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United States
Environmp"*-'
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       Office of Research and
       Development
       Washington, DC 20460
              EPA/600/R-01/042
              June 2001
              www.epa.gov/ncerqa
             lings
     'CDC Centers for
  _.  nvironmental Health
       Prevention Research
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         United States
         Environmental Protection
         Agency
   Office of Research and
   Development
   Washington, DC 20460
EPA/600/R-01/042
June 2001
www.epa.gov/ncerqa
x>EPA Proceedings
         EPA/NIEh
S/CDC Centers for
         Children's Environmental Health
         and Disease Prevention Research
         Progress Review Workshop
         November 5-7,
2000
         University of California, Berkeley
         Clark-Kerr Center

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


                                          Table of Contents

Introduction  	 vii

University of Southern California

Overview and Description of the Southern California Children s Environmental Health Center
        Respiratory Disease and Prevention	3
        H. Gong, Jr., J. Peters, R. McConnell, D. Diaz-Sanchez, F. Gilliland, C. Jones, A. Hricko

Environmental Tobacco Smoke Alters the In Vivo Allergic Response in the Human Upper Airway	5
        David Diaz-Sanchez,

Determinants of Childhood Lung Susceptibility to Air Pollution	6
        Frank Gilliland


Columbia University

Overview and Description of the Columbia Center for Children s Environmental Health	9
        Frederica Perera

Pesticide Exposure During Pregnancy Among Minority Women Residing in Northern Manhattan
        and the South Bronx	 12
        R.M. Whyatt, D.E. Camann, D.B. Barr, P.L. Kinney, A. Reyes, J. Ramirez, J. Dietrich,
        D. Diaz, F.P. Perera

Indoor and Outdoor Environmental Exposures In Minority Children of Northern Manhattan
        and the South Bronx	 13
        P.L. Kinney, M. Aggarwal, G. Chew, R.M. Whyatt, F.P.  Perera


Mount Sinai School of Medicine

Overview and Description of the Mount Sinai Center for Children s Environmental Health
        and Disease Prevention: Inner-City Toxicants and Neurodevelopmental Impairment	 17
        Mary S. Wolff

Kinetic PCR on Pooled DNA:  A High-Throughput, High-Efficiency Alternative in Genetic
        Epidemiologic Studies	 18
        J. Chen, R. Higuchi, S. Germer, J.G. Wetmur
University of Washington

Overview and Description of the Center for Child Envi onmental Health Risks Research	21
        Elaine M. Faustman

Eifects of Human Paraoxanase (PONf) Single Nucleotide Polymorphisms on Susceptibility
        to Specific Organophosphate Insecticides  	22
        Clement E. Furlong, Wan-Fen Li, Lucio G. Costa, Victoria H. Brophy, Rebecca J. Richter,
        Diana M. Shih, Aaron Tward, Aldon J. Lusis
The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop	


                                 Table of Contents (continued)

Preliminary Baseline Results From the Randomized Community Trial To Assess the Efficacy of
        a Community-Wide Program To Reduce Children s Exposure to Pesticides	23
        Beti Thompson, Gloria Coronado, Cam Solomon, John Kissel

Summer Project for Minority High School Student Interns: Effects of Home Parties
        on Farmworkers	25
        R. Godinci, G. Martinez, I. Islas , G.D. Coronado, B. Thompson
University of California, Berkeley

Overview and Description of the Center for the Health Assessment of the Mothers and Children
        of Salinas (CHAMACOS): A Community/University Partnership 	29
        Brenda Eskanazi

Preliminary Approaches To Assessing Organophosphate Pesticide Exposure and Potential Health
        Risks to Pregnant Women Living in the Salinas Valley, California	30
        Brenda Eskanazi

CHAMACOS Laboratory Core: Challenges of Biological Sample Collection and Processing	32
        Nina Holland
Johns Hopkins University

Overview and Description of the Johns Hopkins University Center for Childhood Asthma in
        the Urban Environment  	35
        Peyton Eggleston

Ambient Urban Pard'culate-Induced Airway Hyperresponsiveness and Inflammation in Mice	."	38
        D. Walters, P.N. Breysse, M. Wills-Karp

Air Pollution and Allergen Exposure Among Asthmatic Children in Inner-City Baltimore 	39
        T.J. Buckley, P.N. Breysse, C. Beck, A. Escamillia,  P.A. Eggleston


University of Iowa

Overview and Description of the Children s Environmental Airway Disease Center  	43
        Gary W. Hunninghake

A Multicomponent Intervention Study of Asthma in Children From Rural Communities 	45
        E. Chrischilles, J. Merchant, A. Kuehl, R. Ahrens, S. Reynolds, L. Burmeister, P. Pomrehn,
        P. Thorne

lipopolysaccharide Responsiveness and the Development of Subchronic Grain Dust-Induced
        Airway Injury	46
        C.L.S. George, H. Jin, C.L. Wohlford-Lenane, J.N.  Kline, D.A. Schwartz

IL-10 Reduces Grain Dust-Induced Airway Inflammation and Airway Hyperreactivity	47
        T.J. Quinn, S. Taylor, C.L. Wohlford-Lenane, D.A.  Schwartz
iv                                The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


                                 Table of Contents (continued)

TLR4 Mutation Is Associated With Endotoxin Hyporesponsiveness in Humans	48
        N.C. Arbour, E. Lorenz, B.C. Schutte, J. Zabner, J.N. Kline, M. Jones, K. Frees, J.L. Watt,
        DA. Schwartz

Bronchial Hyperreactivity Is Associated With Enhanced Grain Dust-Induced Airflow Obstruction	49
        J.N. Kline, P.J. Jagielo, J.L. Watt, D.A. Schwartz

TNF-Alpha and IL-lBeta Are Not Essential to the Inflammatory Response in LPS-Induced
        Airway Disease	50
        J.G. Moreland, R.M. Fuhrman, C.L. Wohlford-Lenane, T.J. Qulnn, E. Benda,
        J.A. Pritessner, D.A. Schwartz

Endotoxin Responsiveness and Subcnronic Grain Dust-Induced Airway Disease	51
        C.L.S.  George, H. Jin, C.L. Wohlford-Lenane, M.E. O Neill, J.C. Phipps, P.O Shaughnessy,
        J.N. Kline, P.S. Thome, D.A. Schwartz

Mechanisms That Initiate, Promote, and Resolve Grain Dust-Induced Inflammation	52
        Kevin Leidal, William Nauseef, Gerene Denning

Activation of ERK Kinase Activity by Respiratory Syncytial Virus in A549 Cells Is Linked
        to the Production of Interleukin 8	53
        W. Chen, MM. Monick, A.B. Carter, G.W. Hunninghake

Effects of Ragweed and Th-2 Cytokines on the Secretion of IL-8 by Human Airway Epithelial Cells  	54
        W. Chen, G. W. Hunninghake

Respiratory Syncytial Virus Infection Results in Activation of Multiple Protein Kinase C Isoforms
        Leading to Activation of     Mitogen-Activated Kinase	55
        MM. Monick, J.M. Staber, G. W. Hunninghake

Endotoxin Augments Viral Replication and the Inflammatory Response in Respiratory Syncytial
        Virus-Infected Epithelium 	56
        A.B. Carter, M.R. Donahue, K.L. Knudtson, G. Gudmundsson, MM. Monick,
        G.W. Hunninghake
University of Michigan

Overview and Description of the Michigan Center for the Environment and Children s Health  	59
        Barbara Israel

Use of a Screening Questionnaire To Estimate Prevalence of Diagnosed and Undiagnosed
        Asthma Among Minority Children in Detroit	61
        T.G. Robins, E.A. Parker, B.A. Israel, R.W. Brown, T.C. Lewis, CAAA  Steering Committee

Detroit School Children With Symptoms of Persistent Asthma Are Sensitized to Both
        Indoor and Outdoor Allergens	62
        T.C.  Lewis, T.G. Robins, E.A. Parker, R.W. Brown, W.R. Solomon, T.R. Trestyn, B.A. Israel,
        CAAA Steering Committee

Evaluation of Murine Model of Asthma Induced by Exposure to House Dust Extract
        Containing High Levels of Cockroach	63
        Jiyoun Kim, Andrew C. Merry,  Jean A. Nemzek, Daniel G. Remick


The Office of Research and Development s National Center for Environmental Research                                 v

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


                               Table of Contents (continued)

Measurements of Indoor, Outdoor, and Personal Exposure to Paniculate Matter Among
        Asthmatic Children in Detroit, Michigan	64
        Fuyuen Y. Yip, J. Timothy Dvonch, Thomas G. Robins, Edith Parker, Masako Morishita,
        Gerald J. Keeler

Field Comparison of PM2, TEOM and PM,, Manual Filter-Based Measurement Methods
        in Urban Atmospheres	65
        J. Timothy Dvonch, Frank J. Marsik, Gerald J. Keeler, Thomas G. Robins, Fuvuen Yip,
        Masako Morishita

Index of Authors	67

Appendix A: Agenda	69

Appendix B: Report From Pesticide Breakout Session and List of Breakout Session Participants	73

Appendix C: List of Participants	77
                                The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
                                           Introduction
    Nearly 3 years ago, the United States Environmental Protection Agency* (EPA) in partnership with the
National Institute for Environmental Health Sciences (NIEHS) and the Centers for Disease  Control and
Prevention (CDC) created the first  federally  funded research program devoted exclusively to children s
environmental health and disease prevention. The spirit of partnering nurtured by these  three agencies has
rippled  into every aspect  of this extramural  grant program. These center grants were awarded  to eight
university/community partnerships that intricately and  wisely merge the expertise and  resources  of com-
munity-based organizations with  those  of competitive  research  institutions.  The Centers are proving the
concept that basic and applied research is made more meaningful when the public has input and access to
scientists, and vice  versa. Because  information exchange between researchers and  local citizens was a
requirement for grant award, public outreach and participation are a cornerstone of this program. In total, the
Centers are conducting a  wide breadth of research  targeting the etiology and risk factors  for respiratory
ailments, such as asthma and developmental effects from  environmental toxins in the urban and rural areas.

    The Centers for Children s Environmental Health  and Disease Prevention Research Annual  Meeting
offers the Center investigators an  opportunity to present their findings to date, discuss advances in research
and practice, and share their insights with other scientists,  federal and state officials, policymakers,  commu-
nity-based organizations, and others  interested in pediatric environmental health. The  1998 meeting in Re-
search Triangle Park, NC, and the 1999 meeting in Atlanta, GA,  were successful in engaging the university
and community investigators in forward-thinking discussions  about children s vulnerability and susceptibility
to toxins and the challenges of conducting clinical and household interventions in communities fraught with
environmental justice concerns.

    However, to address  these critical issues in a more focused setting, it was decided that in 2000, two
separate meetings would commence to allow investigators  ample time to share and discuss their findings and
chart future directions. Thus, in October 2000, the community-based intervention/prevention research com-
ponents of the eight  Centers met in Seattle, WA, with other like researchers, federal partners, and nonprofit
organizations. For more information regarding the October meeting and the published proceedings, please
contact Fred Tyson via e-mail at tyson2@niehs.nih.gov or call (919) 541-0176.

    This proceedings booklet documents the  meeting in  November 2000 in Berkeley,  CA. The  meeting
brought together the basic  scientists,  exposure  teams, biostatisticians, and clinicians of the eight Centers for
2 days of presentations, dialogue, focused group discussions,  and a field visit  in the Salinas Valley. The
researchers presented preliminary results, highlighted their published data, and advanced  new hypotheses in
childhood susceptibility and analytical  methodologies.  These studies are of critical importance to EPA s
Office of Research and Development, as they have the potential to strengthen the scientific basis for risk
assessment and culturally  appropriate  risk  management  practices  while addressing uncertainties in  the
supporting science. For more information about the 2000  Centers for Children s Environmental Health and
Disease Prevention Research  Annual Meeting, please contact program administrators Nigel Fields via e-mail
at fields.nigel@epa.gov or Gwen Collman at collman@niehs.nih.gov.
*The mission of EPA is to protect public health as well as safeguard and improve the natural environment.  Achievement of this
mission requires the  application of sound science to the assessment of enviromental problems and to the evaluation of possible
solutions. The National Center for Environmental Research at EPA is committed to providing the best products in high-priority areas
of scientific research through significant support for long-term research.
The Office of Research and Development s National Center for Environmental Research                              vii

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University of Southern California
       Los Angeles, CA

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Overview and Description of the
Southern California Children s Environmental
Health Center;  Respiratory Disease and Prevention
H. Gong, Jr., J. Peters, R. McConnell, D. Diaz-Sanchez, F. Gilliland, C. Jones, and A. Hricko
University of Southern California and University of California at Los Angeles, Los Angeles, CA

The Southern California Children s Environmental Health Center (CEHC) involves multi-institutional and
multidisciplinary participation in an administrative core and three  related projects with the common goal of
examining how host susceptibility and environmental exposure determine children s respiratory disease. The
following describes the key cores and their progress to date:

>  The Administrative Core supervises  and coordinates the Center s research and outreach efforts in
    children s environmental health issues. Highlights include:

    +  Funds will be committed for a new faculty member (new scientist) specializing in children s  environ-
       mental health.

    *  A Web  site is currently under development.

    +  Research and educational interactions with the NIEHS-supported Southern California Environmental
       Health Sciences Center (SCEHSC) continue to grow.

    *  The Community  Outreach and Education Program has  facilitated numerous collaborations with
       community-based and public health organizations in Southern California.

>  Dr. Gilliland is evaluating the effect of outdoor air pollution on the susceptibility to respiratory disease
    in  a large cohort of school children, and the potential protective effects of dietary intake and genetic
    polymorphisms involved in lung defenses.  Highlights include:

    +  Buccal cell collections (n = 419) have  continued for DNA assays.  One challenge has been  children
       who have moved or refused participation.

    +  During  Year 2, more than 2,100 food  frequency questionnaires  were collected.  Two papers on
       nutrients and lung function are being prepared.

    *  A Taqman was purchased to increase the throughput of DNA assays for genotyping target enzymes.

>  Dr. Diaz-Sanchez is evaluating the effect of environmental tobacco smoke (ETS) on allergic responses
    in children with smokers in  the home  and in chronic animal  models. Highlights include:

    +  Preliminary  studies  show  that ETS  and ragweed nasal challenge potentiate  local ragweed-specific
       IgE and IgG4 as compared to ragweed challenge alone.

    *  ETS synergizes with allergen to produce both Th2  interleukins (IL-4, IL-5, IL-13) and eosinophil
       influx in the nose.

    *  Animal  models (challenged with ovalbumin) were developed with BALB/c and CB57/Black/6 mice
       for chronic ETS exposures. This  allows longitudinal immunologic  evaluation according to genetic
       background and age.
The Office of Research and Development s National Center for Environmental Research

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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


>-  Drs.  McConnell and Jones  are evaluating  the  efficacy  of  a community-based health education
    intervention to control indoor allergen exposure and to reduce the severity of asthma in the homes of
    children who are allergic to cockroach or house dust mite allergen. Highlights include:

    *  A new set of health education materials on reducing exposure to indoor allergens was developed for
       use in training inner city families in the study.

    *  Community-based educators (supported by several community partners) are being trained for the
       intervention group.

    *  The CEHC provided staff support for subject recruitment, protocol development, and data analysis
       for a pilot study of professionally cleaned homes of children allergic to cockroaches. This experience
       and pilot data will assist the current project.

