TP54; SeParations SeParations Group, Group, dichloromethane; dichloromethane; polyurethane polyurethane foam foam plugs plugs ultrasonically ultrasonically
18 18
by by reverse-phase reverse-phase liquid liquid chromatography chromatography on on C a column column dichloromethane; dichloromethane; XAD-2 XAD-2 resin resin by by Soxhlet Soxhlet extraction extraction with with
Analysis Analysis for for phenanthrene phenanthrene was was performed performed isocratically isocratically tracted tracted by by their their own own protocols protocols (filters (filters ultrasonically ultrasonically with with
resin resin samples, samples, and and polyurethane polyurethane in in foam foam amber amber plugs plugs vials vials were were at at each each - 20°C. 20°C. ex
pling pling centrated centrated trains trains for for to to approximately approximately eight eight hours hours at at 5 5 1.7 1.7 mL mL Lpm. Lpm. under under The The nitrogen nitrogen filters, filters, and and XAD-2 XAD-2 stored stored
stream. stream. Clean Clean air air was was drawn drawn through through tions, tions, each each one one aliphatic, aliphatic, of of the the 10 10 two two sam aromatic, aromatic, and and one one polar, polar, were were con
sampling sampling trains trains with with polyurethane polyurethane from from the the foam foam column column plugs plugs into into down the the fraction fraction collector. collector. The The four four frac
XAD-2 XAD-2 tubes tubes downstream, downstream, and and five five methyl methyl were were ether; ether; assembled assembled this this step step into into backflushed backflushed the the polar polar compounds compounds
til til analysis; analysis; five five were were assembled assembled and and into into sampling sampling the the mobile mobile trains trains phase phase with with changed changed to to 100 100 percent percent tert-butyl tert-butyl
to to 15 15 filters. filters. Five Five spiked spiked filters filters were were a a ChronTrol ChronTrol stored stored in in the the timer timer freezer freezer (Lindburg (Lindburg un Enterprises, Enterprises, San San Diego, Diego, CA), CA),
phenanthrene, phenanthrene, tochrome tochrome (Milford, (Milford, pyrene, pyrene, chrysene, chrysene, MA) MA) solenoid solenoid and and benzo[ benzo[ valve valve programmed programmed ]pyrene ]pyrene a a (BaP) with with
of of standard standard the the column column solutions solutions was was of of reversed reversed six six PARs-naphthalene, PARs-naphthalene, automatically automatically with with fluorene, fluorene, an an Au
pared pared teen teen with with minutes minutes that that of of after after XAD-2 XAD-2 the the sample sample resin resin by by injection, injection, adding adding known known the the flow flow amounts amounts through through
ficiency ficiency of of collection collection by by polyurethane polyurethane eluted eluted foam foam with with plugs plugs varying varying was was retention retention com times times over over the the next next 20 20 mL. mL. Fif
terial terial recovered recovered in in each each extraction extraction and and was was 4 4 determined. determined. mL mL after after injection; injection; The The ef olefinic olefinic and and aromatic aromatic compounds compounds
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
11 11
creatinine creatinine For For site. site. collection collection the the at at ice ice on on placed placed mediately mediately in in bacteria bacteria concentrated concentrated of of mL mL 0.1 0.1 and and DMSO; DMSO; in in extract extract
im were were bottles bottles The The bottles. bottles. plastic plastic polyethylene polyethylene or or glass glass urine urine of of mL mL 0.005 0.005 mix; mix; S9 S9 of of mL mL 0.1 0.1 ice: ice: on on tubes tubes culture culture
amber-colored amber-colored sterile, sterile, washed, washed, in in collected collected was was Urine Urine glass glass sterile sterile 75-mm 75-mm 12- to to stated, stated, x x order order the the in in added, added, were were
ingredients ingredients following following the the assay, assay, microsuspension microsuspension the the For For
Methods Methods Extraction Extraction and and Collection Collection Urine Urine
(1978). (1978). Barr Barr and and Ohnishi Ohnishi of of method method Biuret Biuret
modified modified the the using using determined determined as as protein/mL, protein/mL, of of mg mg 52.4 52.4
shift. shift.
work work
the the
during during
contained contained fraction fraction This This throughout. throughout. used used was was 1254 1254 Aroclor Aroclor
mutagens mutagens
to to
exposure exposure of of
index index individual individual
an an
provide provide
to to
with with pretreated pretreated rats rats Sprague-Dawley Sprague-Dawley male male from from S9 S9 The The
possible, possible, if if
thus, thus, and and
times times
sampling sampling
short short
relatively relatively
these these
(1975). (1975). colleagues colleagues and and Ames Ames of of procedure procedure the the to to according according
for for measured measured
be be
could could
filters filters
these these
from from
mutagenicity mutagenicity
not not
prepared prepared were were mix mix S9 S9 the the and and S9 S9 enzyme enzyme metabolic metabolic The The
or or whether whether determine determine to to wanted wanted We We activity. activity. mutagenic mutagenic for for
nm. nm. 550 550 at at density density optical optical the the and and ria ria
tested tested and and NJ) NJ) Springfield, Springfield, Scientific, Scientific, Fisher Fisher grade, grade, (HPLC (HPLC
bacte surviving surviving of of number number the the by by determined determined as as cells/mL, cells/mL,
dichloromethane dichloromethane with with extracted extracted also also were were workers workers shop shop
1 1 x x approximately approximately of of concentration concentration a a to to 7.4) 7.4) pH pH 10
10 10 and and clerks clerks from from filters filters personal personal of of number number selected selected A A
M, M, 0.15 0.15 (PBS, (PBS, saline saline phosphate-buffered phosphate-buffered ice-cold ice-cold in in pended pended
1650.) 1650.)
No. No.
Sample Sample
resus were were Cells Cells minutes). minutes). 10 10 4°C, 4°C, (10,000 (10,000 x x trifugation trifugation g, g,
Reference Reference Standard Standard Standards, Standards, of of Bureau Bureau
National National the the
from from
cen by by harvested harvested and and cells/mL, cells/mL, 10 2 2 to to 1 1 x x approximately approximately
9 9 purchase purchase
for for
available available
also also are are
particles particles
same same (The (The
engines. engines.
to to England) England) Hants, Hants, Ltd., Ltd., (Oxoid (Oxoid No.2 No.2 Broth Broth Nutrient Nutrient Oxoid Oxoid
diesel diesel
heavy-duty heavy-duty
of of
number number
a a from from
exhaust exhaust of of
posite posite
in in overnight overnight grown grown were were bacteria bacteria modification, modification, our our For For
com a a was was obtained obtained was was extract extract the the which which matterfrom matterfrom ulate ulate
mixtures. mixtures. complex complex and and gens gens
partic diesel diesel heavy-duty heavy-duty The The directly. directly. tested tested and and MI), MI), born, born,
muta standard standard of of number number a a using using
tested tested been been
has has
method method
(Dear Co. Co. Motor Motor Ford Ford the the of of Schuetzle Schuetzle from from obtained obtained D. D.
The The response. response. mutagenic mutagenic
specific specific a a for for
required required
mutagen mutagen
was was extract extract The The matter. matter. particulate particulate diesel diesel heavy-duty heavy-duty from from
of of amounts amounts absolute absolute by by judged judged as as 1975), 1975), al. al. et et (Ames (Ames test test
extract extract an an for for determined determined also also were were curves curves Dose-response Dose-response
incorporation incorporation plate plate standard standard the the than than sensitive sensitive more more times times
1983). 1983).
melstein melstein
10 10 least least at at is is method method The The study. study. the the throughout throughout used used was was
Mer and and (Rosenkranz (Rosenkranz (nitro-PAHs) (nitro-PAHs)
hydrocarbons hydrocarbons
aromatic aromatic
1986), 1986), 1983, 1983, al. al. et et (Kado (Kado nonsmokers nonsmokers and and smokers smokers arette arette
nitropolycyclic nitropolycyclic certain certain to to sensitive sensitive less less
be be
to to
reported reported
been been
cig of of urine urine the the in in mutagens mutagens detect detect to to developed developed previously previously
has has strain strain this this because because NR, NR, TA98 TA98 strain strain tester tester in in
tested tested were were
procedure, procedure, preincubation) preincubation) (micro (micro microsuspension microsuspension A A
extracts extracts The The curves. curves. dose-response dose-response the the for for used used were were NR NR
TA98 TA98 and and TA98 TA98 strains strains tester tester Bacterial Bacterial assay. assay. screening screening nary nary Method Method Assay Assay Microsuspension Microsuspension
prelimi the the in in values values revertant revertant highest highest the the demonstrated demonstrated
that that extracts extracts the the of of number number selected selected a a for for established established were were MO). MO). Louis, Louis, (St. (St. Co. Co. Chemical Chemical Sigma Sigma from from purchased purchased
curves curves Dose-response Dose-response activation. activation. metabolic metabolic S9 S9 without without was was J3-Glucuronidase J3-Glucuronidase purification. purification. further further without without used used
and and with with TA100, TA100, and and TA98 TA98 strains strains tester tester bacterial bacterial using using and and WI), WI), (Milwaukee, (Milwaukee, Co. Co. Chemical Chemical Aldrich Aldrich from from chased chased
activity activity mutagenic mutagenic for for screened screened were were facilities, facilities, railroad railroad ent ent pur were were Label) Label) Gold Gold (DMSO, (DMSO, dimethylsulfoxide dimethylsulfoxide and and
differ three three at at matter matter particulate particulate collecting collecting for for used used filters, filters, (4-NQO), (4-NQO), 4-nitroquinoline-N-oxide 4-nitroquinoline-N-oxide (2-NF), (2-NF), nitrofluorene nitrofluorene
high-volume high-volume of of extracts extracts dichloromethane dichloromethane of of number number A A 2- (2-AF), (2-AF), 2-aminofluorene 2-aminofluorene Label), Label), (Gold (Gold Benzo[a]pyrene Benzo[a]pyrene
Extracts Extracts Diesel Diesel Chemicals Chemicals
MUTAGENICITY MUTAGENICITY
photooxidation. photooxidation. potential potential minimize minimize to to F40Go) F40Go) (G.E.N. (G.E.N.
lights lights fluorescent fluorescent yellow yellow with with fitted fitted room room a a in in out out ried ried
1). 1). (Table (Table categories categories these these in in exposure exposure exhaust exhaust diesel diesel tial tial car were were procedures procedures All All experiment. experiment. each each for for determined determined
poten maximize maximize to to trips trips weather weather cold cold the the in in oversampled oversampled were were markers markers Strain Strain hours. hours. 48 48 for for dark dark the the in in 37°C 37°C at at cubated cubated
were were engineers engineers and and workers workers Shop Shop group. group. exhaust-exposed exhaust-exposed in were were Plates Plates plates. plates. glucose glucose minimal minimal onto onto poured poured and and
diesel diesel our our as as workers workers shop shop mostly mostly and and group, group, unexposed unexposed vortex-mixed vortex-mixed were were solutions solutions combined combined The The immediately. immediately.
our our as as carmen, carmen, and and clerks clerks as as such such locomotives, locomotives, operating operating added added was was biotin biotin and and histidine histidine of of nmol nmol 90 90 containing containing 1975) 1975)
from from away away worked worked who who workers workers choose choose to to was was strategy strategy pling pling al. al. et et (Ames (Ames agar agar top top molten molten of of mL mL 2 2 and and time, time, a a at at one one out out
sam our our Therefore, Therefore, exposures. exposures. of of range range wide wide a a had had who who ers ers taken taken then then bath, bath, ice ice an an in in placed placed were were tubes tubes the the minutes, minutes,
work of of that that with with exposure exposure exhaust exhaust diesel diesel no no with with workers workers 90 90 After After shaking. shaking. rapid rapid with with 37°C, 37°C, at at dark, dark, the the in in incubated incubated
of of mutagenicity mutagenicity urinary urinary the the contrast contrast to to was was goal goal Our Our was was mixture mixture The The extract. extract. urine urine the the of of place in in volume, volume, mL mL
0.005 0.005 at at added, added, and and DMSO DMSO in in dissolved dissolved also also were were extracts extracts 50°F. 50°F. proximately proximately
filter filter air air and and standards standards Mutagen Mutagen PBS). PBS). of of /mL /mL 10 x x (1 (1 PBS PBS
ap below below is is temperature temperature outdoor outdoor when when higher higher are are posures posures 10
al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers
determination, a 5-mL aliquot of urine from each sample own extraction control using PBS (0.15 M, pH 7.0). The was dispensed into a scintillation vial fitted with a Teflon mutagens 2-AF and HaP served as positive controls for + S9, lined screw cap. Both the main collection bottle and the and 2-NF and 4-NQO served as positive controls for - S9. scintillation vials were frozen at - 20°C. Frozen samples All positive control mutagens were tested at three doses in were then shipped in dry ice to the University of California, triplicate and were included in every experiment. Four to Davis, Department of Environmental Toxicology, for extrac five plates were routinely used for the DMSO standard that tion and testing. Urine samples were stored at - 20°C until provided the zero concentration. processed and tested. Extracts were stored in amber glass The order of the samples was as follows for the + S9 test: vials, sealed with nitrogen, at - 20°C. Extraction and test (1) batch samples at two or three concentrations; (2) PBS ing of urine samples were delayed because of the need to blank; (3) + S9 positive controls (HaP and 2-AF); and (4) validate the sensitivity of the methods used (see Appendix DMSO blank (zero concentration for the batch). The same B). The approximate time from collection to extraction order (1 through 4) was repeated for each batch. After the ranged from 1 to 10 months, with a median time of 8.5 + S9 samples, the - S9 tests were carried out in the same months (n = 29 separate extraction sets); the approximate order, except that the positive controls for - S9 were 2-NF time from extraction to testing ranged from 1 to 16 weeks, and 4-NQO. Because the DMSO blank was located last in with a median time of 6 weeks (n = 13). each batch, it was not possible to test for a drift within the The urine samples were extracted using the methods of batch. Yamasaki and Ames (1977), incorporating the modification The number of revertants per milliliter equivalent of ofPutzrath anc ,workers (1981), excE-.:>t that we eliminated urine extracted was normalized for urinary concentration of the use of sodium sulfate to dry the extract. The sorbent mutagens by dividing by the concentration of creatinine. XAD-2 (Applied Science Laboratories, Inc., State College, Creatinine was determined in each urine sample using a PA) was repurified by Soxhlet extraction (24 hours acetone, standard clinical chemistry kit (Sigma Chemical Co., St. 24 hours methanol) before use. Concentrated urine was ob Louis, MO). Normalization by creatinine concentration was tained by redissolving the acetone extract (eluted from the considered since the urinary concentration of compounds XAD-2 column) in water (20 mL/100 mL of original urine), could vary depending on each worker's fluid intake for the neutralizing the solution to pH 7.0 with 0.2 N sodium day. This approach has been reported to improve the ac hydroxide, and extracting three times with dichloro curacy of the analysis oftoxic compounds in the urine (Bar methane (7 mL/mL of original urine each time). The ex ber and Wallis 1986). tracts were combined, dried in vacuo at 45°C, capped with (See the Clinical Chemistry section below for further dis nitrogen, and stored at - 20°C until testing. Deionized cussion of urine creatinine measurement and its use for glass-distilled water that had been passed through a char urine marker normalization.) Graphs of the mean number coal adsorption column was used throughout, and an ex of revertants per micromole of creatinine of urine plotted traction control in which PBS (0.15 M, pH 7.0) was substi against concentration frequently show downward curvature tuted for urine was routinely included for every extraction. at the higher concentrations, presumably as a result of com Extracts were reconstituted with DMSO to make a 1,000- pounds in the urine extract that are toxic to bacteria or fold concentrate just before testing. Urine was extracted in of competitively binding metabolic enzymes (data not lots of 11 samples, and each lot had its own PBS extraction shown). In Salmonella testing, particularly with the stan control. dard plate incorporation test, it is not common practice to The XAD-2 resin was a good initial candidate for use in test the toxicity of the sample preparations directly. There extracting mutagens from urine samples, since much of the fore, it became necessary to estimate indirectly the number work on urine mutagenicity, especially from cigarette of revertants per micromole of creatinine of urine. smokers, has been done using this adsorbent. Belisario and The number of revertants per milliliter equivalent of associates (1985) found that XAD-2 used in series with urine was calculated as follows: least-squares regression fit XAD-4 was most efficient in extracting mutagens from the ted the number of revertants per plate to a constant, a linear urine of rats that were injected with diesel particles. term in plate concentration, and a squared term in plate concentration. If the p value for the squared term was less Mutagenicity Testing of Urine than or equal to 0.15 and the coefficient of the squared term All urine samples were tested at a minimum of three was negative, indicating a downward curvature at high urine equivalent doses of 2.5, 5, and 10 mL, in duplicate, doses, samples with the highest dose (usually 10.0-mL both with and without S9 mix. A single experiment could equivalents) were dropped and the process was repeated test three batches of urine (11 samples/batch), each with its until the squared term for nonlinearity was "nonsignifi-
12
13 13
tested. tested.
routinely routinely were were urine urine as as use use for for suggested suggested been been has has tobacco, tobacco, in in components components anide anide
of of
samples samples
Triplicate Triplicate MO). MO).
Louis, Louis, St. St. Co., Co., Chemical Chemical (Sigma (Sigma cy from from derived derived compound compound a a Thiocyanate, Thiocyanate, measured. measured. were were
kit kit
diagnostic diagnostic a a
using using
determined determined were were concentrations concentrations nine nine absorption absorption and and inhalation inhalation smoke smoke tobacco tobacco for for Markers Markers
Creati minute. minute. per per 25°C 25°C at at 275°C 275°C to to 90° 90° of of program program perature perature
CHEMISTRY CHEMISTRY
CLINICAL CLINICAL
tem a a at at Packard) Packard) Hewlett Hewlett silicon, silicon, phenylmethyl phenylmethyl percent percent
(5 (5 column column capillary capillary diameter diameter inner inner 0.31-mm 0.31-mm x x 25-m 25-m a a on on
results.) results.) CA) CA) Sunnyvale, Sunnyvale, Packard, Packard, Hewlett Hewlett detector, detector, gen-phosphorus gen-phosphorus
the the for for
D D
Appendix Appendix and and
details, details, for for section section Form Form Model Model nitro with with 580, 580, (model (model chromatography chromatography gas gas by by analyzed analyzed
Methods, Methods,
Statistical Statistical
(See (See 2-AF. 2-AF. control control mutagenic mutagenic tive tive was was extracts extracts cotinine cotinine and and nicotine nicotine of of each each microliter microliter One One
posi the the to to response response batch batch differential differential for for adjust adjust do do tivity tivity
analysis. analysis. chromatographic chromatographic gas gas for for
ac mutagenic mutagenic predicting predicting for for
analyses analyses
regression regression
multiple multiple
vial vial sampling sampling a a into into poured poured layer layer top top the the and and again, again, frozen frozen
the the However, However, experiments. experiments. the the of of
course course the the
over over
strains strains
vortex-mixed, vortex-mixed, was was solution solution resulting resulting The The added. added. were were (9:1) (9:1)
bacterial bacterial the the of of sensitivity sensitivity differential differential for for
adjusted adjusted
not not
toluene:butanol toluene:butanol of of mL mL 0.4 0.4 and and hydroxide) hydroxide)
ammonium ammonium N N
are are monograph monograph this this in in
statistics statistics
mutagenicity mutagenicity
Descriptive Descriptive
0.2 0.2 (in (in carbonate carbonate potassium potassium percent percent 50 50 of of mL mL 0.5 0.5 discarded; discarded;
variability. variability. observed observed the the distorted distorted have have layer layer top top the the and and frozen frozen again again was was solution solution The The N). N). (1.0 (1.0 acid acid
would would
this this
because because
zero, zero, to to
forced forced not not were were results results Negative Negative sulfuric sulfuric of of mL mL 0.5 0.5 containing containing tube tube new new a a into into poured poured was was
blank. blank. the the
for for that that
than than lower lower was was result result sample sample a a if if values values layer layer top top the the frozen; frozen; was was layer layer bottom bottom the the until until bath bath acetone acetone
negative negative in in
resulted resulted
calculation calculation This This batch. batch. each each in in samples samples and and dry-ice dry-ice a a in in placed placed then then were were tubes tubes The The vortex-mixed. vortex-mixed.
individual individual for for slopes slopes the the
from from subtracted subtracted was was blank blank PBS PBS was was solution solution the the and and added added then then were were (7:3) (7:3) toluene:butanol toluene:butanol
the the
for for slope slope the the
differences, differences, batch batch possible possible for for correct correct To To of of milliliters milliliters Three Three N). N). (0.2 (0.2 hydroxide hydroxide ammonium ammonium in in N) N) (2 (2
hydroxide hydroxide urn urn i i sod sod of of mL mL 0.5 0.5 by by followed followed throughout. throughout. used used cotinine), cotinine), were were methyl methyl 5-
slopes slopes and and nicotine nicotine point-deletion point-deletion the the (5-methyl (5-methyl slopes, slopes, standards standards tangent tangent the the internal internal in in of of mL mL 0.1 0.1 variability variability added added
was was greater greater which which to to the the of of tube, tube, Because Because culture culture 0.82. 0.82. was was 100-mm 100-mm slopes slopes x x 13 13 a a S9 S9 - to to the the of of added added that that was was and and
0.73, 0.73, was was urine urine of of slopes slopes milliliter milliliter One One point-deletion point-deletion the the and and modifications. modifications. tangent tangent with with S9 S9 (1981), (1981), the the + + of of leagues leagues
col correlation correlation and and rank rank Jacobs Jacobs by by < < Spearman Spearman The The described described as as 0.01.) 0.01.) at at analyses, analyses, p p test test sum sum rank rank chromatographic chromatographic
gas gas Wilcoxon Wilcoxon by by CA) CA) paired paired the the by by Francisco, Francisco, (San (San significant significant are are Benowitz Benowitz results results Neal Neal (All (All and and Yu, Yu, culation. culation. Lisa Lisa
cal Jacobs, Jacobs, Peyton Peyton by by point-deletion point-deletion the the determined determined from from were were those those than than nicotine) nicotine) of of higher, higher, were were metabolite metabolite slopes slopes
(a (a tangent tangent the the of of cotinine cotinine errors errors and and standard standard nicotine nicotine of of individual individual the the and and concentrations concentrations greater, greater, Urinary Urinary
was was slopes slopes tangent tangent the the of of
variability variability between-sample between-sample The The smoking. smoking.
results. results.
S9 S9
the the + + for for
true true
especially especially was was This This C.1). C.1). Table Table and and cigarette cigarette for for control control to to analyses analyses final final the the in in used used not not
C C
Appendix Appendix (see (see fit fit
linear linear a a
obtain obtain to to observations observations of of tion tion were were results results thiocyanate thiocyanate the the measurements, measurements, cotinine cotinine and and
dele
after after
calculated calculated
those those than than higher, higher, much much sometimes sometimes nicotine nicotine specific specific and and sensitive sensitive more more the the With With equivalents:' equivalents:'
higher, higher,
were were fit fit
quadratic quadratic
the the from from slopes slopes tangent tangent The The "thiocyanate "thiocyanate reflect reflect may may measurement measurement our our and and cyanate, cyanate,
thio for for is is assay assay the the specific specific how how know know not not zero. zero. at at do do we we months, months,
curve curve two two fitted fitted least least the the at at to to for for 4°C 4°C tangent tangent at at the the of of stable stable slope slope were were the the as as standards standards estimated estimated was was thiocyanate thiocyanate
urine urine of of Although Although milliliter milliliter per per added. added. was was revertants revertants of of nitrate nitrate ferric ferric number number the the after after and and values values sample, sample, sorbance sorbance
ab each each from from for for doses doses subtracted subtracted all all to to was was fitted fitted nitrate nitrate was was ferric ferric of of model model addition addition quadratic quadratic the the a a before before Second, Second,
recorded recorded tested. tested. 600 600 than than Absorbance Absorbance more more nm. nm. of of 460 460 out out at at S9, S9, read read was was without without and and with with absorbance absorbance both both and and ity, ity,
mutagenic for for vortex-mixed, vortex-mixed, and and samples samples dozen dozen added added a a was was than than fewer fewer solution solution to to dose dose nitrate nitrate higher higher ferric ferric a a M M 0.5 0.5
added added of of mL mL This This 1 1 0.10. 0.10. standards, standards, stringent stringent and and more more the the samples samples to to all all To To changed changed was was 500 500 trend trend ~-tmol. ~-tmol. and and 250, 250,
125, 125, 62.5, 62.5, at at nonlinear nonlinear of of triplicate triplicate in in detection detection for for prepared prepared value value were were the the p p mL) mL) first, first, (2 (2 the the In In thiocyanate thiocyanate tempted. tempted.
at potassium potassium were were of of relation relation Standards Standards nm. nm. 460 460 dose-response dose-response at at the the read read was was calculating calculating of of absorbance absorbance the the methods methods
other other and and two two added, added, chosen, chosen, was was water water of of procedure procedure mL mL 1 1 exact exact tube, tube, the the to to culture culture results results 100-mm 100-mm our our x x a a 13 13
of of in in sensitivity sensitivity placed placed the the was was evaluate evaluate To To triplicate) triplicate) (in (in doses. doses. sample sample higher higher the the each each of of from from mation mation milliliter milliliter One One
infor of of rpm). rpm). 2,000 2,000 elimination elimination the the of of approximately approximately because because and and minutes, minutes, (15 (15 here) here) (0.15 (0.15 centrifuge centrifuge vature vature table-top table-top
cur of of clinical clinical a a detection detection in in for for chosen chosen centrifuged centrifuged level level was was urine urine the the significance significance and and the the of of tubes, tubes, nature nature 100-mm 100-mm
x x subjective subjective 13 13 in in the the of of placed placed because because were were sample sample criticized criticized be be urine urine can can it it each each of of but but milliliters milliliters literature, literature, Four Four
toxicology toxicology the the in in modifications. modifications. slight slight practice practice with with common common (1974), (1974), is is coworkers coworkers procedure procedure and and This This Butts Butts and and
(1944) (1944)
Bowler Bowler by by
outlined outlined procedures procedures the the using using determined determined points. points. dose dose remaining remaining the the
were were
concentrations concentrations
thiocyanate thiocyanate Urinary Urinary 1979). 1979). Junge Junge and and to to fit fit linear linear simple simple a a from from estimated estimated was was urine urine of of milliliter milliliter
(Borgers (Borgers
exposure exposure
smoke smoke tobacco tobacco
of of measure measure convenient convenient a a per per revertants revertants of of number number > > the the Finally, Finally, 0.15. 0.15. p p with with cant;' cant;'
al. al. et et Schenker Schenker B. B. M. M.
14 14
deleted deleted once once created. created. Tape Tape backups backups job job groups. groups. were were regularly regularly made. made.
vertent vertent modification, modification, no no merged merged analysis analysis data data set set could could be be dose-response dose-response effect effect only only within within diesel diesel exhaust-exposed exhaust-exposed
merged merged for for final final analyses. analyses. To To guard guard against against possible possible inad posure. posure. In In addition, addition, some some analyses analyses attempted attempted to to detect detect a a
SAS SAS data data sets sets were were produced produced for for each each data data type type and and and and work work at at jobs jobs with with known known potential potential for for diesel diesel exhaust exhaust ex
the the amount amount of of respirable respirable particles particles and and nicotine nicotine in in the the air, air, to to the the standard standard order. order.
effect effect of of "exposure" "exposure" was was expressed expressed as as an an interaction interaction between between find find out-of-sequence out-of-sequence samples. samples. Those Those found found were were relocated relocated
multiple-regression multiple-regression dummy-variable dummy-variable model model in in which which the the Frequency Frequency counts counts and and other other checks checks were were used used primarily primarily to to
amount amount of of diesel diesel particulate particulate matter matter in in the the air; air; and and (2) (2) a a full full histograms histograms were were used used to to detect detect possibly possibly unlikely unlikely values. values.
struction struction of of ordered ordered groups, groups, based based on on an an index index of of the the were were corrected corrected in in the the raw raw data data file. file. Subsequently, Subsequently, plots plots and and
exhaust exhaust exposure, exposure, two two different different methods methods were were used: used: (1) (1) con and and laboratory laboratory technician. technician. Any Any errors errors detected detected at at this this stage stage
For For assessing assessing the the response response of of urine urine mutagenicity mutagenicity to to diesel diesel against against the the original original forms forms by by the the study study research research associate associate
Printouts Printouts of of all all transferred transferred laboratory laboratory data data were were checked checked
Indices Indices of of Diesel Diesel Exhaust Exhaust Exposure Exposure
management. management.
BMDP BMDP Statistical Statistical Software Software (1983) (1983) were were used used for for all all data data liers liers and and unduly unduly influential influential observations. observations.
The The
Statistical Statistical
Analysis Analysis System System (SAS (SAS Institute Institute 1985) 1985) and and (Cook (Cook 1977) 1977) were were regularly regularly examined examined for for evidence evidence of of out
puter puter tape tape
to to the the Occupational Occupational
Health Health
Program Program VAX VAX 11/750. 11/750. interactions. interactions. Regression Regression residuals residuals and and Cook's Cook's D-statistic D-statistic
onto onto
a a computer computer
disk. disk. The The data data were were transferred transferred by by com tion, tion, urine urine cotinine, cotinine, thiocyanate, thiocyanate, and and nicotine nicotine levels, levels, and and
where where
they they were were
entered entered
independently independently
two two operators operators by by smoked, smoked, tar tar content content of of cigarettes, cigarettes, depth depth of of smoke smoke inhala
before before
being being sent sent to to the the University University Keypunch Keypunch Service, Service, predictive predictive models models for for factors factors such such as as numbers numbers of of cigarettes cigarettes
checked checked against against this this list. list. All All data data forms forms were were checked checked by by eye eye ple ple regression regression techniques techniques examined examined a a large large number number of of
master master
file file was was created created by by hand, hand, and and all all entries entries were were had had not not been been exposed exposed to to diesel diesel exhaust. exhaust. "All-subsets" "All-subsets" multi
To To guard guard
against against errors errors in in the the key key identifier identifier variables, variables, a a to to choose choose factors factors predictive predictive of of mutagenicity mutagenicity in in smokers smokers who who
naire naire was was labeled labeled by by the the field field staff. staff. Data Data on on cigarette cigarette smoking smoking characteristics characteristics were were examined examined
tocol tocol were were also also included. included. Each Each sample sample vial vial and and question
Preliminary Preliminary Smoking Smoking Analyses Analyses
trip, trip, date date of of sample, sample, and and code code number number of of the the two-day two-day pro
first first or or second second day day of of the the protocol. protocol. For For quality quality control, control,
the the
and and scatter scatter plots plots were were used. used.
protocols protocols the the worker worker had, had, and and an an "a" "a" or or "b" "b" designating designating the the
butions butions of of two two or or more more variables variables at at a a time, time, rank rank correlations correlations
a a single single digit digit designating designating how how many many repeat repeat two-day two-day field field
approximately approximately equal equal numbers numbers in in each. each. For For studying studying distri
sisted sisted of of three three digits digits unique unique to to each each participant participant in in the the study, study,
continuous continuous variables variables into into two, two, three, three, or or four four categories categories with with
field field staff staff at at the the time time of of enrollment. enrollment. The The identifier identifier (ID) (ID) con
and and 75th 75th percentiles). percentiles). A A frequent frequent technique technique was was to to group group
mation. mation. The The five-digit five-digit key key identifier identifier was was assigned assigned by by
the the
means, means, standard standard deviations, deviations, medians, medians, and and quartiles quartiles (25th (25th
Data Data input input forms forms were were designed designed for for each each source source of of infor
For For simple simple descriptive descriptive techniques, techniques, we we relied relied upon upon
DATA DATA MANAGEMENT MANAGEMENT generalized generalized techniques techniques could could be be employed. employed.
data data were were transferred transferred to to BMDP BMDP files, files, where where the the appropriate appropriate
variables variables and and models models was was chosen chosen for for intensive intensive analysis, analysis, the the
dependence dependence of of repeat repeat observations. observations. After After a a reduced reduced set set of of
daily daily samples samples as as the the analytic analytic unit, unit, without without allowance allowance for for urine urine markers. markers.
Preliminary Preliminary screening screening for for variable variable selection selection used used the the smoker, smoker, two two smoker) smoker) were were excluded excluded from from analyses analyses that that used used
creatinine creatinine concentration concentration of of less less than than 0.3 0.3 g/L g/L (six (six non displayed. displayed.
(Barber (Barber and and Wallis Wallis 1986). 1986). Therefore, Therefore, all all samples samples with with urine urine results results by by two-day two-day protocol protocol or or by by individual individual worker worker were were
ples ples 0.3) 0.3) were were outliers outliers (too (too high) high) (< (< from from predicted predicted levels levels propriate propriate to to use use the the daily daily sample sample as as the the reporting reporting unit, unit,
that that all all cigarette cigarette smoking smoking markers markers in in the the dilute dilute urine urine sam for for most most of of the the descriptive descriptive tables. tables. When When it it was was clearly clearly inap
used used the the previously previously described described cutoff cutoff of of 0.3 0.3 giL giL and and found found strategy strategy was, was, generally, generally, to to ignore ignore observation observation dependence dependence
creatinine) creatinine) may may overcorrect overcorrect for for the the marker marker of of interest. interest. We We problems problems of of dependent dependent observations observations arose arose at at the the outset. outset. Our Our
tion tion for for excessively excessively dilute dilute urine urine (very (very low low concentration concentration of of Since Since multiple multiple samples samples were were collected collected from from each each worker, worker,
One One potential potential problem problem with with this this adjustment adjustment is is that that correc descriptive descriptive procedures procedures and and tables tables to to more more complex complex models. models.
for for toxic toxic substances substances in in the the urine urine (Barber (Barber and and Wallis Wallis 1986). 1986). The The statistical statistical analysis analysis generally generally proceeded proceeded from from simple simple
cedure cedure has has been been shown shown to to improve improve the the accuracy accuracy of of assays assays
General General Strategy Strategy
ity) ity) were were corrected corrected for for creatinine creatinine concentration. concentration. This This pro
All All urine urine concentrations concentrations (smoking (smoking markers, markers, mutagenic STATISTICAL STATISTICAL METHODS METHODS
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
15 15
unex- were were
who who
study study this this in in
nonsmokers nonsmokers and and smokers smokers in in rewritten: rewritten: be be can can equation equation
data data on on
validated validated
was was
relationship relationship This This constant. constant. different different a a the the fumes, fumes, diesel diesel to to exposed exposed potentially potentially jobs jobs In In jobs. jobs. posed posed
with with
smoke smoke
tobacco tobacco
environmental environmental
for for true true holds holds equation equation unex in in particles particles diesel diesel no no are are there there assumption, assumption, By By
same same the the uniform, uniform, is is particles particles respirable respirable of of level level ground ground
Remainder. Remainder.
+ +
Particles Particles
Diesel Diesel + +
Smoke Smoke
Tobacco Tobacco
back the the studies studies chamber chamber the the in in that that assumed assumed is is If If it it
Particles Particles Respirable Respirable = =
[NICOTINE]. [NICOTINE].
8.6 8.6
x x + +
Constant Constant
equation: equation:
primary primary the the on on based based
Chamber/m
the the in in
Particles Particles
Respirable Respirable
3 3
was was exposure exposure diesel diesel of of index index an an of of construction construction The The
Construction Construction Index Index that that
approximately approximately showed showed section) section) 1 1 Aim Aim Specific Specific Results, Results, the the
(see (see smoke smoke tobacco tobacco 2). 2). (Table (Table environmental environmental exhaust exhaust of of diesel diesel studies studies to to chamber chamber exposure exposure First, First, actual actual the the of of
estimate. estimate. estimates estimates this this for for better better obtain obtain available available to to were were possible possible sets sets was was data data it it different different population, population, Two Two study study
this this
on on
used used
were were study study this this in in developed developed methods methods the the smoke. smoke. tobacco tobacco environmental environmental estimate estimate to to necessary necessary
When When section). section).
Exposure Exposure Exhaust Exhaust Diesel Diesel Discussion, Discussion, the the became became it it Therefore, Therefore, (NICOTINE). (NICOTINE). nicotine nicotine vapor-phase vapor-phase
(see (see
exhaust exhaust diesel diesel
reflect reflect not not but but exposures exposures matter matter ticulate ticulate directly, directly, measured measured been been had had smoke smoke tobacco tobacco vironmental vironmental
par
the the
increase increase would would that that sand, sand, and and dirt dirt as as such such en of of matter, matter, constituents constituents the the of of one one Only Only remained: remained: still still error error
particulate particulate
nondiesel nondiesel to to exposed exposed were were groups groups both both cause cause of of source source another another exclusions, exclusions, these these with with even even However, However,
be make make to to difficult difficult were were carmen carmen and and
brakers brakers
the the of of
posures posures
analysis. analysis. the the from from excluded excluded were were welders welders men men
ex exhaust exhaust diesel diesel the the
of of
estimates estimates
However, However,
exposure. exposure. of of
car and and brakers brakers both both Therefore, Therefore, measured. measured. directly directly been been
estimates estimates accurate accurate more more
develop develop
to to
able able
were were
we we
method, method,
had had particle particle of of type type neither neither as as possible, possible, not not were were fumes fumes
sampling sampling the the in in
improvements improvements
through through
addition, addition, In In ing. ing.
welding welding or or dust dust of of presence presence the the for for Corrections Corrections centrations. centrations.
test mutagenicity mutagenicity for for
collected collected
was was
urine urine his his
that that
day day the the
con high high in in present present were were fumes fumes welding welding welders, welders, carmen carmen
on on exposure exposure
individual's individual's
an an of of
measurement measurement actual actual
volved volved
the the for for substantial; substantial; occasionally occasionally was was air air the the in in dust dust brakers, brakers,
in project project this this for for
design design
study study the the
1988a,b), 1988a,b), al. al. et et
(Woskie (Woskie
the the For For section). section). Mutagenicity Mutagenicity Methods, Methods, the the (see (see large large
day day given given a a on on exposure exposure worker's worker's a a
of of
estimate estimate an an
as as
group group
systematically systematically be be to to apt apt was was term term remainder remainder the the welders, welders,
job job a a of of exposure exposure average average
the the
use use
than than
Rather Rather
unexposed. unexposed.
carmen carmen the the and and brakers brakers the the workers, workers, of of subgroups subgroups two two
as as classified classified were were carmen carmen and and
workers workers
clerical clerical
and and
exhaust, exhaust,
in in First, First, reasons. reasons. two two for for fashion fashion unbiased unbiased an an in in estimated estimated
diesel diesel of of
concentrations concentrations
lower lower
to to exposed exposed
were were
engineers engineers
be be not not could could particles particles diesel diesel of of quantity quantity the the form, form, this this In In
and and Brakers Brakers particles. particles. diesel diesel to to exposure exposure potential potential highest highest
Remainder. Remainder.