In addition to regular meetings, the projects investigators collaborated by using certain assays (e.g., geno-
typing, IgE levels), as well as with the SCEHSC s Facility Cores  (e.g., DNA storage, biostatistics,  ETS
generation).
                                 The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Environmental Tobacco Smoke Alters
the In Vivo Allergic Response in the Human Upper Airway
David Diaz-Sanchez
University of Southern California, Los Angeles, CA

We have focused on experiments  to  determine how environmental tobacco smoke (ETS) interacts with
allergen and demonstrated our hypothesis  that acute challenge  with ETS will  exacerbate allergen-induced
nasal allergic responses. We recruited  10 volunteers with a positive skin test to  short ragweed and a history
of allergic rhinitis. The antigen used in  our challenge studies, short ragweed, is not present in the Los Angeles
area. Subjects were challenged by spraying the nose with a symptom active dose of allergen. After a space
of at least 4 weeks, the subjects were recalled and challenged with their active dose of allergen following ETS
exposure. We have used our previously established human ETS exposure model performed in collaboration
with Dr. Gong at the Los Amigos Research and Education Institute. In this model,  subjects are exposed to
the side-stream smoke of five Kentucky Reference cigarettes in a 2-hour period. Production of antibody of
the Immunoglobulin E class (IgE) is the hallmark of allergy. Ragweed-specific IgE levels  measured in nasal
lavages performed 4 days after exposure  to ragweed  plus ETS were significantly higher than following
challenge with ragweed alone (mean =  58.4 U/mL vs. 2.2 U/mL, p<0.001  paired t-test). Similarly, ragweed-
specific IgG4 levels were enhanced by ETS exposure (mean = 26.7 U/mL vs. 5.6 U/mL). ETS also poten-
tiated levels of total IgE and IgG4. Histamine levels were measured in nasal lavages performed before and
10 minutes following allergen challenge. As expected,  in these ragweed allergic subjects, challenge with
ragweed caused a significant rise in histamine levels from baseline levels (3.8 nM vs.  0.6 nM, p<0.01).
However, this increased more than fourfold to 16.1 nM when subjects  were exposed with ETS immediately
before allergen. When the above experiments were repeated, but instead of ETS, exposure was  to Carbon
Black, elemental carbon essentially devoid of chemicals, no  increase in histamine levels  was apparent.
Similarly, ETS synergised  with allergen produced  a local Th2 cytokine milieu. This cytokine response is
characteristic of an enhanced allergic response and is critical to allergic inflammation.

We also have successfully established  an animal model of chronic exposure to ETS mice. In our model, mice
receiving the protein ovalbumin (OVA) alone did not produce either OVA-specific  allergic antibody at any
time. In contrast, animals receiving both ETS and OVA had significantly elevated  levels of both total  and
OVA-specific  allergic antibodies  12 days after the initial exposure. The levels  of these antibodies were
significantly higher in young rather than adult mice.

These results demonstrate that ETS can synergize with allergen to exacerbate the allergic response in both
human and murine models. They demonstrate for the first time in humans a direct  causal effect of ETS on
the allergic  response. In addition, the murine model suggests that the effects of ETS also are relevant and,
indeed, may be of greater importance in the young.
The Office of Research and Development s National Center for Environmental Research

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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Determinants of Childhood
Lung Susceptibility to Air Pollution
Frank Gilliland
University of Southern California, Los Angeles, CA

In the last year, efforts  have focused on examining the effects of diet on children s lung health. An emerging
body of evidence supports an adverse effect of low antioxidant vitamin  intake on adult lung function. The
respiratory health effects of low antioxidant vitamin  during childhood have yet to be fully defined. To in-
vestigate the effects of  dietary intake of three antioxidant vitamins, vitamins A, E, and C, on children s lung
function, we examined  cross-sectional dietary data and pulmonary function tests from 2,566 participants in
the Children s Health Study. At followup visits during the  1998-1999 school year, each student completed
a health update questionnaire, a validated food frequency questionnaire (FFQ), and spirometric lung function
testing. To assess the effects of vitamins A, E, and C on lung function, regression  splines that account for the
nonlinear relationship between pulmonary function,  height, and age in children were used. Low vitamin C
intake was associated with deficits in measures of airway flow that were larger in girls (FEV, -3.3%  [95%
CI -6.0, -0.5], FEF2,.7,-5.5% [95% CI -10.5, -0.3]) than boys (FEV, -2.3% [95%  CI -4.8, 0.3], FEF2,.7, -2.4%,
[95% CI -7.4, 2.8]). Children with low vitamin E intake had lower FEF25.7, (boys -8.9 [95% CI -14.2,  -3.3],
girls, -2.5% [95% CI -8.3,3.7]). Low vitamin A intake also was associated with FEV, only among boys with
asthma (-6.3% [-12.4,0.3]).

We also investigated the effects of dietary magnesium (Mg) on children s  lung function. Girls with low Mg
intake had lower FEF2,75 (-4.8%, 95% CI -9.8, 0.4) and FEF75 (-8.3%, 95% CI -14.8, -1.4) than  girls with a
higher intake; and the reductions were larger in girls with asthma (FEF,5.75 [-16.2%, 95% CI -22.7, -9.1] and
FEF7, [-24.9%, 95% CI -32.8, -16.1]) than in girls without asthma (FEF,,.75 [-2.0%, 95% CI -7.4, 3.8] and
FEF75 [-4.1%, 95% CI -11.3, 3.7]). Boys with low Mg intake showed deficits in FEV, (-2.7%, 95% CI -5.4,
0.1) and FVC (-2.8%), 95% CI -5.4, -0.2) compared  to boys with higher intake. In summary, children with
low antioxidant  vitamin intake had lower lung function. The effects of low Mg intake did not vary  sub-
stantially in boys with  and without asthma.  Because children s magnesium intake is generally inadequate,
small individual deficits in lung function associated  with low Mg intake may be important on a population
level, especially among  girls with asthma.
                                The Office of Research and Development s National Center for Environmental Research

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Columbia University
  New York, NY

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Overview and Description of the
Columbia Center for Children s Environmental Health
Frederica Perera
Columbia University,  New York, NY

The long-term objective of the Columbia Center for Children s Environmental Health (CCCEH) is to prevent
developmental damage, asthma, and cancer risk in African-American and Latino infants and children living
in Northern Manhattan and the South Bronx, as well as children elsewhere. These communities are dispro-
portionately exposed  to various environmental neurotoxic, asthmagenic, and carcinogenic pollutants; and they
have disproportionately high rates of low  birth weight, other developmental disorders, and childhood asthma.
The main hypotheses of the Center are:  (1)  prenatal and/or postnatal environmental exposures  to airborne
particulate matter (PM), including diesel exhaust particulate (DEP), polycyclic aromatic hydrocarbons (PAH),
environmental tobacco smoke  (ETS), pesticides,  and home allergens,  increase  the risk of developmental
impairment and/or asthma and cancer in  these  disadvantaged communities, controlling for known physical
(e.g., PCBs and lead) and  psychosocial  risk factors; (2) inadequate nutrition and family  and community
stressors exacerbate the impact  of these  environmental toxicants;  and (3) community  and individual-level
interventions to reduce these toxic exposures and improve nutritional status can reduce the risk  of disease.
The structure of the CCCEH is shown in the Figure below.
                                                      Partners/ Community Advisors
                               Administrative Core
                           F. Perera, Director
                           V. Raul), Deputy Director
                           R. Whyatt Co-Deputy Director
                           S. Illman, Program Coordinator
                                  Harlem & NY Presbyterian Hospitals,
                                    NYSDOH, CDC, WEACT, Alianza
                                     Dominicans, Best Beginnings,
                                   Community Assoc. of Progressive
                                  Dominicans, ECHO, Harlem Dowlinj-
                                   W. Side, Heart of Harlem, No. Mann.
                                  Perinatal Partnership, St. Mary's, So.
                                      Bronx Clean Air Coalition
Scientific
Advisory
 Board
                                                                      Community
                                                                       Advisory
                                                                        Board
                                 Community Outreach & Education Program (COEP)
                                     R. Fullilove, Director & P. Shepard, Co-Director
                                                                       Data Management,
                                                                      Statistics, Community,
                                                                      Impact Modeling Core
Exposure Assessment
      Core
                  P. Kinney, Director
                  HAggarwal. Co-lnv.
                  G. Chew, Co-inv.
                                                 H. Andrews, Director
                                                  W.y. Tsai, Deputy Director
                                                 R.Wallace, Deputy Director
                                                           Research Project
                                                             on Cancer
                                                          Community-Based
                                                          Intervention Project
                       Research Project
                         on Asthma
   Research Project
Growth and Development
F. Perera, P.I.
V. Rauh, Co-lnv.
R. Whyatt Co-lnv.
                                                          F. Perera, P.L
                                                          V. Rauh, Co-lnv.
                                                          R.Santel la. Co-lnv.
                                                          R. Whvatt. Co-lnv.
                                                         D. Evans, P.I.
                                                         M. Fullilove, Co-lnv
                                                         P. Shepard, Co-lnv.
                                                                Co-lnv.
                     J. Ford, P.I.
                     R. Miller, Co-lnv.
                     I. Meyer, Co-lnv.
The research team is investigating these hypotheses in a molecular epidemiologic prospective cohort study
of 600  pregnant women,  following the infants  and their  mothers for 2  years postnatally.  Questionnaire,
monitoring, biomarker, and clinical data are collected. A multilevel analysis is examining relationships be-
tween environmental and susceptibility factors and adverse  health outcomes at the individual and community
levels. An  intervention research project is testing whether a community campaign,  Healthy  Home Healthy
Child,   can be  effective  in heightening awareness about environmental risks  and measures individuals can
take to  protect their children. An individual/household intervention will test whether reduction in levels of
The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


allergens and nutritional supplementation can reduce the risk of asthma among infants at high risk. Through
its Community Outreach and Education Program, the CCCEH is working in partnership with the community
in all phases of the etiologic research, as well as in communicating the research results and their implications
for policy.

Enrollment: As of September 12,2000,498 women had been enrolled, 357 of whom had undergone prenatal
monitoring. A total of 334 babies had been delivered, of whom 27 had reached their second birthday. The
ethnic distribution was 43 percent African American and 57 percent Latina.

Summary of Preliminary Findings: (1) Monitoring and biomarkers show variable, widespread, and in some
cases, high exposure during pregnancy to PAH, pesticides, ETS, and allergens (Kinney, Chew, Whyatt, Perera
et al.); (2) In utero allergic sensitization to multiple indoor allergens is common (Miller et al.); (3) There is
a high prevalence of respiratory problems (Meyer, Ford et al.) and developmental disorders (Rauh et al.) by
age 1; (4) Community-level analyses of NYC show geographic concordance for high rates  of childhood
asthma hospitalization, low birth weight, and social/economic stressors (Wallace et al.; Rauh and Andrews);
and (5) A survey of 556 women showed awareness of environmental health  threats to be high, but practical
knowledge about preventing harmful exposures was low (Evans, Fullilove, Shepard et al.).

Personal Monitoring: Personal  exposure  monitoring (48-hour) to determine the pregnant  woman s in-
halation exposure  to pollutants showed that all 157 samples initially tested  had detectable levels of one or
more carcinogenic PAH.  Total PAH exposures averaged 3.94 ng/m3 and  varied significantly among the
women, with a range of 0.02 44.81 ng/m 3 (Kinney et al., in preparation).

Analysis of Pesticides: Analysis of pesticides  in personal  air samples from an  initial subset (72) of the
pregnant women showed detectable concentrations of at least three neurotoxic pesticides (chlorpyrifos,
diazinon, and propoxur) that are widely used to control cockroaches and other pests in urban homes (Whyatt
et al., in preparation). The concentrations were similar to those reported in Jacksonville, Florida, an area with
high household pesticide use (Whitmore et al., 1994).

Home Monitoring/Exposure Assessment: Eighty-five percent of the homes had detectable concentrations
of cockroach allergen, with 46 percent above 8 mg/g; 70 percent had detectable mouse allergen, with 30
percent above 8 mg/g (Chew, Aggarwal, and Kinney). These concentrations  are comparable to those seen in
a major national study (NCICAS) and show that exposure  to these allergens is prevalent, in some  cases
ranging up to levels associated with health effects.

Immune Changes: Immune biomarkers have been analyzed in the initial subset of mothers and newborns,
including total IgE levels, allergen-specific IgE levels, mitogen or allergen-induced lymphocyte proliferation,
and cytokines. In  the first group of babies  enrolled, there was a  high prevalence of in utero allergic sen-
sitization as evidenced by the lymphocyte proliferation  assay (>50% to cockroach allergen) (Miller et al,
submitted). In many cases, cord blood allergen-induced proliferation occurred in the absence of maternal
blood allergen-induced proliferation.

Respiratory Health: Preliminary analysis of data on the initial 158 infants whose mothers were interviewed
shows a higher rate of health care utilization for respiratory complaints in the study population (Meyer et al.,
manuscript in preparation) compared to  both New York City and the entire  United States. The 3-month
prevalence of breathing symptoms at ages 3,6,9, and 12 months were 46 percent, 54 percent, 50 percent, and
43 percent with correspondingly high rates of Emergency Room (20%, 22%,  17%, and 9%, respectively) and
doctor s office visits for these symptoms (37%, 39%, 28%, and 20%, respectively). Ten percent of the infants
had a physician-diagnosed asthma by the age of 12 months.

Fetal Growth and Child Developmental Assessments: Measures of fetal growth  and gestational age
(abstracted from medical records) showed that 9 percent were low birth weight. The ethnic difference in birth
 10                      The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s EnvironmentaLHealth and Disease Prevention Research Progress Review Workshop

weight is explained by the larger proportion of African American preterm infants  (p=.02). Cognitive and
motor developmental testing at infant age 6 months (Denver II Developmental Test) showed that 14.9 percent
of infants scored in the at-risk  range. Twelve-month Bayley scores show that 15.9 percent of infants scored
in the delayed range for cognitive development, and 25.8 percent in the delayed range for motor development
(Rauhetal.).

Community Participation, Outreach, and Education: The CCCEH has an active working relationship with
local community organizations. Specifically,  the Center has worked closely with  West Harlem Environ-
mental Action (WEACT) on  many projects, including the development of materials for an environmental
health education campaign and the  organization of a large community  conference in  March 2000, which
engaged more than 500 health care providers, scientific researchers, grassroots and community organization
representatives, environmentalists, and local and national policymakers in a discussion about children s health
and the urban  environment. The Center has formed a Community Advisory Board (CAB) comprised of
representatives from nine different organizations working in Washington  Heights,  Harlem, and  the South
Bronx in the areas of health, education, environment, and community development.  The advice of the CAB
is sought on issues such as how the Center can better meet the needs of the communities in which it works,
the development and distribution of educational materials, and the effective communication of research results
to the study subjects and the community.
The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Pesticide Exposure During Pregnancy Among Minority
Women  Residing in Northern Manhattan and the South Bronx
R.M. Whyatt', D.E. Camann \ D.B. Barr3, P.L, Kinney ', A. Reyes ', J. Ramirez ', J. Dietrich ', D. Diaz ',
and P.P. Perera '
'Columbia Center for Children s Environmental Health, Mailman School of Public Health, Columbia University,
New York, NY,  2Southwest Research Institute, San Antonio, TX, 3Centers for Disease Control and Prevention,
Atlanta, GA

Residential  pesticide use  is widespread in the United  States,  with  the  nonpersistent  pesticides  (NPP)
(organophosphates, carbamates, and pyrethroids) generally used for insect  control. However, little data are
available specific to pesticide use among minority populations. As part of the prospective cohort study being
conducted by the Columbia Center for Children s Environmental Health, we have gathered questionnaire data
on pesticide use during pregnancy from 131 African American and Dominican women residing in  Washing-
ton Heights, Harlem, and  the South Bronx. Additionally, 72 of the women underwent personal ambient air
monitoring  for 48 hours  during the third trimester to  determine NPP exposure levels. Of  the women
questioned, 34 percent report that their homes were sprayed by an exterminator during the pregnancy,  with
47 percent  of those saying the spraying was done once per month.  One-third of the women also report use
of can sprays.  Ninety percent of the pesticide  use was for cockroach control. Of the  women monitored, all
(100%) were simultaneously exposed to two organophosphates, diazinon (range in air concentrations 2.0-
6,000 ng/m') and chlorpyrifos (range 0.7-193 ng/m1), and to the carbamate, propoxur (range 3.8-l,380ng/m\
These results  are of concern in light of experimental evidence linking prenatal organophosphate exposure to
adverse  neurocognitive development.  We  recently have begun to validate the measurements of NPP in
meconium  as a biomarker of cumulative prenatal exposure to facilitate evaluation  of the effects of this
exposure on newborns in the cohort. Pilot results on 20 meconium samples found that two organophosphate
metabolites, diethyltphosphate and diethylhiophosphate, which are common to both chlorpyrifos and dia-
zinon, were detected in 95  percent and 100 percent of the samples, with  a range of 0.8-3.2 mg/g  and 2.0-5.6
mg/g, respectively.
 12                             The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Indoor and Outdoor Environmental Exposures
in Minority Children of Northern Manhattan and the South Bronx
P.L. Kinney, M. Aggarwal, G. Chew, R.M. Whyatt, and P.P. Perera
Columbia Center for Children s Environmental Health, Columbia University, Mailman School of Public Health,
New York, NY

African-American and Latino residents of New York City experience some of the highest rates of asthma
morbidity and mortality in the United States.  Children  and  individuals with preexisting respiratory diseases
may be especially sensitive to the effects of airborne particles and other toxic air pollutants.  As a part of the
Children s Environmental Health Center, exposure to indoor allergens (cockroaches, dust mites, and mice)
and air pollutants  (PM2   NOX, PAHs,  and diesel exhaust particles) is assessed in mothers and newborns.
During the 32nd week of pregnancy, each participant s home is visited to collect  dust samples from  the
woman s bed and  kitchen. At this time point, women also carry a portable personal exposure monitor for 48
hours to determine their  inhalation exposure to polycyclic  aromatic hydrocarbons (PAHs).  The home is
revisited when the child turns 12 months of age,  when four separate dust samples are collected from  the
mother s bed, the child s bed, the floor surrounding the child s bed, and the kitchen. Also, a 25 percent sub-
sample of homes undergo air pollution monitoring within and immediately outside the home at the  12-month
time point. Air monitors are placed in subjects  homes for a 2-week period to collect PM2 5, NOX, elemental
carbon, and indoor airborne allergens. At each of these homes, traffic also is counted for two 15-minute
periods on the street  segment nearest to the apartment.  Traffic  counts are segregated as diesel buses, diesel
trucks, and cars. Currently, 159 samples of PAH have been  analyzed and all were found to have detectable
levels of one or more carcinogenic PAHs. Total PAH exposures for these 159 women averaged 3.94 ng/m'
and ranged from  0.02-44.81 ng/m\ Results  for allergen analysis from prenatal dust samples are  shown in
Table  1.
                                      Table 1. Prenatal samples.