-
Smoke Smoke
Tobacco Tobacco - Particles Particles Respirable Respirable the the have have to to known known were were shop shop railroad railroad the the in in employed employed
Particles Particles Diesel Diesel
= = workers workers 1988a), 1988a), al. al. et et (Woskie (Woskie work work previous previous our our From From
welding. welding. or or burning, burning, cutting, cutting, doing doing those those excluding excluding workers workers repair repair
Car Car d d
tasks. tasks. and and
locations locations
job job of of
observation observation and and
1988a,b) 1988a,b)
al. al. et et (Woskie (Woskie work work previous previous on on based based assignment assignment exposure exposure exhaust exhaust diesel diesel priori priori c c A A
text}. text}.
(see (see
concentration) concentration) nicotine nicotine x x (8.6 (8.6 - particles particles respirable respirable concentration concentration = = particle particle respirable respirable Adjusted Adjusted b b
8.6. 8.6. x x
concentration concentration nicotine nicotine smoke smoke tobacco tobacco = = Environmental Environmental
8 8
163) 163)
113 113 (71, (71,
94 94
± ±
127 127
156 156 14) 14)
69 69 (0, (0, ± ±
29 29 158 158 200) 200) (87, (87, 99 99 132 132 ± ± 153 153 156 156 workers workers Shop Shop
exposure exposure High High
88) 88) 73 73
(63, (63,
85 85 ± ± 99 99
± ±
27 27 83) 83) 5 5 (0, (0,
83 83 48 48 ± ± 30 30 190) 190) (75, (75, 91 91 168 168 156 156 33 33
Brakers Brakers
78) 78) 54 54
(10, (10,
± ± 73 73
133 133
± ±
12 12 257) 257) (0, (0, 46 46
143 143 114 114 ± ± 14 14 244) 244) (61, (61, 157 157 182 182 183 183 14 14 Engineers Engineers
exposure exposure Moderate Moderate
± ±
189) 189) 89 89
(44, (44,
101 101
20 20 104 104 ± ± 157) 157) (2, (2,
64 64
108 108 132 132 20 20 264) 264) (102, (102, 115 115 201 201 ± ± 194 194
21 21
nonweldersd nonweldersd
Carmen Carmen
± ±
319) 319)
(75, (75,
178 196 196 178 210 210 ± ±
14 14
131) 131) (6, (6, 52 52
88 88 108 108 14 14 445) 445) (137, (137, 251 251 ± ± 285 285 170 170 14 14
welders welders
Carmen Carmen
98) 98)
(25, (25,
± ± 60 60
54 54 54 54 ± ±
44 44 101) 101) 58 58 (17, (17,
69 69 71 71 44 44 164) 164) (70, (70, 115 115 ± ± 119 119 71 71 57 57 Clerks Clerks
Unexposed Unexposed
Percentiles Percentiles
Median Median SD SD
n n
Percentiles Percentiles Median Median n n SD SD Percentiles Percentiles SD SD Median Median n n Group Group Job Job and and
75th 75th
Mean± Mean± 75th 75th
Mean Mean ± ± 75th 75th Mean± Mean±
Assignmentc Assignmentc
and and
25th 25th and and 25th 25th
and and 25th 25th Exposure Exposure
(~g/m3) (~g/m3)
(~g/m3) (~g/m3) (~g/m3) (~g/m3)
Concentrationb Concentrationb
Particle Particle Smokea Smokea Tobacco Tobacco Environmental Environmental Concentration Concentration Particle Particle Respirable Respirable
Respirable Respirable Adjusted Adjusted
Group Group Job Job
by by
Exposures Exposures Particle Particle Respirable Respirable Adjusted Adjusted and and Smoke, Smoke, Tobacco Tobacco Environmental Environmental 2. 2. Particle, Particle, Table Table Respirable Respirable
al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers
posed to diesel exhaust. A regression of respirable particles Formally, we included as predictors in the multiple on nicotine in these samples (excluding brakers and carmen regression model a dummy variable for potential exposure welders) yielded the same nicotine coefficient, 8.6 (SE = to diesel exhaust (EXPOSED = 1 for exposed jobs, EX 1.3), as the estimate from the chamber studies. POSED = 0 for unexposed), and two interaction variables: These considerations, and the concordance of the two EXPOSED with respirable particles (Respirable Particles x methods for estimating the nicotine coefficient, led us to EXPOSED) or with air nicotine (NICOTINE x EXPOSED). the following index of exposure to diesel particles: Any differential effect of respirable particles and nicotine in exposed jobs in this model is expressed through the sig Adjusted Respirable Particles = nificant interaction terms. The test of the three interaction Respirable Particles - 8.6 x [NICOTINE]. terms has three degrees of freedom. This approach bypasses entirely the need to construct an index. It is not necessary to subtract a constant term in this equa tion, because any regression model will include a constant Phenanthrene Models term. The adjusted respirable particle index indicates the Phenanthrene was measured in fewer than half the study excess of respirable particles over the amount to be expected air samples. Although it was used as an indicator for diesel based on the nicotine concentration in the air of men not exhaust in some of the multiple regression analyses, we do exposed to diesel particles. It accounts for diesel exposure not report the results. However, for a subset of observations only in men with potential diesel exhaust exposure. on nonsmokers with repeated observations on different This index of diesel particles is subject to error. After ex days, mutagenicity is plotted against phenanthrene. clusion of the brakers and carmen welders, the error should be less systematically related to job category. One conse Other Predictive Factors for Regression Models quence of the (random) error for estimating diesel particles For preliminary analyses, we screened a number of is that the observed association between mutagenicity and predictive factors for inclusion with the grouped and inter adjusted respirable particles will be of lower magnitude action exposure models. These analyses showed very differ than any true association, as will the probability of detect ing an association (Judge et al. 1980). This is known in ent effects in smokers and in nonsmokers, and so we report results separately by smoking category. Besides the expo statistics as the "errors in variables" problem. sure-definition variables discussed above, the following To study both unexposed and exposed men in the same predictive factors are included in one or more of the analysis, and to reduce the effect of error in measuring reported models: (1) urinary cotinine (control for internal diesel exhaust exposure, we grouped air samples according dose of tobacco smoke) for all models; (2) "protective food" to concentration of adjusted respirable particles. All sam consumption (cabbage, Brussels sprouts, or fish) (1 = yes, ples taken from men in unexposed jobs constituted the first 0 = no) for nonsmoker models; (3) cigarettes smoked on group. Samples from exposed men were grouped roughly study day for smoker models. into thirds, for a total of four groups. The samples from ex We also considered including dummy variables for posed workers with the highest exposure to adjusted respi differential effects of the six trips made to the railroad facil rable particles (group 4) should have higher true diesel ex ity (a compounding of batch, handling, seasonal, storage, haust exposure than the samples in the unexposed group and processing effects). However, the samples from most (group 1) and those in group 2. However, the random error visits were from workers who were all exposed or all unex in adjusted respirable particles makes it likely that some posed (Table 1). Therefore, exposure status was inextricably overlap and misclassification in true diesel exhaust ex confounded with visit effect, and controlling for visit posure remains. The differences among the four exposure differences would have made the finding of exposure effects groups are tested with an For chi-squared test using three impossible. degrees of freedom in the regression models. Urinary thiocyanate was also entered into preliminary Interaction Model for Exposure Effects models as an indicator of active and passive smoking. How ever, it was not as successful as urinary cotinine and the As an alternative to the adjusted respirable particle index number of cigarettes actually smoked. Urinary nicotine was discussed above, we also used a multiple regression model also considered, but was rejected as less predictive than the to detect a differential effect of measured respirable parti combination of cotinine alone and the number of cigarettes cles and nicotine in exposed jobs. smoked on the study day.
16
17 17
As As
standards. standards. the the
all all of of
deviations deviations standard standard smallest smallest the the in in
varying varying be be to to appeared appeared what what for for
analyses analyses
regression regression the the
ing ing
resulted resulted
mutagen mutagen
standard standard the the as as 2-AF 2-AF of of use use Furthermore, Furthermore,
correct of of ways ways sought sought we we findings, findings,
ofthese ofthese
basis basis the the
On On
above. above. (2), (2), form form of of models models the the did did than than
deviations deviations
standard standard
4-NQO. 4-NQO.
and and 2-NF 2-NF controls, controls, S9 S9 - the the were were than than
residual residual smaller smaller give give to to
appeared appeared
model model
of of form form This This
S. S. by by
results results
S9 S9 - the the
with with
correlated correlated highly highly more more were were and and BaP, BaP,
multiplied multiplied are are terms terms error error
the the (1), (1),
type type
model model in in
However, However,
2-AF 2-AF
controls, controls, S9 S9
two two + + the the
surprisingly, surprisingly, Somewhat Somewhat casion. casion.
(4). (4). type type model model for for form form
the the
to to
identical identical
is is
mean mean
response response
oc
that that
on on
assayed assayed
results results
sample sample study study the the with with occasion occasion
the the for for form form the the by by S, S, multiplied multiplied
are are
(1) (1)
model model for for
equation equation
each each
on on
means means
standard standard the the of of correlation correlation rank rank Spearman Spearman
the the of of sides sides both both If If similar. similar. very very
are are
(4) (4)
and and (1) (1)
Models Models
the the shows shows E.3 E.3 Table Table percent. percent. 20 20 than than greater greater were were and and BaP BaP
fitted.) fitted.)
2-AF 2-AF for for variation variation of of coefficients coefficients
between-experiment between-experiment the the
0
be be a
say, say,
, ,
intercept, intercept, separate separate a a that that required required section], section],
seen, seen, be be can can As As E.2. E.2. and and E.1 E.1
Tables Tables E, E,
Appendix Appendix in in
scribed scribed
next next
[see [see
BMDP3V BMDP3V used, used, we we program program computer computer the the fact, fact, (In (In
de are are experiments experiments control control these these for for statistics statistics Descriptive Descriptive
(4) (4)
Terms. Terms. S9. S9. - Error Error for for 4-NQO 4-NQO + + and and 2-NF 2-NF and and S9, S9, + +
2
2 2 1
0 0 1 1
Zk)] Zk)] x x (bk (bk + + ...... Z + + x x ) ) (b Z + + for for x x ) ) and and (b + + 2-AF 2-AF BaP BaP [b x x S S sensitivity: sensitivity: strain strain bacterial bacterial for for control control
positive positive a a as as occasion occasion Response Response each each on on = = evaluated evaluated were were mutagens mutagens
standard standard
different different
four four above, above, section section Mutagenicity Mutagenicity the the in in
transform): transform): its its
(or (or
mutagenicity mutagenicity for for
model model
described described
As As
progressed. progressed. study study the the as as occasions occasions different different
linear linear following following the the using using
to to
amounted amounted
model model
This This
13 13 on on processed processed were were samples samples mutagenicity mutagenicity urine urine The The
(3) (3) mean). mean). standard standard one one by by covariates covariates
Strains Strains Bacterial Bacterial of of
original original (multiply (multiply Rate Rate Mutagenicity Mutagenicity
Response Response
= =
Sensitivity Sensitivity Differential Differential for for Accounting Accounting Form: Form: Model Model
reported. reported. not not are are approach approach
this this of of results results the the Therefore, Therefore, below. below. reported reported mode, mode, of of type type
analysis. analysis.
any any
from from
servations servations
third third the the from from differ differ not not did did conclusions conclusions the the and and better better no no
ob two two than than more more no no of of
elimination elimination
the the
to to led led
it it subjective, subjective,
were were fits fits model model the the addition, addition, In In response. response. no no give give standards standards
somewhat somewhat was was criterion criterion
this this
Although Although
remained. remained.
vations vations
the the when when even even response response positive positive a a extrapolates extrapolates it it because because
obser influential influential unduly unduly no no until until
recalculated recalculated
was was statistic statistic
unsatisfactory unsatisfactory was was formulation formulation this this however, however, view, view, of of point point
D Cook's Cook's and and refit refit was was model model the the
elimination, elimination, of of
round round
biological biological a a From From analysis. analysis. any any in in influential influential unduly unduly as as
each each After After values. values. lower lower with with
observations observations
from from
separated separated
identified identified were were observations observations two two than than
more more no no and and sponse, sponse,
clearly clearly appeared appeared generally generally
these these > >
because because 0.5, 0.5,
with with D D tions tions
re for for used used were were values values original original the the approach, approach, this this lowing lowing
observa eliminated eliminated We We
1977). 1977).
(Cook (Cook
calculated calculated
was was
tion, tion,
Fol time. time. one one at at included included be be could could standard standard one one than than more more
observa an an omitting omitting of of
effect effect
the the measures measures
which which
statistic, statistic,
of of activity activity mutagenic mutagenic because because attractive attractive was was model model This This
D Cook's Cook's observations, observations, influential influential unduly unduly detect detect To To
(2) (2) covariates). covariates). as as
added added
response. response. a a as as chosen chosen was was creatinine, creatinine, of of
means means (standard (standard Rate Rate
Mutagenicity Mutagenicity
Response Response
= =
milliliter milliliter per per revertants revertants in in rate, rate, mutagenicity mutagenicity untransformed untransformed
original, original, the the only only
considerations, considerations, these these of of result result a a As As sible. sible. reported. reported. not not are are results results the the and and abandoned, abandoned,
implau
also also were were
4, 4, as as A A such such 1, 1, than than = = greater greater power power with with was was then, then, approach, approach, This This high. high. too too seemed seemed that that number number
transformations transformations
power power
Suggested Suggested component. component. background background a a analyses, analyses, some some for for influential influential unduly unduly as as identified identified were were
strong strong the the of of face face
the the in in
unappealing unappealing was was phenomenon phenomenon
observations observations 65 65 of of 5 5 to to up up -%),but -%),but example, example, p p (for (for sible sible = =
This This
counts. counts.
low low
very very
these these to to emphasis emphasis strongest strongest implau transformations transformations power power suggested suggested the the were were only only Not Not
gave gave
diagnostic diagnostic
the the
by by
suggested suggested sometimes sometimes -1), -1), (A (A = = unsatisfactory. unsatisfactory. were were fits fits the the but but model, model, this this in in tried tried were were
inverse inverse or or 0.5), 0.5),
= = (A (A
root root
square square 0), 0), = = A A
to to
2 (corresponding (corresponding 1 standards standards four four All All covariates. covariates. are are Zk Zk Z ., ., Z Here Here , , , ,
logarithm logarithm the the as as such such transformations, transformations, Familiar Familiar negative. negative.
(1) (1)
Terms. Terms.
Error Error
+ +
were were many many indeed, indeed, counts; counts; (blank) (blank) background background the the than than
2
2 2 1 1 1
Zk) Zk) x x (bk (bk + + ...... Z + + x x ) ) (b Z + +
x x ) )
(b + + bo bo
higher higher little little were were responses responses observed observed the the of of Most Most factory. factory.
= = Rate/S Rate/S
Mutagenicity Mutagenicity = =
Response Response
unsatis proved proved results results the the but but 1986), 1986), (Atkinson (Atkinson tried tried were were
analyses: analyses: nary nary
Mutagenicity" Mutagenicity" Variable Variable Response Response = =
prelimi in in data data the the to to fit fit were were models models regression regression of of types types
form form the the of of 1964) 1964) Cox Cox and and different different three three following following The The date. date. experimental experimental an an on on
(Box (Box
transformation transformation
power power a a for for tests tests diagnostic diagnostic Formal Formal standard standard a a of of mutagenicity mutagenicity mean mean the the indicate indicate LetS LetS casions. casions.
oc
experimental experimental different different on on strain strain TA98 TA98 the the of of
sensitivity sensitivity
Diagnostics Diagnostics Model Model and and Transformations Transformations
al. al. et et Schenker Schenker B. B. M. M.
18 18
freedom. freedom. This This was was also also true true of of the the test test for for the the three three interac- ration ration on on a a reverse-phase reverse-phase column. column. Although Although gradient gradient chro-
respirable respirable particles, particles, the the resulting resulting test test had had three three small small degrees degrees aromatic aromatic of of fraction fraction was was then then analyzed analyzed by by isocratic isocratic sepa
formed formed on on the the basis basis of of diesel diesel exhaust exhaust exposure exposure and and adjusted adjusted larger larger aromatic aromatic compounds; compounds; and and polar polar compounds. compounds. The The
parameters parameters in in the the subset. subset. For For the the analyses analyses with with aliphatic aliphatic strata strata compounds; compounds; two-ring two-ring or or three-ring three-ring aromatics; aromatics;
bution bution with with degrees degrees of of freedom freedom equal equal to to the the number number of of extract extract solutions solutions into into the the following following chemical chemical fractions: fractions:
in in the the log-likelihoods log-likelihoods was was referred referred to to a a chi-squared chi-squared distri methods methods first first fractionated fractionated the the filter filter and and polyurethane polyurethane foam foam
models models by by the the computer computer program. program. Two Two times times the the difference difference the the polyurethane polyurethane foam foam samples. samples. Liquid Liquid chromatographic chromatographic
sumption sumption of of normality) normality) were were computed computed for for full full and and reduced reduced A A two-step two-step analysis analysis was was developed developed to to measure measure PAHs PAHs in in
model"). model"). Log-likelihood Log-likelihood statistics statistics (based (based on on a a nominal nominal as with with the the collection collection of of phenanthrene. phenanthrene.
the the subset subset
("full ("full
model") model")
and and one one
without without
it it ("reduced ("reduced
and and those those without. without. Thus, Thus, the the treated treated filter filter did did not not interfere interfere
For For testing testing
a a subset subset
of of variables, variables,
one one model model
was was
fit fit with with
samplers samplers with with the the treated treated filter filter (which (which collects collects nicotine) nicotine)
is is that that it it requires requires the the fitting fitting of of a a separate separate intercept. intercept. tent tent of of the the polyurethane polyurethane foam foam plugs plugs was was found found between between
components components are are less less biased. biased. One One drawback drawback to to this this program program tobacco tobacco smoke smoke demonstrated demonstrated that that no no difference difference in in PAH PAH con
Sampson Sampson 1976; 1976; Harville Harville 1977), 1977), and and (2) (2) estimated estimated ments ments variance variance in in the the environmental environmental chamber chamber with with environmental environmental
Gaussian Gaussian distributions distributions for for the the random random terms terms (Jennrich (Jennrich and and polyurethane polyurethane foam foam sections, sections, and and none none on on the the third. third. Experi
cients cients are are likely likely to to be be robust robust to to the the formal formal assumption assumption of of plugs. plugs. All All of of the the phenanthrene phenanthrene was was found found on on the the first first two two
hood hood estimates estimates are are presented presented because because (1) (1) estimated estimated coeffi three three quarters quarters as as much much fluorene fluorene as as the the polyurethane polyurethane foam foam
Statistical Statistical Software Software 1983). 1983). The The restricted restricted maximum maximum likeli XAD-2 XAD-2 resin resin collected collected only only half half as as much much phenanthrene phenanthrene and and
Analysis Analysis of of Variance Variance was was used used for for these these calculations calculations fluorene fluorene (BMDP (BMDP and and phenanthrene phenanthrene that that was was lost lost from from the the filter, filter, the the
The The computer computer program program BMDP3V BMDP3V (General (General Mixed Mixed Model) Model) While While the the polyurethane polyurethane foam foam collected collected virtually virtually all all of of the the
ficiently; ficiently; both both collected collected the the pyrene pyrene that that the the had had independence independence been been lost. lost. model. model.
the the polyurethane polyurethane foam foam plugs plugs collected collected the the naphthalene naphthalene ef and and so so are are also also somewhat somewhat different different from from the the effects effects under under
of of sampling sampling at at 1.7 1.7 Lpm. Lpm. Neither Neither the the XAD-2 XAD-2 resin resin tubes tubes nor nor regression regression effects effects are are weighted weighted to to have have minimum minimum variance, variance,
of of the the naphthalene naphthalene were were lost lost from from the the filter filter after after eight eight hours hours are are independent independent (Judge (Judge et et al. al. 1980). 1980). The The point point estimates estimates for for
phenanthrene, phenanthrene, 66 66 percent percent of of the the fluorene, fluorene, and and 100 100 percent percent ple ple regression regression model, model, which which assumes assumes that that all all observations observations
pling, pling, but but 35 35 percent percent of of the the pyrene, pyrene, 62 62 percent percent of of the the der der the the generalized generalized model model than than under under the the ordinary ordinary multi
the the chrysene chrysene and and BaP BaP remained remained on on the the filter filter during during sam Statistical Statistical values values p p and and standard standard errors errors are are more more valid valid un
ther ther
XAD-2 XAD-2 resin resin
or or polyurethane polyurethane foam) foam) showed showed that that all all of of person'' person'' or or "error" "error" variance variance component. component.
ments ments
with with the the
different different
sampling sampling
schemes schemes
(filter (filter
plus plus ei taken taken from from the the same same person, person, and and is is called called the the "within
PAHs PAHs
were were
found found in in
the the third third
extraction. extraction.
Laboratory Laboratory experi term term represents represents the the differences differences between between daily daily samples samples
with with PAHs PAHs removed removed
nearly nearly
all all the the PAHs PAHs
added, added,
and and no no "between-person" "between-person" variance variance component. component. The The second second random random
posure. posure.
One One extraction extraction
of of a a polyurethane polyurethane
foam foam plug plug
spiked spiked
trip, trip, varying varying between between subjects subjects with with a a variance variance called called the the
railroad railroad
workers workers
as as
a a possible possible
marker marker
for for
diesel diesel
exhaust exhaust
ex to to all all the the samples samples taken taken from from the the same same person person on on the the same same
cient cient
at at collecting collecting
phenanthrene phenanthrene
from from
the the breathing breathing
zone zone of of regression. regression. The The first first random random term term is is assumed assumed to to be be common common
The The
polyurethane polyurethane
foam foam
plugs plugs were were
determined determined
to to
be be effi fixed fixed mean mean is is the the same same as as would would be be fit fit by by ordinary ordinary multiple multiple
regression regression
mean mean
and and
two two
independent independent
random random terms. terms. The The Phenanthrene Phenanthrene as as a a Marker Marker for for Diesel Diesel Exhaust Exhaust
model, model, the the response response was was modeled modeled as as the the sum sum of of a a fixed fixed
SPECIFIC SPECIFIC AIM AIM
1 1
least-squares least-squares model model was was fit fit (Judge (Judge et et al. al. 1980). 1980). In In this this
that that were were recorded recorded in in the the study, study, an an estimated estimated generalized generalized
To To accommodate accommodate the the multiple multiple observations observations per per person person RESULTS RESULTS
Generalized Generalized Least Least Squares Squares
of of Sin Sin model model (4). (4). fits. fits.
tants/ng tants/ng (Table (Table
E.1). E.1).
This This was was
used used throughout throughout
as as the the value value mum mum likelihood likelihood fits, fits, not not on on restricted restricted maximum maximum likelihood likelihood
overall overall
study study
mean mean for for mutagenicity mutagenicity of of 2-AF, 2-AF, 3.46 3.46 rever ordering. ordering. As As a a result, result, all all likelihood likelihood tests tests are are based based on on maxi
mated mated coefficients coefficients
from from
the the model model
were were multiplied multiplied by by the the restricted restricted maximum maximum likelihood likelihood fits fits do do not not always have have always this this
To To calculate calculate
predicted predicted effects effects
from from model model ( ( 4). 4). the the esti ing ing the the analysis, analysis, it it was was discovered discovered that that log-likelihoods log-likelihoods from from
Results Results for for model model type type ( ( 4) 4) are are reported. reported. duced duced model model is is always always less less than than that that for for a a full full model. model. Dur
servations servations were were detected detected and and eliminated eliminated from from any any maximum maximum analysis. analysis. likelihood likelihood problems, problems, the the log-likelihood log-likelihood for for a a re
with with model model type type (2), (2), only only one one or or two two tion tion unduly unduly factors factors influential influential in in the the exposure-interaction exposure-interaction ob model. model. For For regular regular
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
19 19
with with reaction reaction by by respirable respirable collection collection adjusted adjusted during during the the of of stabilized stabilized percent percent is is 50 50 Nicotine Nicotine approximately approximately average, average,
on on that, that, suggesting suggesting 0.003, 0.003, was was samples samples personal personal these these for for nicotine. nicotine. of of j.!g j.!g 5 5 for for percent percent 98 98 averaging averaging plete, plete,
ratio ratio mean mean The The true. true. be be to to this this showed showed 62) 62) made made ~ ~ (n (n were were com was was nicotine nicotine of of extraction extraction filter, filter, the the wet wet help help to to anol anol
measurements measurements both both which which on on sample sample personal personal each each for for eth of of addition addition the the With With variable. variable. highly highly was was it it but but percent, percent,
concentration concentration particle particle respirable respirable to to phenanthrene phenanthrene of of ratio ratio 75 75 approximately approximately was was efficiency efficiency extraction extraction the the traction, traction,
the the of of Calculation Calculation SD). SD). 2 2 (mean (mean + + 0.0076 0.0076 or or 0.006 0.006 than than less less ex initial initial the the during during ethanol ethanol of of addition addition the the Without Without tine. tine.
or or to to equal equal be be should should samples samples personal personal workers' workers' exposed exposed nico of of amounts amounts varying varying with with spiked spiked were were Filters Filters nicotine. nicotine.
exhaust diesel diesel the the in in concentration concentration particle particle respirable respirable of of recovery recovery inefficient inefficient to to led led base base less less but but solution, solution, the the to to
to to phenanthrene phenanthrene of of ratios ratios the the then then activities, activities, mechanical mechanical added added was was hydroxide hydroxide sodium sodium N N 10 10 of of mL mL 2 2 when when recoveries recoveries
and and sanding, sanding, grinding, grinding, through through as as such such particles, particles, borne borne good good yielded yielded extraction extraction liquid/liquid liquid/liquid The The heptane. heptane. into into
air of of sources sources other other to to exposures exposures have have workers workers repair repair if if and and extraction extraction liquid/liquid liquid/liquid by by concentrated concentrated be be can can it it that that so so
true, true, is is this this If If particles. particles. respirable respirable and and phenanthrene phenanthrene of of put put nicotine nicotine the the neutralizes neutralizes and and deprotonates deprotonates hydroxide hydroxide dium dium
out constant constant approximately approximately an an had had source source the the and and source source so of of addition addition The The stages. stages. several several in in developed developed were were filters filters
same same the the from from came came samples samples the the if if expected expected be be would would This This sample sample from from recovery recovery nicotine nicotine for for conditions conditions Optimal Optimal
0.006. 0.006. of of particles particles respirable respirable to to phenanthrene phenanthrene of of ratio ratio stant stant
Smoke Smoke Tobacco Tobacco Environmental Environmental for for Marker Marker a a as as Nicotine Nicotine
con surprisingly surprisingly a a showed showed exhaust, exhaust, diesel diesel all all virtually virtually are are
which which shops, shops, repair repair the the from from samples samples area area The The established. established.
be be to to had had concentration concentration particle particle respirable respirable to to phenanthrene phenanthrene exhaust. exhaust. diesel diesel for for marker marker a a as as pursued pursued be be should should
of of ratio ratio the the exhaust, exhaust, diesel diesel for for marker marker a a as as phenanthrene phenanthrene that that phenanthrene phenanthrene suggest suggest data data the the However, However, analyzed. analyzed.
use use To To detection. detection. of of limit limit the the above above were were workers workers samples samples shop shop area area few few were were there there because because cautiously cautiously interpreted interpreted
from from samples samples the the of of most most while while detection, detection, of of be be limit limit the the should should low low data data These These particles. particles. respirable respirable to to phenanthrene phenanthrene
be or or at at were were carmen carmen and and clerks clerks from from samples samples the the of of Most Most of of ratio ratio the the to to similar similar was was samples samples area area versus versus personal personal
samples). samples). blank blank the the of of SD SD 2 2 mean mean + + ~ ~ detection detection of of (limit (limit in in (0.003) (0.003) particles particles respirable respirable adjusted adjusted to to phenanthrene phenanthrene
detection detection of of limit limit the the
near near and and , , j.!g/m 1 1 than than less less of of were were ratio ratio ples ples the the 4), 4), (Table (Table low low were were workers workers shop shop nonsmoking nonsmoking 3
sam field field from from concentrations concentrations phenanthrene phenanthrene of of the the of of All All exposures exposures smoke smoke tobacco tobacco environmental environmental Because Because levels. levels.
phenanthrene phenanthrene to to ng). ng). 30 30 (53 (53 significantly significantly ± ± variable variable contribute contribute quite quite not not were were do do that that values values the the cles cles and and threne, threne,
parti smoke smoke phenan tobacco tobacco more more much much environmental environmental had had plugs plugs include include may may foam foam particles particles polyurethane polyurethane blank blank
respirable respirable Field Field because because 9), 9), Figure Figure phenanthrene. phenanthrene. (see (see of of ng ng 5 5 about about phenanthrene phenanthrene to to had had filters filters particles particles blank blank Field Field
respirable respirable adjusted adjusted of of ratio ratio ng/mL. ng/mL. the the 5 5 examined examined also also approximately approximately We We was was samples.) samples.) detection detection of of limit limit the the tions, tions,
individual individual for for solu ratios ratios standard standard observed observed For For the the not not method. method. and and categories, categories, job job for for spectrophotometric spectrophotometric ultraviolet ultraviolet
8] 8] Figure Figure [see [see standard standard the the to to phenanthrene phenanthrene and and relative relative 4] 4] through through 2 2 phenanthrene phenanthrene for for [Tables [Tables tions tions sensitivity sensitivity and and ity ity
concentra particle particle specific respirable respirable increase increase of of to to used used values values was was average average give give which which spectrofluorimeter spectrofluorimeter a a Finally, Finally,
data, data, involved. involved. cumulative cumulative samples samples the the of of from from number number result result large large not not the the does does analyze analyze ratio ratio to to similar similar suming suming
(A (A exhaust. exhaust. diesel diesel not not was was time-con samples samples too too was was it it nonsmokers' nonsmokers' in in separation, separation, particles particles better better achieved achieved matography matography
parts. parts. metal metal of of welding welding and and burning, burning, cutting, cutting, includes includes "Welding" "Welding" a a
164) 164) (71, (71, 94 94 112 112 ± ± 149 149 + + 129 129 200) 200) (86, (86, 129 129 100 100 152 152 149 149 nonwelders nonwelders worker worker
Shop Shop
± ±
161) 161) (57, (57, ± ± 147 147 102 102 100 100 7 7 251) 251) (138, (138, 163 163 71 71 173 173 7 7 weldersa weldersa
worker worker Shop Shop
± ±
189) 189) (44, (44, 89 89 ± ± 102 102 104 104 20 20 264) 264) (102, (102, 201 201 115 115 194 194 21 21 nonwelders nonwelders
Carmen Carmen
± ±
319) 319) (75, (75, 196 196 ± ± 178 178 14 14 210 210 445) 445) (137, (137, 251 251 170 170 285 285 14 14 weldersa weldersa Carmen Carmen
Percentiles Percentiles SD SD Median Median n n Percentiles Percentiles Median Median SD SD n n Group Group Job Job
± ± 75th 75th and and 25th 25th ± ± Mean Mean 75th 75th and and 25th 25th Mean Mean
) ) (!lg/m (!lgfm3) (!lgfm3) 3
Concentration Concentration Concentration Concentration
Particle Particle Respirable Respirable Adjusted Adjusted Particle Particle Respirable Respirable
Nonwelders Nonwelders and and Welders Welders Worker Worker Shop Shop
and and Carmen Carmen for for Exposures Exposures Particle Particle Respirable Respirable Adjusted Adjusted and and Particles Particles Respirable Respirable 3. 3. Table Table
al. al. et et Schenker Schenker B. B. M. M.
20 20
with with varying varying tar tar and and nicotine nicotine contents contents in in mainstream mainstream ulate ulate matter matter was was calculated calculated by by multiplying multiplying the the ambient ambient
(Rickert (Rickert et et al. al. 1984). 1984). Several Several different different brands brands of of cigarettes, cigarettes, The The environmental environmental tobacco tobacco smoke smoke contribution contribution to to partic
regardless regardless of of their their degree degree of of mainstream mainstream smoke smoke ventilation ventilation
variation variation 50 50 percent). percent). = =
smoke smoke is is found found
among among
different different
brands brands
of of cigarettes, cigarettes,
poor poor compared compared to to that that of of the the slope slope (SE (SE 4.4, 4.4, coefficient coefficient of of = =
smoke. smoke. Little Little variability variability in in the the nicotine nicotine levels levels
of of sidestream sidestream
percent. percent. Thus, Thus, the the precision precision of of the the intercept intercept estimate estimate is is
the the air, air, is is the the
major major component component
of of
environmental environmental
tobacco tobacco
intercept intercept had had a a SE SE 125 125 for for a a coefficient coefficient of of variation variation of of 250 250 = =
which which is is emitted emitted
from from
the the
tip tip of of
the the
cigarette cigarette
directly directly into into
tine tine level level of of zero. zero. By By jackknife jackknife resubstitution, resubstitution, the the estimated estimated
(Schlotzhauer (Schlotzhauer
and and
Chortyk Chortyk
1983). 1983).
Sidestream Sidestream
smoke, smoke,
tainty tainty in in extrapolating extrapolating the the chamber chamber data data to to an an airborne airborne nico
and and by by other other changes changes
in in
the the
design design
of of
the the cigarette cigarette
The The value value of of this this intercept intercept reflects reflects the the statistical statistical uncer
achieved achieved by by filtering filtering the the smoke, smoke, by by increasing increasing ventilation, ventilation,
Respirable Respirable
Particles Particles
8.6 8.6 [NICOTINE] [NICOTINE]
x x
50.5. 50.5.
+ + = = Reduction Reduction of of nicotine nicotine content content in in mainstream mainstream smoke smoke is is
lation: lation: would would have have a a limit limit of of detection detection of of 2 2 Jlg/m . .
3
A A principal principal components components analysis analysis yielded yielded the the following following re of of detection detection of of nicotine nicotine of of 0.2 0.2 Jlg/m ; ; a a one-hour one-hour
sample sample 3
nicotine nicotine and and particles particles from from environmental environmental tobacco tobacco smoke. smoke. ples ples collected collected at at 1.7 1.7 Lpm Lpm for for eight eight hours hours would would have have a a limit limit
experiments) experiments) were were used used to to calculate calculate the the relationship relationship of of flow flow rates rates and and sampling sampling times; times; in in these these studies, studies, field field sam
ticulate ticulate concentration concentration was was less less than than 1,200 1,200 Jlg/m (11 (11 such such
limit limit of of detection detection of of nicotine nicotine in in air air would would depend depend on on the the 3 3
lower lower levels, levels, only only those those chamber chamber experiments experiments in in which which par formed formed with with as as little little as as 0.2 0.2 Jlg Jlg of of nicotine nicotine added. added. The The overall overall
smoke smoke exposures exposures of of the the railroad railroad workers workers were were at at much much Jlg Jlg of of nicotine nicotine per per treated treated filter. filter. Recovery Recovery studies studies were were per
around around 1,000 1,000 Jlg/m . . Because Because the the environmental environmental tobacco tobacco chromatography chromatography with with a a nitrogen-selective nitrogen-selective detector detector was was 0.06 0.06 3
conducted conducted at at relatively relatively high high particle particle concentrations, concentrations, of of chromatographic chromatographic analysis. analysis. The The limit limit of of detection detection by by gas gas
Most Most of of the the environmental environmental chamber chamber experiments experiments were were efficiently efficiently and and desorb desorb the the nicotine nicotine quantitatively quantitatively for for gas gas
method method of of treated treated filters filters was was found found to to collect collect nicotine nicotine vapor vapor marker marker for for environmental environmental cigarette cigarette smoke. smoke.
cent cent of of the the nicotine nicotine found found in in the the first first treated treated filter. filter. Thus, Thus, the the brands. brands. Thus, Thus, nicotine nicotine would would appear appear to to be be an an appropriate appropriate
treated treated filters. filters. All All of of these these filters filters contained contained less less than than 1 1 per mental mental tobacco tobacco smoke smoke did did not not vary vary significantly significantly among among the the
placed placed in in the the cigarette cigarette exposure exposure chamber chamber contained contained backup backup ber. ber. The The ratio ratio of of nicotine nicotine to to particulate particulate matter matter in in environ
nicotine nicotine from from treated treated filters filters during during sampling, sampling, 13 13 samplers samplers tested tested under under similar similar conditions conditions in in the the environmental environmental cham
the the bisulfate bisulfate ion. ion. To To check check for for possible possible breakthrough breakthrough of of smoke smoke (Federal (Federal Trade Trade Commission Commission Report Report 1985), 1985), were were
b b Work Work in in large, large, indoor indoor repair repair facility. facility.
a a Clerks. Clerks.