                                Mother s Bed Samples
Kitchen Samples
Allergen
Cockroach (Bla g 1 )
Cockroach (Bla g 2)
Mouse Urinary Protein
n
80
117
96
% > 2 Jlg/g*
31%
27%
17%
Max Cone.
46U/g
148 U/g
20|J,g/g
n
72
108
89
% > 2 |lg/g*
58%
63%
17%
Max Cone.
781 U/g
798 U/g
1478|ig/g
  Cockroach allergens are expressed in U/g.
The Office of Research and Development s National Center for Environmental Research
                       13

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Mount Sinai School of Medicine
       New York, NY

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Overview and Description of the Mount Sinai
Center for Children s Environmental Health and Disease
Prevention:  Inner-City Toxicants and Neurodevelopmental Impairment
Mary S. Wolff
Mount Sinai School of Medicine, New York, NY

Children in America s cities  are  at  risk of exposure to multiple known and  potential developmental
toxicants pesticides,   polychlorinated biphenyls (PCBs), and lead. The goal of the Mount Sinai Center  for
Children s Environmental Health and Disease Prevention  will be to identify, elucidate, and prevent devel-
opmental deficits that result from exposures to environmental toxicants in the inner city.

Project 1, Barbara Brenner, Principal Investigator (PI), is a community-based prevention program being  un-
dertaken in East Harlem, New York City, in partnership with Boriken Health Center. At Boriken, we  are
recruiting expectant mothers and implementing an intervention to reduce exposures to pesticides and other
developmental toxicants in their homes. A nonintervention comparison group consists of expectant mothers
in similar housing enrolled in Project 2 and at Settlement Health, a nearby community health center. Pesticide
levels and roach infestation levels are being assessed in  both groups. In Years 3 and 4, IPM  will be gen-
eralized to housing and schools throughout East Harlem, using community intervention strategies.

Project 2, Trudy Berkowitz, PI, is a prospective epidemiologic study of an ethnically diverse birth cohort of
infants born at Mount Sinai. More than 400  mothers have been recruited so far, with more than 100 births.
The ultimate aim is to assess whether in utero exposures to pesticides and other toxicants are associated with
developmental delays in children in New York City.

Project 3, Jim Wetmur, PI, is studying polymorphisms in the enzymes that activate and detoxify organo-
phosphates and other pesticides in the population of mothers and infants enrolled in Project 2.

Project 4, Tom Matte, PI, is a retrospective study  of African-American men enrolled in the Collaborative
Perinatal Project. PCBs have been measured in  154 maternal sera to assess whether in utero exposures to
PCBs are associated with disordered neuropsychological function in adolescent or adult life.

Project 5, supports Dr. Andrea Gore as a newly recruited Center scientist. She is examining the mechanisms
by which environmental toxicants affect neuroendocrine development. Experiments in a female rat model will
characterize interactions between toxicants and hypothalamic GnRH neurosecretory neurons, key regulators
of reproductive development. Using a neuronal cell line, effects  of chlorpyrifos, PCBs, and methoxychlor
have been assessed in vitro and found to alter GnRH expression in GT1 cells.

The Center contains Facilities Cores in Exposure Assessment and Biostatistics/Data Management as well as
an Administration Core.
The Office of Research and Development s National Center for Environmental Research                             17

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Kinetic PCR on Pooled DNA: A High-Throughput,
High-Efficiency Alternative in Genetic Epidemiologic Studies
/. Chen, R. Higuchi, S. Germer, andJ.G. Wetmur
Mount Sinai School of Medicine, New York, NY

The ideal technology for screening SNPs requires high-throughput with minimal cost per sample, minimal
usage of valuable DNA resources, and maximal flexibility for introduction of new polymorphisms. Array
hybridization that relies on the difference between hybridization of  matched and mismatched  products to
allele-specific oligonucleotides on the array is powerful, although not  foolproof for detecting heterozygosity.
Other competing methods such as  Taqman  and molecular beacons probes require fluorescent labeling of
probes, which increases the expense. We will describe a new technology, kinetic allele-specific PCR with
DNA pooling, developed at Roche  Molecular Systems (S. Germer, M.J. Holland, and R. Higuchi. High-
throughput SNP allele-frequency determination in pooled DNA samples by kinetic PCR. Genome Res 2000;
10:258-266), which  satisfies all of these criteria and  offers  a powerful new tool for detecting meaningful
polymorphic differences in candidate gene association studies  and genome-wide linkage disequilibrium scans.

We will present data from two blinded tests of the technology. We had three individuals prepare pooled DNA
samples from (A) 252 individuals  and (B) 271 individuals separated  into three ethnic groups. We had pre-
viously determined their PON1 Q191R genotypes by PCR-RFLP.  These 12 pooled DNA  samples were
genotyped by our collaborative investigators at Roche Molecular Systems using kinetic allele-specific PCR.
For test A, with 3 pools and a total  of 12 PCR measurements, the allele frequency was 0.449-0.010, com-
pared to 0.448 determined by PCR-RFLP. For test B, with 9 pools and  a total of 36 PCR measurements, the
allele frequencies for Caucasians (n=56), Latinas (n=127), and African-Americans (n=86) were 0.266-0.011,
0.386-0.011, and 0.617-0.010, respectively, compared to 0.267,0.409, and 0.610 determined by PCR-RFLP.
These results demonstrate a powerful new technology for determining frequencies  of SNPs in  an epide-
miological study.
 18                             The Office of Research and Development s National Center for Environmental Research

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University of Washington
      Seattle, WA

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Overview and Description of the
Center for Child Environmental Health Risks Research
Elaine M. Faustman
University of Washington, Seattle, WA

In 1998, the U.S. Environmental Protection  Agency awarded grants to create eight centers across die Nation
to examine children s environmental health.  The Center for Child Environmental Health Risks Research, part
of the Department  of Environmental Health  at the University of Washington, is one of these centers. Our re-
search aims to understand the mechanisms  defining children s susceptibility to pesticides, and Ifce implica-
tions of this susceptibility to development and learning.

Researchers at the  Center for Child Environmental Health Risks Research represent multiple  disciplines and
several  different institutions. Our work spans the continuum of environmental health  research ftom the
cellular level to community-wide intervention projects. Our  research takes place in the laboratory, and in the
field.

Laboratory-based researchers are working to identify the cellular, biochemical, and molecular mechanisms
of developmental neurotoxicity of pesticides. The emphasis of these studies is to assess the potential for
functional impacts  on neurodevelopment.  Researchers also  will evaluate the impact of genetic variations on
how people metabolize pesticides, particularly organophosphate pesticides.

Field-based researchers are working to identify the critical pathways of pesticide exposure for children. Our
Center includes a partnership with communities in the Yakima Valley, a key Washington State sgricultural
area. Researchers are working to develop culturally appropriate  interventions to reduce occupatioiial take-
home exposure. The goal is to ultimately reduce exposure of farmworker families to pesticides.

To learn more about the Genetic and Cellular, Behavioral, Exposure Assessment, and Community Inter-
vention and Communication projects at the  Center for Child Environmental Health Risks Research, please
contact Tiffany Potter-Chiles at 206-616-9133.
The Office of Research and Development s National Center for Environmental Research                             21

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Effects of Human Paraoxanase (P6W7) Single
Nucleotide Polymorphisms on Susceptibility
to Specific Organophosphate  Insecticides
Clement E. Furlong', Wan-Fen Li', Lucio G. Costa ''2, Victoria H. Brophy ', Rebecca J. Richter', Diana M.
Shih 3, Aaron Tward3, and Aldon J. Lusis3
'University of Washington, Seattle, WA; : University of Roma La Sapienza, Roma, Italy; University of California at
Los Angeles School of Medicine, Los Angeles, CA

Paraoxonase (PON1) is tightly associated with HDL particles and appears to be involved in the metabolism
of oxidized lipids; however, its role in the metabolism of organophosphate insecticides has been investigated
much more extensively. Human PON1  exhibits a substrate dependent polymorphism determined  by the
R192Q polymorphism. The P6W7RI92 isoform hydrolyzes paraoxon at a tenfold higher catalytic efficiency
than does the PON1QW2 polymorphism; however, it is much less efficient  at hydrolyzing sarin. A second
polymorphism, M55L, also present in human populations, has been associated with lower serum PON1 levels;
however, this association may be due to  linkage with inefficient promoter polymorphisms.

For many years, it has been assumed that individuals homozygous for the PON1RW2 isoform would be much
more resistant to paraoxon/parathion exposures than individuals with the PON1Q[92 polymorphism. Develop-
ment of the PON1 knockout mouse model has allowed us to examine directly this assumption. Surprisingly,
PON1 knockout mice, which are devoid of both plasma and YwerPONl, were not more sensitive to paraoxon
than wild type mice. They were, however, very sensitive to both diazoxon and chlorpyrifos oxon. Injection
of the knockout mice with either human PON1192 isoform allowed for the testing of the efficacy of each  in
the detoxication of paraoxon, diazoxon, and chlorpyrifos oxon under physiological conditions. Both PONl^
isoforms afforded equivalent protection against diazoxon exposure, while the PONlm92 isoform provided
better protection against chlorpyrifos oxon exposure. Neither PON119, isoform provided protection against
paraoxon exposure, nor did expression of a high level of human -PCW/R19, in atransgenic mouse line. In vitro
catalytic efficiency of PON1 for specific substrate hydrolysis was closely correlated with the  protection
against exposure to specific  OPs afforded by injection into knockout mice of each PON1W2 isoform. The
relatively low efficiency of paraoxon hydrolysis  by PON1  explains the lack of increased sensitivity  to
paraoxon observed in the PON1 knockout mice. Exposure of the knockout mice to the parent  compounds,
diazinon and chlorpyrifos, indicates that PON1 provides protection mainly against the oxon forms of these
OPs, which can be significant in actual  exposures. These findings also raise the important point that safety
testing of diazinon and chlorpyrifos should include oxon levels comparable to those experienced in actual
exposures. Previous testing has been done with very pure compounds.

Plotting the rates of diazoxon hydrolysis versus paraoxon hydrolysis for human population samples provides
an accurate inference ofPONll92 genotype as well as the level ofPONl expressed in each individual, which
can vary by  as much as fifteenfold among individuals.  Polymorphisms in the PON1 promoter region are
responsible for a large part of the observed variability in serum PON1 levels among individuals. Newborns
have very low PON1 levels, which increase to their respective adult levels at about 1  year of age. Thus, three
factors appear to govern an individual s  sensitivity to diazoxon and chlorpyrifos oxon: age, position 192 ge-
notype, and the level of serum PON1.
22                     The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Preliminary Baseline Results From
the Randomized Community Trial To Assess the Efficacy
of a Community-Wide Program To Reduce Children s Exposure to Pesticides
Beti Thompson, Gloria Coronado, Cam Solomon, John Kissel
University of Washington, Seattle, WA

Introduction
For Healthy Kids! is a community-based, randomized  trial taking place in the Lower Yakima Valley of
Washington State. After baseline data collection that included an in-person  interview, urine collection from
a farmworker and a child aged 2 to 6 in the same household, a dust sample from the referent household, and
a dust sample from a vehicle used by a farmworker in the referent household, 24 communities  were ran-
domized to an intervention or control condition.

A Community Advisory Board, made up of representatives from all of the constituents interested in pesticide
use or control, is extremely active in the project. The Board was formed prior to the baseline assessment and
was able to contribute to questionnaire development. The Board selected the  local Project Coordinator and
staff.  The Board reviews data and progress,  makes recommendations, and suggests strategies for interven-
tion components. The Board also has opened many doors to other gatekeepers in the community.

Methods
Using randomly selected census blocks and randomly selected households within blocks, an in-person survey
was conducted  with agricultural workers in  20  communities and  8  labor camps in the Yakima Valley of
Washington State. The survey included questions about job tasks,  pesticide exposure, workplace  character-
istics, and home practices. A total of 571 respondents completed the interviews.

Results
Baseline Results: Response Rates. Responses were received from 571 farmworkers, constituting 89.6 percent
of the sample drawn or 93.1 percent of known eligibles. Of farmworkers with an eligible child in  the house-
hold (N=231), 92.2 percent participated in the urine collection part of the trial; 91.3 percent of the children
participated. Of eligible households, 96.3 percent participated in the house dust collection and of those who
had vehicles, 97.2 percent participated in the vehicle dust collection.

Baseline Results: Interview. The farmworkers were overwhelmingly Hispanic (88%), had low  education
levels (74% of Hispanics had an 8th grade education or less), the vast majority of respondents lived in  the
Valley year-round (95%), and respondents tended to be male (76%).

Respondents were asked about the job tasks they had performed in the past 3 months and perceived exposure
to pesticides during those job tasks. Of four job tasks  harvesting, weeding, thinning, and pruning 56.7
percent of farmworkers reported being exposed while thinning. For all tasks combined,  50 percent of farm-
workers reported exposure to pesticides.

Applicators and mixer/loaders were more likely to use personal protective equipment while performing their
job tasks than farmworkers who reentered the fields before the approved interval. Among nonapplicator
farmworkers, less than 50 percent reported wearing protective boots (42%), gloves (40%), or protective lenses
(23%) when working in a pesticide-exposed area.

There was considerable variation in decontamination facilities at workplaces. Most had  bathrooms, and 90
percent had drinking water available. About  78 percent of workplaces had  water for washing hands; how-
ever, only 64 percent provided soap or towels. Showers onsite were reported in  25 percent of workplaces.
Only 40 percent of workplaces had eyewash stations onsite.
The Office of Research and Development s National Center for Environmental Research                             23

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Farmworker home practices  to prevent pesticide transmission to their children and the home environment
varied by practice. The vast majority removed hats (88%) when entering the home. Eighty-three percent
reported washing work clothes separately from the rest of the family s laundry. Fewer farmworkers reported
removing their boots before  coming into the home (57%), changing out  of work clothes soon after entering
the home (52%), and washing work clothes after one wearing (58%). About 49 percent reported bathing or
showering within 1 hour after coming from their work, and only 50 percent reported that they never held their
children while wearing  their dirty work clothes after coming in from the field.

Baseline Results: Urine  Data. The overwhelming majority of  adults in the study (N=213) were Hispanic
(97%). The total family income for 90 percent of respondents was $25,000 or less per year. Dimethyl com-
pounds were  the only compounds found in measurable quantity in both the adult and child urine. Among
workers who  reported harvesting, 60.5 percent had detectable  urinary  metabolites of pesticides (0.1 mg/L or
greater) compared to 47.4 percent of those who did not harvest. The difference was not detectable among
weeders, but was significant  among pruners and thinners.

In comparing the relationship between adult urinary metabolite concentrations and child concentrations, there
is a direct relationship between the amount of adult metabolite concentrations and those of children in  the
household. Sixty-seven percent of children in households with farmworkers in the highest quartile  of meta-
bolite concentrations also had detectable concentrations. Conversely, among adults in the lowest quartile of
concentrations, only 28.9 percent of children had a detectable level of urinary metabolites.