108 108 Nonsmokers Nonsmokers 110 110 113 113 1 1 (72, (72, 161) 161) 5) 5) 108 108 110 110 (0, (0, (71, (71, 155) 155)
42 42 233 233 Smokers Smokers 42 42 94 94 (145, (145, 305) 305) 42 42 (3, (3, 131) 131) 136 136 (59, (59, 213) 213)
Shop Shop workersb workersb
Nonsmokers Nonsmokers 9 9 8 8 143 143 1 1 (56, (56, 203) 203) 3) 3) 8 8 116 116 (0, (0, (51, (51, 204) 204)
24 24 24 24 247 247 96 96 Smokers Smokers (124, (124, 348) 348) (39, (39, 160) 160) 24 24 138 138 (53, (53, 259) 259)
Carmen Carmen
17 17 18 18 83 83 Nonsmokers Nonsmokers 0 0 90) 90) (65, (65, 2) 2) 16 16 74 74 (0, (0, (64, (64, 85) 85)
Smokers Smokers 14 14 12 12 190 190 (93, (93, 312) 312) 124 124 (35, (35, 158) 158) 11 11 73 73 (60, (60, 292) 292)
Brakers Brakers
Outside Outside workers workers
40 40 33 33 104 104 Nonsmokers Nonsmokers 59 59 (45, (45, (18, (18, 164) 164) 106) 106) 33 33 44 44 (23, (23, 96) 96)
17 17 11 11 133 133 Smokers Smokers 53 53 (103, (103, 169) 169) (13, (13, 63) 63) 11 11 66 66 (38, (38, 111) 111)
Inside Inside workersa workersa
Job Job Group Group n n n n Median Median Median Median Percentiles Percentiles Percentiles Percentiles Percentiles Percentiles n n Median Median
25th 25th and and 25th 25th and and 75th 75th 75th 75th 25th 25th and and 75th 75th
(Jlg/m ) ) (Jlg/m3) (Jlg/m3) (Jlg/m3) (Jlg/m3) 3
Concentration Concentration Smoke Smoke Concentration Concentration Concentration Concentration
Adjusted Adjusted Respirable Respirable Particle Particle Respirable Respirable Particle Particle Environmental Environmental Tobacco Tobacco
Table Table Workplace Workplace Exposures Exposures by by Job Job 4. 4. Location Location and and Cigarette Cigarette Smoking Smoking Status Status
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
21 21
the the extracts. extracts. exceeding exceeding railroad railroad frequency, frequency, diesel diesel mutation mutation increased increased an an as as fined fined
the the in in present present were were compounds compounds related related or or de was was nitro-PARs nitro-PARs if if assay assay mutation mutation forward forward the the in in response response (positive) (positive)
examine examine to to wanted wanted We We 1983). 1983). Mermelstein Mermelstein significant significant and and A A 5). 5). (Table (Table (Rosenkranz (Rosenkranz 1978) 1978) al. al. et et (Skopek (Skopek assay assay tation tation
nitro-PARs nitro-PARs certain certain to to insensitive insensitive is is strain strain NR NR mu TA98 TA98 the the forward forward Salmonella Salmonella the the using using group, group, Thilly's Thilly's William William
because because S9 S9 without without used used was was NR, NR, by by TA98 TA98 strain, strain, Technology Technology of of tester tester Institute Institute other other Massachusetts Massachusetts the the at at tested tested ously ously
An matter. matter. particulate particulate diesel diesel in in previ mutagens mutagens were were the the to to extracts extracts same same sensitivity sensitivity These These procedure. procedure. micro micro the the using using
historical historical on on and and procedure, procedure, screening screening activity, activity, the the in in found found mutagenic mutagenic for for sitivity sitivity tested tested were were facilities facilities railroad railroad of of
sen strain strain ofthe ofthe basis basis the the on on variety variety a a at at determinations determinations collected collected dose-response dose-response particles particles of of samples samples filter filter high-volume high-volume
the the for for chosen chosen was was S9, S9, without without and and of of with with extracts extracts TA98, TA98, strain strain Tester Tester dichloromethane dichloromethane extracts, extracts, diesel diesel for for method method
3). 3). (Figure (Figure facilities facilities railroad railroad two two from from extracts extracts microsuspension microsuspension the the particle particle of of the the sensitivity sensitivity the the of of validation validation a a As As
of of some some for for determined determined were were relationships relationships
Dose-response Dose-response
Extracts Extracts Diesel Diesel for for Sensitivity Sensitivity Assay Assay Microsuspension Microsuspension
Extracts Extracts Diesel Diesel of of Relationships Relationships Dose-Response Dose-Response
2 2 AIM AIM SPECIFIC SPECIFIC
5). 5). (Table (Table
S9 S9 of of addition addition the the without without especially especially activity, activity, mutagenic mutagenic section). section). Methods Methods Statistical Statistical
detecting detecting in in sensitive sensitive less less was was TA100 TA100 Strain Strain assay. assay. sion sion (see (see analyses analyses some some from from excluded excluded were were welders welders men men
microsuspen the the using using S9, S9, without without and and with with TA98, TA98, strain strain car and and brakers brakers particles, particles, nondiesel nondiesel of of excess excess systematic systematic a a
in in positive positive be be to to determined determined were were assay assay mutation mutation ward ward with with samples samples from from contributed contributed bias bias minimize minimize To To particles. particles.
for the the in in negative negative tested tested that that samples samples the the of of all all However, However, respirable respirable adjusted adjusted the the to to contribute contribute also also may may pollution) pollution)
S9. S9. without without and and with with TA100, TA100, and and TA98 TA98 strains strains tester tester both both air air fumes, fumes, welding welding gravel, gravel, (sand, (sand, particles particles other other although although
in in assay assay microsuspension microsuspension the the in in positive positive also also were were S9, S9, out out exposure, exposure, exhaust exhaust diesel diesel of of marker marker a a as as used used be be can can ticles ticles
with and and with with both both assay, assay, mutation mutation forward forward the the in in positive positive par respirable respirable adjusted adjusted the the and and smoke smoke tobacco tobacco vironmental vironmental
tested tested that that samples samples The The revertants. revertants. of of number number ground ground en to to exposure exposure of of marker marker a a as as used used be be can can concentration concentration
back the the of of doubling doubling a a as as defined defined was was method method suspension suspension nicotine nicotine The The concentration. concentration. particle particle respirable respirable adjusted adjusted an an
micro the the in in response response positive positive A A cultures. cultures. control control treated treated yield yield to to concentration concentration particle particle respirable respirable the the from from tracted tracted
un the the of of limit limit confidence confidence upper upper percent percent 99 99 historical historical sub then then was was value value This This 8.6. 8.6. by by concentration concentration nicotine nicotine
1978). 1978). a!. a!. et et (Skopek (Skopek Technology Technology of of Institute Institute Massachusetts Massachusetts at at performed performed assay assay the the of of SD SD 3 3 x x ~ ~ Positive Positive c c
diameter. diameter. aerodynamic aerodynamic 3.5 3.5 at at ~m ~m efficiency efficiency collection collection percent percent 50 50 with with device device
preselection preselection cyclone cyclone a a with with filter filter ~ ~ a a on on collected collected particles particles Respirable Respirable device. device. size-selective size-selective no no with with filter filter a a in in collected collected ~ ~ matter matter particulate particulate total total Total Total b b
1988a). 1988a). a!. a!. et et (Woskie (Woskie exhaust exhaust diesel diesel railroad railroad of of study study previous previous to to Refers Refers • •
Blank Blank
± ± ± ±
± ± 106 106 ± ± 5 5 9 9 13 13 5 5 130 130 16 16 26 26 Solvent Solvent DMSO DMSO
filter filter
19 19 32 32 Blank Blank III III D0691 D0691
Negative Negative 141 141 279 279 78 78 110 110 T T II II D0134C D0134C
Negative Negative 179 179 336 336 196 196 384 384 R R IV IV D1085A D1085A
Positive Positive 453 453 774 774 1,012 1,012 1,388 1,388 T T IV IV D1080D D1080D
130 130 Negative Negative 195 195 159 108 108 159 T T III III D0690A D0690A
S9) S9) (- Negative Negative
S9) S9) ( ( Positive Positive + + 103 103 348 348 54 54 448 448 R R III III D0689A D0689A
198 198 Positive Positive 820 820 400 400 1,290 1,290 R R III III D0682 D0682
Positive Positive 199 199 686 686 717 717 1,280 1,280 T T III III D0681 D0681
S9 S9 Assayc Assayc + + S9 S9 - S9 S9 S9 S9 + + - No. No. (R)b (R)b Sample Sample No.a No.a
Mutation Mutation
Respirable Respirable
Study Study
TA100 TA100 TA98 TA98
Forward Forward or or Railroad Railroad
Salmonella Salmonella extract) extract) of of J.Lg J.Lg (revertants/100 (revertants/100 (T) (T) Total Total Diesel Diesel
for for Reported Reported Assay Assay Microsuspension Microsuspension from from Activity Activity Mutagenic Mutagenic
Previously Previously
Activity Activity
Filters Filters Railroad Railroad from from Exhaust Exhaust Diesel Diesel of of
Activity Activity Mutagenic Mutagenic of of Assays Assays Mutation Mutation Forward Forward and and Microsuspension Microsuspension of of Comparison Comparison 5. 5. Table Table
al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers
15 Heavy-Duty Diesel Extract (Jlg/plate) Figure 4. Mutagenic activity of heavy-duty diesel engine exhaust extracts
1000~------~ (National Bureau of Standards Reference No. 1650). Each data point repre sents the mean ( ± SD) of triplicate plates. Filter Sample 689A
800
600 in automobile and diesel exhaust (National Research Coun
400 cil 1981; Pitts et al. 1982; Schuetzle et al. 1982; Xu et al. 1982; Nakagawa et al. 1983). The chemical composition of the complex mixture of diesel extract tested with strains 200 TA98 NR-s9 TA98 and TA98 NR can be inferred by the response of the strains to known compounds such as 2-NF and 1-nitropy rene. Generally, other nitro-substituted PAHs (such as the 40 60 80 100 120 0 20 dinitropyrenes) respond differently in that the responses of the two strains do not decrease in potency (Rosenkranz et Diesel Extract (Jlg/plate) al. 1981). Figure 3. Mutagenic activity using the microsuspension procedure for total particulate matter collected at two railroad facilities. Each data point Dose-response curves were determined for the extracts represents the mean ( ± SD) of triplicate plates. (from D. Schuetzle, Ford Motor Co., Dearborn, MI) from the heavy-duty diesel engines using bacterial tester strains TA98 and TA98 NR (Figure 4). The dose-response curves follow the pattern of response reported for other diesel ex tracts (Claxton 1981; National Research Council 1981) in Extract from a single railroad (Filter Sample 1080A in Fig which mutagenic activity, with and without S9, is similar ure 3) was derived from the respirable sized particles and and the activity with strain TA98 NR is substantially de was mutagenic with and without S9 liver homogenate. The creased. The dose-response curves for the extracts from activity greatly decreased with strain TA98 NR. The other railroad-diesel-particulate matter are similar to those for railroad sample (Filter Sample 689A in Figure 3) was from heavy-duty diesel engines. Direct comparisons of muta the total suspended particles collected by a high-volume genic activity between railroad diesel and heavy-duty sampler, and was also mutagenic with and without S9. diesel engine particulate matter (per milligram of particu Again, as in the case of the first sample, the mutagenic ac late matter collected) are difficult, because the conditions tivity in strain TA98 NR was significantly less than that in of collection for the particles are quite different. The parti strain TA98. The insensitivity of tester strain TA98 NR to the cles from heavy-duty diesel engines (the extracts from Ford railroad diesel extracts indicates that the compounds in the Motor Co. were tested) were direct emissions collected from diesel mixtures are nitro-substituted compounds such as a dilution tunnel, whereas the particles collected in the 1-nitropyrene, which is highly mutagenic and has been work areas of diesel railroad employees were airborne sam identified with several other nitro-substituted compounds ples, which are considerably more dilute.
22
23 23
investigation investigation
field field
multidisciplinary multidisciplinary
large, large, this this in in source source railroad, railroad, this this within within population population the the of of size size limited limited the the of of
each each
from from
excluded excluded
or or
missing missing were were samples samples Some Some Because Because railroad. railroad. the the on on years years 10.8 10.8 16.1 16.1 for for ± ± worked worked had had
they they and and years, years, ± ± 9.7 9.7 47.2 47.2 was was 7. 7. subjects subjects of of Table Table age age in in mean mean given given The The are are ect. ect.
study study the the for for proj the the samples samples of of during during number number studied studied total total were were and and subjects subjects male male prevalence, prevalence, ing ing Eighty-seven Eighty-seven
smok ages, ages, mean mean categories, categories, job job The The 6). 6). and and 1 1 (Tables (Tables base base
Population Population Study Study
data data total total the the represent represent 4)] 4)] 151) 151) x x + + [(2 [(2 samples samples 306 306 The The
day. day. single single a a for for data data usable usable provided provided but but 3 3 AIM AIM collection, collection, day's day's SPECIFIC SPECIFIC
second second the the finish finish not not did did subjects subjects four four Another Another subjects. subjects.
87 87 the the studies. studies. by by completed completed epidemiological epidemiological were were in in protocols protocols methodology methodology two-day two-day this this 151 151 of of evaluate evaluate total total A A
and and
characterize characterize to to
necessary necessary be be will will samples samples of of numbers numbers exposure. exposure. exhaust exhaust diesel diesel highest highest
larger larger with with
studies studies Further Further
possible. possible. are are level level sampling" sampling" the the had had who who workers, workers, shop shop among among done done was was resampling resampling
"personal "personal
the the
at at
activity activity
mutagenic mutagenic
of of measurements measurements that that most most The The subjects. subjects. study study same same the the on on sampling sampling repeat repeat
indicates indicates
pumps pumps
sampling sampling
low-volume low-volume to to attached attached filters filters do do to to project project the the in in early early decided decided was was it it workers, workers, exposed exposed
on on
collected collected
matter matter
particulate particulate the the of of study study pilot pilot this this here, here, exhaust diesel diesel highest highest the the oversample oversample to to desire desire the the and and
measured measured 8) 8) (n (n samples samples filter filter = = personal personal of of number number small small
the the with with evaluated evaluated be be not not could could activity activity mutagenic mutagenic urine urine
the the and and airborne airborne the the between between
association association
an an
Although Although
2. 2. day day or or 1 1 day day sampling sampling identical identical on on lected lected
col
were were samples samples
Personal Personal
clerks. clerks. and and
workers workers
shop shop
both both from from
filters filters sonal sonal
samples. samples.
of of
number number
small small
the the of of
because because
made made
per on on particles particles respirable respirable of of extracts extracts from from
activity activity Mutagenic Mutagenic 5. 5.
Figure Figure
be be can can generalizations generalizations no no
but but workers, workers, shop shop
from from filters filters
Worker Worker
ID ID
the the for for than than higher higher was was
clerk clerk the the
from from filters filters the the
of of
tivity tivity
4529 4529
4285 4285
4206 4206
4170 4170
ac
mutagenic mutagenic The The period. period. sampling sampling two-day two-day same same the the from from 0 0
were were filters filters The The 5). 5). (Figure (Figure workers workers shop shop diesel diesel three three from from
filters filters six six of of activities activities the the to to compared compared were were clerk clerk single single 500 500
a a from from acquired acquired filters filters two two of of f- activities activities Mutagenic Mutagenic lished. lished.
<( <(
Ol Ol estab be be could could mutagenicity mutagenicity
urine urine or or 1000 1000 cotinine, cotinine, and and tine tine co co
a: a: nico urine urine
nicotine, nicotine, airborne airborne with with relationship relationship no no small, small,
(]) (])
> >
1500 1500
was was set set
data data the the Since Since m 0.8 0.8 by by . . meter meter
cubic cubic per per revertants revertants
3 (]) (])
t: t:
the the multiplying multiplying by by calculated calculated be be Cll Cll can can filter filter per per activity activity
genic genic
c c
2000 2000
muta approximate approximate
The The here. here. tested tested filters filters all all for for m
0.8 0.8 (f) (f)
3 3 .§ .§
approximately approximately
was was
filter filter per per sampled sampled air air of of volume volume The The
('") ('")
2500 2500
El El -S9 -S9
1987). 1987). al. al. et et
(Louis (Louis S9 S9 with with than than
+S9 +S9 lml lml
2 2
Day Day
S9 S9 without without activity activity greater greater have have air air ambient ambient from from lected lected 3000 3000
col matter matter particulate particulate of of extracts extracts which which in in example, example, for for
reports, reports, are are There There unprecedented. unprecedented. not not is is S9 S9 with with that that over over
S9 S9 without without activity activity mutagenic mutagenic elevated elevated The The samples. samples. borne borne
air personal personal collected collected in in activity activity mutagenic mutagenic examining examining
toward toward effort effort early early an an is is study study pilot pilot Our Our 1987). 1987). al. al. et et (Ling (Ling
assay assay
microsuspension microsuspension the the using using smoke, smoke, cigarette cigarette stream stream Worker Worker ID ID
main and and sidestream sidestream
of of studies studies
pilot pilot
in in seen seen
pattern pattern mal mal 3529 3529 3285 3285 3206 3206 3170 3170
nor the the not not is is This This S9). S9). ( ( + + activation activation metabolic metabolic with with 0 0 than than
S9) S9) (- activation activation metabolic metabolic without without higher higher was was extracts extracts
filter filter these these in in activity activity
mutagenic mutagenic 500 500 The The air. air. of of revertants/m
3 3
f-
2,000 2,000 to to 750 750 approximately approximately was was S9 S9 of of addition addition the the without without
<( <(
Ol Ol
1000 1000 activity activity of of range range The The sampled. sampled. air air
of of
revertants/m 1,500 1,500 co co
3 3
a: a:
to to 250 250
approximately approximately was was
S9) S9) (TA98 (TA98 + + activity activity of of range range the the
(]) (])
> >
1500 1500
workers), workers),
shop shop from from 6 6 and and
clerks clerks from from (6 (6 filters filters 12 12 of of study study (]) (])
t: t:
Cll Cll
pilot pilot this this In In 5). 5). (Figure (Figure periods periods sampling sampling
two-day two-day identical identical c c
2000 2000
in in collected collected filters filters ()j ()j personal personal few few a a on on particles particles respirable respirable
E E
of of extracts extracts from from determined determined was was
activity activity ('") ('") Mutagenic Mutagenic
2500 2500
fZI fZI -S9 -S9
Samplers Samplers
Personal Personal from from I'm I'm
+S9 +S9
1 1 Day Day
3000 3000
Particles Particles Respirable Respirable of of Mutagenicity Mutagenicity
al. al. et et Schenker Schenker B. B. M. M.
24 24
ples ples were were collected. collected. However, However,
8 8 were were missing missing particulate particulate
± ± 87 87 306 306 47.1 47.1 Total Total 51 51 9.7 9.7
A A total total of of 303 303 of of the the possible possible 306 306 personal personal exposure exposure sam
± ±
Shop Shop 23 23
workers workers 158 158 43.2 43.2 16 16 10.2 10.2
Respirable Respirable Particle Particle Exposure Exposure
± ± 8 8 16 16 Engineers Engineers 55.7 55.7 4 4 5.2 5.2
± ± 21 21 60 60 Clerks Clerks 51.0 51.0 15 15 7.6 7.6
± ± tively tively correlated correlated with with reported reported 18 18 depth depth of of 35 35 smoke smoke Carmen Carmen inhalation. inhalation. 45.3 45.3 5 5 9.9 9.9
± ±
Brakers Brakers 17 17 45.3 45.3 37 37 9.7 9.7 11 11 depth depth of of smoke smoke inhalation, inhalation, but but tar tar per per cigarette cigarette was was posi
of of cigarettes cigarettes smoked smoked on on the the study study
shift shift
and and self-reported self-reported
Job Job Category Subjects Subjects Category (Mean± (Mean± Samples Samples smokers smokers SD) SD)
than than 5 5 mg). mg). No No association association was was seen seen between between the the number number Age Age Non-
cigarettes cigarettes with with "ultra-low" "ultra-low" levels levels of of tar tar per per cigarette cigarette (less (less
Table Table 7. 7. Subject Subject and and Sample Sample
Numbers Numbers
by by
Job Job Category Category
However, However, only only two two samples samples were were from from subjects subjects who who smoked smoked
eral eral Trade Trade Commission Commission Report Report 1985), 1985), range range 1.2 1.2 to to 23.3. 23.3.
The The mean mean tar tar per per cigarette cigarette was was 13.1 13.1 5. 5. 2 2 mg mg 98) 98) (Fed ± ± (n (n
= =
rette rette smokers. smokers. Three Three nonsmokers nonsmokers and and two two smokers smokers used used
ter ter the the markers markers present present in in the the urine urine at at the the end end of of the the shift. shift.
Thirty-seven Thirty-seven percent percent of of the the subjects subjects 32) 32) were were ciga = = (n (n
prior prior to to urine urine collection collection was was used, used, because because it it reflected reflected bet
analyses, analyses, the the number number of of cigarettes cigarettes smoked smoked during during the the day day Cigarette Cigarette Smoking Smoking
prior prior to to urine urine sample sample collection collection was was 14.4 14.4 ± ± 8.1. 8.1. For For most most
The The mean mean number number of of cigarettes cigarettes smoked smoked for for the the entire entire day day ing ing the the maximum maximum number number of of samples samples for for separate separate analyses. analyses.
of of cigarettes cigarettes smoked smoked during during the the study study shift shift was was 11.8 11.8 ± ± 6.8. 6.8. power. power. Therefore, Therefore, we we elected elected to to use use the the data data subsets subsets provid
percent percent of of samples). samples). Among Among smokers, smokers, the the average average number number mation mation would would have have seriously seriously reduced reduced the the study study size size and and
smoking smoking were were present present for for the the other other job job categories categories (27 (27 to to 40 40 variables, variables, analyzing analyzing only only the the data data set set with with complete complete infor
the the carmen carmen studied studied (68 (68 percent percent of of samples). samples). Lower Lower rates rates of of cause cause data data were were missing missing for for different different subjects subjects on on different different
The The highest highest rate rate of of cigarette cigarette smoking smoking was was found found among among visit, visit, when when the the method method had had not not yet yet been been developed. developed. Be
in in 12 12 percent percent of of the the samples, samples, primarily primarily were were excluded excluded from from the the or or first first considered considered field field separately. separately.
urine urine was was inadequate. inadequate. Ambient Ambient nicotine nicotine was was not not analyses analyses measured measured of of nonsmokers nonsmokers who who used used other other tobacco tobacco products products
samples; samples; in in most most cases cases this this was was smokers, smokers, because because and and the the they they quantity quantity were were of of analyzed analyzed together. together. Results Results for for urine urine
in in 6 6 percent percent of of samples samples and and without without tobacco tobacco S9 S9 in in products products 10 10 percent percent were were of of similar similar to to those those for for cigarette cigarette
urine urine remaining. remaining. Mutagenicity Mutagenicity was was four four not not subjects. subjects. measured measured Results Results with with for for S9 S9 cigarette cigarette smokers smokers who who used used other other
urine urine assays, assays, and and there there was was often often an an insufficient insufficient quantity quantity percent). percent). of of Smoking Smoking status status was was unknown unknown on on the the study study day day for for
latter latter measurements measurements because because they they were were done done after after ples ples all all for for other other nonsmokers nonsmokers who who used used other other tobacco tobacco products products (4 (4
nicotine nicotine and and cotinine. cotinine. Missing Missing samples samples were were high high samples samples for for these these for for nonusers nonusers of of tobacco tobacco (62 (62 percent), percent), and and 12 12 sam
percent percent for for urinary urinary thiocyanate thiocyanate to to 12 12 percent percent for for smokers smokers urinary urinary who who used used other other tobacco tobacco products products (3 (3 percent), percent), 189 189
variables. variables. The The number number of of missing missing cigarette cigarette samples samples smokers smokers ranged ranged (32 (32 from from percent), percent), 1 1 10 10 samples samples for for cigarette cigarette
(Table (Table 6). 6). The The reasons reasons for for this this were were other other different different forms forms for for of of the the tobacco. tobacco. specific specific This This yielded yielded 97 97 urine urine samples samples for for
Total Total Number Number Possible Possible 189 189 95 95 10 10 12 12 306 306
Nicotine Nicotine 175 175 85 85 10 10 10 10 8 8 280 280
Respirable Respirable particles particles 181 181 92 92 10 10 12 12 4 4 295 295
Personal Personal Sampling Sampling Pump Pump
Cotinine Cotinine 168 168 84 84 8 8 10 10 12 12 270 270
Nicotine Nicotine 167 167 84 84 8 8 10 10 12 12 269 269
Thiocyanate Thiocyanate 188 188 93 93 10 10 11 11 1 1 302 302
Mutagenicity Mutagenicity 170 170 - S9 S9 89 89 9 9 8 8 10 10 276 276
S9 S9 Mutagenicity Mutagenicity + + 178 178 92 92 9 9 9 9 6 6 288 288
Urine Urine
Tobacco Tobacco Smokers Smokers Users Users Tobacco Tobacco Users Users (%) (%) Total Total
Nonusers Nonusers of of Cigarette Cigarette Tobacco Tobacco Only Only Other Other Missing Missing
Only Only and and Other Other
Cigarette Cigarette
Table Table 6. 6. Number Number of of Samples Samples Analyzed Analyzed by by Smoking Smoking Group Group (Before (Before Exclusions) Exclusions)
Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers
25 25
repair. repair. during during locomotives locomotives house house that that does does than than fume fume metal metal more more substantially substantially generates generates ting ting
garages garages
large large in in
work work
workers workers shop shop the the and and outdoors, outdoors, work work cut Also, Also, 3). 3). (Table (Table minutes minutes 15 15 to to 1 1 of of periods periods short short for for weld weld
to to tend tend
brakers brakers
and and
carmen carmen areas, areas, indoor indoor small small in in work work generally generally who who workers, workers, shop shop locomotive locomotive among among smaller smaller
both both
engineers engineers
and and
Clerks Clerks outdoors. outdoors. work work and and areas; areas; open open much much was was difference difference This This carmen. carmen. the the among among welders welders
large large in in
indoors indoors
work work
spaces; spaces; enclosed enclosed small small relatively relatively in in non and and welders welders the the of of exposures exposures average average the the in in seen seen
indoors indoors
work work
characteristics: characteristics: enclosure enclosure on on based based categories categories was was difference difference large large A A day. day. per per hours hours four four to to hour hour one-half one-half
three three
to to
groups groups
job job
assigned assigned We We area. area. work work the the of of closure closure occupied occupied activity activity of of type type this this repairs; repairs; during during cars cars train train of of
en of of
degree degree the the
is is
exposure exposure in in factor factor important important Another Another chassis chassis the the burning burning and and cutting cutting for for torch torch acetylene acetylene an an using using
by by or or welding welding by by generated generated matter matter particulate particulate to to groups. groups. job job exposed exposed
were were the the them them of of between between some some seen seen because because part, part, difference difference in in the the of of all all resulted, resulted, This This explain explain posure. posure. cannot cannot this this
ex exhaust exhaust However, However, diesel diesel Jlg/m no no or or 30 30
little little about about . . of of have have they they concentration concentration though though air air even even cles, cles, background background 3
a a to to parti exposed exposed are are respirable respirable to to workers workers all all exposures exposures that that highest highest the the assuming assuming had had carmen carmen locomotives, locomotives, The The
diesel diesel
operating operating
with with have have
groups groups these these contact contact relative relative the the groups. groups. job job most most in in samples samples of of number number small small
by by
expected expected
that that to to close close
was was
carmen, carmen, welding welding excluding excluding 2), 2), relatively relatively the the to to due due be be may may which which distributions, distributions, skewed skewed
(Table (Table
particles particles respirable respirable
adjusted adjusted of of concentration concentration by by jobs jobs of of pattern pattern clear clear or or consistent consistent a a show show not not did did centrations centrations
the the of of
ranking ranking
relative relative
The The
apparent. apparent. more more are are exposures exposures con particle particle respirable respirable The The nicotine. nicotine. no no or or little little contained contained
exhaust exhaust
diesel diesel
relative relative the the
value, value, particle particle respirable respirable justed justed that that nonsmokers nonsmokers from from samples samples of of number number large large tively tively
ad
an an give give to to
subtracted subtracted
is is value value particle particle respirable respirable each each rela the the by by part part in in caused caused was was which which smoke), smoke), tobacco tobacco tal tal
to to
smoke smoke
tobacco tobacco
environmental environmental
of of contribution contribution the the If If (environmen concentrations concentrations nicotine nicotine ambient ambient in in tribution tribution
dis smokers. smokers. of of skewed skewed showed showed percentage percentage groups groups highest highest job job the the had had ofthe ofthe All All carmen carmen 7). 7). The The (Figure (Figure clerks. clerks. workers workers
the the all all and and for for engineers engineers the the concentration concentration by by smoke smoke followed followed tobacco tobacco carmen, carmen, the the environmental environmental among among bient bient
am the the in in found found and and were were 6), 6), smoke smoke (Figure (Figure tobacco tobacco workers workers shop shop environmental environmental of of locomotive locomotive the the of of concentrations concentrations
mean mean highest highest concentrations concentrations The The 4). 4). particle particle (Table (Table respirable respirable smoke smoke of of tobacco tobacco histograms histograms the the in in vironmental vironmental seen seen
en by by be be can can contributed contributed mass mass skewness skewness This This particle particle exposures. exposures. respirable respirable the the to to airborne airborne of of largely largely distributions distributions
the the groups groups in in job job observed observed between between commonly commonly concentrations concentrations skewness skewness particle particle the the reveal reveal respirable respirable of of quartiles quartiles
and and similarity similarity this this medians medians The The attribute attribute 2. 2. We We Table Table in in 1988a). 1988a). al. al. et et shown shown are are (Woskie (Woskie smoke) smoke) railroads railroads tobacco tobacco
U.S. U.S. from from environmental environmental findings findings minus minus previous previous our our with with concentration concentration consistent consistent particle particle is is which which (respirable (respirable
mass mass similar, similar, particle particle were were 2) 2) respirable respirable (Table (Table adjusted adjusted groups groups job job and and the the of of smoke, smoke, tobacco tobacco tal tal concentrations concentrations cle cle
parti respirable respirable the the environmen clerks, clerks, mass, mass, the the and and particle particle workers workers respirable respirable shop shop of of the the tween tween concentrations concentrations the the
be for for exposure exposure statistics statistics exhaust exhaust Summary Summary diesel diesel 6). 6). in in (Table (Table range range wide wide nicotine nicotine of of the the Despite Despite measurements measurements
280 280
and and
particles particles
respirable respirable to to exposure exposure personal personal of of ments ments 2). 2). (Table (Table high high still still although although groups, groups, other other
measure 295 295 was was
set set data data final final The The values. values. nicotine nicotine ing ing the the of of range range the the within within are are carmen carmen the the for for concentrations concentrations
miss
were were 23 23
and and
damage, damage,
filter filter of of result result a a as as values values mass mass particle particle respirable respirable the the group, group, carmen carmen the the from from removed removed are are
work) work) torch torch doing doing those those (including (including welders welders the the If If welding. welding.
status. status. smoking smoking by by workers, workers, shop shop locomotive locomotive among among
concentrations concentrations particle particle respirable respirable of of distribution distribution Frequency Frequency 6. 6. Figure Figure
8.6. 8.6.
x x
nicotine nicotine as as
lated lated
Midpoint Midpoint
(J.Lg/m
) )
Concentration Concentration
Particle Particle
Respirable Respirable
calcu is is concentration concentration smoke smoke Tobacco Tobacco status. status. 3 smoking smoking by by subjects, subjects, all all for for
exposures exposures smoke smoke
tobacco tobacco
environmental environmental of of
distribution distribution
Frequency Frequency
7. 7.
Figure Figure
525 575 575 525 425 475 475 425 325 375 375 325 275 275 225 225 125 175 175 125 75 75 25 25
(J.Lg/m ) ) Concentration Concentration Smoke Smoke
Tobacco Tobacco
3
525 525 475 475 425 425 325 375 375 325 275 275 225 225 175 175 125 125 25 25 75 75
0 0
10 10
u.. u..
~ ~
50 50
u.. u..
0" 0"
~ ~
::l ::l
0" 0"
(]) (])
c c ::l ::l 20 20
u u (]) (])
>. >. c c
u u
100 100 >. >.
Smokers Smokers II II
II II Smokers Smokers
30 30 D D Nonsmokers Nonsmokers
D D Nonsmokers Nonsmokers
150 150
al. al. et et Schenker Schenker B. B. M. M.
26 26
provided provided a a more more precise precise estimate estimate of of concentration; concentration; the the phenanthrene. phenanthrene. Only Only a a weak weak relationship relationship was was found found be-
phenanthrene phenanthrene than than any any of of the the personal personal samplers, samplers, and and so so be be
only only ng/m and and 11 11 would would not not , , be be a a significant significant source source of of 3
volume volume sampler sampler collected collected substantially substantially more more total total mass mass of of in in 11g/m environmental environmental 120 120 tobacco tobacco
smoke smoke particles particles would would 3 3
one one high-volume high-volume sample) sample) also also were were analyzed. analyzed. The The high level level found found in in the the offices; offices; therefore, therefore, the the phenanthrene phenanthrene level level
polyurethane polyurethane
foam foam plug plug samples samples personal personal samples samples (16 (16 and and proximately proximately 11g/m 1,200 1,200 which which was was about about
times times the the , , 10 10 3
clerks, clerks, carmen, carmen, and and shop shop workers. workers. Seventeen Seventeen field field blank blank respirable respirable particle particle concentrations concentrations in in the the chamber chamber were were ap
sonal sonal samples samples were were chosen chosen from from three three major major job job groups: groups: tained tained phenanthrene phenanthrene above above the the limit limit of of detection, detection, the the
area area samples samples from from the the repair repair shop shop were were analyzed. analyzed. The The per though though exposure exposure chamber chamber cigarette cigarette smoke smoke samples samples con
samples, samples, one one set set of of high-volume high-volume samples, samples, and and three three sets sets of of posures posures that that were were just just above above the the limit limit of of detection. detection. Al
were were analyzed. analyzed. Polyurethane Polyurethane foam foam plugs plugs from from personal personal 105 105 amounts amounts of of phenanthrene; phenanthrene; and and most most of of the the clerks clerks had had ex
maining maining field field samples samples only only the the polyurethane polyurethane foam foam plugs plugs 8); 8); most most of of the the carmen carmen were were exposed exposed to to less less than than detectable detectable
polyurethane polyurethane foam foam plugs plugs rather rather than than the the filters, filters, so so for for all all re shop shop workers workers were were well well above above the the limit limit of of detection detection (Figure (Figure
(more (more
than than 90 90 percent) percent) of of the the phenanthrene phenanthrene was was on on the the 66 66 + + ng/m 2(38) 2(38) The The 142 142 exposures exposures of of nearly nearly
all all the the = = . . 3
of of the the first first or or so so field field 20 20 samples samples showed showed that that virtually virtually all all tection tection was was defined defined as as the the value value for for the the blank blank mean mean SD: SD: + + 2 2
the the polyurethane polyurethane foam foam plugs plugs after after sampling, sampling, the the analyses analyses air, air, m as as the the 0.8 0.8 personal personal filter filter
samples). samples). , , The The limit limit of of de 3
cent cent of of the the phenanthrene phenanthrene spiked spiked onto onto filters filters was was found found on on (expressed (expressed as as if if they they were were samples samples of of the the same same amount amount of of
the the repair repair shop. shop. As As in in the the laboratory laboratory tests, tests, in in which which per 68 68 of of these these groups groups and and for for the the blank blank polyurethane polyurethane foam foam plugs plugs
large-diameter large-diameter polyurethane polyurethane foam foam plugs plugs were were collected collected in in range range of of phenanthrene phenanthrene concentration concentration (mean (mean SD) SD) for for each each ± ±
few few high-volume high-volume samples samples Llminute) Llminute) (430 (430 that that used used three three (nonwelders), (nonwelders), and and repair repair shop shop workers. workers. Figure Figure 8 8 shows shows the the
used used to to collect collect area area samples samples in in the the repair repair shop; shop; finally, finally, a a 94 94 were were taken taken in in the the three three major major job job groups: groups: clerks, clerks, carmen carmen
breathing breathing zone; zone; second, second, the the same same sampling sampling devices devices were were Of Of the the personal personal samples samples 104 104 analyzed analyzed for for phenanthrene, phenanthrene,
hundred hundred
personal personal samples samples were were collected collected in in the the workers' workers' aged aged diesel diesel exhaust. exhaust.
phenanthrene phenanthrene
exposure exposure
in in the the railroad railroad
jobs. jobs. First, First,
several several represent, represent, predominantly, predominantly, the the phenanthrene phenanthrene content content of of
Three Three
types types
of of samples samples
were were collected collected
to to determine determine
the the
ble ble particles particles (with (with mean mean = = and and SD SD 0.006 0.006 = = and and 0.0008), 0.0008),
samples samples
had had very very
similar similar ratios ratios of of phenanthrene phenanthrene to to respira Phenanthrene Phenanthrene Exposure Exposure
threne threne and and J.tg/m respirable respirable 34 34 particles. particles. These These limited limited
3 3
third third day day of of the the same same week week collected collected ng/m phenan
176 176
smokers. smokers. 3 3
J.tg/m
respirable respirable
particles, particles,
while while
a a personal personal
sampler sampler
on on
the the
particle particle exposures exposures were were also also 3 3 similar similar for for smokers smokers and and non
repair repair
shed, shed, averaged averaged
ng/m phenanthrene phenanthrene 230 230
and and 40 40
their their nonsmoking nonsmoking colleagues, colleagues, but but the the adjusted adjusted respirable respirable 3 3
day day
other other than than the the
high-volume high-volume
collection collection
day day
in in the the same same
approximately approximately double double the the respirable respirable particle particle exposures exposures of of
samples, samples,
collected collected
with with the the
personal personal
sampling sampling
train train on on a a smokers smokers and and nonsmokers. nonsmokers. Outside Outside and and shop shop smokers smokers had had
respirable respirable
particle particle
concentration concentration
was was
J.tg/m
Two Two 75 75 area area
. .
the the adjusted adjusted respirable respirable particle particle values values were were similar similar for for 3
phenanthrene phenanthrene concentration concentration
was was ng/m
420 420 and and the the
, ,
reduced reduced these these differences differences considerably. considerably. For For inside inside workers, workers, 3
ticle ticle values values by by correcting correcting for for environmental environmental tobacco tobacco smoke smoke
than than those those of of the the nonsmokers. nonsmokers. Adjustment Adjustment of of respirable respirable par
sana! sana! samples. samples.
Blanks Blanks are are expressed expressed as as air air concentrations concentrations respirable respirable in in the the same same particle particle volume volume exposures exposures as as the the per· per· of of the the smokers smokers were were higher higher
the the respirable respirable particle particle concentrations concentrations of of job job categories categories
(see (see text text and and Table Table 4). 4).
posures posures in in smokers smokers and and nonsmokers. nonsmokers. As As expected, expected,
the the
respirable respirable particle particle concentration concentration 1,200 1,200 = = 11g/m or or approximately approximately 10 10 times times
3 3
anthrene anthrene values values are are given given in in means means 2 2 SD. SD. For For chamber chamber ence ence ± ± cigarette cigarette was was smoke, smoke, present present for for environmental environmental tobacco tobacco smoke smoke ex
Figure Figure 8. 8. Distribution Distribution of of phenanthrene phenanthrene concentrations concentrations by by job job group. group.
Phen· Phen·
large large spaces spaces (outdoors (outdoors or or in in repair repair shops), shops), a a distinct distinct differ
Job Job
Category Category
left left in in ashtrays ashtrays for for long long periods periods of of time. time. Among Among Smoke Smoke workers workers in in
Cigarette Cigarette Wor1 tics tics of of indoor indoor workers; workers; for for example, example, burning burning cigarettes cigarettes are are Chamber Chamber Shop Shop Carmen Carmen Cler1 0 0 areas. areas. This This may may be be due, due, in in part, part, to to the the smoking smoking characteris n=3 n=3 n=62 n=62 centrations centrations for for smokers smokers in in these these same same small small enclosed enclosed work work a.. a.. .<:: .<:: side side were were equivalent equivalent to to environmental environmental tobacco tobacco smoke smoke con 100 100 Q) Q) <:: <:: environmental environmental tobacco tobacco smoke smoke for for Ill Ill nonsmokers nonsmokers working working in c c .<:: .<:: smoker's smoker's particle particle exposure exposure (Table (Table The The 4). 4). concentrations concentrations for for ~ ~ 200 200 <:: <:: small small enclosed enclosed spaces spaces and and significantly significantly contribute contribute to to a a non Q) Q) -S -S strating strating that that environmental environmental tobacco tobacco smoke smoke can can build build up up in in 0, 0, E E posures posures of of either either 300 300 the the outside outside or or shop shop nonsmokers, nonsmokers, demon "'~ "'~ door, door, small-space small-space nonsmokers nonsmokers were were higher higher than than the the ex The The environmental environmental tobacco tobacco smoke smoke exposures exposures of of the the in 400 400 Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers 27 27 < < < < 0.05. 0.05. p p 0.01 0.01 b b < < 0.001. 0.001. p p a a 0.47a 0.47a cotinine cotinine Urinary Urinary -0.06 -0.06 0.46a 0.46a thiocyanate thiocyanate Urinary Urinary 0.58a 0.58a 0.08 0.08 0.38a 0.38a sample sample of of day day on on smoked smoked Cigarettes Cigarettes 0.18 0.18 0.07 0.07 0.15 0.15 0.15b 0.15b S9 S9 - 0.54a 0.54a 0.51a 0.51a o.2ob o.2ob 0.38a 0.38a 0.52a 0.52a S9 S9 + + mutagenicity mutagenicity Urinary Urinary Nicotine Nicotine Cotinine Cotinine Thiocyanate Thiocyanate Smoked Smoked S9 S9 S9 S9 - + + Urinary Urinary Urinary Urinary Urinary Urinary Cigarettes Cigarettes Mutagenicity Mutagenicity Smokers Smokers Cigarette Cigarette from from Samples Samples for for Coefficients Coefficients Correlation Correlation Order Order Rank Rank Spearman Spearman 8. 8. Table Table sub- the the in in used used not not was was thiocyanate thiocyanate mutagenicity, mutagenicity, urinary urinary only. only. workers workers shop shop for for concentration concentration particle particle respirable respirable justed justed or or dose dose smoking smoking and and thiocyanate thiocyanate of of correlation correlation lower lower the the ad and and concentration concentration particle particle respirable respirable versus versus Phenanthrene Phenanthrene 9. 9. Figure Figure of of Because Because nicotine. nicotine. of of half-life half-life the the than than longer longer are are cotinine cotinine (llg/m ) ) Particles Particles Respirable Respirable Adjusted Adjusted 3 and and thiocyanate thiocyanate of of half-lives half-lives the the that that reflecting reflecting perhaps perhaps > > 0.05), 0.05), p p 0.06, 0.06, (r (r nicotine nicotine 600 600 with with none none < < but but - = 400 400 0.001) 0.001) p p 200 200 0.46, 0.46, 0 0 -200 -200 0 0 (r (r cotinine cotinine with with = = correlation correlation high high a a show show did did Thiocyanate Thiocyanate .. .. > > . . 0.05) 0.05) p p 0.08, 0.08, (r (r thiocyanate thiocyanate = = urinary urinary and and day day study study the the ~ ~ ...... on on smoked smoked cigarettes cigarettes of of number number between between association association no no was was ...... , , 1- 200 200 .. .. a.. a.. there there however, however, smokers, smokers, among among nonsmokers; nonsmokers; and and smokers smokers ,.a: 1-tf 1-tf ~ ~ .. .. .. .. .. .. Q) Q) between between discriminated discriminated thiocyanate thiocyanate Urinary Urinary nicotine. nicotine. c c t1! t1! .. .. E E .. .. urinary urinary of of half-life half-life biologic biologic shorter shorter the the < < reflecting reflecting 0.01), 0.01), p p 400 400 ...... ~ ~ = = 0.38, 0.38, ~ ~ (r (r cotinine cotinine urinary urinary for for < < than than 0.001) 0.001) p p = = 0.58, 0.58, (r (r .. .. c c nicotine nicotine urinary urinary for for stronger stronger was was association association This This ure ure 11). 11). Q) Q) s s 600 600 .. .. Fig 8; 8; (Table (Table cotinine cotinine and and nicotine nicotine urinary urinary and and day day study study 0, 0, E E the the on on smoked smoked cigarettes cigarettes of of number number the the between between seen seen also also M~ M~ was was relationship relationship strong strong A A 10). 10). Figure Figure < < 8; 8; (Table (Table .. .. 0.05) 0.05) p p 0.15, 0.15, BOO BOO S9 S9 (r (r without without = = mutagenicity mutagenicity urine urine with with association association cant cant signifi but but weak weak a a < < and and 0.001), 0.001), p p 0.38, 0.38, r r (Spearman (Spearman S9 S9 = = 1000 1000 with with mutagenicity mutagenicity urine urine with with relationship relationship dose-response dose-response strong strong a a showed showed studied studied shift shift work work the the during during smoked smoked (llg/m ) ) Particles Particles Respirable Respirable 3 cigarettes cigarettes of of number number the the smokers, smokers, cigarette cigarette Among Among 600 600 500 500 400 400 300 300 0 0 200 200 100 100 -100 -100 Smoking Smoking Cigarette Cigarette Active Active of of Markers Markers 04-~--r-~~--~,--r-,--~-r~----~~ 04-~--r-~~--~,--r-,--~-r~----~~ .. .. 4 4 AIM AIM SPECIFIC SPECIFIC ...... . . ., ., ...... 200 200 .. .. •: •: I I ...... Jl. Jl. a.. a.. ...... •(A! •(A! ~· ~· .. .. .. .. paper. paper. .. .. ~ .. .. Q) Q) .. .. c c log-probability log-probability on on plots plots ...... linear linear .. .. nearly nearly gave gave data data the the and and t1! t1! .. .. .. .. E E 400 400 .. .. ...... log-normal, log-normal, approximately approximately were were concentrations concentrations anthrene anthrene ~ ~ ~ ~ phen of of distributions distributions The The values. values. particle particle .. .. respirable respirable low low bly bly c c Q) Q) unreasona had had three three all all and and SD, SD, 2 2 mean mean + + sample sample area area the the 600 600 s s .. .. than than larger larger ratios ratios had had samples samples three three Only Only 0.003. 0.003. averaged averaged 0, 0, E E workers workers shop shop nonsmoking nonsmoking of of samples samples personal personal the the in in ticles ticles M"' M"' .. .. 800 800 par respirable respirable adjusted adjusted to to phenanthrene phenanthrene of of ratio ratio The The 9). 9). ure ure (Fig concentrations concentrations particle particle respirable respirable adjusted adjusted or or particle particle respirable respirable either either and and concentration concentration phenanthrene phenanthrene tween tween 1000 1000 al. al. et et Schenker Schenker B. B. M. M. 28 28 was was used used as as an an indicator indicator of of passive passive smoking smoking to to evaluate evaluate they they did did the the not not achieve achieve statistical statistical significance. significance. Nicotine Nicotine concentration concentration on on the the personal personal sampling sampling filters filters ables ables were were entered entered in in the the regression regression full models models even even when when particle particle concentration concentration in in the the regression regression models. models. Both Both vari Markers Markers of of Passive Passive Cigarette Cigarette Smoking Smoking of of environmental environmental tobacco tobacco smoke smoke to to measured measured respirable respirable posures. posures. Ambient Ambient nicotine nicotine also also adjusts adjusts for for the the contribution contribution the the models models before before testing testing for for an an effect effect of of diesel diesel exhaust. exhaust. dose, dose, including including nonworkplace nonworkplace and and previous-work-shift previous-work-shift ex ber ber of of cigarettes cigarettes smoked smoked on on the the study study day day were were entered entered in in flects flects a a longer-term longer-term average average of of environmental environmental tobacco tobacco smoke smoke mutagenicity mutagenicity among among smokers, smokers, both both cotinine cotinine and and the the num work work shift, shift, and and urinary urinary cotinine cotinine a1llong a1llong nonsmokers nonsmokers re To To control control maximally maximally for for cigarette cigarette smoking smoking in in models models of of environmental environmental tobacco tobacco smoke smoke exposure exposure during during the the study study urinary urinary mutagenicity mutagenicity ( ( S9) S9) in in addition addition to to urinary urinary + + cotinine. cotinine. cotinine. cotinine. Ambient Ambient nicotine nicotine may may be be considered considered a a marker marker of of study study day day remained remained a a significant significant independent independent predictor predictor for for trolled trolled for for by by entering entering both both ambient ambient nicotine nicotine and and urinary urinary smokers. smokers. However, However, the the number number of of cigarettes cigarettes smoked smoked on on the the markers markers among among nonsmokers, nonsmokers, passive passive smoking smoking was was con just just for for smoking smoking when when testing testing for for a a diesel diesel effect effect among among In In the the multivariate multivariate modeling modeling of of urinary urinary mutagenicity mutagenicity achieved achieved with with cotinine, cotinine, which which was was used, used, therefore, therefore, to to ad tine tine measured measured on on the the work work shift. shift. highest highest predictive predictive of of value the the urine urine markers markers was was generally generally show show the the same same strength strength of of association association with with ambient ambient nico achieved achieved the the same same level level of of statistical statistical significance. significance. The The posures posures prior prior to to the the study study work work day day and, and, therefore, therefore, not not regression regression model, model, the the other other smoking smoking markers markers no no longer longer with with its its longer longer half-life, half-life, may may reflect reflect nonoccupational nonoccupational ex When When urinary urinary cotinine cotinine (or (or nicotine) nicotine) was was added added to to the the day day up up until until the the urine urine sample sample collection. collection. Urine Urine cotinine, cotinine, cant cant independent independent predictors predictors in in these these models. models. number number of of cigarettes cigarettes smoked smoked by by coworkers coworkers during during the the work work (Federal (Federal Trade Trade Commission Commission Report Report were were not not 1985) 1985) signifi logic logic half-life half-life of of nicotine, nicotine, and and appears appears to to correspond correspond to to the the mutagenicity mutagenicity without without S9. S9. Tar Tar and and nicotine nicotine per per cigarette cigarette shown). shown). This This association association consistent consistent is with with the the shorter shorter bio association association was was also also present present for for these these two two variables variables and and of of the the smaller smaller sample sample size size (r (r 0.26, 0.26, p p data data not not 0.13; 0.13; = = = = independent independent predictors predictors of of mutagenicity mutagenicity with with S9. S9. A A weak weak not not achieve achieve the the same same level level of of statistical statistical significance significance because because of of cigarettes cigarettes smoked smoked and and smoke smoke inhalation inhalation were were significant significant tween tween ambient ambient and and urinary urinary nicotine, nicotine, but but the the association association did did were were excluded excluded from from the the prediction prediction equations, equations, the the number number mental mental tobacco tobacco smoke, smoke, a a stronger stronger association association was was seen seen be minants minants of of urinary urinary mutagenicity. mutagenicity. When When urinary urinary markers markers workers, workers, the the group group with with the the highest highest exposure exposure to to environ smoke smoke inhalation) inhalation) were were next next tested tested as as independent independent deter response response associations associations were were not not present: present: Among Among indoor indoor study study day, day, tar tar per per cigarette, cigarette, nicotine nicotine per per cigarette, cigarette, and and highest highest environmental environmental tobacco tobacco smoke smoke exposure, exposure, but but dose All All of of the the smoking smoking variables variables (cigarettes (cigarettes smoked smoked on on the the Urinary Urinary cotinine cotinine and and thiocyanate thiocyanate showed showed increases increases for for the the posure posure with with urinary urinary mutagenicity. mutagenicity. for for urinary urinary nicotine nicotine (r (r = = = = p p 0.15, 0.15, (Table (Table 9; 9; 0.06) 0.06) Figure Figure 12). 12). cigarette cigarette smoking smoking in in testing testing for for an an association association of of diesel diesel ex response response relationship relationship was was seen seen for for nicotine nicotine on on the the filter filter and and sequent sequent multivariate multivariate models models that that controlled controlled for for the the effect effect of of various various urinary urinary markers. markers. Among Among nonsmokers, nonsmokers, a a weak weak dose did did not not smoke smoke on on the the study study shift. shift. the the study study shift. shift. on on number number of of cigarettes cigarettes smoked smoked on on the the study study shift shift and and exclude exclude smokers smokers who who smoked smoked on on the the study study shift shift and and exclude exclude smokers smokers who who did did not not smoke smoke during during creatinine) creatinine) x x 10; 10; cotinine, cotinine, nicotine nicotine l!gll!mol l!gll!mol of of creatinine. creatinine. ~ ~ Data Data are are based based (revertants/l!mol (revertants/l!mol of of creatinine). creatinine). The The data data are are based based on on number number of of cigarettes cigarettes tile tile concentration concentration in in urine urine calculated calculated as as follows: follows: thiocyanate thiocyanate (mg/l!mol (mg/l!mol of of ~ ~ shift. shift. Urine Urine mutagenicity mutagenicity is is expressed expressed as as median median and and quartile quartile mutagenicity mutagenicity smoked smoked on on study study shift. shift. Urine Urine mutagenicity mutagenicity is is expressed expressed as as median median and and quar Figure Figure 10. 10. Urine Urine mutagenicity mutagenicity by by number number of of cigarettes cigarettes smoked smoked on on study study Figure Figure 11. 11. Urinary Urinary markers markers of of cigarette cigarette smoking smoking by by number number of of cigarettes cigarettes TA98 TA98 +89 +89 Thiocyanate Thiocyanate Cotinine Cotinine TA98-S9 TA98-S9 Nicotine Nicotine 0 0 0 0 ::J ::J ·c ·c ::J ::J c c 100 100 ·c ·c ctl ctl c c c c 2 2 ctl ctl c c () () 200 200 3 3 0 0 () () c c 0 0 () () 4 4 c c Q) Q) () () c c 300 300 Q) Q) c c 5 5 ~ ~ c c II II C'") C'") ~ ~ 0 0 6 6 0 0 B B B B + + 11 11 Cigarettes/Shift Cigarettes/Shift 11+ 11+ Cigarettes/Shift Cigarettes/Shift c c 400 400 0 0 1::::1 1::::1 7 7 1::::1 1::::1 1-1 1-1 0 0 Cigarettes/Shift Cigarettes/Shift c c 1-10 1-10 Cigarettes/Shift Cigarettes/Shift c c ~ ~ D D D D Nonsmokers Nonsmokers II II Nonsmokers Nonsmokers 8 8 C'") C'") Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers 29 29 of of effect effect the the models, models, these these In In D.2). D.2). and and D.1 D.1 (Tables (Tables similar similar inine. inine. creat of of Jlg/Jlmol Jlg/Jlmol are are values values nicotine nicotine and and cotinine cotinine creatinine; creatinine; of of mg/11mol mg/11mol are are conclusions conclusions the the models, models, exposure-interaction exposure-interaction the the For For are are values values Thiocyanate Thiocyanate quartiles. quartiles. upper upper and and medians medians as as given given are are urine urine in in Concentrations Concentrations pump. pump. sampling sampling personal personal on on error. error. nicotine nicotine of of strata strata standard standard by by its its ers, ers, of of half half than than more more not not was was group group posed posed nonsmok among among smoking smoking cigarette cigarette passive passive of of markers markers Urinary Urinary 12. 12. Figure Figure unex the the from from difference difference mean mean estimated estimated the the group group posed posed Cotinine Cotinine Nicotine Nicotine Thiocyanate Thiocyanate ex every every for for and and association, association, dose-response dose-response of of evidence evidence no no showed showed means means predicted predicted The The respectively. respectively. S9, S9, without without and and :::l :::l with with > > mutagenicity mutagenicity for for 0.05) 0.05) (p (p 0.21 0.21 and and 2.09 2.09 are are groups groups ·c: ·c: "' "' c: c: exposure exposure the the among among differences differences for for freedom) freedom) of of degrees degrees 2:- 10 10 () () (3 (3 statistics statistics chi-squared chi-squared ratio ratio likelihood likelihood The The 11). 11). (Table (Table 0 0 c: c: ~ ~ respectively respectively J.Lg/m 181.4 181.4 and and , , 110.0, 110.0, 50.2, 50.2, were were models models the the 3 E E ~ ~ in in used used exposure exposure exhaust exhaust diesel diesel of of strata strata high high and and medium, medium, 5 5 20 20 low, low, the the in in particles particles respirable respirable adjusted adjusted of of levels levels median median The The 10). 10). (Table (Table tobacco tobacco of of nonusers nonusers the the among among S9, S9, without without or or with with mutagenicity, mutagenicity, on on effect effect an an of of evidence evidence no no showed showed ~ ~ Nicotine Nicotine • • variable variable particle particle respirable respirable adjusted adjusted the the of of Stratification Stratification fa fa Nicotine Nicotine 0.1~<5 0.1~<5 [J [J Nonsmokers Nonsmokers variability). variability). assay assay + + Concentration Concentration Nicotine Nicotine Air Air between for for normalize normalize (to (to 2-AF 2-AF control, control, mutagen mutagen standard standard to to response response and and smoke), smoke), tobacco tobacco environmental environmental (for (for nine nine coti urinary urinary food, food, protective protective for for controlled controlled and and variances, variances, smokers smokers from from samples samples for for Similarly, Similarly, 0.07. 0.07. 0.51 0.51 was was it it ± ± 153), 153), between-person between-person and and within- for for allowed allowed models models regression regression foods foods same same (n (n the the consume consume not not = = did did who who nonsmokers nonsmokers The The D.3). D.3). and and D.2, D.2, 10, 10, (Tables (Tables S9 S9 without without or or with with either either from from samples samples for for and and ± ± 0.12, 0.12, 0.20 0.20 was was 25) 25) collection collection (n (n = = mutagenicity mutagenicity urinary urinary of of predictor predictor independent independent significant significant sample sample to to prior prior days days two two the the in in foods foods protective protective consumed consumed a a not not was was exposure exposure exhaust exhaust diesel diesel samples, samples, these these For For S9. S9. who who nonsmokers nonsmokers from from samples samples for for SE) SE) ( ( ± ± mutagenicity mutagenicity without without mutagenicity mutagenicity for for analyzed analyzed be be could could samples samples 121 121 of of level level mean mean The The nonsmokers. nonsmokers. and and smokers smokers among among S9 S9 and and S9, S9, with with mutagenicity mutagenicity for for analyzed analyzed be be could could tobacco tobacco of of with with mutagenicity mutagenicity urinary urinary lower lower with with associated associated was was study study nonusers nonusers from from samples samples 124 124 exclusions, exclusions, After After Nonsmokers. Nonsmokers. the the to to prior prior days days two two the the in in fish) fish) or or sprouts, sprouts, Brussels Brussels bage, bage, cab is, is, (that (that foods" foods" "protective "protective of of ingestion ingestion self-reported self-reported Exposure Exposure Exhaust Exhaust Diesel Diesel Only Only exposures). exposures). chemical chemical with with hobbies hobbies dyes, dyes, hair hair ing, ing, weld pesticides, pesticides, example, example, (for (for exposures exposures nonoccupational nonoccupational nonsmokers. nonsmokers. or or and and meat), meat), grilled grilled example, example, (for (for days days two two previous previous the the smokers smokers in in S9 S9 without without mutagenicity mutagenicity of of predictive predictive not not was was during during intake intake dietary dietary psoriasis), psoriasis), cancer, cancer, example, example, (for (for tions tions variable variable food food protective protective The The D.2). D.2). and and D.1 D.1 Tables Tables D, D, pendix pendix condi clinical clinical of of consisted consisted They They predictors. predictors. independent independent (Ap nonsmokers nonsmokers among among only only S9 S9 with with mutagenicity mutagenicity urinary urinary as as tested tested were were and and priori priori a a mutagenicity mutagenicity urinary urinary of of minants minants lower lower of of determinant determinant significant significant statistically statistically a a remained remained tor tor deter potential potential considered considered were were variables variables other other Several Several fac food food protective protective this this models, models, multivariate multivariate the the In In ± ± 0.46. 0.46. 5.23 5.23 was was not, not, it it did did who who those those for for and and ± ± 0.55, 0.55, 3.58 3.58 was was ity ity Mutagenicity Mutagenicity Urinary Urinary of of mutagenic mean mean 17), 17), foods foods (n (n protective protective = = consumed consumed who who Predictors Predictors Exhaust Exhaust Non-Diesel Non-Diesel Other Other < < < < p p 0.01. 0.01. 0.001 0.001 b b < < p p 0.001. 0.001. a a 0.15 0.15 nicotine nicotine Urinary Urinary 0.07 0.07 0.66a 0.66a cotinine cotinine Urinary Urinary 0.05 0.05 0.3oa 0.3oa 0.19b 0.19b thiocyanate thiocyanate Urinary Urinary --0.03 --0.03 -0.06 -0.06 0.00 0.00 -0.12 -0.12 S9 S9 - 0.08 0.08 0.09 0.09 0.13 0.13 -0.06 -0.06 S9 S9 0.45a 0.45a + + mutagenicity mutagenicity Urinary Urinary Nicotine Nicotine Nicotine Nicotine Cotinine Cotinine Thiocyanate Thiocyanate S9 S9 S9 S9 - + + Ambient Ambient Urinary Urinary Urinary Urinary Urinary Urinary Mutagenicity Mutagenicity Urinary Urinary Tobacco Tobacco of of Nonusers Nonusers from from Samples Samples for for Coefficients Coefficients Correlation Correlation Order Order Rank Rank Spearman Spearman 9. 9. Table Table al. al. et et Schenker Schenker B. B. M. M. 30 30 0.05). 0.05). are are very very sensitive sensitive to to the the assumption assumption of of normally normally distributed distributed degrees degrees of of freedom) freedom) are are > > 1.87 1.87 and and 0.82, 0.82, respectively respectively (p (p errors errors (Tables (Tables D.1 D.1 and and D.2). D.2). However, However, these these standard standard errors errors haust haust exposure exposure term term and and the the two two interaction interaction terms terms (3 (3 ance ance components components are are both both more more than than twice twice their their standard standard and and without without S9, S9, the the chi-squared chi-squared values values for for the the diesel diesel ex tions tions taken taken from from the the same same worker. worker. The The between-person between-person vari tion tion and and with with respirable respirable nicotine. nicotine. For For mutagenicity mutagenicity with with This This indicates indicates a a definite definite lack lack of of independence independence in in observa haust haust exposure exposure status status with with respirable respirable particle particle concentra genicity, genicity, both both with with and and without without S9 S9 (Tables (Tables D.1 D.1 and and D.2). D.2). exposure exposure status status (yes (yes or or no) no) and and the the interaction interaction of of diesel diesel ex person person (between-sample) (between-sample) variance variance is is about about 1:2.6 1:2.6 for for muta exposure exposure was was measured measured by by three three variables: variables: diesel diesel exhaust exhaust The The ratio ratio of of the the between-person between-person variance variance to to the the within Models Models for for smokers smokers also also included included d d the the number number of of cigarettes cigarettes smoked smoked on on the the study study day. day. c c Nonsmoker Nonsmoker models models also also included included protective protective foods foods dummy dummy variable. variable. study study mean mean for for the the mutagenicity mutagenicity of of 2-AF. 2-AF. each each exposed exposed group group to to base base line. line. Differences Differences and and standard standard errors errors were were obtained obtained by by multiplying multiplying dummy-variable dummy-variable effects effects and and standard standard errors errors by by 3.46, 3.46, the the Covariates Covariates for for all all models: models: b b 2-AF 2-AF (standard (standard mutagen mutagen revertants/ng), revertants/ng), 2-AF 2-AF x x urinary urinary cotinine, cotinine, 2-AF 2-AF x x airborne airborne nicotine, nicotine, 2-AF 2-AF x x dummy dummy variables variables comparing comparing a a Mutagenicity Mutagenicity is is expressed expressed in in revertants/~mol revertants/~mol of of creatinine. creatinine. Chi-squared Chi-squared for for exposure exposure (3 (3 degrees degrees of of freedom) freedom) 1.10. 1.10. = = High High adjusted adjusted respirable respirable particles particles 19 19 0.300 0.300 0.238 0.238 - 0.010 0.010 (0.114) (0.114) Medium Medium adjusted adjusted respirable respirable particles particles 11 11 0.178 0.178 0.158 0.158 -0.090 -0.090 (0.118) (0.118) Low Low adjusted adjusted respirable respirable particles particles 13 13 0.282 0.282 0.251 0.251 0.003 0.003 (0.156) (0.156) Diesel Diesel exhaust-exposed exhaust-exposed Diesel Diesel exhaust-unexposed exhaust-unexposed 16 16 0.248 0.248 0.248 0.248 - S9 S9 Chi-squared Chi-squared for for exposure exposure (3 (3 degrees degrees of of freedom) freedom) 0.48. 0.48. = = High High adjusted adjusted respirable respirable particles particles 19 19 4.298 4.298 3.977 3.977 0.398 0.398 (0.644) (0.644) Medium Medium adjusted adjusted respirable respirable particles particles 11 11 3.908 3.908 3.755 3.755 0.176 0.176 (0.668) (0.668) Low Low adjusted adjusted respirable respirable particles particles 13 13 4.553 4.553 3.946 3.946 0.367 0.367 (0.917) (0.917) Diesel Diesel exhaust-exposed exhaust-exposed Diesel Diesel exhaust-unexposed exhaust-unexposed 20 20 3.579 3.579 3.579 3.579 S9 S9 + + Smokersd Smokersd Chi-squared Chi-squared for for exposure exposure (3 (3 degrees degrees of of freedom) freedom) 0.21. 0.21. = = High High adjusted adjusted respirable respirable particles particles 22 22 0.105 0.105 0.092 0.092 -0.017 -0.017 (0.104) (0.104) Medium Medium adjusted adjusted respirable respirable particles particles 38 38 0.112 0.112 0.130 0.130 0.021 0.021 (0.090) (0.090) Low Low adjusted adjusted respirable respirable particles particles 30 30 0.057 0.057 0.119 0.119 0.010 0.010 (0.111) (0.111) Diesel Diesel exhaust-exposed exhaust-exposed Diesel Diesel exhaust-unexposed exhaust-unexposed 29 29 0.109 0.109 0.109 0.109 - S9 S9 Chi-squared Chi-squared for for exposure exposure (3 (3 degrees degrees of of freedom) freedom) 2.09. 2.09. = = High High adjusted adjusted respirable respirable particles particles 22 22 0.343 0.343 0.326 0.326 0.052 0.052 (0.125) (0.125) Medium Medium adjusted adjusted respirable respirable particles particles 0.369 0.369 40 40 0.395 0.395 0.121 0.121 (0.093) (0.093) Low Low adjusted adjusted respirable respirable particles particles 0.266 0.266 32 32 0.392 0.392 0.118 0.118 (0.121) (0.121) Diesel Diesel exhaust-exposed exhaust-exposed Diesel Diesel exhaust-unexposed exhaust-unexposed 30 30 0.274 0.274 0.274 0.274 S9 S9 + + Nonsmokersc Nonsmokersc n n Mean Mean (SE) (SE) Meanb Meanb Observed Observed Predicted Predicted Unexposed Unexposed from from Difference Difference Adjusted Adjusted Respirable Respirable Particle Particle Concentrationsa Concentrationsa Table Table 10. 10. Observed Observed and and Predicted Predicted Mutagenicity Mutagenicity from from Multivariate Multivariate Regression Regression Models Models by by Strata Strata of of Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers 31 31 magni- absolute absolute in in equal equal almost almost are are terms terms main-effect main-effect The The cigarettes cigarettes of of number number the the by by and and creatinine) creatinine) for for (corrected (corrected nicotine: nicotine: ambient ambient of of interactions interactions and and effects effects main main the the and and cotinine cotinine urinary urinary by by for for controlled controlled was was smoking smoking Cigarette Cigarette particles particles respirable respirable ofthe ofthe fit fit the the for for noted noted was was phenomenon phenomenon models. models. S9 S9 - the the from from excluded excluded were were three three and and models, models, interesting interesting < < An An 0.25). 0.25). 6.15, 6.15, p p level level (x 0.05 0.05 S9 S9 the the the the from from + + = = excluded excluded were were outliers outliers Two Two analysis. analysis. tivariate tivariate 2 2 at at significant significant not not is is exposure-interaction terms terms exposure-interaction three three the the mul for for the the of of part part were were S9 S9 without without samples samples 59 59 and and S9 S9 with with test test overall overall The The D.3. D.3. Table Table in in shown shown are are smokers smokers in in genicity genicity samples samples smokers' smokers' 61 61 exclusions, exclusions, After After Smokers. Smokers. Cigarette Cigarette muta S9 S9 for for model model + + exposure-interaction exposure-interaction the the for for Results Results results. results. regression regression multiple multiple the the for for support support > > additional additional provides provides 0.05). 0.05). observation observation (p (p This This 14). 14). respectively respectively and and 13 13 (Figures (Figures mutagenicity, mutagenicity, S9 S9 - and and association association S9 S9 positive positive for for a a of of 1.10 1.10 + + and and 0.48 0.48 suggestion suggestion no no are are show show which which values) values) threne threne statistics, statistics, phenan chi-squared chi-squared and and particles particles log-likelihood log-likelihood the the by by respirable respirable confirmed confirmed (adjusted (adjusted are are exposure exposure diesel diesel of of These These markers markers carmen). carmen). two two the the (clerks, (clerks, versus versus group group unexposed unexposed creatinine) creatinine) of of the the to to compared compared (revertants/J.lmol (revertants/J.lmol S9, S9, without without or or S9 S9 with with without without either either and and with with mutagenicity, mutagenicity, in in mutagenicity mutagenicity increase increase cant cant urinary urinary of of workers workers signifi three three statistically statistically a a these these has has from from group group samples samples exposed exposed repeated repeated individual individual the the No No for for 10). 10). plots plots Simple Simple (Table (Table particles particles respirable respirable adjusted adjusted of of level level and and S9) S9) without without determined. determined. were were workers workers these these of of exposures exposures phenanthrene phenanthrene or or (with (with mutagenicity mutagenicity between between relation relation dose-response dose-response a a and and days, days, 10 10 to to 8 8 on on sampled sampled been been had had workers workers these these of of found found was was evidence evidence no no groups, groups, exposure exposure for for particles particles ble ble of of Each Each samplers. samplers. personal personal their their in in levels levels nicotine nicotine ambient ambient respira adjusted adjusted of of strata strata using using models models regression regression the the In In the the on on based based smoke, smoke, tobacco tobacco environmental environmental much much to to posed posed ex not not were were who who and and smoke smoke not not did did who who identified identified were were exposure. exposure. smoking smoking recent recent workers workers shop shop repair repair Three Three reflects reflects analysis. analysis. day day study study detailed detailed the the for for on on selected selected smoked smoked cigarettes cigarettes of of number number the the was was exposure exposure exhaust exhaust while while day, day, diesel diesel study study high high the the to to prior prior potentially potentially and and smoking smoking smoke smoke cigarette cigarette by by weighted weighted tobacco tobacco environmental environmental low low is is had had half-life, half-life, who who long long and and a a days days with with different different Cotinine, Cotinine, significance. significance. biological biological in in on on studied studied been been had had who who differ differ workers workers also also of of They They subset subset a a D.3). D.3). (Table (Table excretion, excretion, S9 S9 with with mutagenicity mutagenicity for for tion tion mutagen mutagen urinary urinary in in differences differences equa individual individual < < regression regression for for the the to to control control 0.05, 0.05, To To at at p p contributed, contributed, each each they they 0.38), 0.38), (r (r correlated correlated are are variables variables two two these these = = Although Although tion. tion. guide. guide. rough rough a a as as only only taken taken be be can can therefore, therefore, and, and, effects) effects) collec sample sample of of time time the the to to up up day day study study the the on on smoked smoked regression regression the the of of errors errors standard standard the the (unlike (unlike terms terms random random text]. text]. (see (see nicotine nicotine ambient ambient smoke smoke tobacco tobacco 8.6 8.6 x x ~ ~ Environmental Environmental a a 95.5) 95.5) 50.7 50.7 (2.6, (2.6, 339.0) 339.0) 305.0 305.0 (257.0, (257.0, 305.1) 305.1) 9, 9, (190. (190. 222.3 222.3 19 19 Smokers Smokers 2.6) 2.6) 0.9 0.9 (0.0, (0.0, 311.0) 311.0) 200.0 200.0 (177.0, (177.0, 311.0) 311.0) (166.0, (166.0, 181.4 181.4 27 27 Nonsmokers Nonsmokers particles particles respirable respirable adjusted adjusted High High 110.5) 110.5) 81.7 81.7 (14.6, (14.6, 241.0) 241.0) (139.0, (139.0, 183.0 183.0 136.0) 136.0) (93.0, (93.0, 115.8 115.8 13 13 Smokers Smokers 7) 7) 6. 6. 3.4 3.4 (0.0, (0.0, 139.5) 139.5) (103.3, (103.3, 118.5 118.5 129.0) 129.0) (96.0, (96.0, 110.0 110.0 52 52 Nonsmokers Nonsmokers particles particles respirable respirable adjusted adjusted Medium Medium 244.2) 244.2) (78.3, (78.3, 119.5 119.5 267.0) 267.0) 147.0 147.0 (80.0, (80.0, 32.0) 32.0) 10.0 10.0 26.1, 26.1, (- 15 15 Smokers Smokers 3.4) 3.4) 0.9 0.9 (0.0, (0.0, 68.5) 68.5) (26.8, (26.8, 51.5 51.5 65.9) 65.9) (25.6, (25.6, 50.2 34 34 Nonsmokers Nonsmokers particles particles respirable respirable adjusted adjusted Low Low .8) .8) 117 117 54.2 54.2 (28.4, (28.4, 253.0) 253.0) 162.0 162.0 (111.0, (111.0, 141.6) 141.6) (43.0, (43.0, 78.8 78.8 23 23 Smokers Smokers 80.0) 80.0) 48.2 48.2 (4.3, (4.3, (66, (66, 182) 182) 107.0 107.0 99.3) 99.3) 51.4 51.4 (26.7, (26.7, 39 39 Nonsmokers Nonsmokers Unexposed Unexposed Percentiles Percentiles Median Median Percentiles Percentiles Median Median Percentiles Percentiles Median Median n n Exposure Exposure Exhaust Exhaust Diesel Diesel 75th 75th and and 25th 25th 75th 75th and and 25th 25th 75th 75th and and 25th 25th (!lg/m ) ) (!lg/m ) ) (!lg/m ) ) 3 3 3 Smoke Smoke Tobacco Tobacco Concentration Concentration Concentration Concentration Environmental Environmental Particle Particle Respirable Respirable Particle Particle Respirable Respirable Adjusted Adjusted Smokea Smokea Tobacco Tobacco Environmental Environmental and and Particles Particles Respirable Respirable of of Concentrations Concentrations Associated Associated with with Models Models Regression Regression Index Index in in Used Used Particles Particles Respirable Respirable Adjusted Adjusted of of Strata Strata 11. 11. Table Table al. al. et et Schenker Schenker B. B. M. M. 32 32 counterintuitive counterintuitive ways. ways. For For mutagenicity mutagenicity with with S9, S9, Table Table D.3 D.3 shows shows that that both both uri- related related variables, variables, the the individual individual coefficients coefficients can can behave behave in in sponse sponse of of 3.46 3.46 revertants/ng). revertants/ng). cigarettes cigarettes smoked, smoked, and and urinary urinary cotinine. cotinine. With With such such cor ~g/m of of respirable respirable particles particles (assuming (assuming a a 2-AF 2-AF control control re 3 3 tions tions among among total total respirable respirable particles, particles, ambient ambient nicotine, nicotine, more more of of creatinine creatinine revertants/~mol revertants/~mol for for each each 1,000 1,000 extra extra ticles. ticles. This This puzzling puzzling finding finding is is probably probably due due to to the the correla nine nine of of respirable respirable particles particles among among exposed exposed samples samples is is 0.76 0.76 which which also also showed showed a a negative negative coefficient coefficient for for respirable respirable par respirable respirable particles particles in in terms terms of of of of creati revertants/~mol revertants/~mol This This was was borne borne out out by by analyses analyses in in unexposed unexposed men men alone, alone, els els of of Table Table 10. 10. In In fact, fact, the the predicted predicted effect effect of of increasing increasing creasing creasing respirable respirable particles particles is is to to decrease decrease mutagenicity. mutagenicity. actly actly what what was was observed observed in in the the three three exposure-group exposure-group mod 39.72 39.72 Ambient Ambient Nicotine. Nicotine. That That is, is, the the predicted predicted effect effect of of in dose-response dose-response association association in in exposed exposed samples. samples. This This is is ex and and ambient ambient nicotine nicotine is is (- 2.66 2.66 Respirable Respirable x x Particles) Particles) + + tine. tine. These These coefficients coefficients are are negligible, negligible, corresponding corresponding to to no no In In unexposed unexposed men, men, the the joint joint effect effect of of respirable respirable particles particles tine tine 0.31 0.31 Respirable Respirable x x Particles Particles 0.93 0.93 Ambient Ambient x x Nico + + = = nicotine nicotine are are 39.724 39.724 and and - 38.788, 38.788, respectively. respectively. x x Respirable Respirable Particles Particles (39.72 (39.72 - 38.79) 38.79) x x + + Ambient Ambient Nico tively; tively; the the main-effect main-effect and and interaction interaction terms terms for for ambient ambient posed posed men, men, the the corresponding corresponding joint joint effect effect is is (- 2.66 2.66 2 2 .97) .97) + + terms terms for for respirable respirable particles particles are are - 2.656 2.656 and and 2 2 .971, .971, respec adding adding the the main-effect main-effect and and interaction interaction terms. terms. Thus, Thus, in in ex opposite opposite in in sign. sign. Thus, Thus, the the main-effect main-effect and and interaction interaction of of respirable respirable particles particles and and ambient ambient nicotine nicotine are are found found by by tude tude to to the the corresponding corresponding exposure-interaction exposure-interaction terms, terms, but but In In men men with with potential potential diesel diesel exhaust exhaust exposure, exposure, the the effects effects repeated repeated measurements. measurements. respirable respirable particle particle concentration concentration for for three three nonsmoking nonsmoking shop shop workers workers with with threne threne for for three three nonsmoking nonsmoking shop shop workers workers with with repeated repeated measurements. measurements. Figure Figure 13. 13. Urine Urine mutagenicity mutagenicity S9 S9 (top) (top) and and Figure Figure - S9 S9 14. 14. (bottom) (bottom) Urine Urine + + versus versus mutagenicity mutagenicity adjusted adjusted S9 S9 (top) (top) and and - S9 S9 (bottom) (bottom) + + versus versus phenan- (~g/m ) ) Adjusted Adjusted (ng/m~ (ng/m~ Respirable Respirable Particles Particles Phenanthrene Phenanthrene 3 0 0 100 100 600 600 200 200 -100 -100 400 400 500 500 300 300 1000 1000 BOO BOO 600 600 0 0 200 200 400 400 ::a: ::a: -1 -1 -1 -1 ::a: ::a: ~ ~ "' "' s s Cl Cl Cl Cl Q) Q) ·c: ·c: Q) Q) ·c: ·c: "l3 "l3 "l3 "l3 ~ ~ • • c c .. .. ~ ~ c c c c • • c c I I c c "&. "&...... 0 0 ~ ~ I I .. ltl ltl ...... 0 0 . . (/) .. .. .c .c .. .. (/) (/) ... loc loc m m .. .. m m .. .. c• c• • • ~ ~ c c .. .. g g > > ~ ~ ~ ~ E E E E 0 0 0 0 c c 0 0 0 0 10 10 2B5 2B5 c c 102B5 102B5 c c ~ ~ (.) (.) ~ ~ .. .. .. .. 10206 10206 10206 10206 ~ ~ -~ -~ ·c: ·c: . . ·c: ·c: . . 10170 10170 10170 10170 c:: c:: c:: c:: 2 2 Q) Q) Q) Q) 2 2 Phenanthrene Phenanthrene (ng/m ) ) (~g/m ) ) Adjusted Adjusted Respirable Respirable Particles Particles 3 3 600 600 0 0 200 200 100 100 600 600 1000 1000 BOO BOO 200 200 400 400 300 300 0 0 500 500 -1 -1 00 00 400 400 ::a: ::a: ·1 ·1 -1 -1 ::a: ::a: s s .. .. ~ ~ ...... g g .. .. Q) .. .. ·c: ·c: Q) Q) ...... ·c: ·c: "l3 "l3 ·13 ·13 ~ ~ >. >. + + c c c c c c c c + + . . 0 0 0 0 (/) (/) (/) (/) m m m m .. .. I I .. .. c c .. .. c c 1 1 ~ ~ ~ ~ c c c c > > c c > > ...... ll ll • • ...... .. .. c c ::::l. ::::l. ::::l. ::::l. E E E E c c c c 0 0 0 0 .. .. 0 0 c c c c 0 0 102B5 102B5 c c 10 10 2B5 2B5 c c (.) (.) (.) (.) .. .. ~ ~ 10206 10206 ~ ~ .. .. 10 10 206 206 ...... ~ ~ "15 "15 . . 10170 10170 ·c: ·c: ·c: ·c: . . 