An examination of the child-based urinary metabolite concentration (creatine-based) dose estimates, assuming
the samples are the result of guthion (azinphosmethyl) exposure, indicates that 23 percent of the  children
exceed the RfD  (for volume-based, the percentage is 40%). If the samples are the result of exposure to
phosmet, then only  1.5 percent (creatine-based) or 2.6 percent (volume-based) of children exceed  the RfD.
The highest estimated child dose was 31 times (creatine-based) or 137 times (volume-based) higher than the
azinphosmethyl RfD.

Baseline Results: Dust Data. House dust samples were assessed for six  different chemicals  (diazinon, mala-
thion, chlorpyrifos, methyl-parathion, phosmet, and azinphosmethyl). Azinphosmethyl was detected in  the
greatest proportion of households (63.8%). The median concentration of azinphosmethyl was 0.53 mg/g. Less
than one-half of  the households had detectable levels of the other chemical types. The median concentration
of azinphosmethyl in vehicles was 0.85 mg/g.

There were few  differences  in median child urinary metabolite concentrations in homes with detectable dust
residues compared with those without detectable dust residues. Similarly, there were few differences in child
urinary metabolites and vehicle dust; however, in households where vehicle dust contained measurable levels
of methylparathion, phosmet, or azinphosmethyl, the median levels of child urinary metabolites of pesticide
were consistently higher than in households without detectable levels of those compounds in vehicle dust.

Conclusion
The baseline data indicate that there is reason to be concerned about pesticide exposure in the Lower Yakima
Valley. The intervention study as conducted in a randomized community  trial should provide insights about
the value of community-wide intervention on this issue.
24                              The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Summer Project for Minority High School
Student Interns:  Effects of Home Parties on Farmworkers
R. Godina*, G. Martinez*, I. Islas, G.D. Coronado, and B. Thompson
University of Washington, Seattle, WA

Background
Agricultural workers and their children are thought to experience high levels of exposure to pesticides. Little
is known about the effectiveness of strategies to change agricultural workers practices and beliefs related
to pesticides. In a large randomized, community trial, a variety of community events are being used to make
farmworkers aware of things that can be done to  reduce the amount of pesticides taken home from their
workplaces. By reducing this  take-home  pathway, it is thought  that children in the homes are less likely
to be exposed to pesticides. One intervention strategy used in this project is  home parties, where family
members and neighbors gather in a farmworker s  home and participate in an interventionist-led discussion
about pesticides and prevention techniques.

Purpose
The purpose of this  student project was  to conduct a process evaluation of the efficacy of the home parties.
This was done by characterizing the beliefs and home practices of agricultural workers who attended in-home
pesticide education sessions (home parties) and comparing these with baseline community survey findings.

Methods
Telephone  interviews were conducted among a random  sample  of 150 home party  attendees  from two
communities in the Lower Yakima Valley. A number of telephones were disconnected, and some individuals
were unable to be reached, leaving 92 known eligible agricultural workers.

Results
Ninety-eight percent of known eligible  attendees responded to the survey. The educational and income
distribution of the sample matched those of the baseline survey. Home party attendees were more familiar
with the factual information of pesticide exposure than those in the baseline survey. They reported engaging
in more pesticide protective home practices compared to community baseline survey respondents.  Attendees
reported having learned important  key  messages, including general pesticide information, washing  work
clothing separately from family clothing,  and removing shoes before entering the home.

Conclusion
Home parties appear to be an efficacious way to present messages regarding pesticides to agricultural work-
ers. This study compared attendees to baseline respondents and, therefore, has methodological limitations.
Future research should evaluate home parties using control groups  for comparison.
*Minority High School Student Interns
The Office of Research and Development s National Center for Environmental Research                            25

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University of California, Berkeley
         Berkeley, CA

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Overview and Description of the Center
for the Health Assessment of the Mothers and Children
of Salinas (CHAMACOS):  A Community/University Partnership
Brenda Eskanazi
University of California at Berkele\, Berkele\, CA

In the last few years, several studies have demonstrated pesticide contamination in the homes of young
children living in both agricultural and suburban areas. However, to  date,  only a few studies have been
conducted to assess the extent of children s exposure to pesticides and no studies have examined  whether
low-level chronic exposure can lead to adverse health consequences. Our goal is to investigate in utero and
postnatal pesticide and  allergen exposures and their potential health effects on young children living in the
Salinas Valley, an agricultural community in Monterey County, California. Ultimately, we aim to translate
research findings into sustainable strategies to reduce environmental exposures to children, and thus reduce
the incidence  of potentially  environmentally  related childhood disease. Our Center also will generate in-
formation  critical for implementation of the Food Quality Protection Act,  which requires  that pesticide
tolerance levels in food take into account the special vulnerabilities of children.

Our specific aims are to:

>   Estimate  sources,  pathways,  and levels of in utero and postnatal pesticide exposures of children living
     in an agricultural community  by  measuring biological and environmental  samples.

>   Determine whether exposure to pesticides is associated  with poorer neurodevelopmental functioning and
     behavioral problems, delayed growth, and increased respiratory symptoms and disease.

>   Determine whether  exposure to environmental allergens  and respiratory irritants is associated with
     increased respiratory symptoms and disease. We will determine whether pesticide exposure modifies
     the relationship of allergen exposure and respiratory outcomes.

>   Evaluate  the impact  of  Healthy  Homes  interventions  on the reduction  of pesticide exposure to farm-
     worker children.

Enrollment ended in  October 2000, after a full year of recruitment through community clinics. We have
succeeded in enrolb'ng the targeted number of pregnant  women (n=550) by the expected date. Overall, we
have been able to meet our goals and are developing a repository of samples that we will be  able to utilize
for future studies (e.g., endocrine-disrupting  pesticides). As of October 5,  2000,  we  have  collected  and
processed approximately  1,100 urine, 650 maternal and cord blood samples, and 118 breast milk samples.
Preliminary  pesticide urinary metabolite data  are available for 93 women.  We also have completed 395
prenatal home inspections and successfully collected 790 mini-burkard  for airborne pollen and mold, 1,106
dust samples for allergen testing, and  395 dust samples for pesticide analysis. A total of 202 babies have been
born; Brazelton assessments have been completed on 136.  To date, we have been able to locate more than
90 percent of these women when their child reached 6 months. More than 80 percent have participated at 6
months, and we have been able  to collect urine  samples from all children  participating in  the 6 months
neurobehavioral assessments (17 children to date). Currently, we are entering the data from prenatal ques-
tionnaires and  home  inspections, and we are  working with collaborators at  the California Department of
Health Services to begin  utilizing  the 7999 Pesticide Use  Reports, which recently became  available. Sta-
tistical analyses of prenatal and delivery data will  begin later this fall as data  entry progresses and the CDC
provides additional testing results. We also have maintained  close contacts with our community partners and
advisory board. Pending notification of funding from the California Endowment  Foundation, we will plan
and implement a community-based education and intervention program to reduce environmental exposures
to children.
The Office of Research and Development s National Center for Environmental Research                             29

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Preliminary Approaches To Assessing
Organophosphate Pesticide Exposure and Potential
Health Risks to Pregnant Women Living in the Salinas Valley, California
Brenda Eskanazi
University of California at Berkelev, Berkeley, CA

Unique in the Nation,  California requires reporting of all agricultural pesticide use. Pesticide use reporting
(PUR) data indicate widespread use of organophosphate pesticides in California agriculture. In  the Salinas
Valley, which  is intensively farmed  for vegetables and fruit, approximately 500,000 pounds are  used
annually, raising concerns  about exposures to farmworkers, other members of the  community, and  their
families. Organophosphate (OP) metabolite data are available for 93 of approximately 550 pregnant women
participating in the  Center  for the Health Assessment of the Mothers and Children of Salinas study. These
urine  samples were collected between October 1999 and January 2000, when fewer agricultural pesticides
are used compared  to the spring and summer. The Centers for Disease Control and Prevention (CDC) tested
the samples for six  dialkylphosphate  metabolites: dimethylphosphate  (DMP); dimethyldithiophosphate
(DMDTP); dimethylthiophosphate (DMTP); diethylphosphate (DEP); diethyldithiophosphate (DEDTP); and
diethylthiophosphate (DETP). These metabolites derive from approximately 40 OP compounds, falling into
the general categories of dimethoxy and diethoxy organophosphate pesticides.

Median concentrations for the three dimethyl metabolites were 27 percent to 240 percent higher than the
median reference range reported by the CDC for the National Health and Nutrition Examination Survey III
(NHANES III) (D.  Barr, personal communication) DMP:  2.3 vs. 1.8 mg/L; DMTP:  16.5 vs. 4.8 mg/L; and
DMDTP:  1.4 vs. 0.55 mg/L. Median concentrations for the three diethyl metabolites, however, were lower
than or very similar to the  reference values DEP: 1.0 vs.  4.5   |ig/L; DETP: 1.1 vs. 1.2  [ig/L; and DEDTP:
0.09 vs. Not detected above LOD.

We estimated women s OP pesticide doses  from urinary OP metabolite concentrations using a deterministic
steady-state model described by Fenske et al.' Preliminary dose estimates  were calculated assuming that
diethyl urinary metabolites were attributable to either chlorpyrifos or diazinon, and that dimethyl metabolites
were attributable to malathion or oxydemeton-methyl. These compounds are four of the most heavily used
OP compounds in the Salinas Valley (see Table 1).

    Table 1.  Preliminary risk assessment of organophosphate exposures to study participants, n=93
             (assumes all exposure due to single parent compound1).
Pesticide
Chlorpyrifos
Diazinon
Malathion
Oxydemeton-methyl
1999 Use (pounds)2
59,000
104,000
67,000
69,000
Proportion of Women Exceeding
Reference Dose (%)
19
2
0
56
  Fenske RA, Kissel JC, Lu C, Kalman D, Simcox NJ, Allen E, Keifer MC. Biologically based pesticide dose
  estimates for children in an agricultural community. Environ Health Perspect 2000; 108(6):515-520.
  Monterey County. Source: California Department of Pesticide Regulation PUR system.
30
The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


These findings suggest the possibility that a significant proportion of pregnant women participating  in the
study experience exposures that exceed EPA chronic dietary reference doses, which may not account for the
special sensitivity of the fetus. Our exposure calculations may be an underestimate because pesticide use and
potential exposures are higher during the growing season. Additional risk evaluations will be completed when
data for all participants are available from the CDC. We also are developing modeling approaches to estimate
fetal exposures and risk. Finally,  we are exploring  the possibility of metabolite-specific chemical analyses
to quantitatively  attribute risk to specific OP compounds. Planned statistical analyses will investigate  deter-
minants of pesticide metabolite  levels in urine, such  as nearby  pesticide use (PUR data) and occupational
status.
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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


CHAMACOS Laboratory Core:  Challenges
of Biological Sample Collection and Processing
Nina Holland
University of California at Berkeley, Berkeley, CA

Children differ from adults in size, types and extent of exposures, diet, and potential long-term health effects
of environmental exposure. To  learn more about  differences between adults and children, and to predict
potential adverse effects, new molecular  and cytogenetic methods need to be developed and employed to
identify biological markers  or biomarkers. Biological  measurements, such as changes in the  structure  of
chromosomes, DNA polymorphisms, cellular characteristics, activities of enzymes, and levels of pesticides
or other chemicals in blood or other body, can be used as biomarkers. The main advantage of biomarker
studies is the  ability to detect effects long before clinical manifestation of  disease, thus allowing  for in-
tervention and prevention. A far smaller  population is needed to  obtain an informative conclusive  study.
Mechanistic information  can be acquired, including factors that affect  interindividual  variability; in other
words, biomarkers may help us to understand not only why children are  different from  adults but also how
children differ from one another in response to diet, environmental pollution, or infectious diseases. The main
focus of our study is to establish potential effects  of pesticide exposure, by obtaining comprehensive epi-
demiological information and environmental and biological samples from  pregnant women and their children
during early development.

Specific aims of the Laboratory Core of the CHAMACOS project include development of standard operating
procedures for sample collection and processing, optimization of sample banking, database management, and
implementing  quality control and quality  assurance (QA/QC)  procedures. An additional aim of the Labora-
tory Core is to obtain consent in a manner  that would allow future use of these specimens. Collection,
processing, and storage of samples is coordinated between the Natividad Hospital field office in Salinas, the
Children s Hospital of Oakland Research  Institute  in Oakland, and the School of Public Health at the Uni-
versity of California at Berkeley. Samples are sent for analysis to the Centers for Disease Control and Pre-
vention and  other collaborators.

As of October 5, 2000, we have collected and processed approximately  1,100 urine, 650 maternal and cord
blood, and 118 breast milk samples; 790  mini-Burkard slides  for airborne pollen and mold analysis; 1,106
dust samples for allergen and endotoxin testing; and 395 dust samples for pesticide analysis. Each  individual
blood sample processing results in at least 18 different aliquots, including serum, clot,  plasma, buffy coat,
red blood cells, blood smears, and stabilized samples for folate and cholinesterase analysis. Urine and breast
milk are aliquoted for assessment of organophosphate pesticide exposure, as well as other potential environ-
mental pollutants. Dust is being processed for allergen and endotoxin analysis, and for potential pesticide
measurement.  The total number of expected samples will  reach approximately  60,000, to be completed within
the next year.

Quality Assurance/Quality Control (QA/QC)  procedures  have been developed and maintained for storage,
processing, and analysis of various specimens.  For example, blank and  spiked samples and internal  control
replicates were created for both urine  and dust samples. All samples are barcoded, and all further analyses
are performed blindly. Currently, provisions have already been made for the analysis of a number  of bio-
markers of exposure and susceptibility.

The main goal for the future is  to develop and implement a comprehensive plan for the most effective and
informative  use  of the valuable  biological and environmental samples collected from this unique population
of agricultural workers  and their children exposed to pesticides.  To accomplish this important task, dis-
cussions with  prospective collaborators from other Children s Environmental Health Centers and other or-
ganizations are underway. It will require the acquisition of significant additional resources through extramural
funding.
32                              The Office of Research and Development s National Center for Environmental Research

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Johns Hopkins University
     Baltimore, MD

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Overview and Description of the Johns Hopkins
University Center for Childhood Asthma in the Urban Environment
Peyton Eggleston
Johns Hopkins University, Baltimore, MD

The long-term goal of the Center for Childhood Asthma in the Urban Environment is to understand how
exposures to environmental pollutants and allergens may relate to airway  inflammation and respiratory
morbidity in children with asthma living in the inner city of Baltimore, and to develop effective strategies to
reduce morbidity by changing these exposures.

The immediate goal of these studies is twofold: (1) test currently available recommendations for modifying
environmental pollutant and indoor allergen exposures, and (2) develop data that will allow us to  create new
strategies that may combine other interventions or target them at genetically susceptible hosts. Within  the
Center, the research projects will function in an integrated way, directed toward a single goal and holding
frequent conferences to discuss our progress and to increase our understanding of relevant scientific progress
in each component s field.  At the same time, we will provide this information to the Baltimore community
and involve them in an ongoing dialog to evaluate our progress and plan our intervention efforts. Finally, we
will have involved a large number of families in scientific studies of asthma and will have developed rela-
tionships that will allow trials  of these newly developed interventions to be conducted efficiently. Our ulti-
mate goal  remains to  develop the scientific  understanding that  will allow us to recommend effective
intervention strategies.

To accomplish these goals,  we have created a multidisciplinary program that includes both basic and applied
research programs in combination with a community-based prevention research project.

Genetic Mechanisms of Susceptibility to Inhaled Pollutants.  Steven R. Kleeberger, Ph.D., Principal
Investigator (PI). This research project is  designed to examine the genetic basis for susceptibility to an in-
flammatory response in airways to  reactive oxidant species generated as a result of exposure to ozone (O,).
Using O,-susceptible C57BL/6J (B6) and O,-resistant C3H/HeJ (C3) mouse strains, this project is generating
high-resolution linkage  maps of the regions of mouse chromosomes 17 and 11 carrying O, susceptibility
quantitative trait loci (QTL). Candidate genes in the two QTLs have been identified, and we currently  are
searching for sequence polymorphisms  in two genes, tumor necrosis factor alpha (TNF-a)  and  heat shock
protein 70 (Hsp-70), located in the chromosome 17 QTL. Using BXH recombinant inbred (BXH RI) strains
of mice, a strong susceptibility QTL was identified on chromosome 4, and minor QTLs were identified on
chromosomes 3 and 11. Within the chromosome 4 QTL,  we identified toll-like receptor 4 (Tlr4) as a strong
candidate gene. Significantly greater injury in wild type  C3H/HeOuJ mice compared to the Tlr4 mutant
C3H/HeJ strain, strongly supported a role for  Tlr4 in  responsivity to O,. A manuscript that reports  our
findings has been published  (Am J Respir Cell Mol Biol 2000;22:620-627). In the coming year, we will con-
tinue to pursue our objectives to understand the role(s) of susceptibility genes in pulmonary responses to  O,.