10170 10170 c:: c:: c:: c:: Q) Q) Q) Q) 2 2 2 2 Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers 33 33 in- Also Also area. area. the the from from data data limited limited of of basis basis the the on on pler, pler, detected. detected. exposure exposure exhaust exhaust diesel diesel of of effect effect an an was was analyses analyses sam dichotomous dichotomous a a of of fraction fraction respirable respirable or or fine fine the the in in be be alternative alternative these these of of none none In In tried. tried. also also was was urine urine of of lent lent would would percent percent 55 55 approximately approximately which which of of matter, matter, ticulate ticulate equiva milliliter milliliter per per revertants revertants of of number number the the of of culation culation par total total of of ~g/m 25 25 of of mean mean geometric geometric annual annual an an showed showed cal the the for for fit fit quadratic quadratic a a to to line line tangent tangent The The removed. removed. were were 3 3 sampled sampled we we where where area area the the from from data data quality quality Air Air air. air. bient bient effects effects food food protective protective and and trip trip The The observations. observations. outlier outlier am the the in in found found matter matter particulate particulate respirable respirable background background possible possible including including and and excluding excluding of of effects effects the the includ<:::d includ<:::d include include samples samples All All matter. matter. particulate particulate of of sources sources other other analyses analyses other other Still Still section. section. Methods Methods Statistical Statistical the the in in rized rized include include still still because they they because exhaust exhaust diesel diesel of of measure measure rect rect summa as as considered, considered, were were forms forms model model Alternative Alternative di a a not not are are exposures exposures particle particle respirable respirable adjusted adjusted The The analysis. analysis. sion sion 4). 4). (Table (Table exposures exposures diesel diesel regres for for samples samples few few too too had had 2 2 day day on on mutagenicity mutagenicity urine urine highest highest the the have have to to expected expected be be would would and and locomotives, locomotives, predict predict to to 1 1 day day of of results results mutagenicity mutagenicity urine urine the the using using running running with with space space enclosed enclosed partially partially or or enclosed enclosed an an in in are are Models Models observed. observed. was was exposure exposure exhaust exhaust diesel diesel of of effect effect workers workers shop shop The The exhaust. exhaust. diesel diesel diluted diluted to to exposed exposed are are so so no no particles; particles; respiratory respiratory of of effect effect independent independent an an for for ined ined and and trains, trains, the the near near outside outside or or inside inside are are either either conductors, conductors, exam were were jobs jobs exposed exposed in in working working men men of of Subgroups Subgroups and and brakers, brakers, engineers, engineers, the the of of up up made made crew, crew, train train The The lates. lates. results. results. regression regression the the affected affected changes changes these these of of ther ther particu other other to to exposed exposed are are they they but but circumstances, circumstances, rare rare Nei cotinine. cotinine. urine urine of of place place in in used used also also was was nicotine nicotine Urine Urine in in except except trains, trains, the the from from exhaust exhaust diesel diesel to to unexposed unexposed are are measured. measured. not not was was cotinine cotinine which which in in samples samples those those for for carmen carmen and and clerks clerks the the sampled, sampled, we we groups groups job job the the of of terns terns used used was was regressions regressions these these from from cotinine cotinine urinary urinary of of value value pat work work the the of of observation observation by by judged judged As As exposure. exposure. exhaust exhaust predicted predicted The The thiocyanate. thiocyanate. urinary urinary and and inhalation, inhalation, smoke smoke of of diesel diesel worker's worker's a a estimating estimating in in contaminants contaminants interfering interfering the the degree degree smoked, smoked, cigarettes cigarettes on on cotinine cotinine urine urine regressed regressed we we of of one one remove remove to to used used be be can can it it how how and and exposure exposure smoke smoke data, data, full full with with samples samples all all In In measured. measured. been been not not had had nine nine tobacco tobacco environmental environmental the the measuring measuring for for method method a a strated strated coti urine urine which which in in samples samples utilizing utilizing for for technique technique stage stage demon have have We We exhaust. exhaust. diesel diesel and and smoke, smoke, tobacco tobacco mental mental two a a employed employed We We out. out. carried carried were were analyses analyses other other of of ber ber environ work, work, during during generated generated matter matter particulate particulate other other anum tables, tables, the the in in displayed displayed analyses analyses the the to to addition addition In In or or dirt dirt dust, dust, matter, matter, particulate particulate ambient ambient of of mixture mixture plex plex Analyses Analyses Other Other com a a are are exposures exposures particle particle respirable respirable workers' workers' Railroad Railroad smoking. smoking. for for time time of of availability availability and and density density worker worker the the on on depend depend also also they they smoking, smoking, cigarette cigarette of of case case the the In In ment. ment. environ work work the the of of characteristics characteristics ventilation ventilation and and size, size, (0.011). (0.011). component component layout, layout, the the on on and and source source contaminant contaminant the the of of rate rate output output the the within-person within-person the the than than larger larger is is (0.070) (0.070) component component variance variance on on depend depend area area work work a a in in particles particles of of concentrations concentrations Air Air between-person between-person the the smokers, smokers, in in S9 S9 without without mutagenicity mutagenicity For For 0.079). 0.079). 0.006 0.006 0.085 0.085 (- negative negative + + is is men men posed posed - = exhaust. exhaust. diesel diesel ex in in coefficient coefficient predicted predicted the the fact, fact, In In men. men. unexposed unexposed or or in in PAHs PAHs abundant abundant most most the the of of one one phenanthrene, phenanthrene, to to posure posure exhaust-exposed exhaust-exposed diesel diesel either either in in particles particles respirable respirable of of fects fects ex measuring measuring and and particles), particles), respirable respirable (adjusted (adjusted mass mass ef separate separate no no again, again, shows, shows, parameters parameters interaction interaction the the of of to to contribution contribution smoke smoke tobacco tobacco environmental environmental the the tracting tracting > > Inspection Inspection 0.05). 0.05). 5.39, 5.39, p p model model the the to to (x significantly significantly = = sub concentration, concentration, particle particle respirable respirable yield yield to to sampling sampling 2 2 contribute contribute not not did did also also parameters parameters exposure exposure exhaust exhaust diesel diesel size-selective size-selective included included These These sources. sources. contaminant contaminant borne borne the the D.4), D.4), (Table (Table model model interaction interaction exposure- regression regression air other other of of interference interference positive positive the the minimized minimized that that haust haust > > multiple full full the the In In 0.05). 0.05). 1.10 1.10 (p (p was was statistic statistic test test squared squared ex diesel diesel for for markers markers develop develop to to used used were were strategies strategies several several chi ratio ratio log-likelihood log-likelihood The The 10). 10). (Table (Table particles particles respirable respirable directly, directly, measured measured be be not not could could exhaust exhaust diesel diesel Since Since gravel. gravel. of of strata strata exposure exposure within within association association dose-response dose-response a a of of and and sand sand from from dust dust generated generated mechanically mechanically and and sources, sources, found found was was evidence evidence no no also, also, S9 S9 without without mutagenicity mutagenicity For For combustion combustion other other from from particles particles smoke, smoke, tobacco tobacco mental mental environ exhaust, exhaust, diesel diesel include include They They workers. workers. railroad railroad by by regression. regression. multiple multiple least-squares least-squares nary nary breathed breathed air air the the in in particles particles of of sources sources several several are are There There ordi to to corresponds corresponds least-squares least-squares generalized generalized case, case, this this In In known. known. are are covariates covariates all all once once person, person, same same the the from from taken taken Exposure Exposure Smoke Smoke Tobacco Tobacco samples samples of of independence independence complete complete to to corresponds corresponds ponent ponent Environmental Environmental and and Particle Particle Respirable Respirable com between-person between-person zero zero A A 0.794). 0.794). (SE (SE 3.152 3.152 of of variance variance = = EXPOSURE EXPOSURE within-person within-person EXHAUST EXHAUST the the to to DIESEL DIESEL compared compared zero, zero, or or negligible negligible 0.644), 0.644), (SE (SE 0.235, 0.235, was was component component variance variance = = between-person between-person The The mutagenicity. mutagenicity. of of predictive predictive individually individually were were study study the the of of DISCUSSION DISCUSSION day day the the on on smoked smoked cigarettes cigarettes of of number number and and cotinine cotinine nary nary al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers eluded in the adjusted respirable particle measurement are posure among railroad workers (Woskie et al. 1988b). These dirt and dust generated by the work itself. The carmen low levels may reflect the decline in railroad activity in this spend much of their time outdoors along the dusty tracks country, with associated reductions in diesel engine use where the freight cars are repaired. The brakers also work and exposure to diesel exhaust. outside along the train tracks, where they are exposed to the Willems and associates (1989) have reported no increase dirt and dust produced by their own and the locomotives' in fecal or urinary mutagenicity attributable to diesel ex activities, one of which is to apply sand to the tracks fre haust exposure among car mechanics. The Salmonella quently in the winter to increase traction. In our previous plate incorporation assay (Ames et al. 1975) was used to study, we found that freight brakers' respirable particle sam measure mutagenicity. ples were only 23 percent extractable in dichloromethane, The absence of an association of mutagenic activity in the while other job groups' samples were about 40 to 45 percent urine with diesel exhaust exposure does not necessarily extractable, which implies that a substantial portion of the mean that diesel workers did not excrete diesel-associated sample mass may be inorganic dirt and dust (Hammond et mutagens. First, the level of mutagenic activity could be be al. 1988). Nevertheless, even though the adjusted respirable low the level of detection. For example, a worker who was particle exposure may overestimate the diesel exhaust exposed to a mutagen concentration of 1,000 revertant exposure of these workers, it is a useful marker of the rela equivalents per cubic meter of air and who breathed 7 to tive diesel exhaust exposure found among these railroad 10m3 of air during the seven-hour exposure period re workers. ceived a total mutagen dose of approximately 7,000 to 10,000 revertant equivalents (1,000 revertants/m3 x m3/hour x 7 Phenanthrene Exposure to 10 hours). The value of 1,000 revertant equivalents per cu Our observation of a constant ratio of phenanthrene to bic meter of air was estimated from a diesel shop worker's respirable particles on area samples from the diesel personal filters and was measured using the microsuspen exhaust-exposed repair shops suggests that phenanthrene sionprocedure (Figure 5). If we assume that a small fraction should be pursued as a marker for diesel exhaust. Many of mutagens is excreted in the urine in a mutagenic form, other markers have been suggested for diesel exhaust, none then the measureable revertant equivalents can be approxi of which is yet established as a "gold standard;' and even mated. This may not be an unreasonable assumption, since less was known about these alternatives at the time this the mutagens adsorbed on diesel particulate matter will study was begun. A complete discussion of alternative have to be desorbed in physiologic fluids. The bioavailabil markers is beyond the scope of this report, but a discussion ity of mutagens on diesel particulate matter has been of why phenanthrene was selected as a marker of diesel ex reported to be a relatively slow desorption process (King et haust exposure is contained in Appendix F. al. 1981). The work by Belisario and colleagues (1984), who We selected phenanthrene in part because our earlier injected rats with whole diesel particles and measured uri analyses of railroad diesel exhaust found it to be the PAH nary mutagenic activity, indicated that only a small fraction present in the highest concentration. This was an important of measurable mutagens is excreted in a 24-hour period. consideration in designing a study that used personal sam For illustrative purposes, if 1 percent of the total revertant pling pumps that collect less than 1 percent of the volume equivalents is excreted in the afternoon urine sample col of air collected by the high-volume samplers. lected, 70 to 100 revertant equivalents would be in 100 to 200 mL of urine (0.01 x 7,000 to 10,000 revertant equivalents). The concentration would therefore add, at the most, 0.7 to URINARY MUTAGENICITY 1 revertant equivalent per milliliter of urine (70 to 100 rever tant equivalents/loa mL of urine). This is the level of activ Diesel Exhaust ity generally determined in the urine of nonsmokers (Kado We did not observe an effect of diesel exhaust exposure et al. 1983). on urinary mutagenicity. Several factors may have con Two important considerations in evaluating these results tributed to this negative result. Despite our attempt to max are the unknown kinetics of diesel exhaust absorption and imize diesel exhaust exposure, the median level of respira excretion in humans, and the limited ability to evaluate ble particle concentration from diesel exhaust among them in the field setting. For example, it was feasible to col nonsmoking subjects in our highest exposed job group lect only a single spot urine at the end of the work shift. Al (shop workers) was only 113 ~g/m 3 , and 75 percent of sam though the spot urine samples showed a good dose ples were below 161 ~g/m 3 . These concentrations are lower response relation with smoking dose on the work day, if than many historical measurements of diesel exhaust ex- diesel mutagens were excreted more slowly than mutagens 34 35 35 (1988) (1988) colleagues colleagues and and Hayatsu Hayatsu particles. particles. emission emission diesel diesel promising. promising. was was approach approach this this that that felt felt from from extracts extracts including including mixtures, mixtures, complex complex of of number number a a of of we we filters, filters, such such testing testing of of feasibility feasibility the the examine examine to to study study mutagenicity mutagenicity the the inhibited inhibited chlorophyllin chlorophyllin that that found found (1986) (1986) pilot pilot a a As As analyses. analyses. of of frame frame time time the the in in us us to to available available made made coworkers coworkers and and Ong Ong 1985). 1985). Hoeven Hoeven der der van van and and (Terwel (Terwel were were filters filters personal personal few few very very only only Second, Second, time. time. pling pling chlorophyllin chlorophyllin or or riboflavin, riboflavin, acid, acid, ellagic ellagic either either by by creased creased sam of of hours hours eight eight approximately approximately in in filters filters personal personal the the de also also was was condensate condensate smoke smoke cigarette cigarette and and BaP BaP of of ity ity on on available available is is sample sample limited limited very very a a only only since since point, point, cant cant mutagenic the the and and products, products, pyrolysis pyrolysis mutagenic mutagenic highly highly of of signifi a a is is This This done. done. be be could could samples samples filter filter personal personal on on activity activity mutagenic mutagenic the the inhibited inhibited compound compound protein-bound protein-bound mutagenicity mutagenicity of of measurement measurement that that show show to to was was data data these these of of a a that that reported reported (1978) (1978) colleagues colleagues and and Kada Kada mutagenicity. mutagenicity. purpose purpose the the First, First, workers. workers. shop shop and and clerk clerk the the from from samples samples 3-methylcholanthrene 3-methylcholanthrene inhibited inhibited chlorophyll chlorophyll that that ported ported filter filter personal personal the the for for case case the the not not was was this this S9), S9), (- vation vation re (1980) (1980) associates associates and and Lai Lai example, example, For For test. test. poration poration acti metabolic metabolic without without than than higher higher usually usually is is TA98) TA98) strain strain incor plate plate Salmonella Salmonella the the in in activity activity antimutagenic antimutagenic their their (using (using extracts extracts smoke smoke tobacco tobacco environmental environmental of of S9) S9) ( ( tion tion + + for for tested tested been been have have foods foods in in naturally naturally found found Compounds Compounds activa metabolic metabolic with with activity activity mutagenic mutagenic the the Although Although 1988). 1988). Compounds Compounds Occurring Occurring Santella Santella 1988; 1988; (Perea (Perea reviewed reviewed been been have have monitoring monitoring man man Naturally Naturally of of Activity Activity Antimutagenic Antimutagenic hu for for methods methods other other and and this this of of significance significance and and cation cation appli The The (1984). (1984). associates associates and and ofRanderath ofRanderath methods methods ing ing estimate. estimate. to to difficult difficult are are meats meats fried fried eating eating from from P-postlabel the the as as such such humans humans of of monitoring monitoring biological biological 32 urine urine workers' workers' railroad railroad in in be be to to anticipated anticipated levels levels mutagenic mutagenic for for used used been been have have methods methods of of number number A A exhaust. exhaust. diesel diesel the the meats, meats, fried fried involving involving study study a a in in used used been been yet yet not not has has in in compounds compounds reactive reactive the the of of adducts adducts protein protein or or DNA DNA tual tual procedure procedure microsuspension microsuspension the the Since Since test. test. incorporation incorporation ac measure measure to to is is dose dose monitoring monitoring to to approach approach Another Another plate plate the the on on based based were were meats meats fried fried consumed consumed individuals individuals question. question. this this address address should should studies studies further further but but sibility, sibility, which which in in studies studies the the in in reported reported values values mutagenic mutagenic The The pos this this evaluate evaluate to to mutagenicity mutagenicity particle particle of of measurements measurements (1981). (1981). colleagues colleagues and and Nader Nader by by earlier earlier reported reported enough enough do do not not did did we we Unfortunately, Unfortunately, exhaust. exhaust. diesel diesel of of nent nent was was as as mutagenic, mutagenic, not not was was cooking cooking microwave microwave by by pared pared compo active active biologically biologically the the to to exposure exposure measuring measuring for for pre meat meat (1984), (1984), coworkers coworkers and and Sousa Sousa and and (1982) (1982) coworkers coworkers marker marker potential potential another another offers offers samplers samplers personal personal from from cles cles and and Baker Baker by by studies studies the the In In revertants. revertants. of of level level background background parti respirable respirable of of activity activity Mutagenic Mutagenic present. present. were were one one if if the the times times five five to to two two reached reached activity activity of of level level The The sumption. sumption. mutagenicity, mutagenicity, urinary urinary with with exposure exposure exhaust exhaust diesel diesel of of tion tion con after after hours hours six six to to two two approximately approximately appeared appeared tivity tivity associa an an observe observe to to ability ability the the reduce reduce would would errors errors these these ac peak peak that that and and meal, meal, beef beef fried fried a a after after urine urine the the in in tected tected of of both both that that likely likely is is exhaust. exhaust. It It diesel diesel of of components components genic genic de was was activity activity mutagenic mutagenic that that reported reported (1985) (1985) coworkers coworkers muta all all for for marker marker best best the the be be not not may may and and phase phase vapor vapor and and Sousa Sousa environment:' environment:' work work the the in in chemicals chemicals mutagenic mutagenic the the in in compounds compounds mutagenic mutagenic represent represent may may it it but but exhaust, exhaust, or or smoke smoke cigarette cigarette to to exposure exposure after after reported reported that that than than diesel diesel of of marker marker improved improved an an be be to to seemed seemed Phenanthrene Phenanthrene lower lower much much and and marginal, marginal, is is meat meat to to exposure exposure to to related related ity ity above. above. noted noted as as markers, markers, tobacco tobacco for for correction correction the the in in sion sion activ mutagenic mutagenic "urinary "urinary that that concluded concluded and and activity activity genic genic impreci to to due due be be may may error error this this particles, particles, respirable respirable justed justed muta lower lower much much reported reported but but results, results, these these confirmed confirmed ad For For remained. remained. certainly certainly assessment assessment exposure exposure in in error error (1984) (1984) associates associates and and Dolara Dolara bacon. bacon. or or pork pork fried fried cluding cluding some some but but estimates, estimates, exposure exposure exhaust exhaust diesel diesel our our of of accuracy accuracy in meals meals consumed consumed who who volunteers volunteers of of urine urine the the in in elevated elevated the the improved improved workers) workers) of of subset subset a a (on (on phenanthrene phenanthrene of of and and was was mutagenicity mutagenicity that that reported reported (1982) (1982) colleagues colleagues and and Baker Baker smoke smoke tobacco tobacco environmental environmental for for adjusted adjusted concentrations concentrations example, example, For For published. published. been been have have mutagenicity mutagenicity urinary urinary on on particle particle respirable respirable for for measurements measurements personal personal of of use use The The consumption consumption food food of of effects effects the the on on reports reports of of number number A A mutagen. mutagen. the the of of form form lized) lized) Factors Factors Dietary Dietary (unmetabo parent parent the the releasing releasing in in result result not not may may steps steps tion tion Deconjuga polar. polar. are are they they since since XAD, XAD, by by extracted extracted ciently ciently exposure. exposure. effi be be not not may may they they or or mutagenic, mutagenic, be be not not may may mutagens mutagens the the exhaust exhaust diesel diesel of of levels levels higher higher with with occurred occurred likely likely and and ing, ing, form, form, conjugated conjugated this this In In sulfates. sulfates. or or glucuronides glucuronides as as such such smok cigarette cigarette from from risk risk the the of of fraction fraction a a only only is is 1988) 1988) 1987, 1987, molecules, molecules, polar polar to to conjugated conjugated be be may may mutagens mutagens associated associated al. al. et et Garshick Garshick 1983; 1983; al. al. et et (Howe (Howe studies studies some some in in observed observed exhaust diesel diesel The The urine. urine. the the in in mutagenic mutagenic not not is is that that workers workers railroad railroad exhaust-exposed exhaust-exposed diesel diesel among among cancer cancer lung lung form form a a in in excreted excreted are are or or feces, feces, the the as as such such route, route, another another of of risk risk increased increased The The present. present. were were one one if if mutagenicity, mutagenicity, by by excreted excreted part, part, in in are, are, mutagens mutagens diesel diesel the the that that possible possible urinary urinary on on exhaust exhaust diesel diesel of of effect effect the the observe observe to to enough enough also also is is It It mutagens. mutagens. exhaust exhaust diesel diesel the the detect detect to to enough enough tive tive great great been been have have not not may may study study the the of of power power the the Finally, Finally, sensi been been have have not not might might assay assay the the smoke, smoke, cigarette cigarette from from al. al. et et Schenker Schenker B. B. M. M. 36 36 ranged ranged from from 0 0 to to 15 15 revertants/Jlmol revertants/Jlmol of of creatinine, creatinine, with with 75 75 issue, issue, and and are are more more appropriate appropriate to to evaluate evaluate such such an an associa- For For smokers, smokers, observed observed S9 S9 urinary urinary mutagenic mutagenic activity activity + + ers ers (Garshick (Garshick et et al. al. 1987, 1987, 1988) 1988) have have directly directly addressed addressed this this exposure. exposure. Other Other epidemiological epidemiological studies studies of of railroad railroad work cent cent probability probability in in this this study. study. sions sions regarding regarding cancer cancer risks risks associated associated with with diesel diesel exhaust exhaust These These differences differences would would have have been been detected detected with with 93 93 per This This study study provides provides no no basis basis on on which which to to draw draw conclu um, um, and and high) high) were were 0.1, 0.1, 0.3, 0.3, and and 0.4, 0.4, respectively. respectively. + + + + + + strata strata of of exposed exposed adjusted adjusted respirable respirable particles particles (low, (low, medi ity ity in in personal personal filter filter samples samples showed showed promise. promise. pose pose that that the the true true differences differences from from base base line line for for the the three three tectable tectable form form in in the the urine. urine. The The measurement measurement of of mutagenic difference difference of of about about 0.1 0.1 revertants/Jlmol revertants/Jlmol of of creatinine. creatinine. Sup of of detection. detection. This This assumes assumes that that mutagens mutagens are are excreted excreted in in de The The predicted predicted S9 S9 means means in in Table Table 10 10 had had a a maximum maximum + + would would require require higher higher exposure exposure levels levels or or greater greater sensitivity sensitivity tween tween about about 0.12 0.12 and and 0.50 0.50 revertants/Jlmol revertants/Jlmol of of creatinine. creatinine. nary nary mutagenicity mutagenicity for for epidemiologic epidemiologic studies, studies, therefore, therefore, nine, nine, with with the the middle middle 75 75 percent percent of of observations observations be falling as as cotinine cotinine is is a a marker marker for for cigarette cigarette smoke. smoke. The The use use of of uri ranged ranged roughly roughly from from -0.9 -0.9 to to 2.4 2.4 revertants/Jlmol revertants/Jlmol of of creati currently currently no no specific specific biological biological marker marker for for diesel diesel exhaust, exhaust, For For nonsmokers, nonsmokers, S9 S9 urinary urinary mutagenicity mutagenicity in in the the study study + + association association exist, exist, as as we we have have noted. noted. Unfortunately, Unfortunately, there there is is tectable. tectable. Several Several possible possible explanations explanations for for this this absence absence of of an an exposure exposure are are generated generated to to calculate calculate the the power. power. gens gens associated associated with with these these levels levels of of exposure exposure were were unde Synthetic Synthetic data data based based on on each each alternative alternative hypothesis hypothesis about about environmental environmental tobacco tobacco smoke smoke found found in in our our study, study, the the muta standard standard deviations deviations are are also also fixed fixed at at their their estimated estimated values. values. suggests suggests that that at at the the levels levels of of exposure exposure to to diesel diesel exhaust exhaust and and values values of of the the covariates covariates observed observed in in the the study. study. The The residual residual haust haust exposure exposure with with urinary urinary mutagenicity. mutagenicity. Our Our research research study study data. data. This This method method estimates estimates power power conditional conditional on on the the sociation sociation of of environmental environmental tobacco tobacco smoke smoke or of of or diesel diesel ex (1982) (1982) was was modified modified for for post post hoc hoc calculations calculations with with the the mutagens. mutagens. However, However, we we did did not not detect detect an an independent independent as tions tions were were treated treated as as independent. independent. The The method method of of O'Brien O'Brien of of cigarette cigarette smoking smoking and and of of certain certain foods foods with with urinary urinary ity, ity, between-person between-person variability variability was was ignored ignored and and all all observa In In the the full-regression full-regression models, models, we we observed observed an an association association group group means, means, after after adjustment adjustment for for other other factors. factors. For For simplic test test (with (with three three degrees degrees F F of of freedom) freedom) of of differences differences in in rette rette smoking smoking by by comparison comparison with with other other smoking smoking markers. markers. of of exposure, exposure, were were used. used. Power Power was was calculated calculated dated dated the the dose-response dose-response for for the the partial partial characteristics characteristics of of the the assay assay for for ciga in in Table Table 10, 10, with with the the four four adjusted adjusted respirable respirable assay assay for for urinary urinary particle particle mutagens mutagens strata strata in in the the field field study study and and have have vali smokers smokers and and nonsmokers. nonsmokers. The The multiple multiple regression regression models models particles particles and and phenanthrene). phenanthrene). We We have have also also used used a a sensitive sensitive Study Study power power was was evaluated evaluated for for S9 S9 analyses analyses of of both both estimating estimating + + diesel diesel exhaust exhaust exposure exposure (adjusted (adjusted respirable respirable studies studies and and have have demonstrated demonstrated two two improved improved methods methods for for Study Study Power Power issues issues in in measuring measuring exposure exposure to to diesel diesel exhaust exhaust in in field field railroad railroad workers. workers. We We have have addressed addressed some some of of the the complex complex necessary necessary to to evaluate evaluate this this association. association. diesel diesel exhaust, exhaust, and and to to use use these these methods methods in in a a field field study study of of suggests suggests a a causal causal association. association. Further Further controlled controlled studies studies are are and and for for using using urinary urinary mutagenicity mutagenicity to to evaluate evaluate exposure exposure to to consistency consistency of of the the findings findings with with the the results results of of other other studies studies mating mating diesel diesel exhaust exhaust exposure exposure in in epidemiological epidemiological studies studies take take was was based based on on self-report self-report and and was was not not validated, validated, the the The The aims aims of of this this study study were were to to develop develop methods methods for for esti in in fried fried bacon bacon meals. meals. Although Although our our measure measure of of dietary dietary in IMPLICATIONS IMPLICATIONS OF OF THE THE FINDINGS FINDINGS fruits fruits and and vegetables vegetables on on the the urinary urinary clearance clearance of of mutagens mutagens however, however, reported reported that that there there were were no no effects effects of of ingested ingested Brussels Brussels sprouts, sprouts, and and broccoli. broccoli. Baker Baker and and associates associates (1986), (1986), who who frequently frequently ingest ingest raw raw vegetables, vegetables, been been especially especially missed. missed. cabbage, cabbage, that that there there was was a a decreased decreased risk risk of of detected, detected, colon colon cancer cancer although although in in people people smaller smaller differences differences might might well well have have 1982). 1982). For For example, example, Graham Graham and and colleagues colleagues such such large large (1978) (1978) exposure-related exposure-related reported reported effects effects would would have have gone gone un tive tive fects fects effect effect must must of of these these be be considered considered foods foods (National (National possible. possible. Academy Academy Thus, Thus, it it of of is is Sciences Sciences unlikely unlikely that that sprouts sprouts Given Given is is consistent consistent the the ranges ranges with with in in which which numerous numerous the the observed observed reports reports on on data data the the fell, fell, protec such such ef diesel diesel workers workers who who consumed consumed fish, fish, cabbage, cabbage, or or Brussels Brussels have have been been declared declared 84 84 percent percent of of the the time. time. genic activity activity genic was was decreased decreased in in the the urine urine of of nonsmoking nonsmoking In In this this case, case, statistical statistical significance significance at at the the 0.05 0.05 level level would would in in the the urine urine would would be be informative. informative. Our Our finding finding that that muta low, low, medium, medium, and and high high adjusted adjusted respirable respirable particle particle groups. groups. these these "modulators" "modulators" on on excretion excretion of of mutagenic mutagenic compounds compounds from from the the base-line base-line mean mean were were 0.2, 0.2, 1.5, 1.5, and and 2.0, 2.0, for for the the + + + + + + the the activity activity of mutagens, mutagens, of a a study study examining examining the the effect effect of of posure posure groups groups was was about about 0.4. 0.4. Suppose Suppose the the true true differences differences Since Since some some naturally naturally occurring occurring compounds compounds modulate modulate difference difference between between predicted predicted means means among among the the four four ex ogenesis. ogenesis. revertants/Jlmol revertants/Jlmol of of creatinine. creatinine. In In Table Table 10, 10, the the maximum maximum have have percent percent reviewed reviewed of of the the dietary dietary observations observations inhibitors inhibitors occurring occurring of of mutagenesis mutagenesis between between and and 1.9 1.9 carcin and and 5.4 5.4 Markers Markers of of Exposure Exposure to to Diesel Diesel Exhaust Exhaust in in Railroad Railroad Workers Workers 37 37 M. M. Lodovici Lodovici L, L, Tringle Tringle M, M, Salvadori Salvadori G, G, Caderni Caderni P, P, Dolara Dolara mutagenicity mutagenicity Urine Urine S. S. 1986. 1986. Truswell Truswell M, M, Woolton Woolton C, C, waite waite Braith A, A, Arlauskas Arlauskas AM, AM, Bonin Bonin I, I, Darnton-Hill Darnton-Hill RSU, RSU, Baker Baker 19:15-18. 19:15-18. Technometrics Technometrics regression. regression. ear ear lin in in observations observations influential influential of of Detection Detection RD. RD. 1977. 1977. Cook Cook Lett Lett 16:81-89. 16:81-89. 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Mutagenicity and Mutagen 6:476. genotoxicity of nitroarenes: All nitro-containing chemicals were not created equal. Mutat Res 114:217-267. Sousa J, Nath J, Tucker JD, Ong T. 1985. Dietary factors af fecting the urinary mutagenicity assay system: I. Detection Santella RM. 1988. Application of new techniques for the of mutagenic activity in human urine following a fried beef detection of carcinogen adducts to human population meal. Mutat Res 149:365-374. monitoring. Mutat Res 205:271-282. Terwel L, van der Hoeven JCM. 1985. Antimutagenic activ SAS Institute Inc. 1985. SAS User's Guide: Version 5 Edi ity of some naturally occurring compounds towards ciga tion. Cary, NC. rette-smoke condensate and benzo[a]pyrene in the Salmo nella/microsome assay. Mutat Res 152:1-4. Schenker MB. 1980. Diesel exhaust exposure: An occupa tional carcinogen? JOM 22:41-46. Vainio H. 1985. Current trends in the biological monitoring of exposure to carcinogens. Scand J Work Environ Health Schenker MB, Smith T, Munoz A, Woskie S, Speizer FE. 11:1-6. 1984. Diesel exposure and mortality among railroad work ers: Results of a pilot study. Br J Ind Med 41:320-327. Venitt S. 1988. The use of short-term tests for the detection of genotoxic activity in body fluids and excreta. Mutat Res Schenker MB, Speizer FE. 1980. A retrospective cohort 205:331-353. study of diesel exhaust exposure in railroad workers: Study design and methodologic issues. In: Health Effects of Diesel Willems MI, deRat WK, Wesstra JA, Bakker GL, Dubois G, Engine Emissions: Proceedings of an International Sympo vanDokkum W. 1989. Urinary and fecal mutagenicity in car sium Held at Cincinnati, OH, on December 3-5, 1979 mechanics exposed to diesel exhaust and 4 unexposed of (Pepelko WE, Danner RM, Clarke NA, eds.) pp. 1114-1126. fice workers. Mutat Res 222:375-391. U.S. Environmental Protection Agency. EPA-600/9-80-057. Government Printing Office, Bethesda, MD. Wong 0, Morgan RW, Kheifets L, Larson SR, Wharton MD. 1985. Mortality among members of a heavy construction Schlotzhauer WS, Chortyk OT. 1983. Effects of varied smok equipment operators union with potential exposure to ing machine parameters on deliveries of total particulate diesel exhaust emissions. Br J Ind Med 42:435-448. matter and selected smoke constituents from an ultra low tar cigarette. J Anal Toxicol 7:92-95. Woskie SR, Smith TJ, Hammond SK, Schenker MB, Gar shick E, Speizer FE. 1988a. Estimation of the diesel exhaust Schuetzle D. 1983. Sampling of vehicle emissions for chem exposures of railroad workers: I. Current exposures. Am J ical analysis and biological testing. Environ Health Perspect Ind Med 13:381-394. 47:65-80. Woskie SR, Smith TJ, Hammond SK, Schenker MB, Gar Schuetzle D, Riley TL, Prater TJ, Harvey TM, Hunt DF. 1982. shick E, Speizer FE. 1988b. Estimation of the diesel exhaust 40 41 41 48:121-129. 48:121-129. Res Res Mutat Mutat Salmonella. Salmonella. on on N-nitrosamines N-nitrosamines of of Mutagenicities Mutagenicities 1977. 1977. M. M. Okado Okado T, T, Sugimura Sugimura T, T, Matshushima Matshushima Y, Y, Seino Seino M, M, Nagao Nagao T, T, Yahagi Yahagi 18:330-333. 18:330-333. Technol Technol Sci Sci Environ Environ foam. foam. polyurethane polyurethane with with vapors vapors hydrocarbon hydrocarbon aromatic aromatic polycyclic polycyclic of of lection lection 136:163-174. 136:163-174. col the the on on volatility volatility of of Influence Influence 1984. 1984. T. T. Bidleman Bidleman S, S, You You Acta Acta Chim Chim Anal Anal particulates. particulates. exhaust exhaust diesel diesel in in nitroarenes nitroarenes mutagenic mutagenic of of identification identification and and Isolation Isolation 1982. 1982. AL. AL. game game 74:3555-3559. 74:3555-3559. USA USA Burlin SM, SM, Rappaport Rappaport ET, ET, Wei Wei Jin Jin JP, JP, ZL, ZL, Nachtman Nachtman XB, XB, Xu Xu Sci Sci Acad Acad Natl Natl Proc Proc urine. urine. mutagenic mutagenic have have smokers smokers Cigarette Cigarette XAD-2: XAD-2: resin resin nonpolar nonpolar the the with with absorption absorption by by urine urine from from 13:395-404. 13:395-404. Med Med Ind Ind J J Am Am posures. posures. mutagens mutagens of of Concentration Concentration 1977. 1977. BN. BN. Ames Ames E, E, Yamasaki Yamasaki ex historical historical and and National National II. II. workers: workers: railroad railroad of of exposures exposures al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers If YES, please answer the following questions (a-h) APPENDIX A. Health Survey (a) How MANY cigarettes did you smoke each day in the past TWO DAYS? (recall that there are 20 cigar ettes in each package) __Yesterday (all day) CA~D • __Today (all day until ID NUMBER now) TODAY Is DATE {b) What BRAND of cigarettes do you now smoke? (please {:no/day/yrl be as specific as possible, e.g. Marlboro Light 100' s) (c) What SIZE are your cigarettes? Please complete the following set of questions. If you are no1:. (1) regular sure of how to respond to a question. please ask the staff for (2)-kingsize clarification. Your responses are CONFIDENTIAL and will NOT be (3}-100'5 released to anyone except in summary form. Thank you in (4}-120'5 advance for your cooperation. (5)=other, please specify: CARD II ID NUMBER 1. What is your BIRTHDATE? (month)/ _{day)/ _(year) (d) Are your cigarettes 2. What is your current JOB TiTLE? FILTERED TIP? - YES '0 I e) your 3. What are your current JOB DUTIES? Are cigarettes MENTHOL? - YES "0- I f) Are your cigarettes LIGHTS? - YES NO 4. How long have you had this job? __years (g) Are your cigarettes ULTRALIGHTS? - YES '0 5. In the PAST TWO DAYS, have you worked in the presence of diesel (h) Do you INHALE? _YES NO_ exhaust fumes? _YES NO_ If YES, how deeply? (1) _into your mouth only If YES, for each day please indicate the approximate length ( 2) back of your throat of time exposed and the level of intensity. Intensity is ( 3) =top of your lungs classified as: (4)_deep into your lungs LIGHT--no smell or eye irritation ~RATE--occasional odor and/or eye irritation HEAVY--strong odor and intense eye irritation 10. In the PAST TWO DAYS, have you smoked . PIPE? - YES NO - Intensity If YES, how many ?ipefills each day? __Yesterday __Today (a) no exposure {a) light (b) -less than 1 hour (b) -rr.oderate If YES, do you inhale? _YES NO (c) l up to J hours (c)_heavy - (d) 3 up to 5 hours (e) 5 or more hours 11. In the PAST TWO DAYS, have you smoked a CIGAR? _YES NO_ (a) no exposure (a) light If YES, how many cigars each day? __Yesterday (b) -less than l hour (b)-moderate __Today (c) -1 up to 3 hours (cl=heavy (d) -3 up to 5 hours If YES, do you inhale _YES NO_ (e) =5 or more hours 6. Are any of the following HEATING SYSTEMS regularly (i.e. at 12. In the PAST TWO DAYS, have you used CHEWING TOBACCO or least once per week) used in your home and/or working areas? SNUFF? _YES NO_ (please check YES or NO for each type) If YES, how many ounces each day? __Yesterday wood stove __Today coal stove fireplace 13. LA,ST NIGHT, were people smoking in your presence? _YES NO_ 7. What type of COOKING STOVE is regularly used in your home? (please check only one) If YES, how many people? __people (a) electric stove (b) =gas stove 14. In your OFFICE of WORK AREA, does anyone other than yourself (c) wood stove REGULARLY smoke cigarettes? _YES NO_ (d)=other, please specify: If YES, how many people were smoking in your presence B. In the PAST \'lEEK, have you regularly worked with any of the TODAY? __people following mater1als at WORK or at HOME? (clease each part (a-j) by checking NO or YES for each) NO YES Work Hom (a} Raw D1esel Fuel (b) Gasoline. LS. In the PAST TWO DAYS, have you used any of the following? (please check YES or NO for each item) (c) Motor Oil Aspirin...... • YES NO (d) Paint Stripper. Nicotine gum . . ... -YES NO- Vitamins ...... -YES NO- (e) Pa1nt Thinner Any other medication . -YES NO- If YES, please S[)eci!y (inclu~both "?rescription and (f) Welding Fumes over-the-counter medications): (g) Adhesives or Glue (h) Solvents. 