Mechanisms of Particulate-Induced Allergic Asthma. Marsha Wills-Karp, Ph.D., PI. The objective of this
project is to examine the mechanisms by which ambient paniculate matter (PM) may exacerbate  allergic
airways disease,  or play a role in the induction of an asthma-like phenotype in a murine model. Our studies
show that a single exposure to ambient PM (0.5 mg) collected in inner city Baltimore induces sustained
airway  hyperresponsiveness (AHR), and  pulmonary inflammation  (i.e.,  eosinophilia and neutrophilia).
Conversely, these responses are not observed  when animals are exposed to a reference source  of fly ash.
Interestingly, this phenotype is observed in  all murine strains studied to date, although to varying degrees,
suggesting that genetic  susceptibility to  develop allergic airway  responses is  not required. Additionally, in
an ongoing study, our results indicate that repeated exposure to low doses of PM (5 ^g) may have a cumu-
lative effect in the  development of AHR  accompanied by mild,  but persistent eosinophilia. Studies  are
underway to investigate the  role of different cell types such as eosinophils and macrophages in PM-induced
airway responses. One of our most interesting findings is that PM-induced AHR appears to be dependent on
the complement  component (C3). Studies are underway to define  the exact mechanisms by which comple-

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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


merit activation mediates AHR. Taken together, these results indicate that exposure to low levels of ambient
PM alone is sufficient to induce an allergic phenotype and may provide an explanation for the increased
incidence in asthma in inner city environments. Some of these results were published in abstract form at the
1999 and 2000 American Thoracic Society (ATS) meetings, and future findings will be presented at the 2001
ATS meeting. A manuscript on the kinetics of airway responses to ambient PM has been submitted to the
American Journal of Respiratory Cell and Molecular Biology. Additional manuscripts reporting the strain
distribution pattern  of PM responsiveness, the role of eosinophils in PM-induced AHR, and the role of
complement in these responses are in preparation. Dr. Wills-Karp has accepted a position at the University
of Cincinnati, and she hopes to continue as PI of this component. Pending institutional, EPA, and National
Institute of Environmental Health Sciences approval, this Center is enthusiastic about accepting her continued
involvement under  a subcontract as she has been very productive and is willing to continue the goals of the
Center at the University of Cincinnati.

The Relationship of Airborne Pollutants and Allergens to Asthma Morbidity. Gregory Diette M.D.,
M.S., PI. This community-based epidemiologic research will compare ambient and indoor exposures  to a
variety of pollutants and allergens in the home and ambient environment to children s morbidity from asthma.
A case-control study will be conducted among children 6 to 12 years old with 150 prevalent cases of asthma
from several East Baltimore schools and  neighborhood controls. Participants will be evaluated with a ques-
tionnaire regarding demographics, environmental exposures, psychosocial stressors, and health. A home  visit
will be conducted to assess the  home exposure to pollutants and allergens. During a clinic visit, the child will
be  evaluated with allergen skin tests,  spirometry, and serum  cotinine and RAST tests.  Recruitment  and
evaluation procedures for this study are identical to those in the intervention study, and have thus been fully
established at this point. The Principal Investigator for this study, Dr. Jounni Jaakkola, left the University on
March 1, 2000, to take a position at the University of Gothenburg, Sweden. Dr. Eggleston assumed adminis-
trative direction of the study until a new Principal Investigator, Greg Diette M.D., M.S., could be recruited
to head the epidemiologic study. We are  enthusiastic with this choice in that he has already had experience
conducting epidemiologic and intervention research in East Baltimore, has a joint appointment in the Depart-
ments of Medicine and Epidemiology, and is willing to carry the original study into the field.

A Randomized, Controlled Trial of Home Exposure Control in Asthma. Peyton A. Eggleston, M.D., PI.
This community-based prevention research project will conduct a randomized, controlled trial of the effec-
tiveness  of current  intervention methods  to reduce hazardous exposures  and their adverse health effects. It
will test a global intervention that is composed of several components that have proved already to be effective
in middle-class homes. The major outcome of this trial will not be changes in health status, but will be envi-
ronmental exposure, and it will examine the feasibility of modifying inner-city home environments  with
currently recommended procedures.  Data on the  child s health status also will be collected, but  it is anti-
cipated that future studies with interventions that already have been proved effective will be necessary to
provide adequate power to test a health outcome. During Year 2 of the Centers funding, we have established
recruiting, evaluation, and intervention procedures. Recruiting has begun with the presentation of an asthma
self-management curriculum in five schools:  Tench Tilghman, Dr. Bernard Harris, Collington  Square, Dr.
Raynor Brown, and Thomas G. Hayes elementary schools. The curriculum has been presented to 61 children
in 49 famines by the educator and her assistant.  The families were visited a second time and 35 had an
eligible child and were  interested  in participating in the intervention project. Twenty eight (28)  of these
families  have been consented and 24 have been fully evaluated and randomized into the trial. This is an extra-
ordinarily high rate of  recruiting success and is a tribute not  only to the educators but also to the study
interviewer and to the study coordinator. During the coming year, recruitment should be even more rapid as
we expand the asthma curriculum from three to five elementary schools. The treatment group will receive
an  allergen-proof mattress and pillow encasings for the child s bed, a room air cleaner for the child s room,
pest control services if needed, smoking cessation  support, and environmental avoidance education.  The
intervention is being conducted by a community health educator, Mayme Grant, and Karen Callahan, B.S.N.,
M.S.,  and has been well received by nine families. All families will receive two home  visits at  6 and 12
months,  a telephone interview at 3, 6, 9, and 12 months, and a final clinic evaluation.
36                              The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


The project for the newly  recruited young investigator is entitled  Evaluation of Indirect PM l(,  Exposure
Assessment in a Cross-Sectional  Epidemiological Study  of  Asthma Exacerbation in  Urban Children.
Timothy J. Buckley, Ph.D., PI. This project compares the exposure assessment in children as gathered from
home area samples with that measured by diary and continuous personal monitoring over a 3-day period. To
date, 29 children and their homes have been successfully monitored for allergens, PM, nitrogen dioxide, and
ozone. Urinary samples have been collected to assess ETS exposure by cotinine analysis. Questionnaires have
been developed and  administered to acquire information about time activity patterns and housing charac-
teristics. All measures are within the range of previous environmental studies. Results of this study have been
presented at PM 2000: Paniculate Matter and Health:  The Scientific Basis for Regulatory Decision Making
in Charleston, SC. Preliminary results show median PMU) levels of 53, 32, and 22 mg/m1 for the  personal,
indoor, and outdoor measurements, respectively. On average, the personal PMm levels were 24 percent and
134 percent higher than  indoors and outdoors, respectively. Median indoor levels of NO, and O, were 10.2
(n=5) and 5.6 (n=7)  ppb, respectively. Dust allergen concentrations varied by location (bedroom, TV room,
kitchen) from below  detection to 51,000 ng/g of MUP measured in one home s kitchen.

With the completion  of Dr. Buckley s Newly Recruited Young Investigator Project,  we have selected two
young investigators for the position, starting in November 2000. Sekhar Reddy, Ph.D., will conduct a project
entitled Molecular Basis of TLR4 Receptor Regulation by  Endotoxin and Ozone.   Alison Geyh, Ph.D., will
begin a project entitled  Assessing  Early Childhood Exposures to Transition Metals  Associated With Par-
ticulate Matter.

The two community-based applied  research projects will be supported by an experienced Data Management
Core Facility directed by Sukon Kanchanaraksa, Ph.D., and by an Exposure Assessment Core Facility direct-
ed by Patrick Breysse, Ph.D. Dr. Kanchanaraksa has established the study forms on an automated screening
format and has recruited staff who  are contributing to interim data analysis in the center and in several other
asthma-related projects.  Dr. Breysse has established the laboratory procedures to allow three homes to be
evaluated weekly.

The Community Advisory Committee met monthly to review the protocol regarding feasibility and appro-
priateness in the East Baltimore community, review study protocols and progress reports, provide a forum
to discuss community-based issues  concerning the conduct of the trial, and provide  liaison to translate study
findings to the community. Members of the Committee have been chosen from the school system, commu-
nity-based research and service organizations, churches, and from families of asthmatic children. The Com-
mittee has met monthly during the past year and has been invaluable in designing the protocol and the
implementation plan.
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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


Ambient Urban Particulate-Induced Airway
Hyperresponsiveness and Inflammation in Mice
D. Walters, P.N. Breysse, and M.  Wills-Karp
Johns Hopkins University, Baltimore, MD

Airborne paniculate matter (PM) is hypothesized to play a role in increases in asthma prevalence, although
a causal relationship has yet to  be established. To investigate the effects of real-world PM exposure on airway
reactivity and bronchoalveolar lavage (BAL) cellularity, we exposed naive A/J mice 6 7 weeks of age to a
single dose (0.5 mg/mouse) of PM collected in urban Baltimore (AUB) or to coal fly ash. We found that AUB
exposure induced increases in airway responsiveness (AHR) and BAL cellularity, whereas coal fly ash (CFA)
exposure did not elicit significant changes in either of these parameters. We further examined AUB-induced
temporal changes in AHR, BAL cells, and lung cytokine levels over a 2-week period. AUB-induced AHR
was sustained over 7 days  and returned to control levels at 14 days. The increase in AHR was preceded by
dramatic increases in  BAL  granulocytes, particularly eosinophils. The decline in AHR from peak values was
associated with significant  increases  in macrophages. A Th2 cytokine pattern  (IL-5,  IL-13, eotaxin) was
observed early on with a shift toward a Thl pattern  (IFN-g), as AHR and granulocytes returned to normal
levels. Finally, in a leaching experiment, we found that the  active component(s) of AUB are not water-
soluble, but remain particle-bound. We conclude that ambient PM can induce asthma-like parameters in naive
mice suggesting that PM exposure may contribute to increases in asthma prevalence.
38                             The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children's Environmental Health and Disease Prevention Research Progress Review Workshop


Air Pollution and Allergen Exposure
Among Asthmatic Children in Inner-City Baltimore
T.J. Buckley, P.N. Breysse, C. Beck,  A. Escamillia, and P.A. Eggleston
The Johns Hopkins University School of Hygiene and Public Health, Baltimore, MD

The number of asthmatics has more than doubled from 1980 to 1994. It is estimated that 7 percent of children
under 18 years old suffer from asthma. Populations in the inner-city environment are most severely affected.
Air pollution and allergens in the indoor environment are believed to play a prominent etiologic role, although
few studies have been conducted characterizing exposure in this susceptible population. The goal of this study
is to demonstrate the feasibility of assessing allergen and air pollution exposure among inner-city  asthmatic
children and to evaluate the effectiveness of a strategy based on time-weighted indoor and outdoor monitoring
to estimate actual exposure. We also examine the contribution of air pollution of ambient origin on indoor
and personal exposures.

Methods: Monitoring was conducted from April 1999 through August 2000, on a convenient sample of 29
children with reported physician-diagnosed asthma. All were Baltimore City residents and ranged in age from
6 to 12 years. PMI(1 and/or PM^ _, were collected indoors, outdoors, and on the child (personal) using MSP
impactors (St. Paul, MN) over a 3-day sampling period. Indoor and  personal ozone and NO2 were sampled
passively using Palmes tubes. A single urine sample was collected on the third day of monitoring for analysis
of cotinine as an indicator of exposure to environmental tobacco smoke. Indoor allergens, including dust mite
Der p J, cockroach Bla g I, cat Pel d I, and mouse urine protein MUP, were sampled in the settled dust by
vacuum in three indoor home locations (bedroom, kitchen, and child s bedroom) and analyzed by immuno-
chemical assay. Each child was asked to maintain a 24-hour time activity diary for each of the  3 days of
sampling, a home inspection was conducted, and a household questionnaire was administered.

Results:  Preliminary results show median PMln levels of 56, 39, and 22 g/m ' for the personal, indoor, and
outdoor measurements. On average, the personal PMln levels were 24 percent and 134 percent higher than
indoors  and outdoors, respectively. As has  been  observed in the general population, children s personal
exposures were poorly predicted from the outdoor central site monitoring (R2=0.01). On average, the children
reported that they spent 86 percent of their time indoors (home, school, and other). Median indoor levels of
NO2 and O, were 14 and 5.6 ppb, respectively. Dust allergen concentrations varied by location (bedroom, TV
room, kitchen) from below detection to 51,000 ng/g of MUP measured in one home s kitchen.

Conclusions:  PM concentrations and their relationships between personal, indoor, and outdoor locations
have been measured for a population subgroup believed to be susceptible to PM morbidity and mortality.
These concentrations and relationships are within the  range  of what has been observed in previous studies
of the general population. Allergens in house dust have been measured at concentrations shown to result in
sensitization and exacerbation of disease. Therefore, the current  study establishes  procedures  and provides
preliminary data by which to assess exposure to test for synergistic effects of air pollution and common in-
door allergens in a susceptible urban population.
The Office of Research and Development s National Center for Environmental Research                             39

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University of Iowa
  Iowa City, IA

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Overview and Description of the
Children s Environmental Airway Disease Center
Gary W. Hunninghake
Universit\ of Iowa, Iowa Cit\, IA

The theme of the Center is to investigate the etiology and pathogenesis of airway disease in children from rural
communities. We chose this theme for the Center for the following reasons: (1) asthma is the most common
chronic illness in children; (2) the rural setting introduces unique environmental exposures that are known to play
a role in the development of airway disease; (3) environmental models of asthma provide an ideal opportunity
to investigate fundamental issues in childhood asthma such as the biological origin and persistence of airway
disease; and (4) this theme builds  on existing scientific expertise and ensures a highly  interactive program.
Because grain dust and endotoxin  are common in the rural setting and both are associated with acute and
chronic forms of airway disease, we have used these  very relevant environmental exposures to further focus
the projects in the Center. The end result is a highly integrated and focused program that has the potential to
make a number of novel, related observations. In aggregate, the coupled scientific findings from the Center will
substantially enhance our understanding of airway  disease in children. The primary hypothesis unifying this
research program  is that understanding the etiology and pathogenesis of airway disease in children from rural
communities will provide the scientific rationale to develop primary, secondary, and tertiary preventive programs
that reduce the morbidity and mortality of childhood asthma in the rural setting.

The project-specific hypotheses within the Center are:

>-   Project 1: Environmental intervention is an essential component of an asthma intervention program that
     must be coordinated with other improvements in  health care to reduce the prevalence and severity of
     asthma among children from rural communities.

>   Project 2: Many of the biologic features of acute and reversible airway inflammation are fundamental to
     the development of chronic grain dust-induced airway disease.

>-   Project 3: Mechanisms that initiate, promote, and resolve grain dust-induced inflammation may be distinct
     from those mechanisms involved in LPS-induced airway inflammation.

>   Project 4: Respiratory syncytial virus infection upregulates the response of airway  epithelium to LPS.

During Year 2 of the Children s Environmental Airway Disease Center at the University of Iowa, substantial
progress has taken place. The four funded projects  at the Center have made excellent progress and have met
all of their goals for the first year. In this  regard, a number of publications have either been  submitted, accepted,
or published. The  investigators also have received a substantial number of invitations to present their results at
national and international meetings. Finally, the overall  structure and necessary interactions within  the Center
have been formalized to ensure the continued  success  of the program.  Further evidence of the health of the
program is that the  Center has provided some financial support for exciting new programs that were not
described  in the original application but that will  interface closely with existing projects and provide new
directions  for research. In Year 1, we  were able to provide support for Dr. Klekamp, a promising Assistant
Professor in Pediatrics, to pursue studies on the role  of adhesion molecules in airway inflammation. She has
found that ICAM-1 and the B integrins  are essential to neutrophil recruitment. In Year 2, we provided support
for Dr. Carter, who is a promising new investigator in the Department of Medicine. He has found that endotoxin
increases the replication of viruses in airway epithelia. Drs. Hunninghake, Nauseef, and Schwartz are evaluating
the role of toll-4 in lipopolysaccharide signaling.
The Office of Research and Development s National Center for Environmental Research                    43

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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Thus, the Children s Environmental Airway Disease Center at Iowa strengthens our research efforts related to
environmental airway disease. The program is energetic, imaginative, cohesive, and productive. In the following
paragraphs, the highlights of the accomplishments of various components of the program are described. More
detailed progress reports can be found in the individual projects.