16. Has a doctor EVER told you that you have any of the following conditions? (please check YES or NO for each item) ( i} Cutting Oils. Asthma. . (j} Pesticides. Diabetes. Emphysema Psoriasis High Blood Pressure 9. In the PAST l'i'EEK, have you regularly smoked cigarettes (at Kidney Disease. . . least one cigarette per day)? _YES NO_ Bladder Disease .. Cancer ..• If NO, please skip to question 10 on the next page If YES, what type 42 43 43 conclu- same same the the to to led led shown) shown) (not (not results results The The 26. 26. rates rates + + blank blank saline saline phosphate-buffered phosphate-buffered the the of of mean mean the the of of traction traction the the of of logarithms logarithms and and roots roots square- the the for for repeated repeated were were ses ses sub by by corrected corrected were were shown) shown) (not (not values values plate plate dividual dividual analy the the check, check, To To nonsmokers. nonsmokers. and and smokers smokers for for ponents ponents In models. models. components-of-variance components-of-variance by by analyzed analyzed were were com variance variance the the roughly, roughly, equalize, equalize, to to ability ability its its by by judged judged B.1 B.1 Table Table in in data data mutagenicity mutagenicity retest retest and and reextraction reextraction The The as as here, here, suc,cessful suc,cessful was was transformation transformation fifth-root fifth-root The The B.1). B.1). (Table (Table months months component. component. six six of of period period the the over over decrease decrease to to appeared appeared nonsmokers nonsmokers and and retest retest large large the the for for control control to to batch batch test test each each on on block block should should smokers smokers both both for for thiocyanate thiocyanate of of concentrations concentrations the the However, However, treatment treatment statistical statistical The The together. together. analyzed analyzed be be should should significantly. significantly. change change to to appear appear not not did did also also retested retested were were batches batches Sample Sample possible. possible. if if three, three, beyond beyond number number the the that that extracts extracts initial initial The The changes. changes. order-of-magnitude order-of-magnitude no no increase increase to to enough enough substantial substantial is is variation variation plate-to-plate plate-to-plate the the were were There There decreasing. decreasing. or or increasing increasing of of pattern pattern consistent consistent First, First, design. design. study study for for implications implications have have results results These These any any have have not not did did months months six six approximately approximately for for storage storage after after nonsmokers nonsmokers and and smokers smokers of of urine urine the the in in activity activity mutagenic mutagenic effects. effects. plate plate and and the the that that indicate indicate B.1, B.1, Table Table in in retest retest for for summarized summarized components components results, results, the the of of The The that that one-third one-third about about is is variance variance between-person between-person the the nonsmokers, nonsmokers, For For zero. zero. or or negligible negligible day. day. is is sample sample same same the the of of extracts extracts different different between between tion tion collection collection initial initial the the after after months months six six activity activity mutagenic mutagenic varia the the sources, sources, other other for for accounting accounting After After sample. sample. one one for for retested retested was was and and nitrogen, nitrogen, with with sealed sealed vial vial screw-cap screw-cap for for plates plates among among that that to to comparable comparable is is extraction extraction same same the the amber amber an an in in 20°C 20°C - at at stored stored was was portion portion first first the the of of tion tion of of retests retests between between Variability Variability nonsmokers. nonsmokers. and and smokers smokers for for extrac XAD-2 XAD-2 initial initial the the after after remaining remaining material material any any tract, tract, estimation estimation of of methods methods two two the the for for similar similar are are results results The The ex actual actual the the on on storage storage of of effect effect the the for for test test To To centration. centration. con thiocyanate thiocyanate and and mutagenicity mutagenicity for for tested tested and and extracted extracted B.2. B.2. Table Table was was it it day, day, in in collection collection shown shown initial initial are are the the after after estimation, estimation, of of months months six six methods methods both both mately mately for for and and smokers, smokers, approxi frozen; frozen; immediately immediately non was was and and portion portion smokers smokers for for other other The The nate. nate. components components variance variance estimated estimated The The thiocya of of concentration concentration the the for for and and mutagenicity mutagenicity for for tested tested normality. normality. of of assumption assumption nominal nominal a a has has option option and and XAD-2 XAD-2 with with extracted extracted immediately immediately was was portion portion One One maximum-likelihood maximum-likelihood The The procedure. procedure. variance-components variance-components portions. portions. equal equal two two into into split split and and collected collected were were smokers smokers SAS SAS the the in in options options maximum-likelihood maximum-likelihood and and type-I type-I the the non seven seven and and smokers smokers seven seven from from samples samples urine urine smokers, smokers, of of means means by by analyzed analyzed were were data data The The interaction.) interaction.) extraction extraction non and and smokers smokers from from urine urine the the in in concentration concentration cyanate cyanate test a a estimate estimate to to possible possible not not was was it it only, only, extract extract nal nal thio and and mutagenicity mutagenicity on on storage storage of of effect effect the the examine examine To To origi the the on on performed performed was was retest retest a a (Because (Because repeatability. repeatability. analytic analytic the the of of indications indications are are others others the the to to relative relative nents nents Concentration Concentration Thiocyanate Thiocyanate and and Mutagenicity Mutagenicity compo extraction extraction and and test test the the of of sizes sizes The The 1980). 1980). Cochran Cochran Urinary Urinary on on B. B. Storage Storage of of Effects Effects APPENDIX APPENDIX and and (Snedecor (Snedecor variance variance of of components components as as known known are are fects fects ef plate plate and and person, person, extraction, extraction, test, test, the the of of variances variances The The variation. variation. systematic systematic the the reflecting reflecting fixed, fixed, as as treated treated was was effect effect person person each each smokers, smokers, CONFIDENTIAL. CONFIDENTIAL. are are responses responses your your Again, Again, questions. questions. of of the the For For persons. persons. among among varying varying randomly randomly as as treated treated set set preceeding preceeding the the completing completing in in cooperation cooperation your your for for you you Thank Thank was was effect effect person person the the nonsmokers, nonsmokers, the the For For nonsmokers. nonsmokers. and and NO_ NO_ _YES _YES washing)? washing)? with with out out rinse rinse smokers smokers for for separately separately analyzed analyzed were were data data the the Therefore, Therefore, that that dyes dyes count count not not (do (do DYE DYE HAIR HAIR use use PER~ENT PER~ENT currenly currenly you you Do Do 21. 21. smoked. smoked. cigarettes cigarettes of of number number the the with with systematically systematically SURE_ SURE_ NOT NOT NO_ NO_ _YES _YES shampoo? shampoo? TYPE TYPE '!'AR '!'AR a a this this Is Is vary vary rates rates mutagenicity mutagenicity that that shown shown have have we we smokers, smokers, For For use? use? regularly regularly you you do do of of brand brand SHA.~OO SHA.~OO What What 20. 20. (Random) (Random) (Random) (Random) (Random) (Random) (Fixed) (Fixed) Effect Effect Effect Effect + + + + + + Effect Effect describe: describe: Effect Effect please please YES, YES, If If Mean Mean + + y y Plate Plate Person Person Reextraction Reextraction Retest Retest Overall Overall NO_ NO_ _YES _YES sprays? sprays? or or chemicals chemicals of of use use regular regular involves involves that that hobby hobby a a have have currently currently you you 19. 19. Do Do was: was: model model statistical statistical basic basic The The NO_ NO_ _YES _YES . . . . • • • • . . • • , , • • • • • • . . • • . . • • • • . . • • • • . . CABBAGE CABBAGE NO- -YES -YES ...... • • • • SPROUTS SPROUTS BRUSSEL BRUSSEL NO -YES -YES salami) salami) bacon, bacon, bologna, bologna, (e.g. (e.g. MEATS MEATS PROCESSED PROCESSED 26)o.zo. 26)o.zo. + + (x (x NO- -YES -YES ...... SHELLFISH SHELLFISH broiled broiled or or barbequed barbequed Grilled, Grilled, NO- -YES -YES ...... FISH FISH broiled broiled or or barbequed barbequed Grilled, Grilled, y y therefore, therefore, was, was, transformation transformation final final The The variance. variance. the the NO- -YES -YES ...... CHICKEN CHICKEN broiled broiled or or barbequed barbequed Grilled, Grilled, NO NO -YES -YES ...... PORK PORK broiled broiled or or barbequed barbequed Grilled, Grilled, stabilize stabilize to to taken taken was was root root fifth fifth the the 1983), 1983), Felton Felton and and Moore Moore NO NO YES YES ...... BEEF. BEEF. broiled broiled or or barbequed barbequed Grilled, Grilled, in in (referenced (referenced Irr Irr and and Snee Snee of of suggestion suggestion the the Following Following food) food) each each for for NO NO or or YES YES check check (please (please FOODS? FOODS? following following the the of of any any of of serving serving one one least least at at eaten eaten you you have have DAYS, DAYS, TWO TWO PAST PAST the the In In 18. 18. x. x. rate, rate, corrected corrected plate's plate's each each to to 26 26 of of addition addition NO_ NO_ _YES _YES vegetarian? vegetarian? a a currently currently you you 17. 17. Are Are by by removed removed were were which which values, values, negative negative to to led led This This plates. plates. al. al. et et Schenker Schenker B. B. M. M. Table B.l. Effects of Storage on Mutagenic Activity and Thiocyanate Concentrations of Smokers' and Nonsmokers' Urine Extractsa Urine Mutagenicity Thiocyanate Retest of Immediately Immediately Tested Immediately Tested After Six-Month After Six-Month Tested Urine Urine Mutagenicityb Thiocyanate Storagec Storage Mutagenicityd Sample Identifier Smoker/Nonsmoker (TA98 revertants/mL) (nmol/mL) (T A98 revertants/mL) (nmollmL) (TA98 revertants/mL) S-1 Smoker 21.2 248 14.2 73 32.0 S-2 Smoker ND8 596 57.2 290 ND S-3 Smoker 0 106 0 62 2.6 S-4 Smoker 22 106 5.8 87 16.4 S-5 Smoker 69 302 74.6 306 55.2 S-6 Smoker 21.6 128 34.2 96 38.6 S-7 Smoker 92.2 334 ND 198 99.4 NS100 Nonsmoker 0.6 208 4.6 148 4.8 NS101 Nonsmoker 4.4 216 7.6 154 5.8 NS102 Nonsmoker 6.8 120 4 ND 7.6 NS103 Nonsmoker 4.8 90 5.2 48 11.2 NS104 Nonsmoker ND 184 7.6 158 17.2 NS105 Nonsmoker ND 200 2 128 13 NS106 Nonsmoker ND 160 7.2 ND 20.2 OTl Nontobacco smoker 2.8 ND 10.4 133 4.4 PBS Buffer extract blank 2 ND 3.8 5.2 a Each table entry is the mean of duplicate or triplicate determinations. b Urine samples split: half immediately extracted and tested; half frozen at - 20'C. c Frozen urine sample thawed, extracted, and tested. d Remainder of extract from immediately tested sample stored at - ZO'C for six months and retested. 8 ND ~ not determined. 45 45 ~ ~ n n 277. 277. b b an an 289. 289. ~ ~ 10.8 10.8 2.3 2.3 1.3 1.3 0.9 0.9 -0.8 -0.8 16.6 16.6 3.1 3.1 1.9 1.9 1.2 1.2 10.3 10.3 - Difference Difference 4.6 4.6 1.5 1.5 0.9 0.9 0.6 0.6 0.2 0.2 17.5 17.5 2.3 2.3 1.0 1.0 0.7 0.7 0.3 0.3 deletion deletion Point Point 14.9 14.9 3.5 3.5 2.3 2.3 0.6 0.6 1.7 1.7 25.5 25.5 5.2 5.2 3.3 3.3 2.2 2.2 0.8 0.8 line line Tangent Tangent Slopes Slopes Estimated Estimated of of Errors Errors Standard Standard 14.3 14.3 3.5 3.5 0.1 0.1 2.0 2.0 -8.1 -8.1 160.0 160.0 16.7 16.7 2.0 2.0 1.1 1.1 - 3 3 16. 16. - Difference Difference 25.5 25.5 6.2 6.2 2.7 2.7 0.7 0.7 -8.6 -8.6 133.8 133.8 5.6 5.6 16.9 16.9 3.5 3.5 12.4 12.4 - deletion deletion Point Point 39.9 39.9 8.9 8.9 4.4 4.4 1.9 1.9 -8.5 -8.5 258.5 258.5 33.3 33.3 7.6 7.6 3.2 3.2 7 7 6. 6. - line line Tangent Tangent Slopes Slopes Estimated Estimated Maximum Maximum Quartile Quartile Median Median Quartile Quartile Minimum Minimum Maximum Maximum Quartile Quartile Median Median Quartile Quartile Minimum Minimum Third Third First First Third Third First First S9b S9b Without Without S9a S9a With With Methods Methods Point-Deletion Point-Deletion and and Tangent-Slope Tangent-Slope by by Calculated Calculated Urine Urine of of Equivalent Equivalent Milliliter Milliliter per per Revertants Revertants of of Comparison Comparison C.l. C.l. Table Table pro- method method tangent-slope tangent-slope the the that that seen seen be be can can C.l. C.l. It It Table Table (A1) (A1) (Dose-Squared) (Dose-Squared) c c (Dose) (Dose) b b + + a a + + Revertants Revertants in in shown shown are are methods methods two two the the for for statistics statistics Descriptive Descriptive urine: urine: of of 0.81. 0.81. was was correlation correlation rank rank the the S9, S9, without without (dose) (dose) concentration concentration the the to to plate plate per per revertants revertants of of number number samples samples the the for for while while 0.73, 0.73, was was methods methods two two the the between between the the relating relating equation equation quadratic quadratic a a fitting fitting by by began began method method correlation correlation rank rank Spearman Spearman the the S9, S9, with with samples samples the the For For each each section, section, Methods Methods the the in in described described As As urine. urine. of of lent lent tions. tions. equiva milliliter milliliter per per revertants revertants of of number number the the ways ways two two in in observa all all used used method method tangent-slope tangent-slope the the whereas whereas fit, fit, calculating calculating of of results results the the compare compare we we appendix, appendix, this this In In the the for for points points dose dose four four or or three, three, two, two, of of use use the the in in resulted resulted method method This This fit. fit. linear linear this this to to b b slope slope fitted fitted the the by by estimated estimated Urine Urine of of Equivalent Equivalent Milliliter Milliliter per per Revertants Revertants of of then then was was urine urine of of milliliter milliliter per per revertants revertants of of number number The The Number Number the the Computing Computing for for Methods Methods Tangent-Slope Tangent-Slope and and Point-Deletion Point-Deletion C. C. of of Comparison Comparison APPENDIX APPENDIX (A2) (A2) (Dose) (Dose) b b a a + + Revertants Revertants = = servations: servations: ob remaining remaining the the to to fit fit was was model model linear linear a a 0.15, 0.15, than than greater greater was was nonlinearity nonlinearity for for value value p p the the which which at at stage stage earliest earliest the the At At dropped. dropped. also also was was point point dose dose highest highest next next the the 0.15, 0.15, than than nonsmokers. nonsmokers. and and smokers smokers less less again again was was for for term term similar similar squared squared the the variance) variance) for for value value reextraction reextraction p p the the If If negligible negligible points. points. the the of of ception ception dose dose ex the the (with (with remaining remaining the the not not to to fit fit were were was was model model components components quadratic quadratic estimated estimated A A The The dropped. dropped. ponents. ponents. was was point point dose dose highest highest the the com 0.15, 0.15, than than variance variance the the less less of of was was value value p p the the contributions contributions If If relative relative the the about about sions sions squares. squares. least least by by (A1) (A1) curve curve fitting fitting after after c c coefficient coefficient squared squared dose the the of of value value p p the the evaluated evaluated method method second second The The applies. applies. term term variance variance random random no no fixed; fixed; as as treated treated were were Smokers Smokers b b fitted. fitted. is is (A1) (A1) curve curve if if b b parameter parameter the the be be to to this this component. component. zero zero or or small small a a shows shows calculus calculus Simple Simple 0.0. 0.0. is is dose dose when when (A1) (A1) curve curve to to gent gent of of indicative indicative is is estimate estimate negative negative the the negative; negative; be be cannot cannot variance variance true true a a A A tan line line the the of of slope slope the the as as urine urine of of equivalent equivalent milliliter milliliter per per 0.0288 0.0288 0.0344 0.0344 0.0476 0.0476 0.0476 0.0476 plates plates Between Between revertants revertants of of number number the the estimated estimated communication, communication, sonal sonal 0.0179 0.0179 0.0161 0.0161 persons persons Between Between b b b b per a a in in Meier Meier Paul Paul by by suggested suggested calculation, calculation, first first The The 0.0000 0.0000 0.0000 0.0000 0.0025a 0.0025a - 0.0058 0.0058 Reextraction Reextraction 0.0191 0.0191 0.0407 0.0407 0.0324 0.0324 0.0543 0.0543 Retest Retest here. here. compared compared are are toxicity, toxicity, for for test test direct direct a a of of absence absence the the in in relation, relation, dose-response dose-response Likelihood Likelihood I I Type Type Likelihood Likelihood I I Type Type the the of of Maximum Maximum portion portion linear linear the the calculating calculating of of Maximum Maximum methods methods Two Two factors. factors. other other to to or or cells), cells), Smokers Smokers Nonsmokers Nonsmokers fewer fewer in in (resulting (resulting toxicity toxicity to to chance, chance, to to due due be be may may curve curve Storage Storage of of Effects Effects the the to to appearance appearance nonlinear nonlinear Any Any dose. dose. and and revertants revertants tween tween on on Studies Studies in in Variance Variance of of Components Components B.2. B.2. Table Table be relationship relationship linear linear a a suggests suggests theory theory Toxicological Toxicological al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers duced systematically higher values than did the point overestimated the linear slope at the lower dose points. Sec deletion method. Inspection of individual plots in which ond, when the observations showed only slight but consis the discrepancy was large shows two reasons for this. First, tent curvature, the point-deletion method sometimes did where the plot showed strong evidence of toxicity only at not drop one or both of the highest two doses. Yet, the the highest dose point (an "obvious" downward bending of curves appeared to be bending. In this case, the tangent the curve), the point-deletion method dropped the point. slope method appeared to be estimating a true line at zero. Quadratic equation (A1) did not fit the data well and so it APPENDIX D. Urinary Mutagenicity Correlation with Multivariate Regression Variables Table D.l. Multivariate Regression Model of TA98 + S9 Mutagenicity Among Nonusers of Tobaccoa Parameter Estimate SE Estimate/SE Constant -0.407 0.162 -2.51b Standard mutagen 2-aminofluorene (revertants/ng) 0.282 0.063 4.45c 2-Aminofluorene x Cotinine (~g/~mol of creatinine)/1,000 -0.150 1.168 -0.13 Protective food (1 = yes) -0.104 0.034 - 3.09d Respirable particles (~g/m 3 )/1,000 - 0.537 0.359 -1.50 Ambient nicotine (~g/m 3 )/1,000 - 1.175 3.824 -0.31 Diesel exhaust exposure (1 = yes)e - 0.031 0.046 -0.68 Diesel exhaust exposure x respirable particles/1,000 0.471 0.379 1.24 Diesel exhaust exposure x ambient nicotine/1,000 -1.838 7.853 -0.23 Within-person variance 0.098 0.016 Between-person variance 0.040 0.018 a Mutagenicity is expressed as revertants/~mol of creatinine; total number of samples is 124. b 0.01 < p "' 0.05. c p"' 0.001. d 0.001 < p "' 0.01. e Chi-squared (3 degrees of freedom) ~ 1.87 and p > 0.05 for diesel exhaust exposure parameters. Table D.2. Multivariate Regression Model of TA98 - S9 Mutagenicity Among Nonusers of Tobaccoa Parameter Estimate SE Estimate/SE Constant -0.235 0.139 -1.69 Standard mutagen 2-aminofluorene (revertants/ng) 0.126 0.054 2.33b 2-Aminofluorene x Cotinine (~g/~mol of creatine)/1,000 0.020 1.021 0.02 Protective food (1 = yes) -0.014 0.029 -0.48 Respirable particles (~g/m 3 )/1,000 -0.298 0.306 -0.97 Ambient nicotine (~g/m 3 )/1,000 1.035 3.325 0.31 Diesel exhaust exposure (1 = yes)c -0.017 0.039 -0.45 Diesel exhaust exposure x respirable particles/1,000 0.264 0.323 0.82 Diesel exhaust exposure x ambient nicotine/1,000 - 1.457 6.675 -0.22 Within-person variance 0.072 0.013 Between-person variance 0.026 0.013 a Mutagenicity is expressed as revertants/~mol of creatinine; total number of samples is 119. b 0.01 < p"' 0.05. c Chi-squared (3 degrees of freedom) ~ 0.82 and p > 0.05 for diesel exhaust exposure parameters. 46 47 47 parameters. parameters. > > exposure exposure exhaust exhaust ~ ~ diesel diesel for for p p 0.05 0.05 and and 5.39 5.39 freedom) freedom) of of degrees degrees (3 (3 Chi-squared Chi-squared b b 59. 59. is is samples samples of of number number total total creatinine; creatinine; of of revertantsi).lmol revertantsi).lmol as as expressed expressed is is Mutagenicity Mutagenicity a a 0.019 0.019 0.070 0.070 variance variance Between-person Between-person 0.019 0.019 0.011 0.011 variance variance Within-person Within-person -1.56 -1.56 4.050 4.050 6.314 6.314 - nicotine/1,000 nicotine/1,000 ambient ambient x x exposure exposure exhaust exhaust Diesel Diesel 0.02 0.02 0.319 0.319 0.006 0.006 ,000 ,000 particles/1 particles/1 respirable respirable x x exposure exposure exhaust exhaust Diesel Diesel 0.72 0.72 0.055 0.055 0.039 0.039 yes)b yes)b (1 (1 exposure exposure exhaust exhaust Diesel Diesel = = 1.90 1.90 4.050 4.050 7.351 7.351 ,000 ,000 )/1 )/1 (Jlg/m nicotine nicotine Ambient Ambient 3 -0.29 -0.29 0.297 0.297 -0.085 -0.085 )/1,000 )/1,000 (Jlg/m particles particles Respirable Respirable 3 1.48 1.48 0.187 0.187 0.277 0.277 day/100 day/100 on on smoked smoked Cigarettes Cigarettes -1.78 -1.78 0.213 0.213 -0.380 -0.380 creatinine)/1,000 creatinine)/1,000 of of (Jlg/Jlmol (Jlg/Jlmol Cotinine Cotinine x x 2-Aminofluorene 2-Aminofluorene 0.85 0.85 0.072 0.072 0.061 0.061 (revertants/ng) (revertants/ng) 2-aminofluorene 2-aminofluorene mutagen mutagen Standard Standard -0.25 -0.25 0.191 0.191 0.048 0.048 Constant Constant Estimate/SE Estimate/SE SE SE Estimate Estimate Parameter Parameter Smokersa Smokersa Cigarette Cigarette Among Among Mutagenicity Mutagenicity S9 S9 - TA98 TA98 of of Model Model Regression Regression Multivariate Multivariate D.4. D.4. Table Table parameters. parameters. > > exposure exposure exhaust exhaust diesel diesel for for p p 0.05 0.05 and and 6.15 6.15 freedom) freedom) of of degrees degrees (3 (3 Chi-squared Chi-squared d d p"' p"' < < 0.01. 0.01. 0.001 0.001 c c < < p"' p"' 0.05. 0.05. 0.01 0.01 b b 63. 63. is is samples samples of of number number total total creatinine; creatinine; of of revertantsi).lmol revertantsi).lmol as as expressed expressed is is Mutagenicity Mutagenicity a a 0.644 0.644 0.235 0.235 variance variance Between-person Between-person 0.794 0.794 3.152 3.152 variance variance Within-person Within-person 17.056 17.056 2.27b 2.27b - -38.788 -38.788 nicotine/1,000 nicotine/1,000 ambient ambient x x exposure exposure exhaust exhaust Diesel Diesel 1.70 1.70 1.749 1.749 2.971 2.971 particles/1,000 particles/1,000 respirable respirable x x exposure exposure exhaust exhaust Diesel Diesel -0.39 -0.39 0.317 0.317 -0.122 -0.122 yes)d yes)d (1 (1 = = exposure exposure exhaust exhaust Diesel Diesel 14.868 14.868 2.67c 2.67c 39.724 39.724 )/1,000 )/1,000 (Jlg/m nicotine nicotine Ambient Ambient 3 1.67 1.67 - 1.593 1.593 2.656 2.656 - )/1,000 )/1,000 (Jlg/m particles particles Respirable Respirable 3 1.072 1.072 4.53b 4.53b 4.879 4.879 day/100 day/100 on on smoked smoked Cigarettes Cigarettes 3.21 3.21 1.091 1.091 3.502 3.502 ,000 ,000 )/1 )/1 creatinine creatinine of of (Jlg/Jlmol (Jlg/Jlmol Cotinine Cotinine x x 2-Aminofluorene 2-Aminofluorene -0.11 -0.11 0.391 0.391 -0.045 -0.045 (revertants/ng) (revertants/ng) 2-aminofluorene 2-aminofluorene mutagen mutagen Standard Standard 0.17 0.17 1.169 1.169 0.199 0.199 Constant Constant Estimate/SE Estimate/SE SE SE Estimate Estimate Parameter Parameter Smokersa Smokersa Cigarette Cigarette Among Among Mutagenicity Mutagenicity S9 S9 TA98 TA98 of of Model Model + + Regression Regression Multivariate Multivariate D.3. D.3. Table Table al. al. et et Schenker Schenker M. M. B. B. 48 48 dates. dates. same same the the on on [revertants/~mol [revertants/~mol evaluated evaluated samples samples study study of of creatinine) creatinine) of of mutagenicity mutagenicity with with dates, dates, processing processing 13 13 on on Assayed Assayed b b parentheses. parentheses. in in given given is is value value p p the the coefficient; coefficient; correlation correlation the the is is entry entry table table Each Each a a (0.01) (0.01) 0.272 0.272 (0.27) (0.27) 0.120 0.120 - (<0.01) (<0.01) 0.343 0.343 (0.03) (0.03) 0.229 0.229 87 87 S9 S9 - (0.68) (0.68) 0.045 0.045 S9 S9 0.01) 0.01) (< (< (0.64) (0.64) 90 90 0.331 0.331 0.050 0.050 - (0.03) (0.03) + + 0.231 0.231 Smokers Smokers (0.52) (0.52) 0.052 0.052 (0.55) (0.55) 0.049 0.049 - (0.01) (0.01) 152 152 0.204 0.204 (0.01) (0.01) 0.200 0.200 S9 S9 - (0.25) (0.25) 0.092 0.092 (0.92) (0.92) 0.008 0.008 S9 S9 0.01) 0.01) (< (< 0.309 0.309 0.01) 0.01) (< (< + + 0.276 0.276 160 160 Nonsmokers Nonsmokers itroquinoline-N-oxide itroquinoline-N-oxide N N 4- 2-Nitrofluorene 2-Nitrofluorene 2-Aminofluorene 2-Aminofluorene Benzo[a]pyrene Benzo[a]pyrene n n Standards Standards S9 S9 - Standards Standards S9 S9 + + Mutagenb Mutagenb Standard Standard Statusa Statusa Smoking Smoking by by Mutagens Mutagens Standard Standard of of (revertants/ng) (revertants/ng) Mutagenicity Mutagenicity of of Correlation Correlation Rank Rank Spearman Spearman E.3. E.3. Table Table 19.0 19.0 7.06 7.06 + + 75.3 75.3 27.97 27.97 DMSO DMSO 37.12 37.12 13 13 20.4 20.4 7.70 7.70 57.1 57.1 37.68 37.68 21.52 21.52 DMSO DMSO 13 13 Variation Variation SD SD (%) (%) Variation Variation SD SD (%) (%) Mean Mean n n S9 S9 Standard Standard of of Coefficient Coefficient of of Coefficient Coefficient Within-Experiment Within-Experiment Between-Experiment Between-Experiment Revertants/Plate Revertants/Plate Standards Standards Negative Negative in in Activity Activity Mutagenic Mutagenic E.2. E.2. Table Table 0.19 0.19 + + 6.6 6.6 0.84 0.84 28.7 28.7 2.86 2.86 13 13 Benzo[a]pyrene Benzo[a]pyrene 0.25 0.25 + + 7.1 7.1 0.76 0.76 3.46 3.46 22.0 22.0 13 13 2-Aminofluorene 2-Aminofluorene 0.35 0.35 4.8 4.8 0.35 0.35 8.9 8.9 13 13 7.30 7.30 4-Nitroquinoline-N-oxide 4-Nitroquinoline-N-oxide 0.87 0.87 5.1 5.1 1.79 1.79 10.4 10.4 13 13 17.24 17.24 2-Nitrofluorene 2-Nitrofluorene SD SD Variation Variation Variation Variation SD SD (%) (%) (%) (%) n n Standard Standard Mean Mean S9 S9 of of Coefficient Coefficient of of Coefficient Coefficient Experiment Experiment Within- Between-Experiment Between-Experiment Points Points Dose Dose Two Two Zero Zero from from Revertants/ng Revertants/ng + + Standards Standards Positive Positive in in Activity Activity Mutagenic Mutagenic E.l. E.l. Table Table Controls Controls E. E. Activity Activity APPENDIX APPENDIX Mutagenic Mutagenic of of Descriptions Descriptions Statistical Statistical Workers Workers Railroad Railroad in in Exhaust Exhaust Diesel Diesel to to Exposure Exposure of of Markers Markers 49 49 diffi- very very be be would would that that -levels -levels 1-nitropyrene 1-nitropyrene of of ng ng 1 1 than than the the of of mass mass the the one-thousandth one-thousandth than than less less is, is, that that ticles; ticles; less less and and pyrene pyrene of of ng ng 10 10 to to 0.5 0.5 only only expect expect would would we we mg, mg, par respirable respirable of of mg mg 0.1 0.1 than than less less collected collected most most and and mass, mass, 0.1 0.1 of of sample sample exhaust-exposed exhaust-exposed diesel diesel personal personal a a in in Thus, Thus, of of mg mg 0.5 0.5 than than less less collected collected samples samples personal personal 500) 500) than than Council1981). Council1981). Research Research (National (National 1-nitropyrene 1-nitropyrene of of ng ng 10 10 to to more more (of (of all all Almost Almost low. low. quite quite were were concentrations concentrations ticle ticle 6 6 and and pyrene pyrene of of ng ng 100 100 to to 5 5 only only are are there there exhaust, exhaust, diesel diesel par the the that that was was workers workers railroad railroad among among exposure exposure exhaust exhaust automobile automobile of of mg mg 1 1 in in However, However, exhaust. exhaust. diesel diesel in in than than diesel diesel of of study study earlier earlier this this from from finding finding major major second second A A smoke smoke tobacco tobacco environmental environmental in in higher higher be be may may ratio ratio the the more. more. because because mass, mass, particles particles respirable respirable to to smoke smoke tobacco tobacco mental mental or or magnitude magnitude of of order order one one by by lower lower generally generally levels, levels, lower lower environ and and exhaust exhaust diesel diesel of of contributions contributions the the separating separating at at present present be be to to tend tend PAHs PAHs other other All All fluoranthene. fluoranthene. by by lowed lowed of of means means a a as as suggested suggested been been has has 1-nitropyrene 1-nitropyrene to to rene rene fol PAHs, PAHs, major major the the are are derivatives derivatives its its and and phenanthrene phenanthrene py of of ratio ratio The The validated. validated. been been yet yet not not has has it it but but exhaust, exhaust, that that indicated indicated also also trucks trucks and and automobiles automobiles from from haust haust diesel diesel for for marker marker good good a a be be to to prove prove may may It It particles. particles. smoke smoke ex diesel diesel of of studies studies literature literature The The filter). filter). the the by by collected collected cigarette cigarette and and diesel diesel between between separation separation good good shows shows but but efficiently efficiently was was phenanthrene phenanthrene the the of of half half than than less less but but filter, filter, particles, particles, exhaust exhaust diesel diesel for for specific specific not not is is carbon carbon Elemental Elemental the the on on remained remained BaP BaP the the of of percent percent 100 100 which which in in section, section, 1 1 undertaken. undertaken. was was study study this this time time the the at at validated validated or or oped oped Aim Aim Specific Specific Results, Results, the the in in results results experiment experiment laboratory laboratory devel been been not not had had markers markers exhaust exhaust diesel diesel suggested suggested the the of of the the (see (see better better much much mentioned mentioned PAHs PAHs other other the the collect collect do do many many but but exhaust, exhaust, diesel diesel for for suggested suggested been been have have markers markers they they but but well, well, phenanthrenes phenanthrenes collect collect not not do do filters filters that that fact fact candidate candidate other other several several study, study, this this of of completion completion Since Since the the by by reinforced reinforced was was conclusion conclusion This This present. present. ones ones nant nant matrix. matrix. extract extract exhaust exhaust diesel diesel the the in in reactions reactions antibody antibody predomi the the were were PAHs PAHs these these that that reasonable reasonable it it deemed deemed of of specificity specificity the the validate validate to to have have would would One One possible. possible. are are researchers researchers the the derivatives, derivatives, alkylated alkylated its its and and phenanthrene phenanthrene mixture mixture the the in in PAHs PAHs other other with with reactions reactions cross cross and and sitive, sitive, find find did did analyses analyses spectrometry spectrometry mass mass and and matography matography sen more more significantly significantly be be to to likely likely not not are are BaP BaP to to antibodies antibodies chro gas gas the the Because Because 20 20 ~-tg/mL. ~-tg/mL. approximately approximately was was which which Specific Specific sample. sample. per per ng ng 1 1 about about of of detection detection of of limit limit a a with with method, method, the the of of detection detection of of limit limit the the reflects reflects probably probably detection, detection, fluorescence fluorescence with with chromatography chromatography liquid liquid using using This This identified. identified. not not were were fluoranthene) fluoranthene) or or BaP, BaP, nitropyrene, nitropyrene, method method a a to to sample, sample, per per mg mg 4 4 about about of of detection detection of of limit limit (pyrene, (pyrene, exhaust exhaust diesel diesel for for markers markers potential potential as as gested gested a a with with methods, methods, spectrometry spectrometry mass mass and and chromatography chromatography sug PAHs PAHs other other Specifically, Specifically, methylfluorene). methylfluorene). had had sample sample gas gas from from changing changing by by 1985) 1985) al. al. et et (Hammond (Hammond study study earlier earlier (one (one phenanthrenes phenanthrenes alkylated alkylated and and phenanthrene phenanthrene tified tified the the from from improved improved were were PAHs PAHs for for detection detection of of limits limits The The iden samples samples ofthese ofthese analyses analyses spectrometry spectrometry mass mass and and phy phy chromatogra gas gas The The 1985). 1985). al. al. et et (Hammond (Hammond 1987). 1987). spectrometry spectrometry al. al. et et (Hammond (Hammond mass mass and and smoke smoke tobacco tobacco chromatography chromatography gas gas environmental environmental using using of of study study analyzed analyzed were were chamber chamber tions tions the the from from frac samples samples in in the the and and found found were were extract extract BaP BaP raw raw of of the the levels levels both both of of Detectable Detectable aliquots aliquots mass, mass, detection. detection. of of greatest greatest the the with with samples samples the the limit limit of of the the at at several several just just For For or or below below are are values values chromatography. chromatography. these these liquid liquid samples; samples; personal personal by by the the on on compounds) compounds) polar polar collected collected and and be be would would BaP BaP hydrocarbons, hydrocarbons, of of ng ng 0.5 0.5 aromatic aromatic and and 0.05 0.05 carbons, carbons, between between hydro (aliphatic (aliphatic levels, levels, fractions fractions high high into into to to exposed exposed separated separated and and workers workers railroad railroad concentrated concentrated from from that, that, this this from from then then were were extracts extracts predict predict These These could could One One dichloromethane. dichloromethane. exhaust. exhaust. with with diesel diesel extracted extracted automobile automobile from from cles cles was was mass mass parti of of particle particle the the of of milligram milligram per per percent percent found found 40 40 were were BaP BaP of of ng ng Approximately Approximately 5 5 . . and and 0.5 0.5 ~-tg/m tween tween 3 486 486 to to 25 25 from from be ranged ranged which which in in concentrations concentrations Council1981), Council1981), Research Research respirable-mass respirable-mass (National (National data data lished lished The The analysis. analysis. pub chemical chemical with with intensive intensive agreement agreement for for in in are are BaP BaP exposure, exposure, of of haust haust levels levels low low The The filters. filters. ex diesel diesel highest highest the the of of on on ng ng areas areas 4 4 the the and and shops, shops, plugs plugs repair repair foam foam running running the the polyurethane polyurethane the the on on threne threne from from matter matter phenan of of ng ng particulate particulate 230 230 respirable respirable of of approximately approximately had had samples samples samples samples high-volume high-volume these these while while 23 23 BaP), BaP), ng ng 0.5 0.5 collected collected was was we we detection detection addition, addition, of of In In (limit (limit 1988). 1988). BaP BaP al. al. et et of of levels levels (Hammond (Hammond tectable tectable smoke smoke tobacco tobacco de than than less less environmental environmental had had by by samples samples these these interference interference of of for for four four All All marker marker shop. shop. this this repair repair the the correct correct to to from from need need the the samples samples and and two two of of exhaust exhaust extracts extracts diesel diesel plug plug for for foam foam marker marker a a as as cles cles polyurethane polyurethane the the parti respirable respirable of of and and filter filter usefulness usefulness the the the the both both on on examined examined we we performed performed were were particular, particular, BaP BaP for for Analyses Analyses In In here. here. reported reported study study the the in in used used were were that that exhaust exhaust diesel diesel study. study. the the of of scope scope the the beyond beyond was was this this but but PAHs, PAHs, of of range range a a for for markers markers of of selection selection the the guided guided study study that that of of results results for for filters filters and and plugs plugs foam foam polyurethane polyurethane the the analyze analyze to to been been The The rates. rates. cancer cancer lung lung and and exposures exposures exhaust exhaust diesel diesel of of had had plan plan original original The The study. study. this this in in analyses analyses the the of of target target investigation investigation an an conducted conducted authors authors the the study, study, this this to to Prior Prior the the became became phenanthrene phenanthrene above, above, given given reasons reasons the the For For PAHs. PAHs. most most of of nanogram nanogram a a than than less less Exhaust Exhaust Diesel Diesel much much and and PAHs, PAHs, all all of of levels levels nanogram nanogram only only contain contain would would for for Marker Marker a a as as F. F. Phenanthrene Phenanthrene APPENDIX APPENDIX sample sample personal personal each each best, best, at at Thus, Thus, samples. samples. high-volume high-volume al. al. et et Schenker Schenker B. B. M. M. Markers of Exposure to Diesel Exhaust in Railroad Workers cult to measure in the highly exposed workers. This would diesel exhaust exposure. Furthermore, collection of blood result in large uncertainties in the calculated ratios. samples in field epidemiologic studies is not as feasible as Another category of proposed markers includes biologi collection of urine samples, because blood collection is cal markers such as DNA or hemoglobin adducts. However, much more invasive. Attempts to collect blood samples the use of a biological marker of diesel exhaust would alter would have resulted in lower subject-response rates. The the basic study design, namely, measurement of diesel ex situation is very different from using paid volunteers in a haust exposure as a determinant of urine mutagenicity, a laboratory. Similarly, collecting 24-hour urine samples is biological marker. Measurement of another biological also not feasible in this setting, even though it would be marker of diesel exhaust, such as hemoglobin adducts, preferable to using spot samples. would itself have to be validated against some measure of 50 51 51 activation activation environ- exhaust, exhaust, diesel diesel to to exposures exposures of of studies studies include include and and metabolic metabolic without without strain strain tester tester Salmonella Salmonella S9 S9 - TA98 TA98 studies studies epidemiological epidemiological for for assessment assessment exposure exposure on on cus cus fo interests interests research research Hammond's Hammond's Dr. Dr. 1981. 