Project 1:  Multicomponent Interventions Study of Asthma in Children From Rural Communities: The
           Rural Health Study.

The most significant achievement during the past year has been the development of a specific protocol for the
multicomponent intervention and for the assessment of its effect. To date, no protocol is available for a
community-based intervention encompassing both environmental and medical issues related to asthma care in
a rural environment, particularly with a focus on individualized family counseling related to barriers for asthma
care. The most important aspect of the study is that  we have a 90 percent response rate.

Project 2:  A Model To Study the Development of Persistent Environmental Airway Disease.

These investigators have found that subacute exposure to grain dust caused chronic airway lesions that are
associated with airway hyperreactivity and airway remodeling. Moreover, the development of chronic grain
dust-induced airway disease appears to be mediated by endotoxin, because mice genetically hyporesponsive to
endotoxin do not develop chronic grain dust-induced airway disease.

Project 3:  Mechanisms That Initiate, Promote, and Resolve Grain Dust-Induced Inflammation.

Studies related to the pathophysiology of asthma commonly involve either in vivo human and animal systems
or in  vitro systems  with single cell types. Although both of these approaches provide crucially important
insights, it is often difficult to bridge the knowledge that the two systems provide. We are developing co-culture
systems that will allow  us to explore the complex interactions between airway cells that are the basis for the
inflammatory response to grain dust. We have  found that the response to grain dust is very cell specific.

Project 4:  Role of RSV Infection and Endotoxin in Airway Inflammation.

RSV upregulates the p45 ERK kinase and then  is related to IL-8  production by airway epithelium. These
investigators also observed that TH-1 and Th-2 cytokines regulate IL-8 release by airway epithelium. Impor-
tantly, we found that environmental exposures  may increase replication of viruses in the airways.
44                     The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
A Multicomponent Intervention Study of
Asthma in Children From Rural Communities
E. Chrischilles, J. Merchant, A. Kuehl, R. Ahrens, S. Reynolds, L. Burmeister, P. Pomrehn, and P. Thome
University of Iowa, Iowa dry, IA

The complex interrelationship of allergens and nonspecific airway stimuli may amplify the effects of individual
airway stimuli as well as weaken single-agent exposure  reduction interventions. Factors such as physician
awareness of patient symptom control, patient self-management, early response to upper respiratory infections,
parent modification of smoking, and modification  of cleaning behaviors may all influence a child s  asthma-
related quality-of-life. A multicomponent intervention approach appears warranted. This community-based
intervention study will take place in two counties in Iowa. The goal is to test the effect of the multicomponent
intervention by  comparing asthma  health outcomes and change in  environmental exposures between  an
intervention county and a demographically similar, noncontiguous comparison  county.  A target total of 300
children, ages 6 14 years, and their families will be enrolled in three, 1-year waves. Children will be identified
through the schools and mailed a 3-page asthma screening questionnaire. The screening  questionnaire will  be
used to classify children as having no asthma, intermittent asthma, or persistent  asthma. Severity of persistent
asthma also will  be assessed via the screener. Measures include annual spirometry and the following measures
at baseline and 1 year:  in-home administered questionnaires; home inspection environmental  checklist; and
environmental measures. Questionnaire-based measures include asthma symptom control and severity, quality-
of-life, barriers to asthma care, self-management behaviors, adequacy and understanding of asthma care plans,
environmental control issues, and health care utilization. Environmental measurements will include quantitative
assessments of endotoxin and antigen levels via dust sampling; CO, CO2, relative humidity, and temperature
via instantaneous air sampling; and NO, and environmental tobacco  smoke via  passive dosimeter. The
intervention includes family-level, health provider-level, school-level, and community-level components. The
individualized family-level intervention addresses medication management and environmental control issues and
is delivered  by the study asthma counselor in quarterly in-person sessions and monthly interval telephone
followups. Feedback is provided from the counselor to the family s regular asthma physician.
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop



Lipopolysaccharide Responsiveness and the

Development of Subchronic Grain Dust-Induced Airway Injury
C.L.S. George, H. Jin, C.L. Wohlford-Lenane, J.N. Kline, and D.A. Schwartz
Duke University, Durham, NC

Endotoxin is one of the principal components of grain dust that causes acute reversible airflow obstruction
and airway inflammation. To determine if endotoxin responsiveness influences the development of chronic
grain dust-induced airway disease,  physiologic and airway  inflammation/remodeling parameters were
evaluated following an 8-week exposure to corn dust extract (CDE) and again following a 4-week recovery
period, in one strain of mice sensitive to (C3H/HeBFeJ) and one resistant to endotoxin (C3H/HeJ). Following
the CDE exposure, both  strains of mice had equal airway hyperreactivity to a methacholine challenge;
however, airway hyperreactivity persisted only in the C3H/HeBFeJ mice after the recovery period. Only the
C3H/ HeBFeJ mice showed significant inflammation of the lower airway following the 8-week exposure to
CDE. Following the recovery period, this inflammatory response completely resolved. Lung  stereologic
measurements indicate that an 8-week  exposure to CDE results in persistent  expansion of  the airway
submucosal cross-sectional area only in the C3H/HeBFeJ mice. Collagen type III and an influx of cells into
the subepithelial  area participate in the expansion of the  submucosa.  Our findings demonstrate that
subchronic inhalation of grain dust extract results in the development of chronic airway disease only in mice
sensitive to endotoxin, but not in mice that are genetically hyporesponsive to endotoxin, suggesting that
endotoxin  is important in the development of chronic airway disease.
46                     The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s EnvironmenUiLHealth and Disease Prevention Research Progress Review Workshop
IL-10 Reduces Grain Dust-Induced
Airway Inflammation and Airway Hyperreactivity
T.J. Quinn, S. Taylor, C.L. Wohlford-Lenane, and D.A. Schwartz
University of Iowa, Iowa City, IA

To determine if interleukin-10 (IL-10) could alter the development of grain dust-induced airway disease, we
pretreated mice with either saline or IL-10 intravenously, exposed the mice to an inhalation challenge with corn
dust extract (CDE), and measured inflammation and the development of airway hyperreactivity. Pre-treatment
with IL-10, in comparison to saline, reduced the concentration and percentage of polymorphonuclear cells in
the lavage fluid 30 minutes after the inhalation challenge with CDE (p < 0. 05). In comparison to saline-treated
mice, IL-10 did not significantly alter the degree of airway hyperreactivity 30 minutes after the exposure to
CDE. IL-10-treated mice lavaged 18  hours after challenge with CDE also exhibited a lower percentage of
polymorphonuclear cells in the lavage fluid (p < 0.05) and had significantly less airway hyperreactivity than did
mice pretreated with the saline placebo (p < 0.05). These findings indicate that exogenous IL-10 is effective in
reducing airway inflammation and airway hyperreactivity due to the inhalation of CDE.

Publications:

Quinn TJ, Taylor S, Wohlford-Lenane CL, and Schwartz DA. IL-10 reduces grain dust-induced airway in-
flammation and airway hyperreactivity. J Appl Physiol 2000;88:173-179.
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
TLR4 Mutation Is Associated With
Endotoxin Hvporesponsiveness in Humans
N.C. Arbour, E. Lorenz, B.C. Schutte, J. Zabner, J.N. Kline, M. Jones, K. Frees, J.L. Watt, and DA. Schwartz
University of Iowa, Iowa City, IA

There is much variability between individuals in the response to inhaled toxins, but it is not known why certain
people develop disease when challenged with environmental agents and others remain healthy. To address this,
we  investigated whether encoding the toll-like  receptor-4 (TLR4), which  has been  shown  to  affect
lipopolysaccharide (LPS) responsiveness in mice, underlies the variability in airway  responsiveness to inhaled
LPS in humans. In this study, we show that common, cosegregating missense mutations (Asp299Gly and
Thr399Ile) affecting the extracellular domain of the TLR4 receptor are associated with a blunted response to
inhaled LPS in humans. Transfection  of THP-1 cells demonstrates that the Asp299Gly mutation (but not the
Thr399Ile mutation) interrupts TLR4-mediated LPS signalling. Moreover, the wild-type allele of TLR4 rescues
the LPS hyporesponsive phenotype in  either primary airway epithelial cells or alveolar macrophages obtained
from individuals with the TLR4 mutations. Our findings provide the first  genetic evidence that common
mutations in TLR4 are associated with differences in LPS responsiveness in humans, and demonstrate that
gene-sequence changes can alter the ability of the host to respond to environmental stress.

Publications:

Arbour NC, Lorenz E, Schutte BC, Zabner J, Kline JN, Jones M, Frees K, Watt JL, and Schwartz DA. TLR4
mutation is associated with endotoxin  hyporesponsiveness in humans. Nat Genet 2000;25:187-191.
48                    The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Bronchial Hyperreactivity Is Associated With
Enhanced Grain Dust-Induced Airflow Obstruction
J.N. Kline, P.J. Jagielo, J.L. Watt, and D.A. Schwartz
University of Iowa, Iowa City, IA

Bronchial hyperreactivity (BHR) is associated with the presence of airway inflammation in asthma and is seen
in individuals occupationally exposed to grain dust. To better understand the relationship between BHR and
pulmonary inflammation after grain dust exposure, we conducted an inhalation challenge to corn dust extract
(CDE) on seven subjects with BHR, [a 20% or greater decrease in forced expiratory volume in  1 second
(FEV( 1)) compared with diluent FEV( 1) with a cumulative dose of histamine < 47.3 breath units] and compared
their physiological and inflammatory responses with those of seven matched control subjects. BHR subjects
were exposed to nebulized CDE (target dose of 0.16 m[0,g/kgendotoxin) as tolerated; matched controls received
equal amounts of CDE. Subjects with BHR complained of chest tightness  and dyspnea  within 2 hours  after
inhalation of CDE significantly more frequently than controls. Similarly, subjects  with  BHR developed
significantly greater percent declines in FEV( 1) at time points up to 4 hours after exposure to CDE. Significant
increases in total cells, neutrophils, tumor necrosis factor-alpha, interleukin-6, and interleukin-8 were detected
in bronchoalveolar lavage fluid 4 hours after inhalation  of CDE in all subjects, but no differences were detected
between the control and BHR groups. These results suggest that, although subjects with BHR develop a more
precipitous decline in FEV(l) after exposure to CDE,  the inflammatory response to CDE is similar in subjects
with and  without BHR.

Publications:

Kline JN, Jagielo PJ, Watt JL, and Schwartz DA. Bronchial hyperreactivity is associated with enhanced grain
dust-induced airflow obstruction. J Appl Physiol 2000;89:1172-1178.
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
TNF-Alpha and EL-lBeta Are Not Essential to the
Inflammatory Response in LPS-Induced Airway Disease
J.G. Moreland, R.M. Fuhrman, C.L. Wohlford-Lenane, T.J. Quinn, E. Benda, J.A. Pruessner, and D.A. Schwartz
University of Iowa, Iowa City, IA

To determine the role of TNF-OC and IL-1P in the lower respiratory tract inflammatory response following
inhalation of LPS, we conducted inhalation exposure studies in mice lacking expression of TNF-CC and/or IL-1
receptor  1; and in mice with functional blockade of these cytokines using adenoviral vector delivery of soluble
receptors to one  or both cytokines. Alterations in airway physiology were assessed by pulmonary function
testing, prior to and immediately following 4-hour LPS exposure, and the cellular inflammatory response was
measured by whole lung lavage and assessment of inflammatory cytokine protein and mRNA expression.
Airway resistance following LPS exposure was similarly increased in all groups of mice without evidence that
blockade of either  or both cytokines was protective from  this response. Additionally, all groups of mice
demonstrated significant increases in lung lavage cellularity with a complete shift in the population of cells to
a predominantly neutrophilic infiltrate as well as elevation of inflammatory cytokine protein and mRNA levels.
There were no significant differences between the groups in measures of lung inflammation.  These results
indicate that TNF-a and IL-1P  do  not  appear to have an essential  role in mediating the  physiologic  or
inflammatory response to inhaled LPS.

Publications:

Moreland JG, Fuhrman RM, Wohlford-Lenane CL, Quinn TJ, Benda E, Pruessner JA, and  Schwartz DA.
TNF-Alpha and IL-1 Beta are not essential to the inflammatory response in LPS-induced airway disease. Am
J Physiol Lung Cell Mol Biol 2001 ;280( 1 ):L173-180.
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  2001 EPA/N1EHS/CDC Centers for Children s Environmenta.tHealth and Disease Prevention Research Progress Review Workshop
Endotoxin Responsiveness and
Subchronic Grain Dust-Induced Airway Disease
C.L.S. George, H. Jin, C.L. Wohlford-Lenane, M.E. O Neill, J.C. Phipps, P. O Shaughnessy, J.N. Kline,
P.S. Thome, andD.A. Schwartz
University of Iowa, Iowa City, IA

Subchronic exposure to corn dust extract (CDE) in a murine model causes airway "inflammation  and
bronchial hyperreactivity similar to that seen in humans with recurrent occupational exposure to grain dust.
Lipo-polysaccharide (LPS) is one of the principle components of grain dust, which causes acute reversible
airflow obstruction and  airway inflammation.  To  determine  if LPS responsiveness  influences  the
development of chronic grain dust-induced airway disease, physiologic parameters and indicators of airway
inflammation/ remodeling were evaluated following an 8-week exposure to CDE in a strain of mice sensitive
to LPS (C3H/BFeJ) and one resistant to LPS (C3H/HeJ). Airway hyperreactivity, measured by response of
enhanced pause pressure (Penh) to methacholine, was assessed at baseline, after 8 weeks of CDE exposure,
and after a 4-week recovery period. Following the 8-week CDE exposure, the C3H/BFeJ  mice  had a
significantly greater increase in their Penh values compared to the C3H/HeJ mice. This increase persisted
through the 4-week recovery period. The C3H/BFeJ mice, but not the C3H/HeJ mice, showed a significant
increase in the number of total cells, the percentage of neutrophils, and the concentration of TNF-oc, IL-6,
and MIP-2 in the lavage fluid after the 8-week exposure to CDE; however, after the recovery period, the
concentration of cells and cytokines in the lavage fluid returned to preexposure values in both strains of mice.
Morphometric measures indicate that  an  8-week exposure to CDE results in expansion of the airway
submucosa with minimal change in the airway epithelia in the C3H/BFeJ mice, but not the C3H/HeJ mice.
Our results demonstrate that Subchronic inhalation of grain dust extract results in the development of chronic
airway disease in  mice sensitive to  endotoxin but not in mice that are genetically hyporesponsive to
endotoxin; suggesting  that endotoxin is  one of the principle components of grain  dust causing the
development of chronic airway disease.

Publications:

George CLS, Jin H, Wohlford-Lenane CL, O Neill ME, Phipps JC, O Shaughnessy P, Kline JN, Thorne PS,
and Schwartz DA. Endotoxin responsiveness and subchronic grain dust-induced airway disease. Am JPhysiol
Lung Cell Mol Biol 2001 ;280(2):L203-213.
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Mechanisms That Initiate, Promote, and
Resolve Grain Dust-Induced Inflammation
Kevin Leidal, William Nauseef, and Gerene Denning
VA Medical Center and University of Iowa,  Iowa City, IA

The hallmark of the bronchial hyperreactivity observed in children with asthma is airway inflammation. In
contrast to eosinophil-mediated asthma, responses stimulated by grain dust are predominately neutrophilic,
suggesting that grain  dust elicits pro-inflammatory signals largely geared toward stimulating  infiltration of
neutrophils. Our studies use primary human airway cells and cell lines to identify early steps in this neutrophilic
inflammatory response.

Hypothesis: Exposure of alveolar macrophages to grain dust results in the release of macrophage-specific
cytokines. We found that:  (a) grain dust stimulates an increase in the release of the cytokines TNF-CC, IL-8,
ENA-78, and Rantes in a dose-dependent manner; (b) the effect is maximal at ~ 0.4-2 mg/mL of grain dust (2-
14 |J,g/mL of associated LPS); and (c) conversely, higher concentrations of grain dust (10 mg/mL) inhibit release
of all  four cytokines relative to maximal levels. These data support the hypothesis that factors present in grain
dust stimulate cytokine release by human alveolar macrophages.

Hypothesis: LPS contributes to the  effects of grain dust on release of cytokines by human alveolar macro-
phages. In our studies, although both purified LPS and grain dust increase release of TNF-a by human alveolar
macrophages, we have observed striking differences in the characteristics  of these effects. First, the dose
response curves for LPS and grain dust (with comparable LPS concentrations) are different. Namely, at low
concentrations LPS is the more potent agonist,  while at higher concentrations grain dust elicits  a greater
response. Next, the effects of LPS but not of grain dust are enhanced by the addition of LPS-bindingprotein
(LBP). Also, the effects of LPS, but less so of grain dust, are inhibited by the endotoxin inhibitors polymyxin
B and bacterial permeability-increasing protein (BPI). Finally, we observed no correlation between TNF-a
release and  LPS concentrations using multiple grain dust preparations. These data lead to two alternative
hypotheses:  (1) the effects of grain  dust on TNF-a  release  are independent of LPS; and (2) LPS bound to
endotoxin has different biological activity and interacts differently with host proteins and surface receptors when
compared to purified LPS.

Hypothesis: Grain dust activates human neutrophils  and this activation is due, at least in part, to LPS. Pre-
liminary studies measuring chemiluminescence demonstrate differences in the dose response curves between
LPS and grain dust and differences in the effect of LBP. These data further suggest the two  alternative hy-
potheses stated above. Additional studies  currently are underway  to characterize further the inflammatory
effects of grain dust using human airway cells. A thorough understanding of these early inflammatory events
will provide essential insight for developing  interventions to prevent and/or treat grain dust-mediated airway
disease.
52                     The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Activation of ERK Kinase Activity by Respiratory Syncytial
Virus in A549 Cells Is Linked to the Production of Interleukin 8
W. Chen, M.M. Monick, A.B. Carter, and G.W. Hunninghake
University of Iowa, Iowa City, 1A

Respiratory syncytial virus is a major cause of viral bronchiolitis and  pneumonia in children. The airway
inflammation that results from the viral infection is associated with a marked increase in interleukin 8 and
neutrophils in the infected sites of the lung. In this study, the relationship between release  of interleukin 8,
infection of A549 cells (a human lung epithelial carcinoma cell line) by respiratory syncytial virus, and activation
of mitogen-activated protein kinases was investigated.  Infection of A549 cells by the virus caused an increase
in the activity of ERK2 by about tenfold compared with the noninfected cells. The increase  in the activity of
ERK2 during the viral infection was an immediate event and occurred prior to the viral replication process.
PD98059, a synthetic chemical inhibitor that blocks the activation of MEK1, inhibited the increase in the activity
of ERK2 by infection of respiratory syncytial virus by about 50 percent at 10 mM. Pre-treatment of A549 cells
with PD98059 before the  viral infection also inhibited the increase in the production of interleukin 8 by 50
percent. This treatment had little effect on the virus-induced increase in interleukin 8 mRNA. The viral infection
had no effect on the activities of p38 and INK. These observations suggest that activation  of ERK2 by
respiratory syncytial virus infection may be one of the mechanisms that result in the increase of the production
of interleukin 8.

Publications:

Chen W, Monick MM, Carter AB, and Hunninghake GW. Activation of ERK2 by respiratory syncytial virus
in A549 cells is linked to the production of interleukin 8. Exp Lung Res 2000;26(1): 13-26.
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Effects of Ragweed and Th-2 Cytokines on the
Secretion of IL-8 by Human Airway Epithelial Cells
W. Chen and G. W. Hunninghake
Universit\ of Iowa, Iowa Cit\, 1A

Ragweed-induced asthma is a very common pulmonary disease. An important part of asthma is a large increase
in inflammatory cells, including neutrophils and eosinophils, in ragweed-sensitized subjects.  In this study, we
determined  whether ragweed treatment could induce the release of interleukin-8 (IL-8) by human airway
epithelial cells. We found that ragweed induces a substantial increase of the secretion of IL-8 from A549 cells
in a dose- and time-dependent manner. Th-2 cy tokines, IL-4, and IL-13 partially inhibited the ragweed-induced
secretion of IL-8. Our results suggest that airway epithelial cells may be one of the cell sources that provide IL-8
during ragweed-induced asthma. The results also indicate that IL-4 and IL-13 may exert inhibitory effects on
IL-8 secretion by airway epithelium.

Publications:

Chen W and Hunninghake GW. Effects of ragweed and Th-2 cytokines on the secretion of IL-8 by human
airway epithelial cells. Exp Lung Res 2000;26:229-239.
54                    The Office of Research and Development s National Center for Environmental Research

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Respiratory Syncytial Virus Infection Results in Activation of Multiple
Protein Kinase C Isoforms Leading to Activation of Mitogen-Activated Kinase
MM. Monick, J.M. Staber, and G. W. Hunninghake
University of Iowa, Iowa City, IA

Respiratory syncytial virus (RSV) is an important respiratory pathogen that preferentially infects epithelial cells
in the airway and causes a local inflammatory response. Very little is known about the second messenger
pathways involved in this response. To characterize some of the acute response pathways involved in RSV
infection, we utilized cultured human epithelial cells (A549) and optimal tissue culture infective (TCID)50 doses
of RSV.  We have previously shown that RSV-induced IL-8 release is linked to activation of the ERK MAP
kinase pathway. In this study, we evaluated the upstream events involved in ERK activation by RSV. RSV
activated ERK at two time points, an early  time point consistent  with viral binding and a later, sustained
activation consistent with viral replication. We next evaluated the role of PKC isoforms in RSV-induced ERK
kinase activity.  We found  that A549 cells contain  the Ca++ dependent isoforms a and bl, and the Ca++
independent isoforms d, e, h, m, q, and z. Western analysis showed that RSV caused no change in the amounts
of these isoforms. However, kinase activity assays demonstrated activation of z at the early time point and
sustained activation of b 1, d, e, and m later in the infection. A cell-permeable peptide inhibitor specific for the
z isoform decreased early ERK kinase activation by RSV. Downregulation of the other PKC isoforms with
PMA blocked the late sustained activation of ERK by RSV. These studies suggest that RSV activates multiple
PKC isoforms, resulting in  the downstream activation of ERK kinase.

Publications:

Monick MM, Staber JM, Thomas K, and Hunninghake GW. Respiratory syncytial virus infection results in
activation of multiple protein kinase C isoforms leading to activation of mitogen-activated protein  kinase.  J
Immunol 2001 ;166(4):2681-2687.
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Endotoxin Augments Viral Replication and the
Inflammatory Response in Respiratory Syncytial Virus-Infected Epithelium
A.B. Carter, M.R. Donahue, K.L. Knudtson, G. Gudmundsson, MM. Monick, and G. W. Hunninghake
University of Iowa, Iowa City, IA

Respiratory syncytial virus (RSV) is an important cause of lower respiratory tract illness in young children, and
RSV infection often results in increased airway reactivity. Airway reactivity in asthma also is increased following
exposure to endotoxin (LPS), and viral respiratory tract infections are increased in  children exposed to indoor
and outdoor pollutants. Due to the fact that many environmental agents contain LPS, we inquired if LPS
enhanced viral replication. We found that LPS significantly increased the amount of virus in infected cells when
compared to RSV-infected cells alone. To determine if this increase in viral replication was associated with more
inflammation, we used IL-8 gene expression as a marker of epithelial inflammation. As expected, we found that
RSV alone increased IL-8 gene expression in epithelium,  while LPS alone had no significant effect. We also
found that RSV-infected epithelium exposed to LPS had about a twofold increase in IL-8 mRNA accumulation
and IL-8 protein release. These data show  that LPS increases viral replication and inflammation in  infected
epithelium.

Publications:

Carter AB, Donohue MR, Knudtson KL, Gudmundsson G, Monick MM, and Hunninghake GW. Endotoxin
augments viral replication and the inflammatory response in respiratory  syncytial virus-infected epithelium.  J
Clin Invest 2000 (submitted).
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University of Michigan
    Ann Arbor, Ml

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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Overview and Description of the Michigan
Center for the Environment and Children s Health
Barbara Israel
University of Michigan, Ann Arbor, MI

The Michigan Center for the Environment and Children s Health (MCECH pronounced M-Check ) is one
of eight  Centers  of Excellence for  Children s Environmental Health  funded by the National Institute of
Environmental Health Sciences (NIEHS) and the U.S. Environmental Protection Agency (EPA) for 5 years,
beginning in the fall of 1998. The overall goal of MCECH is to investigate the environmental, pathophysio-
logical and clinical mechanisms of childhood asthma, which will translate into risk assessment and com-
prehensive community and household level interventions aimed at increasing knowledge and behaviors to reduce
asthma-related environmental threats to individuals and neighborhoods.

MCECH grew out of the priorities set by the Detroit Community-Academic Urban Research Center (URC),
funded by the Centers for Disease Control and  Prevention (CDC). Since  1995, the URC has conducted
community-based research to identify ways to improve health in selected communities in the southwest and east
sides  of Detroit. Representatives of community-based organizations, public  health agencies,  health care
organizations and academia are  involved in every  major phase  of the research from defining the health
problems to  collecting data and disseminating results, to evaluating plans of action to address issues identified.
One such plan of action  was the creation of MCECH.

Based at the  University of Michigan Schools of Public Health and Medicine, MCECH involves partners working
in the southwest and east sides of Detroit, including Latino Family Services, Community Health and Social
Services (CHASS), Warren-Conner Development Coalition, Butzel Family Center, Detroiters Working for
Environmental Justice, Friends of Parkside, United Community Housing Coalition, Kettering-Butzel Health
Initiative, Henry Ford  Health System, the Detroit Health Department, and the CDC.

MCECH conducts its research in ways that are consistent with the Community-Based Public Health Research
Principles developed initially by the  Detroit-Genesee County Community-Based Public  Health  Consortium,
and subsequently modified for and adopted by the Detroit URC. These principles include an emphasis on the
local relevance of public  health problems and an examination of the social, economic, and cultural conditions
that influence health  status and  the ways in which these  affect lifestyle, behavior,  and community
decisionmaking. Thus, MCECH s projects involve a collaborative, empowering process  that builds upon the
local knowledge of community members,  relies on community resources, and contributes  to community
capacity-building and community control of efforts aimed at understanding and changing the environment to
promote and improve community and family health.

There are three projects of MCECH two of which are being implemented  in the southwest and east sides of
Detroit, and  one of which will be based at the University of Michigan School of Medicine. The three projects
are engaged  in coordinated interdisciplinary research aimed at:

>-  Increasing knowledge and behavior to reduce environmental hazards in households and neighborhoods,
    thereby improving asthma-related health status,  through a community-based household and neighborhood
    level intervention;

>  Examining the effects of daily and seasonal fluctuations in  indoor and outdoor ambient air quality on
    pulmonary function and severity of asthma symptoms; and

>  Determining the effects of allergen-induced local, excessive production of chemokines on redox status and
    innervation of the bronchial tree.
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop

Current MCECH projects are:

Community Action Against Asthma (CAAA): This project consists of two closely integrated components:
(1) a community-based intervention to reduce environmental triggers for asthma among children, and (2) a study
of the role of indoor and outdoor air contaminant exposures in childhood asthma aggravation. A primary aim
of the household and neighborhood level community-based intervention research project is to reduce exposure
of children to environmental contaminants within their homes and neighborhoods that trigger asthma, thereby
improving asthma-related health status and reducing asthma-related medical care utilization. Children ages 6-10
years with moderate to severe asthma will be identified through a screening questionnaire mailed to families.
Families will be asked to enroll in a household intervention in which outreach workers will visit each household
12 times in 2 years. Outreach workers will work with the family to reduce indoor household exposure factors
identified as exacerbating asthma such as cockroach mites, cat dander, environmental tobacco smoke, and mold.
Each household will be supplied with education materials and other resources to reduce indoor asthma triggers
such as vacuum cleaners, bedding covers, cleaning kits, and mats. Community organizers also will work with
neighborhood  groups on asthma  awareness and  reduction of environmental threats to children s respiratory
health.

The key hypothesis of the exposure study is that ambient air contaminants such as particulate matter and ozone
will worsen the health status of asthmatic children by increasing the adverse effects of common indoor air
contaminants,  especially indoor allergens. The same families enrolled in the  intervention also will participate in
this component. Levels of outdoor air contaminants as well as indoor air contaminants in schools and homes
will be measured. Indoor measures will include common allergens in dust. The health status of the children with
asthma will be assessed through the  use of daily  symptom  diaries and portable devices to measure breathing
function.

Chemokines in the Pathogenesis of Asthma:  Recent studies have identified that many asthmatic attacks,
particularly for inner-city children, are triggered by  exposure to cockroaches. It is not exposure to the entire
cockroach, but only small fragments or residue, called allergens that are responsible for the allergic response.
In response to such allergens, the body releases chemicals or mediators, which cause inflammation and thus an
asthmatic episode. Targeted therapy has the potential to improve the treatment of asthma by identifying those
mediators directly responsible for asthma. This research  project will test the hypothesis that asthma-like
pulmonary injury is mediated by the local production of mediators, which are called chemokines. Chemokines
are  small molecular weight proteins, which induce the movement and recruitment of inflammatory cells. The
chemokines are powerful  mediators with long-lasting and potent biological activities. The first specific aim of
this project is to determine the acute and chronic pulmonary inflammation that develops after direct injection
of the chemokines into the  lung.  The second aim is to develop a mouse model of asthma-like pulmonary
inflammation in response to cockroach  allergens. The third aim is to investigate the signals responsible for
inducing the cells to make the chemokines. Finally, the  last specific aim is to rigorously test the central
hypothesis that chemokines are important in causing asthma.

The three projects of the MCECH will  provide the following benefits to the Detroit communities involved as
well as  to the asthma research community:

>-   Identification of previously undiagnosed asthmatic children
>-   Provision of household materials, such as vacuum cleaners and clean bedding, aimed at reducing asthma
     triggers
>   Education on potential asthma triggers and methods for reducing those triggers
>   Assistance to families for negotiation with landlords regardingenvironmental factors in the home associated
     with asthma
>   Referrals to families with asthma for available  and affordable medical care
>*   The collection of detailed multiple daily measures of  ambient and indoor air contaminants
>   Identification of the underlying mechanisms of asthma and potential targets for further intervention.
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Use of a Screening Questionnaire To Estimate Prevalence of
Diagnosed and Undiagnosed Asthma Among Minority Children in Detroit
T.G. Robins, E.A. Parker, B.A. Israel, R.W. Brown, T.C. Lewis, and the CAAA Steering Committee
Uni versitv of Michigan, Ann Arbor, Ml, and community partners, Detroit, Ml

A screening questionnaire designed to identify asthmatic children ages 6 to 11 years was developed, pilot tested,
and mailed (n=7,498) or hand-delivered (n=2,182) to households in two geographic areas of Detroit with
predominantly African-American and Hispanic populations as part of Community Action Against Asthma
(CAAA). CAAA, a project of the Michigan Center for the Environment and Children s Health, is a randomized
staggered-entry cohort study of a household-level and community-level intervention to reduce exposure to
environmental triggers of childhood asthma. Among the 3,226 returned questionnaires, 1,671  (51.8%) were
consistent with probable or known asthma of any severity and 425 (13.2%) with probable or known moderate
to severe asthma based on National Asthma Education and Prevention Program diagnostic guidelines. Calculated
minimum population-based estimates of prevalence for any asthma (17.3%) and moderate to severe asthma
(4.4%) substantially exceed national averages. Among those with known or probable moderate to severe asthma,
more than 30 percent had not been diagnosed by a physician, more than one-half were not taking daily asthma
medication, and approximately one-quarter had not taken any physician-prescribed asthma medication in the
past 12 months. Our data suggest that well-constructed screening surveys conducted with community support
can identify large numbers of children with undiagnosed and/or undertreated moderate to severe asthma. These
children are likely to particularly benefit from interventions to improve their asthma disease status.
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Detroit School Children With Symptoms of Persistent
Asthma Are Sensitized to Both Indoor and Outdoor Allergens
T.C. Lewis, T.G. Robins, E.A. Parker, R.W. Brown, W.R. Solomon, T.R. Trestyn, B.A. Israel, and the CAAA
Steering Committee
University of Michigan, Ann Arbor, MI, and community partners, Detroit, Ml