1981. in in Health Health Public Public enzyme enzyme S9 S9 by by activation activation of of School School Harvard Harvard the the from from Sciences Sciences Health Health vironmental vironmental metabolic metabolic with with strain strain tester tester Salmonella Salmonella S9 S9 TA98 TA98 + + En in in M.S. M.S. an an and and 1976 1976 in in University University Brandeis Brandeis from from istry istry error error standard standard SE SE chem in in Ph.D. Ph.D. a a received received She She MA. MA. Worcester, Worcester, School, School, ical ical Med Massachusetts Massachusetts of of University University the the at at Pharmacology Pharmacology and and deviation deviation standard standard SD SD Medicine Medicine Community Community and and Family Family of of Departments Departments the the in in saline saline phosphate-buffered phosphate-buffered PBS PBS Professor Professor Associate Associate an an is is Ph.D. Ph.D. Hammond, Hammond, Katharine Katharine S. S. hydrocarbon hydrocarbon aromatic aromatic polycyclic polycyclic PAH PAH contaminants. contaminants. borne borne 4-nitroquinoline-N-oxide 4-nitroquinoline-N-oxide 4-NQO 4-NQO air of of mechanisms mechanisms toxicological toxicological and and exposures, exposures, cinogen cinogen 2-nitrofluorene 2-nitrofluorene 2-NF 2-NF car and and mutagen mutagen for for humans humans of of monitoring monitoring include include terests terests in research research Kadds Kadds Dr. Dr. Health. Health. Public Public of of School School Harvard Harvard chromatography chromatography liquid liquid high-pressure high-pressure HPLC HPLC the the at at Biology Biology Cancer Cancer of of Department Department the the in in training training doctoral doctoral dimethylsulfoxide dimethylsulfoxide DMSO DMSO post his his received received and and Berkeley, Berkeley, California, California, of of University University Sciences, Sciences, Health Health Environmental Environmental and and Biomedical Biomedical benzo[a]pyrene benzo[a]pyrene of of ment ment BaP BaP Depart the the from from Toxicology Toxicology and and Sciences Sciences Health Health ronmental ronmental 2-aminofluorene 2-aminofluorene 2-AF 2-AF Envi in in Ph.D. Ph.D. a a received received He He Davis. Davis. California, California, of of University University Toxicology, Toxicology, Environmental Environmental of of Department Department the the in in Professor Professor ABBREVIATIONS ABBREVIATIONS Assistant Assistant Adjunct Adjunct an an and and Board Board Resources Resources Air Air California California the the for for Toxicologist Toxicologist Staff Staff a a is is Ph.D., Ph.D., Kado, Kado, Y. Y. Norman Norman studies. studies. health health reproductive reproductive in in considerations considerations power power and and design design are are interests interests special special His His (1978). (1978). Washington Washington of of sity sity studies. studies. fects fects Univer the the at at program program Biomathematics Biomathematics the the from from Ph.D. Ph.D. his his ef health health for for characterization characterization exposure exposure is is emphasis emphasis special special received received He He Davis. Davis. California, California, of of University University the the at at Gynecology Gynecology Her Her Hygiene. Hygiene. Industrial Industrial of of Board Board American American the the by by giene giene and and Obstetrics Obstetrics of of and and Medicine Medicine Internal Internal of of Departments Departments the the hy industrial industrial of of practice practice comprehensive comprehensive the the in in certified certified in in Professor Professor Adjunct Adjunct J. J. Associate Associate is is Ph.D. Ph.D. Samuels Samuels Steven Steven Board is is Woskie Woskie Dr. Dr. Health. Health. Public Public of of School School University University Harvard Harvard the the from from Hygiene Hygiene Industrial Industrial in in M.S. M.S. her her ceived ceived exposure. exposure. hazardous hazardous of of markers markers biological biological cluding cluding re She She School. School. Medical Medical Massachusetts Massachusetts of of University University the the at at in lung, lung, the the to to hazards hazards environmental environmental and and occupational occupational Medicine Medicine Community Community and and Family Family of of Department Department the the in in ist ist of of epidemiology epidemiology the the on on focus focus interests interests research research His His Health. Health. Hygien Industrial Industrial Research Research a a is is C.I.H., C.I.H., Woskie, Woskie, R. R. Susan Susan Public Public of of School School Harvard Harvard the the from from M.P.H. M.P.H. an an received received and and School, School, Medical Medical Harvard Harvard Laboratory, Laboratory, Channing Channing the the at at ology ology studies. studies. epidemiological epidemiological epidemi in in trained trained He He medicine. medicine. occupational occupational and and disease, disease, in in modeling modeling pharmacological pharmacological of of use use the the is is emphasis emphasis cial cial pulmonary pulmonary medicine, medicine, internal internal in in training training clinical clinical pleted pleted spe His His Minnesota. Minnesota. of of University University the the from from Health Health mental mental com and and Francisco, Francisco, San San at at California California of of University University the the Environ in in degrees degrees Ph.D. Ph.D. and and M.S. M.S. his his both both received received Smith Smith from from degree degree M.D. M.D. his his received received He He Health. Health. Environmental Environmental Dr. Dr. School. School. Medical Medical Massachusetts Massachusetts of of University University gram, gram, and and Occupational Occupational for for Center Center the the of of Component Component Davis Davis the the of of Pro Sciences Sciences Health Health Environmental Environmental Medicine, Medicine, munity munity Director Director and and Medicine, Medicine, Internal Internal of of Department Department the the within within Com and and Family Family of of J. J. Professor Professor is is Ph.D., Ph.D., Smith, Smith, Thomas Thomas Medicine Medicine Environmental Environmental and and Occupational Occupational of of Division Division the the of of Chief Chief is is He He Davis. Davis. at at California California of of University University the the at at cine cine contaminants. contaminants. of of concentrations concentrations air air Medi of of Professor Professor Associate Associate is is M.D., M.D., Schenker, Schenker, B. B. Marc Marc to to exposure exposure of of markers markers biological biological various various of of relationships relationships the the studying studying been been has has research research her her in in theme theme related related A A line. line. AUTHORS AUTHORS THE THE ABOUT ABOUT gaso and and fluids, fluids, machining machining styrene, styrene, smoke, smoke, tobacco tobacco mental mental al. al. et et Schenker Schenker B. B. M. M. 53 53 do do than than hydrocarbons hydrocarbons gaseous gaseous and and monoxide monoxide carbon carbon of of the the of of 6(b)(5) 6(b)(5) Section Section standards. standards. health health and and safety safety tional tional levels levels lower lower emit emit and and fuel-efficient fuel-efficient more more are are engines engines Diesel Diesel occupa of of promulgation promulgation informed informed the the to to relates relates also also haust haust ships. ships. and and machinery, machinery, equipment, equipment, farm farm trains, trains, buses, buses, ex diesel diesel to to exposure exposure of of effects effects health health the the on on Research Research trucks, trucks, in in use use wide wide found found have have engines engines diesel diesel Heavy-duty Heavy-duty vehicles. vehicles. of of categories categories fleet. fleet. automotive automotive European European the the of of fraction fraction significant significant or or a a classes classes from from matter matter particulate particulate of of emissions emissions to to plicable plicable represent represent they they market, market, States States United United the the of of segment segment large large a a ap regulations regulations requires requires similarly, similarly, 202(a)(3)(A)(iii), 202(a)(3)(A)(iii), Section Section captured captured not not have have cars cars diesel-powered diesel-powered although although Today, Today, fort. fort. engines." engines." or or vehicles vehicles heavy-duty heavy-duty of of ...... classes classes from from nitrogen nitrogen ef this this stimulated stimulated 1970s, 1970s, early early the the during during especially especially cles, cles, of of oxides oxides and and hydrocarbons, hydrocarbons, monoxide, monoxide, carbon carbon of of sions sions vehi motor motor in in engines engines diesel diesel of of use use Increased Increased abroad. abroad. and and emis to to applicable applicable ...... regulations regulations "prescribe "prescribe to to ministrator ministrator States States United United the the in in research research intense intense of of focus focus a a been been have have Ad the the directs directs specifically specifically 202(a)(3)(A)(i) 202(a)(3)(A)(i) Section Section fare:' fare:' exhaust exhaust engine engine diesel diesel to to exposure exposure of of effects effects health health The The wel or or health health public public endanger endanger to to anticipated anticipated be be reasonably reasonably may may which which pollution pollution air air to, to, contribute contribute or or cause, cause, judgment judgment BACKGROUND BACKGROUND SCIENTIFIC SCIENTIFIC his his in in which which engines, engines, vehicle vehicle motor motor new new or or vehicles vehicles motor motor new new of of classes classes or or class class any any from from pollutant pollutant air air any any of of sions sions emis the the to to applicable applicable standards standards ...... revise) revise) time time to to time time from from (and (and "prescribe "prescribe to to Administrator Administrator the the directs directs 202(a)(1) 202(a)(1) standards. standards. health health tional tional Section Section 1977. 1977. in in amended amended as as Act, Act, Air Air occupa Clean Clean other other the the and and of of 202 202 these these of of modification modification and and creation creation the the Section Section under under emissions emissions with with other) other) (and (and connection connection diesel diesel in in for for standards standards Administration Administration the the by by making making decision decision sets sets (EPA) (EPA) Agency Agency Protection Protection informed informed Environmental Environmental to to U.S. U.S. The The contribute contribute will will workers, workers, railroad railroad as as such such lation, lation, popu occupational occupational discrete discrete a a within within exhaust exhaust engine engine diesel diesel BACKGROUND BACKGROUND REGULATORY REGULATORY to to exposure exposure of of effects effects health health the the to to relating relating Research Research 1988). 1988). Health Health and and Safety Safety Occupational Occupational for for Institute Institute (National (National section section this this under under carcinogen'' carcinogen'' occupational occupational tial tial public. public. the the to to and and HEI HEI the the of of sponsors sponsors the the "poten a a as as listed listed be be exhaust exhaust diesel diesel whole whole that that mendation mendation to to aid aid an an as as perspective, perspective, in in Report Report Investigators' Investigators' the the place place recom the the in in culminated culminated Health Health and and Safety Safety Occupational Occupational to to intended intended is is Commentary Commentary Committee's Committee's Review Review Health Health for for Institute Institute National National the the by by research research Recent Recent 1990.111[f]). 1990.111[f]). The The Commentary. Commentary. Committee's Committee's Review Review the the in in and and Report Report section section C.F.R. C.F.R. (29 (29 evidence'' evidence'' scientific scientific available available the the of of tent tent Investigators' Investigators' the the in in issues issues clarify clarify to to and and comments comments change change ex and and nature nature the the on on "depending "depending categories categories risk risk into into gens gens ex to to opportunity opportunity the the had had investigators investigators the the and and Committee Committee carcino potential potential of of placement placement the the for for provide provide standards standards Review Review the the Report, Report, Investigators' Investigators' the the of of review the the During During latter latter These These 1990). 1990). part part C.F.R. C.F.R. 29 29 (see (see carcinogens carcinogens tional tional occupa colleagues. colleagues. potential potential his his of of and and regulation regulation Schenker Schenker and and Dr. Dr. classification, classification, cation, cation, by by made made subsequently subsequently were were that that identifi the the for for modifications modifications standards standards pending pending promulgated promulgated has has Administration Administration 1988, 1988, January January in in Committee Committee Review Review Health Health the the the the by by addition, addition, cepted cepted In In agents. agents. harmful harmful potentially potentially to to posure posure ac was was and and 1987 1987 May May in in HEI HEI the the at at ex received received was was Report Report occupational occupational regulating regulating standards standards numerous numerous mulgated mulgated Investigators' Investigators' The The $388,638. $388,638. were were pro has has expenditures expenditures Total Total 1983. 1983. Administration, Administration, Health Health and and Safety Safety cupational cupational October October in in began began project project three-year three-year Oc The The the the Davis. Davis. at at through through California California Secretary, Secretary, the the directive, directive, this this to to Pursuant Pursuant of of University University the the to to moved moved Schenker Schenker Dr. Dr. when when necessary necessary field." field." were were that that changes changes staff staff of of because because 1983 1983 of of summer summer the the in in the the in in data data scientific scientific available available latest latest the the ...... and and periments, periments, modified modified then then was was project project the the proposal; proposal; original original the the proved proved ex demonstrations, demonstrations, "research, "research, including including evidence;' evidence;' able able ap HEI HEI The The Exposure." Exposure." Exhaust Exhaust Diesel Diesel to to Sensitivity man man avail best best the the of of basis basis the the "on "on standards standards such such promulgate promulgate Hu of of Monitoring Monitoring "Biologic "Biologic entitled entitled proposal proposal a a submitted submitted to to Secretary Secretary the the directs directs act act the the of of section section This This life. life. ing ing MA, MA, Boston, Boston, University, University, Harvard Harvard Hospital, Hospital, Women's Women's and and work her her or or his his of of period period entire entire the the for for agent agent harmful harmful the the to to Brigham Brigham the the at at then then Schenker, Schenker, B. B. Marc Marc RFA, RFA, this this to to sponse sponse exposure exposure regular has has employee employee the the if if even even capacity capacity tional tional re In In Populations:' Populations:' Susceptible Susceptible of of "Models "Models for for proposals proposals func or or health health of of impairment impairment material material suffer suffer will will employee employee soliciting soliciting 82-3) 82-3) (RFA (RFA Applications Applications for for Request Request a a issued issued no no that that ensuring ensuring of of goal goal the the with with agents, agents, physical physical harmful harmful (HEI) (HEI) Institute Institute Effects Effects Health Health the the 1982, 1982, of of summer summer the the In In other other and and materials materials toxic toxic to to exposure exposure governing governing standards standards health health and and safety safety occupational occupational create create to to Labor Labor of of Secretary Secretary INTRODUCTION INTRODUCTION the the directs directs 1970 1970 of of Act Act Health Health and and Safety Safety Occupational Occupational Institute Institute Effects Effects Health Health ~ ~ ------~~~~~------fE( fE( COMMENTARY COMMENTARY COMMITTEE'S COMMITTEE'S REVIEW REVIEW HEALTH HEALTH mO:..L ______Health Review Committee's Commentary_ gasoline-fueled engines; however, they produce 30 to 100 culty has been to identify sufficiently large groups of indi times more particles or soot. viduals who have been exposed to diesel engine exhaust Diesel engine exhaust consists of a complex mixture of exclusive of other confounding exposures. Specific popu gaseous compounds and particulate matter. The composi lations identified as being at risk include occupational tion of the exhaust varies considerably, depending on en groups such as transportation workers, operators of heavy gine design, fuel, lubrication, emission control technology, construction equipment, railroad workers, and miners. A and operating conditions (Johnson 1988). The available second difficulty has been to document the actual concen data base, which refers mainly to light-duty engine exhaust, trations of diesel exhaust to which the individuals have was characterized over 10 years ago, and may not be repre been, or are, exposed. sentative of current emissions or of emissions from heavy In epidemiologic studies, exposure can be assigned ei duty engines. Hundreds of chemicals, including polycyclic ther quite broadly, based on job classification and work site, aromatic hydrocarbons, have been identified in the gaseous or more precisely, by sampling ambient air over time. In phase of diesel engine exhaust (International Agency for Re early epidemiology studies of the health risks of diesel en search on Cancer 1989). The particle phase contains ag gine exhaust, past exposures were typically estimated by gregates of small carbon particles with adsorbed organic questionnaire, job description, or, occasionally, by air sam and inorganic compounds. The fact that diesel engine ex pling at current job sites. Exposures that are known to con haust particles are readily respirable (0.1 to 0.5 ~m) and found estimates of diesel exhaust exposure include ciga have genotoxic compounds adsorbed to their surfaces raises rette smoke, coal dust, inorganic dusts, and asbestos (Smith concerns regarding their effects on human health. et al. 1984; Wynder and Higgins 1986; Woskie et al. 1988); Numerous investigators, using molecular and cellular ex cigarette smoke is, by far, the largest confounder. perimental systems, have demonstrated the capacity of con Several recent reviews of epidemiologic investigations stituents of diesel exhaust to act as mutagens, to produce were designed to examine the possible association between chromosomal alterations, and to transform mammalian cancer and occupational exposure to diesel engine exhaust cells (Lewtas 1982; International Agency for Research on (Wynder and Higgins 1986; McClellan 1987; International Cancer 1989). These test systems include the Salmonella Agency for Research on Cancer 1989; McClellan et al. typhimurium mutagenicity assays (Huisingh et al. 1978), 1990). The exposed populations included employees in the the mouse lymphoma assay (Mitchell et al. 1980), and the railroad industry (Kaplan 1959; Howe et al. 1983; Garshick determination of the induction of sister chromatid ex et al. 1987, 1988), transportation workers (Raffle 1957; Silver changes in Chinese hamster ovary cells and primary cul man et al. 1986) and miners (Waxweiler et al. 1973). These tures from the lungs of exposed Syrian hamsters (Pereira investigations focused on the risk of lung and bladder can 1982; Li et al. 1983). It has been shown that extracts from cer. The earlier studies generally yielded negative or diesel particles and individual constituents of diesel emis equivocal results, but because of their many limitations, sions are tumorigenic in the mouse-skin bioassay, suggest particularly problems with exposure assessment and the ing the potential carcinogenicity of diesel exhaust (Kotin et failure to exclude cigarette smoking as a confounding fac al. 1955; Nesnow et al. 1984). An increase in lung tumor in tor, it is difficult to draw definitive conclusions. cidence has been shown in rats chronically exposed to high Some of these limitations were addressed in more recent levels of diesel engine exhaust (Brightwell et al. 1986; studies in which investigators examined the relationship Ishinishi et al. 1986; Iwai et al. 1986; Mauderly et al. 1986; between lung cancer and diesel engine exhaust exposure in Stober 1986). After reviewing the epidemiological, gena a large population of American railroad workers. In the first toxicity, and carcinogenicity data relating to diesel exhaust study (Garshick et al. 1987), a case-control study, diesel ex and its constituents, the International Agency for Research haust exposure in workers who had at least ten years of rail on Cancer evaluated diesel engine exhaust to be "probably road service was estimated from an evaluation of job carcinogenic to humans" (International Agency for Research descriptions and work areas, as provided by the U.S. Rail on Cancer 1989). In the United States, the National Institute road Retirement Board. In a later report, the same group of for Occupational Safety and Health has recommended that investigators (Woskie et al. 1988) provided an analysis of the whole diesel exhaust be regarded as a "potential occupa current levels of exposure to respirable particulate matter as tional carcinogen" (National Institute for Occupational Safe determined from an analysis of personal and area samples ty and Health 1988). collected from four railroad facilities in the United States. There are many difficulties in using epidemiological ap Cause of death was determined by death certificates, and proaches to establish an association between diesel exhaust next of kin were surveyed to obtain information on smoking. exposure and carcinogenicity in humans. The first diffi- The relative hazard of lung cancer attributable to exposure 54 55 55 pres- the the in in tymphimurium tymphimurium S. S. incubating incubating of of consists consists test test xenobiotics. xenobiotics. to to exposure exposure of of index index quantitative quantitative a a obtain obtain entire entire The The system. system. activation activation metabolic metabolic the the as as to to used used is is liver) liver) evaluated evaluated been been have have markers markers different different and and approaches approaches rat rat from from generally generally fraction, fraction, postmitochondrial postmitochondrial x x 9,000 9,000 g different different of of number number A A susceptibility. susceptibility. of of markers markers and and effect, effect, (a (a homogenate homogenate S9 S9 An An colonies. colonies. form form and and divide divide growth growth for for of of markers markers exposure, exposure, of of markers markers as as classified classified be be can can They They histidine histidine require require not not do do that that cells cells revertant revertant Only Only substance. substance. Council1989). Council1989). Research Research (National (National samples samples or or systems systems logic logic test test the the and and histidine histidine of of trace trace a a only only containing containing bio in in medium medium events events of of indicators indicators as as defined defined been been have have markers markers a a in in incubated incubated is is occurs, occurs, (reversion) (reversion) mutation mutation back back a a Biological Biological less less dose. dose. internal internal to to exposure exposure relate relate to to and and posure, posure, un histidine histidine of of absence absence the the in in grow grow cannot cannot therefore therefore and and ex personal personal monitor monitor to to markers markers developing developing in in interest interest ing ing grow, grow, to to histidine histidine requires requires that that typhimurium typhimurium Salmonella Salmonella of of increas been been has has there there field, field, health health environmental environmental the the In In strain strain a a assay, assay, Salmonella Salmonella the the In In class. class. separate separate a a as as nized nized pollutants. pollutants. airborne airborne to to populations populations different different of of posure posure recog been been have have carcinogens carcinogens nonmutagenic nonmutagenic that that however, however, ex actual actual the the on on information information accurate accurate more more provide provide to to noted, noted, be be to to is is carcinogens. carcinogens. It It including including agents, agents, chemical chemical potential potential the the has has air air ambient ambient monitoring monitoring with with combination combination of of variety variety a a to to sensitive sensitive and and inexpensive, inexpensive, rapid, rapid, is is it it cause cause in in exposure exposure personal personal Monitoring Monitoring patterns. patterns. activity activity and and be mutation, mutation, gene gene for for test test vitro vitro in in used used widely widely most most the the is is inspiration, inspiration, of of depth depth rate, rate, respiration respiration in in variations variations dividual dividual 1975) 1975) al. al. et et (McCann (McCann assay assay mutagenicity mutagenicity Salmonella Salmonella The The in of of because because worker worker the the to to dose dose actual actual the the to to relate relate ways ways compounds. compounds. notal unidentified unidentified may may yet yet as as however, however, detect detect also also measures, measures, These These exposure. exposure. of of standing standing and and chemicals chemicals of of under spectrum spectrum our our broad broad a a improve improve to to to to taken taken respond respond been been assays assays have have icity icity that that steps steps are are smoke smoke mutagen compounds, compounds, of of tobacco tobacco classes classes or or environmental environmental and and compounds, compounds, exhaust exhaust specific specific tect tect engine engine diesel diesel of of ents ents de to to tailored tailored usually usually are are assays assays constitu analytical analytical While While identifying identifying and and studies. studies. site site job job each each at at samples samples air air bient bient field field in in exposure exposure estimating estimating for for am tool tool Collecting Collecting powerful powerful a a tially tially terminology. terminology. of of interpretation interpretation imprecise imprecise poten is is mutagenicity mutagenicity urine urine and and measuring measuring nonresponse, nonresponse, xenobiotics, xenobiotics, for for misclassification, misclassification, bias, bias, recall recall include include excretion excretion of of route route major major a a represents represents and and descriptions descriptions job job obtained, obtained, and and easily easily questionnaires questionnaires of of use use the the on on reliance reliance is is urine urine Because Because agents. agents. genotoxic genotoxic to to exposure exposure with with monitoring monitoring apparent apparent are are that that drawbacks drawbacks serious serious The The exposure. exposure. to to approach approach another another is is mutagenicity mutagenicity for for estimate estimate to to urine urine Analyzing Analyzing descriptions descriptions category category job job used used frequently frequently have have above, above, discussed discussed those those as as such such studies, studies, Epidemiological Epidemiological source. source. exposure exposure appropriate appropriate the the to to adducts adducts of of fraction fraction relative relative the the assign assign to to is is researchers researchers 1.89). 1.89). and and 1.11 1.11 of of intervals intervals confidence confidence cent cent to to challenge challenge The The adducts. adducts. DNA DNA of of formation formation the the to to per 95 95 lead lead to to 1.45; 1.45; of of risk risk (relative (relative exhaust exhaust engine engine diesel diesel to to posure posure shown shown been been have have 1986) 1986) al. al. et et erath erath Rand Rand 1986; 1986; al. al. et et ex of of (Everson (Everson period period longest longest the the with with workers workers those those in in elevated elevated smoking smoking cigarette cigarette and and 1986) 1986) al. al. et et Wong Wong 1985; 1985; al. al. et et son son significantly significantly was was which which cancer, cancer, lung lung of of risk risk relative relative the the (Jack exposure exposure exhaust exhaust diesel diesel Both Both urine. urine. or or blood, blood, calculate calculate tissues, tissues, to to used used were were ratios ratios rate rate standardized standardized directly directly and and target target in in detected detected be be can can adducts adducts These These proteins. proteins. or or model model RNA, RNA, hazards hazards proportional proportional A A cancer. cancer. lung lung of of cases cases 1,694 1,694 DNA, DNA, as as such such macromolecules, macromolecules, critical critical with with react react revealed revealed agents agents obtained) obtained) were were percent percent (88 (88 certificates certificates death death the the activated activated metabolically metabolically when when formed formed are are that that of of adducts adducts Examination Examination deaths. deaths. 19,396 19,396 been been had had there there time time which which macromolecular macromolecular are are markers markers biochemical biochemical such such at at of of 1980, 1980, amples amples until until followed followed was was cohort cohort The The obtained. obtained. not not were were Ex carcinogens. carcinogens. especially especially compounds, compounds, certain certain to to histories histories posure posure Smoking Smoking years. years. ten ten least least at at for for time time that that at at vice vice ex monitor monitor to to techniques techniques biochemical biochemical sensitive sensitive ser highly highly railroad railroad in in employed employed been been had had and and 1959, 1959, in in age age of of years years and and developed developed newly newly of of advantage advantage take take approaches approaches 64 64 to to Other Other 40 40 were were who who workers workers railroad railroad 55,407 55,407 of of cohort cohort a a in in cancer cancer lung lung of of risk risk the the assessed assessed (1988) (1988) coworkers coworkers and and shick shick assays. assays. available available of of limits limits detection detection the the within within are are that that Gar study, study, cohort cohort retrospective retrospective a a study, study, second second the the In In amounts amounts in in present present are are that that and and unique, unique, nearly nearly or or unique, unique, are are that that markers markers xenobiotic xenobiotic with with successfully successfully used used been been have have exhaust. exhaust. diesel diesel to to exposed exposed were were approaches approaches analytical analytical traditional traditional Such Such 1988). 1988). al. al. they they et et Jarvis Jarvis where where setting setting job job a a in in working working of of chance chance less less had had have have 1987; 1987; O'Neil O'Neil and and (Wallace (Wallace smoke smoke tobacco tobacco to to to to exposure exposure authors authors with with the the by by considered considered were were they they because because separately separately well well correlate correlate to to shown shown been been has has and and urine urine the the in in evaluated evaluated is excreted excreted is was was group group this this death); death); of of time time the the at at years years Cotinine Cotinine Council1986). Council1986). Research Research (National (National 82 82 smoke smoke to to 65 65 tobacco tobacco (age (age workers workers older older of of group group the the in in seen seen was was posure posure environmental environmental or or smoke, smoke, tobacco tobacco tobacco, tobacco, to to exposure exposure for for ex diesel exhaust exhaust diesel of of effect effect No No exhaust. exhaust. diesel diesel to to posure posure markers markers unique unique are are compounds compounds latter latter The The fluids. fluids. body body in in ex of of more more or or years years 20 20 had had who who and and death death of of time time the the at at cotinine cotinine and and nicotine nicotine analyzing analyzing and and blood, blood, in in levels levels globin globin younger younger or or age age of of years years 64 64 were were who who workers workers among among found found carboxyhemo and and hair hair and and fingernails fingernails in in metals metals heavy heavy was was cancer cancer lung lung for for 1.88) 1.88) and and 1.06 1.06 of of intervals intervals confidence confidence measuring measuring include include Examples Examples fluids. fluids. and and tissues tissues iological iological percent percent (95 (95 1.41 1.41 of of ratio ratio odds odds relative relative A A exposure. exposure. asbestos asbestos phys in in xenobiotics xenobiotics inorganic inorganic and and organic organic quantify quantify to to used used and and smoking smoking for for correct correct to to regression regression logistic logistic conditional conditional been been have have techniques techniques chemistry chemistry analytical analytical Traditional Traditional multiple multiple a a using using calculated calculated was was exhaust exhaust engine engine diesel diesel to to ------~ ------~ ~[ ~[ ~~~ ~~~ I£(------Health Review Committee's Commentary ence and absence of S9, with appropriate controls. The JUSTIFICATION FOR THE STUDY standard S. typhimurium tester strains include the TA98 and TA100 strains, which detect frameshift and basepair Under RFA 82-3, "Models of Susceptible Populations;' substitution mutations, respectively. Resins are frequently the HEI solicited proposals for studies that would describe used to concentrate body fluids and to separate mutagens the existence and importance of susceptible human or ani from histidine and other potentially interfering com mal subpopulations that may be particularly vulnerable to pounds. Although determination of urinary mutagens is a mobile-source emissions. In response to this RFA, Schenker promising technique to measure recent exposure to specific and colleagues proposed to examine markers of exposure to carcinogens, confounding factors such as diet, smoking, diesel engine exhaust in a population of railroad workers. and other environmental exposures need to be carefully The investigators planned to explore the usefulness of as evaluated when interpreting the results. says of urine mutagenic activity, as well as the more conven Human exposure to mutagens can occur by a number of tional analysis of personal air samples, to monitor the ex routes: air, water, food, and dermal contact. The contribu posure of individual workers to diesel engine exhaust. They tion of diet to the total mutagenic burden is complex. Food also proposed to develop new approaches to control for the may contain accidentally-occurring mutagenic contaminants, confounding effects of tobacco smoke and other particulate naturally occurring mutagens, as well as mutagens gener matter. ated during food processing (Sugimura et a!. 1981). The The study population was to be selected from railroad mutagenicity of individual food constituents depends on workers at a single railroad where the investigators had pre the cooking method and can be modulated by other compo viously measured exposures (Woskie eta!. 1988). This oc nents in the diet. The mutagenicity of human urine has cupational group was considered a suitable population for been shown to be related to the composition of the diet con studies of the health effects of exposure to diesel engine ex sumed by the subjects (Doolittle eta!. 1989). Recently Lioy haust because typical occupation-related exposures are and coworkers (1988) evaluated the relative contribution of higher for this group than those encountered by the average the air, water, and food routes of exposure for benzo[a]py individual. In previous surveys, the investigators identified rene and found that, in half of the individuals, diet was the a range of exposures to diesel engine exhaust in different predominant source of benzo[a]pyrene exposure. work areas within individual railroad facilities (Woskie et Several investigators have examined the correlation be a!. 1988). tween exposure to diesel engine exhaust or coal dust and urinary mutagens in experimental animal models. Belisario and colleagues (1984) measured mutagenic activity in the GOALS AND OBJECTIVES urine of rats 24 hours after administration of diesel particles and noted a dose-dependent increase in mutagenicity, as The major objectives of this study were to refine and vali tested with a Salmonella/microsome assay; other groups, date methods for estimating exposure to diesel engine ex however, have not found any increase in urinary mutagens haust in epidemiological studies, and to conduct pilot tests after exposure of rodents to diesel exhaust emissions of these approaches in a population of railroad workers. (Pereira eta!. 1981; Ong eta!. 1985). Mutagenic activity has The investigators pursued their objectives through the been detected in the urine of cigarette smokers using the following specific aims: Salmonella assay (Yamasaki and Ames 1977), but not all studies have produced positive results, possibly because of 1. To use personal air samples to compare respirable parti the lack of dietary controls (International Agency for Re cle concentration, a standard measure of exposure to search on Cancer 1986). diesel engine exhaust and other particles, to the concen tration of phenanthrene, which may be a more specific The development of a method to evaluate exposure to marker for diesel emissions. diesel engine exhaust using urinary mutagens, supple mented by measurements of personal exposure to diesel 2. To refine methods to measure exposure to another major and cigarette particulates, represents an experimental ap source of respirable particles, tobacco smoke. proach that warrants further investigation. The use of bio 3. To develop and validate methods to use assays of muta markers as indicators of exposure to diesel exhaust would genic activity in urine samples to assess exposure to contribute to more accurate estimates of exposure in future diesel engine exhaust. epidemiological studies, both in the workplace and in the 4. To conduct a pilot field survey of both diesel exhaust general environment. exposed and unexposed railroad workers using the meth- 56 57 57 activation. activation. ex- metabolic metabolic diesel diesel S9 S9 without without obscured obscured and and with with partially partially TA100, TA100, and and measurements measurements particle particle the the TA98 TA98 to to smoke smoke Salmonella Salmonella strains strains cigarette cigarette of of tester tester bacterial bacterial contribution contribution the the The The using using particles. particles. analyzed analyzed respirable respirable was was total total samples samples of of urine urine levels levels and and similar similar filters filters had had that that sample sample air air both both in in environments environments in in activity activity worked worked jects jects Mutagenic Mutagenic index. index. sub all all grouping grouping that that job job the the appears appears with with it it data, data, correlated correlated these these and and From From tors tors seen. seen. was was grouping grouping moni personal personal of of job job by by subset subset a a on on measured measured was was estimated estimated exposures exhaust exhaust concentration concentration diesel diesel with with correlation correlation Phenanthrene Phenanthrene some some investigated. investigated. fumes, fumes, was was welding welding of of exposure exposure levels levels exhaust exhaust high high to to diesel diesel exposure exposure of of cause cause and and smoke, smoke, cigarette cigarette be for for excluded excluded adjusted adjusted were were carmen carmen concentration, concentration, welding welding particle particle the the ble ble and and smoke, smoke, tobacco tobacco respira between between of of relationship relationship contribution contribution The The the the for for mutagenicity. mutagenicity. corrected corrected urine urine and and were were levels levels particle particle concentration concentration particle particle respirable respirable the the respirable respirable both both When When to to 6). 6). exposure exposure through through 4 4 smoke smoke Tables Tables (see (see egories egories tobacco tobacco passive passive and and active active cat of of job job five five the the in in contribution contribution the the workers workers estimate estimate among among concentration concentration particle particle ble ble to to used used were were shift shift work work the the during during respira to to smoked smoked exposures exposures in in cigarettes cigarettes of of ber ber differences differences clear-cut clear-cut show show to to pear pear num the the and and ap cotinine, cotinine, not not and and do do Report Report nicotine nicotine of of Investigators' Investigators' the the concentrations concentrations in in urine urine presented presented data data The The samples, samples, air air personal personal the the on on measurements measurements Nicotine Nicotine shifts. shifts. Concentration Concentration Particle Particle Respirable Respirable Total Total work work two two the the of of end end the the at at collected collected were were samples samples urine urine and and shifts, shifts, work work consecutive consecutive two two over over taken taken were were samples samples air air authors. authors. the the by by concentration'' concentration'' ticle ticle Personal Personal particles. particles. respirable respirable and and nicotine nicotine between between relation relation par respirable respirable "adjusted "adjusted the the as as to to referred referred is is smoke, smoke, tobacco tobacco the the and and filters filters the the of of efficiency efficiency collection collection nicotine nicotine the the mine mine for for corrected corrected particles, particles, respirable respirable for for value value The The workers. workers. deter to to out out carried carried were were studies studies chamber chamber Additional Additional shop shop and and engineers, engineers, clerks, clerks, welders, welders, welders), welders), without without phenanthrene. phenanthrene. including including compounds compounds ganic ganic and and (with (with carmen carmen brakers, brakers, categories: categories: job job different different five five in in or volatile volatile collected collected that that plugs plugs foam foam polyurethane polyurethane three three workers workers railroad railroad in in nicotine nicotine ambient ambient of of levels levels and and threne, threne, and and nicotine, nicotine, trapped trapped that that filter filter bisulfate-treated bisulfate-treated a a particles, particles, phenan of of concentration concentration concentration, concentration, particle particle respirable respirable respirable respirable collected collected that that filter filter Teflon Teflon efficiency efficiency high high a a ries: ries: total total of of measurements measurements simultaneous simultaneous obtain obtain to to device device se in in collectors collectors three three through through air air drew drew that that designed designed was was sampling sampling air air personal personal new new a a developed developed investigators investigators The The system system sampling sampling unique unique A A emissions. emissions. diesel diesel to to exposure exposure for MONITORING MONITORING ENVIRONMENTAL ENVIRONMENTAL marker marker specific specific potential potential a a as as evaluated evaluated was was engines, diesel diesel railroad railroad from from exhaust exhaust in in concentrations concentrations high high in in found found carbon carbon differently. differently. collected collected were were hydro aromatic aromatic polycyclic polycyclic a a Phenanthrene, Phenanthrene, smoking. smoking. rette rette sources sources two two the the from from samples samples exhaust exhaust diesel diesel the the because because ciga passive passive and and active active for for concentration concentration particle particle spirable spirable possible possible not not was was activity activity mutagenic mutagenic of of comparison comparison direct direct ly, ly, re the the adjust adjust to to measured measured were were concentrations concentrations nicotine nicotine Unfortunate similar. similar. be be may may exhausts exhausts two two the the in in mutagens mutagens Airborne Airborne phenanthrene. phenanthrene. and and nicotine, nicotine, particles, particles, respirable respirable of of curves curves dose-response dose-response the the of of pattern pattern the the that that suggests suggests measure measure to to designed designed were were samplers samplers Personal Personal days. days. two two 4), 4), and and 3 3 (Figures (Figures study study current current the the in in presented presented as as haust, haust, preceding preceding the the over over items items dietary dietary specific specific of of consumption consumption ex diesel diesel locomotive locomotive and and exhaust exhaust diesel diesel automotive automotive of of ity ity the the about about questions questions and and history, history, smoking smoking history, history, posure posure activ mutagenic mutagenic ofthe ofthe analysis analysis the the However, However, characterized. characterized. ex history, history, medical medical brief brief a a included included that that questionnaire questionnaire well well been been not not has has exhaust, exhaust, diesel diesel automotive automotive to to contrast contrast survey survey health health a a completed completed subjects subjects All All location. location. work work and and in in exhaust, exhaust, diesel diesel Locomotive Locomotive 1989). 1989). Cancer Cancer on on search search category category job job of of basis basis the the on on selected selected were were Subjects Subjects haust. haust. Re for for Agency Agency (International (International used used is is equipment equipment powered powered ex engine engine diesel diesel to to workers workers male railroad railroad male 87 87 of of exposure exposure diesel where where mines mines some some in in found found those those as as high high as as not not the the determine determine to to used used were were samples samples personal personal and and Area Area were were they they high, high, were were workers workers railroad railroad the the of of exposures exposures the the While While 1988). 1988). al. al. et et (Woskie (Woskie shops shops repair repair in in working working ~gfm3) ~gfm3) DESIGN DESIGN STUDY STUDY (134 (134 electriCians electriCians and and ) ) ~g/m (114 (114 mechanics mechanics for for were were means means 3 highest highest the the and and ), ), (17 (17 ~g/m clerks clerks for for was was mean mean lowest lowest 3 the the exhaust: exhaust: diesel diesel from from concentrations concentrations particle particle respirable respirable exposures. exposures. occupational occupational of of range range wide wide a a to to exposed exposed were were population population this this in in groups groups non and and smoking, smoking, cigarette cigarette exposure, exposure, exhaust exhaust diesel diesel sub that that indicated indicated reports reports Previous Previous 1988). 1988). al. al. et et Woskie Woskie with with mutagens mutagens urinary urinary of of concentration concentration the the correlate correlate To To 5. 5. 1988; 1988; 1987, 1987, al. al. et et (Garshick (Garshick workers workers railroad railroad in in posure posure evaluated. evaluated. ex exhaust exhaust diesel diesel of of effects effects health health the the on on study study miological miological be be to to were were also also mutagens mutagens urinary urinary and and particles particles ratory ratory epide large large a a to to follow-up follow-up a a as as undertaken undertaken was was study study This This respi of of sources sources Nondiesel Nondiesel 3. 3. through through 1 1 aims aims specific specific in in developed developed been been had had that that smoke smoke tobacco tobacco and and haust haust EVALUATION EVALUATION TECHNICAL TECHNICAL ex engine engine diesel diesel to to exposure exposure personal personal measuring measuring for for ads ads ------rn ------rn m m ~~~ ~~~ ~------H--ea_I_th__ &_e_fl_e_w~C_o_m_m_•~·tt~e~~~s~C~o~mm==~e=nt~a~ry haust exposure differences among the job groups. The sources to investigate this. In this connection, it should be number of smokers and the "degree of enclosure" of the noted that Cantreels and Van Cauwenberghe (1978) reported work site were found to be important variables that deter a gas-phase phenanthrene concentration of 45 ng/m3 in the mined respirable particle concentration on the filters. ambient atmosphere. In the current study, a comparison of the ratio of phenanthrene to particulate mass suggests that Phenanthrene it is unlikely that the phenanthrene to which the clerks were exposed was derived from cigarette smoke; that is, for Phenanthrene is one of the polycyclic aromatic hydrocar clerks the ratio of phenanthrene (160 ng/m3) to respirable bons present in high concentrations in diesel exhaust parti particles (120 j.!g/m3) was 0.003, whereas in cigarette smoke cles. This compound is also present in cigarette smoke at the ratio of phenanthrene (180 ng/m3) to respirable parti concentrations of 0.08 to 0.62 j.!g/cigarette in mainstream cles (1,200 j.!g/m3) was 0.00015. The source of the phenan smoke (International Agency for Research on Cancer 1983). threne to which the clerks were exposed is not known, and The investigators developed a reliable and convenient is one of the major unresolved questions of this study. method to measure very low concentrations of phenan threne (0.1 j.!g or less) in samples of ambient particles col In summary, the investigators developed a technique for lected on the polyurethane filters in the personal air sam the detection and measurement of phenanthrene in en plers. They used these measurements to make independent vironmental samples; however, the use of phenanthrene as estimates of the concentrations of diesel exhaust particles a marker for exposure to diesel exhaust was not fully vali in the work environment. They controlled for phenan dated. Validation of this technique, and more extensive threne originating from cigarette smoke by determining the studies that use phenanthrene or other chemicals present concentration of phenanthrene and respirable particles in in diesel exhaust, will be necessary to develop a reliable tobacco smoke and by doing subset analysis of personal specific marker for diesel exhaust. filters only from nonsmoking shop workers. Phenanthrene concentration in ambient air showed a gradient of exposure Cigarette Smoke with respect to proximity to sources of diesel emission. The levels of environmental tobacco smoke were esti Thus, the repair shop workers were exposed to more mated on the basis of the levels of nicotine collected on phenanthrene than were clerks and carmen; however, as filters downstream of trapped particulate matter. The mea discussed below, the clerks were exposed to more phenan surement of gas-phase nicotine was facilitated by the de threne than expected. velopment of a filter-trapping technique. This technique A constant ratio of phenanthrene to respirable particles appears to be a good method for determining the concentra (0.006 ng/!lg) was found in particles collected in the repair tion of environmental tobacco smoke constituents relative shop, where diesel exhaust concentrations were expected to to the concentration of respirable particles. The ambient be highest. The investigators assumed that diesel particles concentration of environmental tobacco smoke was calcu generated by similar sources and under similar conditions lated on the basis of the following relationship, as deter would show a constant ratio of phenanthrene to particulate mined in chamber studies: respirable particle concentra mass. Using the phenanthrene:particulate mass ratio from tion = 8.6 x [NICOTINE] + 50.5. The authors interpret the repair shop samples, they calculated that half of the respira value of the intercept (50.5) to reflect the statistical uncer ble particles in the personal samples from nonsmoking tainty in extrapolating the chamber data to an airborne nico shop workers were not derived from diesel exhaust, but tine concentration of zero (the intercept is highly variable, were from other airborne particulate matter such as sand with a standard error of 125). and dust particles. The personal samples from workers in different job categories (clerks, carmen, and repair shop Mutagenicity of Respirable Particles workers) showed only a weak relationship between phenan from Personal Samples threne concentration and total or adjusted respirable parti cles (see Figure 9). During the course of this study the investigators also de The data also show that the clerks, who did not work in veloped a method for measuring the mutagenicity of parti the vicinity of any sources of diesel emissions, were ex cle extracts from personal filter samples. They successfully posed to approximately 160 ng/m3 of phenanthrene, which demonstrated that, despite the limited amount of particu is more than twice the concentration found on the field late matter collected on the filters in an eight-hour sampling blanks (66 ng/m3), which was also quite high. The investi time, the microsuspension modification of the Ames assay gators were aware that the value for field blanks was is sufficiently sensitive to detect mutagenic activity. anomalously high, but had neither the time nor the re- The mutagenicity of the small number of personal parti- 58 59 59 not not may may assumption assumption this this However, However, smoke. smoke. cigarette cigarette of of that that gas- the the identifying identifying before before needed needed are are controls controls priate priate to to pattern pattern similar similar a a follows follows exhaust exhaust diesel diesel from from mutagens mutagens appro systems, systems, chromatography chromatography liquid liquid high-performance high-performance of of bioavailability bioavailability the the that that assumed assumed investigators investigators the the First, First, some some in in cotinine cotinine and and caffeine caffeine with with comigrate comigrate to to shown shown been been has has 3-hydroxycotinine 3-hydroxycotinine Since Since nicotine. nicotine. of of follow. follow. metabolites metabolites the the observations observations all all of of these these percent percent 50 50 for for regarding regarding account account comments comments actually actually Several Several may may compounds. compounds. hydroxycotinine, hydroxycotinine, mutagenic mutagenic 3- to to metabolite, metabolite, converted converted nicotine nicotine another another metabolically metabolically that that are are they they indicates indicates that that (1989) (1989) or or workers workers mutagenic, mutagenic, co and and Thuan Thuan are are by by report report engines engines A A diesel diesel of of nonsmokers. nonsmokers. for for emissions emissions the the exposure exposure that that smoke smoke possibility possibility the the tobacco tobacco out out rule rule fact, fact, in in environmental environmental not, not, of of does does indicators indicators correlation correlation best best of of the the lack lack this this out, out, provided provided point point levels levels correctly correctly cotinine cotinine they they As As urinary urinary and and engines. engines. nicotine nicotine diesel diesel of of ambient ambient that that emissions emissions and and the the to to nine, nine, posure posure coti ex was was and and smokers smokers urine urine in in of of urine urine the the in in mutagenicity mutagenicity activity activity between between mutagenic mutagenic of of correlation correlation a a late late discern discern corre to to highest highest unable unable the the were were that that they they suggested suggested smoking, smoking, analyses analyses cigarette cigarette active active Regression Regression and and count. count. samples samples ac into into urine urine in in taken taken also also was was mutagenicity mutagenicity of of smoked smoked levels levels the the cigarettes cigarettes of of between between number number the the association association an an smokers, smokers, For For found found they they smoke. smoke. although although cigarette cigarette to to that, that, exposure exposure conclude conclude passive passive and and investigators investigators tive tive The The ac estimate estimate to to measured measured were were monitors, monitors, air air personal personal from from activation. activation. metabolic metabolic S9 S9 without without and and with with controls, controls, nicotine nicotine ambient ambient as as well well as as samples, samples, urine urine in in thiocyanate thiocyanate blank blank and and positive positive the the of of responses responses actual actual the the as as well well as as and and cotinine, cotinine, nicotine, nicotine, of of Levels Levels urine. urine. of of mutagenicity mutagenicity samples, samples, urine urine of of mutagenicity mutagenicity the the on on data data primary primary the the both both observed observed the the to to smoke smoke tobacco tobacco to to exposure exposure of of contribution contribution examine examine to to want want undoubtedly, undoubtedly, will, will, emissions emissions diesel diesel of of the the determine determine to to effort effort strong strong a a made made investigators investigators The The genotoxicity genotoxicity the the evaluate evaluate to to seeking seeking Others Others samples. samples. urine urine the the on on data data mutagenicity mutagenicity primary primary critical critical the the of of inclusion inclusion Exposure Exposure Smoke Smoke Cigarette Cigarette the the by by strengthened strengthened been been have have would would report report the the form; form; mary mary sum in in data data their their present present investigators investigators the the report, report, this this In In activation). activation). metabolic metabolic S9 S9 without without and and with with S9. S9. TA100, TA100, and and TA98 TA98 strains strains typhimurium typhimurium S. S. (using (using assay assay Ames Ames without without or or with with either either samples, samples, urine urine in in activity activity mutagenic mutagenic the the of of 1986) 1986) al. al. et et (Kado (Kado modification modification microsuspension microsuspension a a by by of of predictor predictor independent independent significant significant a a not not was was exhaust exhaust tested tested was was Mutagenicity Mutagenicity mutagenicity. mutagenicity. for for and and thiocyanate) thiocyanate) diesel diesel to to exposure exposure nonsmokers, nonsmokers, and and smokers smokers both both In In smoke. smoke. and and cotinine, cotinine, (nicotine, (nicotine, smoking smoking of of markers markers for for tored tored cigarette cigarette to to exposure exposure of of contribution contribution the the for for made made were were moni and and shifts shifts work work consecutive consecutive two two of of end end the the at at lected lected corrections corrections Appropriate Appropriate level. level. phenanthrene phenanthrene and and tration, tration, col were were samples samples Urine Urine exhaust. exhaust. engine engine diesel diesel to to posure posure concen particle particle respirable respirable category, category, job job including including posure, posure, ex of of markers markers as as used used be be could could mutagens mutagens urinary urinary not not or or ex exhaust exhaust diesel diesel of of indicators indicators several several and and mutagens mutagens nary nary whether whether explore explore to to was was study study this this of of objectives objectives the the of of One One uri between between relationship relationship the the explored explored investigators investigators The The MONITORING MONITORING URINE URINE Exposure Exposure Exhaust Exhaust Diesel Diesel known. known. it. it. tect tect un remain remain therefore, therefore, samples, samples, clerks' clerks' the the in in activity activity genic genic de to to sensitive sensitive sufficiently sufficiently not not was was assay assay the the and and small small was was muta high high the the of of source source the the and and relationships, relationships, These These nine. nine. nonsmokers nonsmokers in in mutagenicity mutagenicity urine urine in in increase increase any any that that sible sible coti urinary urinary and and nicotine, nicotine, urinary urinary nicotine, nicotine, airborne airborne and and pos is is It It nonsmokers. nonsmokers. of of urine urine the the in in elevated elevated were were tabolites tabolites mutagenicity mutagenicity between between relationship relationship the the of of investigation investigation atic atic me cotinine cotinine of of levels levels the the although although nonsmokers, nonsmokers, in in samples samples system a a include include not not did did design design study study The The 1987). 1987). al. al. et et Ling Ling urine urine of of mutagenicity mutagenicity and and smoke smoke tobacco tobacco 1986; 1986; Lazaridis Lazaridis environmental environmental and and (Lofroth (Lofroth smoke smoke cigarette cigarette of of position position to to exposure exposure of of markers markers between between detected detected com was was the the about about association association known known is is what what with with consistent consistent is is which which No No 1989). 1989). al. al. et et Malaveille Malaveille 1989; 1989; Lofroth 1988; 1988; al. al. et et Lofroth Lofroth activation, activation, Sg Sg without without than than activation activation S9 S9 with with greater greater be be 1988; 1988; al. al. et et Bryant Bryant 1986; 1986; Lazaridis Lazaridis and and Lofroth Lofroth 1974; 1974; al. al. to to et et smoke smoke cigarette cigarette of of activity activity mutagenic mutagenic the the reported reported have have (Kier (Kier mutagenicity mutagenicity direct-acting direct-acting little little very very contains contains smoke smoke others others observation, observation, this this to to contrast contrast in in However, However, sources. sources. cigarette cigarette that that state state that that literature literature the the in in reports reports other other with with ambient ambient other other or or smoke smoke tobacco tobacco environmental environmental to to tributed tributed consistent consistent are are findings findings These These S9. S9. without without tested tested when when at tion tion tentatively tentatively was was samples samples these these of of mutagenicity mutagenicity the the sions, sions, rela dose-response dose-response weak weak a a and and S9, S9, with with tested tested when when nicity nicity emis diesel diesel of of source source any any of of vicinity vicinity the the in in work work not not do do mutage urine urine with with relation relation dose-response dose-response strong strong a a showed showed clerks clerks the the Because Because S9. S9. with with revertants/m 1,500 1,500 to to 250 250 and and 3 3 shift shift work work the the during during smoked smoked cigarettes cigarettes of of number number The The S9, S9, without without revertants/m 2,000 2,000 to to 750 750 was was worked worked clerks clerks 3 3 provided. provided. not not were were procedures procedures chromatography chromatography the the of of tails tails the the where where areas areas the the from from obtained obtained matter matter particulate particulate of of de and and investigated investigated not not were were nicotine nicotine of of metabolites metabolites other other mutagenicity mutagenicity the the TA98, TA98, strain strain Salmonella Salmonella the the Using Using pret. pret. study, study, present present the the In In cotinine. cotinine. as as peak peak chromatographic chromatographic inter to to difficult difficult is is however, however, study, study, this this in in tested tested samples samples cle cle 1-El 1-El ~ ~ ------~~~ ~~~ 60 60 classification classification of of exposures exposures among among job job categories, categories, and and lack lack be be desirable desirable to to do do a a pilot pilot study, study, perhaps perhaps in in animals, animals, to to learn learn clude clude lower lower than than expected expected exposures exposures to to diesel diesel exhaust, exhaust, mis cause cause of of the the difficulties difficulties encountered encountered in in this this study, study, it it may may posure posure and and mutagenicity mutagenicity in in the the urine urine samples. samples. They They in gine gine emissions emissions under under specific specific exposure exposure conditions. conditions. Be for for the the lack lack of of a a correlation correlation between between diesel diesel exhaust exhaust ex biomarkers biomarkers of of the the biologically biologically effective effective dose dose of of diesel diesel en Several Several other other explanations, explanations, were were Further Further offered offered by by research research the the authors authors is is also also needed needed to to develop develop and and validate validate ruled ruled out. out. cigarette cigarette smoke. smoke. tors tors are are related related to to bioconversion bioconversion of of a a the the much much mutagens mutagens larger larger percentage percentage cannot cannot be be of of elemental elemental carbon carbon than than does does sibility sibility that that the the negative negative results results reported reported lates lates should should by by the the be be explored explored investiga because because diesel diesel particulates particulates contain contain are are much much less less mutagenic mutagenic than than their their the the nitro nitro use use of of analogs. analogs. elemental elemental The The carbon carbon pos as as a a marker marker for for diesel diesel particu decreased decreased mutagenicity, mutagenicity, such such as as the the aminoarenes, aminoarenes, which which higher higher in in cigarette cigarette smoke smoke than than in in diesel diesel exhaust. exhaust. Finally, Finally, cies cies that that are are no no longer longer mutagenic, mutagenic, or or to to species species that that exhibit exhibit haust haust because because the the concentration concentration of of pyrene pyrene is is 20 20 times times tion. tion. Alternatively, Alternatively, some some might might have have been been converted converted to to spe respirable respirable particles particles may may also also be be a a useful useful marker marker of of diesel diesel ex are are expected expected to to react react with with target target molecules molecules prior prior to to elimina Bryant Bryant et et al. al. 1988). 1988). The The ratio ratio of of pyrene pyrene to to 1-nitropyrene 1-nitropyrene in in trophilic trophilic species, species, such such as as N-hydroxylaminoarenes, N-hydroxylaminoarenes, which which Schuetzle Schuetzle 1983; 1983; Tong Tong and and Karasek Karasek 1984; 1984; Daisy Daisy et et al. al. 1986; 1986; gens gens could could have have been been converted converted to to highly highly reactive reactive elec sitive sitive markers markers of of cigarette cigarette smoke) smoke) (Jensen (Jensen and and Hites Hites 1983; 1983; Conney Conney 1982; 1982; Nebert Nebert 1982). 1982). In In this this study, study, some some of of the the muta (hemoglobin (hemoglobin adducts adducts of of 4-aminobiphenyl 4-aminobiphenyl also also serve serve as as sen tion tion with with carbohydrates carbohydrates or or sulfates sulfates (Miller (Miller and and Miller Miller 1977; 1977; nitronaphthalene, nitronaphthalene, 4-nitrobiphenyl, 4-nitrobiphenyl, and and 4-aminobiphenyl 4-aminobiphenyl matic matic hydrocarbons hydrocarbons are are excreted excreted in in that that the the urine urine may may after after serve serve conjuga as as markers markers include include 2-nitrofluorene, 2-nitrofluorene, 2- Finally, Finally, it it has has long long been been known known that that threne, threne, most most other other polycyclic polycyclic compounds compounds aro present present in in diesel diesel engine engine exhaust exhaust bient bient levels levels of of diesel diesel engine engine exhaust. exhaust. In In addition addition to to phenan the the scope scope of of the the present present work. work. needed, needed, however, however, to to develop develop reliable reliable biomarkers biomarkers for for am diesel diesel exhaust. exhaust. Such Such experiments, experiments, however, however, were were beyond beyond smoke smoke exposure exposure have have been been improved. improved. Further Further research research is is been been optimized optimized by by prior prior studies studies with with animals animals exposed exposed to to 1989), 1989), appears appears that that the the reliability reliability of of estimates estimates it it of of cigarette cigarette procedures procedures for for recovery recovery and and screening screening of of the the mutagens mutagens had had cil cil 1986; 1986; U.S. U.S. Department Department of of Health Health and and Human Human Services Services kinetics, kinetics, it it would would have have been been preferable preferable if if experimental experimental the the the work work by by several several other other groups groups (National (National Research Research Coun the the diesel-derived diesel-derived urinary urinary mutagens, mutagens, or or on on their their pharmaco From From the the results results presented presented in in this this report, report, as as well well as as from from Third, Third, because because there there is is little little information information on on the the nature nature of of ber ber of of samples samples makes makes generalization generalization difficult. difficult. IMPLICATIONS IMPLICATIONS FOR FOR FUTURE FUTURE RESEARCH RESEARCH or or of of nonsmokers nonsmokers in in a a consistent consistent direction, direction, the the small small num not not appear appear to to affect affect the the mutagenicity mutagenicity of of the the urine urine of of smokers smokers tions tions (Appendix (Appendix B). B). Although Although storage storage for for six six months months did did dietary dietary variables variables in in the the design design of of epidemiology epidemiology studies. studies. of of storage storage on on mutagenic mutagenic activity activity and and thiocyanate thiocyanate concentra gens gens (Doolittle (Doolittle et et al. al. 1989), 1989), they they suggest suggest a a need need to to consider consider as as well well as as the the results results of of a a pilot pilot study study to to examine examine effect effect the impact impact of of individual individual dietary dietary constituents constituents on on urinary urinary muta storage storage time time for for each each sample sample was was documented documented in in the the report, report, not not always always in in agreement agreement with with other other reports regarding regarding reports the the developing developing some some of of the the methods methods used used in in this this study. study. The The meats) meats) (Doolittle (Doolittle et et al. al. 1989). 1989). Although Although these these findings findings are are urine urine samples samples (median (median time time 8.5 8.5 months) months) while while they they were were foods foods that that are are reported reported to to be be mutagenic mutagenic (broiled (broiled or or grilled grilled Second, Second, it it should should be be noted noted that that the the investigators investigators stored stored urine urine of of subjects subjects who who indicated indicated that that they they had had consumed consumed in in diesel diesel exhaust. exhaust. Also, Also, no no enhancement enhancement of of mutagenicity mutagenicity was was seen seen in in the the whereas whereas nitropolycyclic nitropolycyclic aromatic aromatic hydrocarbons hydrocarbons are are found found smokers smokers in in the the absence absence of of S9, S9, or or for for smokers smokers in in either either assay. assay. aromatic aromatic hydrocarbons hydrocarbons are are found found in in cigarette cigarette smoke, smoke, significant significant effect effect of of protective protective foods foods was was found found for for non rette rette smoke smoke and and diesel diesel emissions; emissions; that that is, is, polycyclic polycyclic smokers smokers in in the the 59-activated 59-activated S. S. typhimurium typhimurium TA98 TA98 assay. assay. No No forced forced by by the the difference difference in in the the nature nature of of mutagens mutagens in in ciga with with decreased decreased mutagenicity mutagenicity of of the the urine urine samples samples of of non posure posure to to diesel diesel engine engine exhaust. exhaust. This This possibility possibility is is rein nicity nicity (such (such as as cabbage cabbage or or Brussels Brussels sprouts) sprouts) was was associated associated power power of of the the study study to to detect detect mutagens mutagens originating originating from from ex tion tion of of those those foods foods that that have have been been shown shown to to inhibit inhibit mutage subjects subjects than than the the investigators investigators calculated, calculated, reducing reducing the the had had some some effect effect on on urine urine mutagenicity. mutagenicity. Self-reported Self-reported inges mutagens mutagens may may be be present present in in the the urine urine of of diesel-exposed diesel-exposed In In addition addition to to exposure exposure to to cigarette cigarette smoke, smoke, dietary dietary factors factors (Zaebst (Zaebst et et al. al. 1988). 1988). is is possible possible that that smaller smaller amounts amounts It It of of diesel diesel exhaust exhaust particulates particulates OTHER OTHER FACfORS FACfORS contain more more than than 60 60 percent percent very very little little elemental elemental (less (less carbon than than 10 10 percent), percent), whereas whereas different different from from cigarette cigarette smoke. smoke. Cigarette Cigarette sonal sonal filter filter smoke smoke samples samples contains contains that that were were tested. tested. constituent constituent particles, particles, diesel diesel engine engine mutagenicity mutagenicity exhaust exhaust is is test test chemically chemically and and the the small small numbers numbers of of urine urine and and per be be valid valid of of study study because because power power in in addition addition due due to to both both to to differences differences the the lack lack of of in in sensitivity sensitivity the the size size of of the the of of the the ~l-~------ m m Health Health Review Review Committee's Committee's Commentary Commentary 61 61 for- by by enzymes enzymes microsomal microsomal of of Induction Induction differ- AH. AH. 1982. 1982. clear-cut clear-cut Canney Canney no no workers, workers, railroad railroad of of study study 87 87 pilot pilot a a In In particles. particles. respirable respirable viron viron 12:1132-1141. 12:1132-1141. total total to to smoke smoke cigarette cigarette of of contribution contribution the the determining determining En Atmos Atmos phase. phase. gas gas corresponding corresponding the the and and matter matter ticulate ticulate for for method method good good a a be be to to appears appears particulates particulates trapped trapped from from par airborne airborne between between pollutants pollutants organic organic of of distribution distribution the the downstream downstream nicotine nicotine gas-phase gas-phase collecting collecting of of technique technique on on Experiments Experiments K. K. 1978. 1978. Cauwenberghe Cauwenberghe Van Van W, W, Cantreels Cantreels The The testing. testing. and and validation validation further further without without exhaust exhaust engine engine diesel diesel to to exposure exposure for marker marker specific specific a a as as compound compound this this of of 85:9788-9791. 85:9788-9791. use use the the preclude preclude emissions, emissions, diesel diesel to to USA USA Sci Sci exposure exposure no no Acad Acad or or Natl Natl little little Proc Proc tobacco. tobacco. of of type type and and status status smoking smoking had had who who clerks clerks from from samples samples in in found found with with levels levels Associations Associations than-expected than-expected amines: amines: aromatic aromatic of of adducts adducts Hemoglobin Hemoglobin higher the the and and blanks, blanks, field field the the on on found found SR. SR. 1988. 1988. phenanthrene phenanthrene Tannenbaum Tannenbaum PL, PL, Skipper Skipper P, P, Vineis Vineis MS, MS, Bryant Bryant of of levels levels high high anomalously anomalously the the however, however, emissions; emissions; diesel diesel NY. NY. York, York, New New Co., Co., ing ing of of sources sources to to proximity proximity to to respect respect with with exposure exposure of of gradient gradient Publish Science Science Elsevier Elsevier pp. pp. eds.) eds.) 471-488. 471-488. W, W, StOber StOber RO, RO, a a reflected reflected categories categories job job different different in in workers workers from from samples samples McClellan McClellan A, A, Koizumi Koizumi N, N, (Ishinishi (Ishinishi Exhaust Exhaust Engine Engine Diesel Diesel personal personal in in concentrations concentrations Phenanthrene Phenanthrene studies. studies. field field of of Effects Effects Mutagenic Mutagenic and and Carcinogenic Carcinogenic In: In: emissions. emissions. haust haust future future for for value value potential potential have have refinement, refinement, further further require require ex engine engine of of inhalation inhalation chronic chronic after after rodents rodents in in changes changes they they although although that, that, nicotine nicotine and and phenanthrene phenanthrene of of trations trations functional functional and and Neoplastic Neoplastic F. F. 1986. 1986. Duchosal Duchosal R, R, Gatz Gatz concen low low measuring measuring for for developed developed were were techniques techniques New New D, D, Bernstein Bernstein A-L, A-L, J, J, Cassano-Zoppi Cassano-Zoppi X, X, Fouillet Fouillet Brightwell Brightwell smoke. smoke. tobacco tobacco and and exhaust exhaust engine engine diesel diesel locomotive locomotive to to exposure exposure personal personal of of markers markers measure measure to to device device sampling sampling a a Res Res 135:1-9. 135:1-9. Mutat Mutat particulate. particulate. exhaust exhaust diesel diesel onto onto sorbed sorbed on on investigations investigations pilot pilot conducted conducted and and developed developed vestigators vestigators ad compounds compounds mutagenic mutagenic of of availability availability Biological Biological 1984. 1984. in The The workers. workers. railroad railroad of of population population characterized characterized well well a a F. F. Lorenzo Lorenzo De De E, E, Marinis Marinis De De V, V, Buonocore Buonocore MA, MA, Belisario Belisario in in exhaust exhaust engine engine diesel diesel of of dose, dose, biological biological the the as as well well as as levels, levels, exposure exposure ambient ambient estimate estimate to to methods methods developing developing of of REFERENCES REFERENCES task task difficult difficult the the undertook undertook coworkers coworkers his his and and Schenker Schenker CONCLUSIONS CONCLUSIONS findings. findings. negative negative the the to to contributed contributed have have could could that that Commentary, Commentary, the the in in and and Report Report Investigators' Investigators' preceding preceding the the in in discussed discussed factors, factors, technical many many are are There There exists. exists. tion tion gines. gines. correla such such no no that that data data these these from from conclude conclude to to however, however, en diesel diesel of of emissions emissions the the to to exposure exposure from from risk risk estimate estimate to to premature, premature, be be would would exhaust. exhaust. 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Properties Properties Barrier Barrier Epithelial Epithelial Alveolar Alveolar on on Dioxide Dioxide Nitrogen Nitrogen of of Effects Effects 13 13 1987 1987 September September Fechter Fechter L.D. L.D. Exposure Exposure Monoxide Monoxide Carbon Carbon Prenatal Prenatal of of Neurotoxicity Neurotoxicity 12 12 Inhibitors Inhibitors Proteinase Proteinase 1987 1987 August August Johnson Johnson D.A. D.A. Lung Lung Human Human on on Dioxide Dioxide Nitrogen Nitrogen and and Ozone Ozone of of Effects Effects 11 11 Species Species Laboratory Laboratory and and Humans Humans in in Particles Particles 1987 1987 July July Yu Yu C.P. C.P. Exhaust Exhaust Diesel Diesel Inhaled Inhaled of of Deposition Deposition for for Models Models Predictive Predictive 10 10 Injury Injury Endothelial Endothelial Induced Induced 1987 1987 June June Patel Patel J.M. J.M. Dioxide Nitrogen Nitrogen to to Response Response Metabolic Metabolic and and Biochemical Biochemical 9 9 Lung Lung Developing Developing 1987 1987 May May Mauderly Mauderly J.L. J.L. on on Exhaust Exhaust Diesel Diesel and and Dioxide Dioxide Nitrogen Nitrogen Inhaled Inhaled of of Effects Effects 8 8 1987 1987 April April Groopman Groopman J.D. J.D. Antibodies Antibodies Specific Specific by by Detected Detected Nitropyrene Nitropyrene of of Adducts Adducts DNA DNA 7 7 Function Function Pulmonary Pulmonary and and Metabolic Metabolic April1987 April1987 Drechsler-Parks Drechsler-Parks D.M. D.M. on on Nitrate Nitrate Peroxyacetyl Peroxyacetyl and and Ozone, Ozone, Dioxide, Dioxide, Nitrogen Nitrogen of of Effect Effect 6 6 Exhaust Exhaust Diesel Diesel in in Mutagens Mutagens Chemical Chemical 1987 1987 January January Bagley Bagley S.T. S.T. the the on on Trap Trap Particle Particle Ceramic Ceramic a a of of Effect Effect the the into into Investigation Investigation An An 5 5 1986 1986 August August Beland Beland F.A. F.A. Dinitropyrenes Dinitropyrenes of of Adducts Adducts DNA DNA and and Activation Activation Metabolic Metabolic The The 4 4 Pollutants Pollutants Air Air of of Deposition Deposition 1986 1986 February February Crocker Crocker T. T. for for Sites Sites Target Target at at Particles Particles and and Macromolecules Macromolecules of of Transport Transport 3 3 Inhalation Inhalation After After Nitropyrenes Nitropyrenes Bond Bond 1986 1986 J. J. February February Particle-Associated Particle-Associated and and Free Free of of Metabolism Metabolism and and Disposition Disposition 2 2 Dioxide Dioxide Nitrogen Nitrogen and and Ozone Ozone to to Cells Cells Red Red 1985 1985 August August Amoruso Amoruso M. M. Dehydrogenase-Deficient Dehydrogenase-Deficient 6-Phosphate 6-Phosphate Glucose Glucose of of Risk Risk of of Estimation Estimation 1 1 Date Date Investigator Investigator Title Title No. No. Publication Publication Principal Principal Report Report Reports Reports Research Research (Supplement) (Supplement) Research Research Future Future for for Recommendations Recommendations and and 1988 1988 January January Information Information Scientific Scientific Existing Existing of of Evaluation Evaluation Cancer: Cancer: Human Human and and Exposure Exposure Vapor Vapor Gasoline Gasoline Research Research Future Future for for Issues Issues and and 1987 1987 May May Information Information Scientific Scientific Existing Existing of of Evaluation Evaluation An An Health: Health: Human Human and and Vapors Vapors Methanol Methanol Automotive Automotive Research Research Future Future for for Recommendations Recommendations and and 1985 1985 September September Information Information Scientific Scientific Existing Existing of of Evaluation Evaluation Cancer: Cancer: Human Human and and Exposure Exposure Vapor Vapor Gasoline Gasoline Date Date Publication Publication Title Title Reports Reports Special Special ------n:: ------n:: ~[ ~[ PUBLICATIONS PUBLICATIONS HEI HEI OF OF LIST LIST Copies Copies of of these these reports reports can can be be obtained obtained by by writing writing to to the the Health Health Effects Effects Institute, Institute, 141 141 Portland Portland Street, Street, Suite Suite 7300, 7300, Cambridge, Cambridge, MA MA 02139. 02139. 32 32 Respiratory Respiratory Carcinogenesis Carcinogenesis of of Nitroaromatics Nitroaromatics R.C. R.C. Moon Moon April April 1990 1990 in in Vivo: Vivo: Effects Effects of of Nitroreductase Nitroreductase Induction Induction 31 31 DNA DNA Binding Binding by by 1-Nitropyrene 1-Nitropyrene and and Dinitropyrenes Dinitropyrenes in in Vitro Vitro and and F.A. F.A. Beland Beland November November 1989 1989 Effects Effects from from Inhaled Inhaled Nitrogen Nitrogen Dioxide Dioxide and and Diesel Diesel Exhaust Exhaust 30 30 Influence Influence of of Experimental Experimental Pulmonary Pulmonary Emphysema Emphysema on on Toxicological Toxicological J.L. J.L. Mauderly Mauderly October October 1989 1989 29 29 Early Early Markers Markers of of Lung Lung Injury Injury J.N. J.N. Evans Evans September September 1989 1989 Pilot Pilot Investigations Investigations 28 28 Nitrogen Nitrogen Dioxide Dioxide and and Respiratory Respiratory Infection: Infection: J.M. J.M. Samet Samet September September 1989 1989 High-Altitude High-Altitude Exposure Exposure 27 27 Cardiovascular Cardiovascular Effects Effects of of Chronic Chronic Carbon Carbon Monoxide Monoxide and and J.J. J.J. McGrath McGrath July July 1989 1989 Tract Tract of of Syrian Syrian Golden Golden Hamsters Hamsters Nitrogen Nitrogen and and Sulfur, Sulfur, and and of of Diesel-Engine Diesel-Engine Exhaust, Exhaust, on on the the Respiratory Respiratory 26 26 Investigation Investigation of of a a Potential Potential Cotumorigenic Cotumorigenic Effect Effect of of the the Dioxides Dioxides of of U. U. Mohr Mohr Heinrich) Heinrich) May May (U. (U. 1989 1989 Coronary Coronary Artery Artery Disease Disease CO CO Study Study Team Team 25 25 Acute Acute Effects Effects of of Carbon Carbon Monoxide Monoxide Exposure Exposure on on Individuals Individuals with with November November 1989 1989 HEI HEI Multicenter Multicenter Exposure Exposure to to Nitrogen Nitrogen Dioxide Dioxide 24 24 Altered Altered Susceptibility Susceptibility to to Viral Viral Respiratory Respiratory Infection Infection During During Short-Term Short-Term R.M. R.M. Rose Rose March March 1989 1989 23 23 Responses Responses of of Susceptible Susceptible Subpopulations Subpopulations to to Nitrogen Nitrogen Dioxide Dioxide February February 1989 1989 P.E. P.E. Morrow Morrow 22 22 Detection Detection of of Paracrine Paracrine Factors Factors in in Oxidant Oxidant Lung Lung Injury Injury A.K. A.K. Tanswell Tanswell February February 1989 1989 Carbon Carbon Monoxide Monoxide at at Several Several Altitudes Altitudes 21 21 Maximal Maximal Aerobic Aerobic Capacity Capacity at at Several Several Ambient Ambient Concentrations Concentrations of of S.M. S.M. Horvath Horvath December December 1988 1988 Bacterial Bacterial Infections Infections by by Acute Acute Exposures Exposures to to Nitrogen Nitrogen Dioxide Dioxide 20 20 Modulation Modulation of of Pulmonary Pulmonary Defense Defense Mechanisms Mechanisms Against Against Viral Viral and and G.J. G.J. Jakab Jakab October October 1988 1988 Aerosols Aerosols 19 19 Factors Factors Affecting Affecting Possible Possible Carcinogenecity Carcinogenecity of of Inhaled Inhaled Nitropyrene Nitropyrene August August 1988 1988 R.K. R.K. Wolff Wolff 18 18 Respiratory Respiratory Infections Infections in in Coal Coal Miners Miners Exposed Exposed to to Nitrogen Nitrogen Oxides Oxides July July 1988 1988 M. M. Jacobsen Jacobsen Diploid Diploid Human Human Fibroblasts Fibroblasts and and Related Related Nitro-polycyclic Nitro-polycyclic Aromatic Aromatic Compounds Compounds in in 17 17 Studies Studies on on the the Metabolism Metabolism and and Biological Biological Effects Effects of of Nitropyrene Nitropyrene V.M. V.M. 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