Environmental factors can be significant triggers of asthma. Community Action Against Asthma (CAAA) is a
randomized staggered-cohort study of a household-level and community-level intervention to reduce exposure
to environmental triggers of childhood asthma among African-Americans and Hispanics in Detroit.  As part of
this effort, we sought to describe the pattern of allergen sensitization in this population. We hypothesized that
sensitization to indoor allergens would be more prevalent than sensitization to outdoor allergens in this inner-city
cohort. Children ages 6-11 years with symptoms consistent with persistent asthma were identified via a
screening questionnaire distributed to parents of elementary school students. After obtaining informed consent,
baseline atopic status was assessed by skin prick test with a panel including indoor allergens (cockroach, dust
mite, cat, dog, mouse, rat), outdoor allergens (ragweed, mixed grasses, Alternaria), and negative and positive
controls. Wheals were scored on a scale of 0-4, with 2 or greater interpreted as a positive reaction. Of 510
children who met entrance criteria, to date 242 have successfully undergone skin testing.  A total of 185 (76%)
responded to at least one allergen; 120 (50%) responded to three or more allergens. Atopy to dust mite and
mixed grasses were most common, followed by cat and ragweed. The proportion of children with positive
response to each allergen is shown:

Indoor allergens:
Mite
49%
Cat
38%
Roach
33%
Rat
17%
Dog
16%
Mouse
13%
Outdoor allergens:
Grass
47%
Ragweed
38%
Alternaria
36%
These allergen sensitivity patterns have implications for exposure reduction strategies.
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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Evaluation of Murine Model of
Asthma Induced by Exposure to House
Dust Extract Containing High Levels of Cockroach
Jiyoun Kim, Andrew C. Merry, Jean A. Nemzek, and Daniel G. Remick
Universit\ of Michigan, Ann Arbor, MI

Asthma represents a serious health problem, particularly for inner city children. Recent studies have identified
that many asthmatic attacks are triggered  by exposure to indoor allergens including cockroach allergens.
However, the mediators within the lung dictating the progression of disease have still not been fully defined.
This study tested the hypothesis that asthma-like pulmonary injury may be induced by house dust containing
high levels of cockroach allergens. Households (n=10) with asthmatic children were identified, and the house
dust was collected and tested for cockroach allergens. The sample, which tested highest for cockroach allergens
(Bla gl=65.8U/mL), was used for subsequent studies.  BALB/c mice were immunized  with serially diluted
aqueous house dust extract and received two additional pulmonary challenges. Control mice received sterile PBS
only. The inflammatory cells in bronchioalveolar lavage and peripheral blood and myloperoxidase activity in the
lung were analyzed. Eosinophil  counts and myloperoxidase activity were significantly increased in a dose-
dependent manner by exposure to the house dust. A kinetics study was then performed using dilutions of the
house dust extract with mice sacrificed every 12 hours after second pulmonary challenge. The inflammatory
response reached the peak at 48 hours. In peripheral blood, there were no meaningful changes in the number
of lymphocytes and  red blood cells while the  number of PMN was augmented in all three immunized groups.
Thus, these mice were successfully sensitized and manifested asthma-like responses after house-dust extract
challenge containing high  levels  of cockroach allergens. This murine model may be used for studying the
mechanisms of indoor house dust-mediated asthma.
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Measurements of Indoor, Outdoor, and Personal Exposure to
Particulate Matter Among Asthmatic Children in Detroit. Michigan
Fuyuen Y. Yip, J. Timothy Dvonch, Thomas G. Robins, Edith Parker, Masako Morishita, and Gerald J. Keeler
University of Michigan, Ann Arbor, Ml

The prevalence of childhood asthma in Detroit is particularly high, reflecting trends found elsewhere among
urban populations. Reasons for this  higher prevalence in urban populations are likely to include both socio-
economic and environmental risk factors. In a number of studies, paniculate matter (PM), generally limited to
measurements in ambient air, has been associated with increases in childhood asthma prevalence and severity.
As part of Community Action Against Asthma (CAAA), a multiyear community-based initiative for which data
collection began in October, outdoor, indoor, and personal exposures to PM are characterized to assess their
impact on childhood asthma.

To study the effects of PM exposure and the  potential role of PM in the aggravation of childhood asthma,
continuous PM2 5 is characterized throughout the study period at two community monitoring sites using a
TEOM. At the same sites, daily filters of PM2 5 and PM,0 are collected for 2  weeks each season over 2 years.
During each seasonal assessment, PM2 5 and PMIO samples also are being collected in the homes of 20 asth-
matic children. Additionally, personal monitoring for PMIO is conducted for each child. This data provides a
basis for assessment of indoor, outdoor, and personal levels of PM exposure; thereby contributing to a better
understanding of the role PM may play  on the exacerbation of childhood asthma.
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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Field Comparison of PM25 TEOM and PM2.5 Manual
Filter-Based Measurement Methods in Urban Atmospheres
/. Timothy Dvonch, Frank J. Marsik, Gerald J. Keeler, Thomas G. Robins, Fuyuen Yip, and Masako Morishita
Universit\ of Michigan, Ann Arbor, MI

As part of a community-based project, the effects of air pollutants and the role they play in exacerbating
childhood asthma are being assessed in Detroit, MI. One specific aim of the project is to identify sources of air
pollutants through chemical characterization of paniculate matter (PM), particularly PM, 5 and PMIO, to which
the children are exposed. In addition to daily filter collection during seasonal intensive measurement campaigns
(each 2 weeks in duration) at three ambient monitoring locations, continuous measurement of PM, 5 is made
at each location with a TEOM. Because the uncertainties in physical characterization of PM may vary between
monitoring sites and between measurement techniques on a seasonal basis (due to large variations in ambient
conditions), a direct field  comparison of measurement techniques is needed in the airshed of interest.

For the first intensive measurement period in autumn 1999 (October 26,1999 November 8, 1999), daily PM2,
levels at the three sites ranged from 2-48 |Ig/m\ with mean values ranging from 17-21 |J.g/m3 across the three
sites.  A comparison  of PM2 5 TEOM measurements and PM, _, manual filter-based  measurements was
conducted at each monitoring site during this period. The PM2 s TEOM  (with sharp cut cyclone, SCC) at the
southwest Detroit site reported relatively lower values than those determined with the manual collection
technique using a classical cyclone, with a regression slope of 0.88 (N=13). This relationship also was observed
at the east Detroit site, as the regression slope was 0.77 (N=l 3). A comparison at the Ann Arbor monitoring site
resulted in a regression slope of 1.02 (N=l 1). In contrast to the two Detroit TEOMs, the Ann Arbor TEOM
utilizes a classical cyclone inlet instead of an  SCC. Also in contrast to the two Detroit sites, the measurements
from the Ann Arbor TEOM were not statistically different from those of the manual filter method, as they both
utilized a classical cyclone inlet. The differences observed between the TEOM and manual method at the two
Detroit sites is likely  due to the sharper particle size cut incorporated by the SCC.

Further intercomparative sampling periods, by season through 2000 and 2001, in addition to chemical charac-
terization of the PM will lend additional insight into the performance of these various PM measurement methods
in an urban environment. In addition, these data will allow  investigations into the sources  of PM in the Detroit
airshed regarding PM exposure assessment and the role of air pollutants in exacerbation of childhood asthma.
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   2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
                                       Index of Authors
        Arbour, N.C., 48
        Buckley, T.J., 39
        Carter, A.B., 56
        Chen, J., 18
        Chen, W., 53, 54
        Chrischilles, E., 45
        Diaz-Sanchez, D., 5
        Dvonch, J.T., 65
        Eggleston, P., 35
        Eskanazi, B., 29, 30
        Faustman, E.M., 21
        Furlong, C.E., 22
        George, C.L.S., 46, 51
        Gilliland, F., 6
        Godina, R., 25
        Gong, H., 3
        Holland, N., 32
Hunninghake, G.W., 43
Israel, B., 59
Kim, J., 63
Kinney, P.L., 13
Kline, J.N., 49
Leidal, K., 52
Lewis, T.C., 62
Monick, M.M., 55
Moreland, J.G., 50
Perera, F., 9
Quinn T.J., 47
Robins, T.G., 61
Thompson, B., 23
Walters, D., 38
Whyatt, R.M., 12
Wolff, M.S., 17
Yip, F.Y., 64
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                                     APPENDIX A

                                       Agenda

               EPA/NIEHS/CDC Centers for Children's Environmental
                      Health and Disease Prevention Research
                                    Annual Meeting

                                 November 5-7, 2000
                           University of California-Berkeley
                                  Clark-Kerr Center
4:00 p.m.      Registration and Poster Set-Up

5:00 p.m.      Welcome and Introduction of Speaker
              Brenda Eskanazi, Ph.D.
              Director,  Center for the Health Assessment of the Mothers and Children of Salinas
              (CHAMACOS), University of California-Berkeley

              Invited Speaker: "Socioeconomic Status and the Health Gradient: Mechanisms of
              Influence"
              Nancy Adler, Ph.D.
              Vice Chair of Psychiatry, University of California-San Francisco

5:45 p.m.      Open Discussion: Opportunities for linking the Centers with other Government-sponsored
              programs and nonprofit organizations.

              Chris Rosheim, Agency for Toxic Substances and Disease Registry (ATSDR)
              Jerome Paulson, M.D., Children's Environmental Health Network Soros Advocacy
              Fellow and Co-Director of ATSDR's Region 3 Pediatric Environmental Health Specialty
              Unit (PEHSU) at George Washington University

6:30 p.m.      Poster Session

9:00 p.m.      Adjourn
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Monday, November 6          Morning Session: Center Updates and Scientific Highlights

8:30 a.m.      Special Guest
              Joy Carlson, M.P.H.
               The California Project

                                   CENTER PRESENTATIONS

9:15 a.m.      University of Southern California
              Columbia University

10:15 a.m.     Break

10:30 a.m.     Mount Sinai School of Medicine
              University of Washington
              University of California-Berkeley

12:00 noon    Lunch
1:30 p.m.      The Johns Hopkins University
              University of Iowa
              University of Michigan

3:00 p.m.      Break

3:30 p.m.      Cross-Center Roundtables

              The objective  of this working session is to share cross-Center concerns to address specific
              issues that pertain to data quality, biological sampling/storage, and the ethical considerations
              of human subjects research. Each group will be facilitated through the process of identifying
              common obstacles and opportunities and encouraged to design an action plan to be presented
              and discussed in the plenary session on Tuesday morning.

              Roundtable 1: Lung Disease Etiology and Prevention Research

              Roundtable 2: Pesticide Exposure and Toxicity Research

6:30 p.m.      Adjourn

7:00 p.m.      Director's Dinner
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
9:00 a.m.       Agenda Review/Repair



9:05 a.m.       Report of Roundtable 1 and Discussion



10:00 a.m.      Report of Roundtable 2 and Discussion



11:00a.m.      Wrap-Up



11:30 a.m.      Adjourn



12:00 noon     Leave for CHAMACOS de Salinas
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop


                                          APPENDIX B

                         Report From Pesticide Breakout Session

                                        Topics Addressed

I.       QA/QC and Inter-Center Collaboration
II.      Human Subjects
III.     Pesticide Metabolism
IV.     Linkages With EPA and NIEHS Laboratories
I.       QA/QC and Inter-Center Collaboration
        Questionnaires
        - Sharing and incorporating into study instruments
        - Validation
        Central Laboratory To Assure Commonality
        - Dana Barr s laboratory at CDC
        - One center to provide analytical service to other centers
        - National laboratory
        Sharing Methods/Protocols
        - Clem Furlong-2D digital  assay
        Taking Initiative
        QA Plan
        - Audit (www.epa.gov/ncerqa) announcements
        - EPA to provide guidance
        Information Database
        - Samples
        - Promote inter-Center interaction
        Common Language/Terminology
        Exposures and Genetics
        - Protocol to work with poison control center to obtain urine samples
        Recommendation
        - Project officers to examine means for inter-Center comparison of outcome data

>*      Human Subjects
        HHS Training Requirement
        - All researchers dealing with human subjects
        - Many venues for training
           *NIH (http://ohsr.od.nih.gov/cbt)
           *University (classroom and Web-based)
        Reporting Findings of Illegal Activities
        - Must address with university lawyers
        Consent Forms and Sample Banking
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   2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
///.     Pesticide Metabolism
        Pesticide Uses and Use Patterns
        - Patterns constantly changing
        - Regulatory action
        Mixtures
        - Challenges
        - Technological advancements
        Metabolite Analysis in Relation to Exposure
        - Creatinine and creatine adjustment
        - Children and pregnant women
        - Metabolic pathways
        Use California PUR Data
        - Assumption of exposures
        Urinary Metabolites
        - Three Centers to examine sharing samples
        - Data presentation and reporting
IV.     Linkages With EPA and NIEHS Laboratories
        Agricultural Health Study
        - Desire for more information
        National Children s Longitudinal Cohort Study
        - Desire for more information
        Two-Way Interaction
        - Centers can offer input on study design and implementation
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
 Liam O Fallon

 Robert Menzer

 Rosana Hernandez

 Kelly Birch

 Robin M. Whyatt

 Chris Lau

 David Dix

 Asa Bradman

 Alex Lee

 Mary Wolff

 Lauren Fenster


 Martha Harnly


 Clement Furlong

 Ruth Woods

 Kristina Dam

 Jim Merchant

 Debbie Bennett

 Rosemary Castorina

 Ray Chavira

 Ricky Perera

 Nina Holland

 Allen Dearry

 Brenda Eskenazi
Pesticide Breakout Session

      List of Participants


 NDEHS

 USEPA

 CHAMACOS

 CHAMACOS

 Columbia University

 USEPA

 USEPA

 CHAMACOS

 University of Washington

 Mount Sinai School of Medicine

 California Department of Health
 Services

 California Department of Health
 Services

 University of Washington

 University of Washington

 University of Washington

 University of Iowa

 CHAMACOS

 CHAMACOS

 EPA/Region 9

 Columbia University

 CHAMACOS

 NIEHS

 CHAMACOS
ofallon@niehs.nih.gov

menzer.robert @ epa.gov

sanie@uclink.berkeley.edu

kbirch@chori.org

rmw5 @columbia.edu

lau.christopher@epa.gov

dix.david@epa.gov

abradman@socrates.berkeley.edu

calu@u. washington.edu

mary.wolff@mssm.edu

lfenster@dhs.ca.gov


mharnly@dhs.ca.gov


clem @u. washington.edu

rwoods@u.washington.edu

kdam@u.washington.edu

james-merchant@uiowa.edu

dhbennett@lbl.gov

rcastori@uclink4.berkeley.edu

cha vira. ray mond @ epa.gov

fppl @columbia.edu

ninah @ uclink4.berkeley.edu

dearry@niehs.nih.gov

eskenazi@uclink4.berkeley.edu
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                                        APPENDIX C

              EPA/NIEHS/CDC Centers for Children s Environmental
                        Health  and Disease Prevention Research
                                      Annual Meeting

                                     List of Participants

         ;   s       :'            	   Invited Guests  !        ":	,  -  ,              :
Nancy Adler
University of California, San Francisco
Box 0844, LHts 465 P
San Francisco, C A 94143
Telephone: (415)476-7759
Fax:  (415)476-7744
E-mail: nadler@itsa.ucsf.edu

Joy E. Carlson
J. Carlson Consulting
1506 Hampel Street
Oakland, CA  94602
Telephone: (510)530-7949
Fax:  (510)530-7943
E-mail: carlsonj@dnai.com
Jerry Paulson
George Washington University
  Medical Center
Department of Environmental and
  Occupational Health
2300 K Street, NW
Washington, DC 20037
Telephone: (202)994-9914
Fax:  (202)994-4861
E-mail: hcsjap@gwumc.edu

Chris Rosheim
Agency for Toxic Substances and Disease
  Registry
1600 Clifton Road, MSE-33
Atlanta, GA  30333
Telephone: (404)639-6243
Fax:  (404)639-6207
E-mail: cxr5@cdc.gov
                                           CEHCs
Henry Gong
Principal Investigator
Southern California Children s
  Environmental Health Center
University of Southern California
2250 Alcazar Street, CSC-219
Los Angeles, CA 90033
Telephone:  (323)442-1096
Fax: (323)442-3272
E-mail: hgong@dhs.co.la.ca.us
David Diaz-Sanchez
Clinical Immunology and Allergy
University of California, Los Angeles School
  of Medicine
10833 Le Conte Avenue, Room 52-175 CHS
Los Angeles, CA 90095-1680
Telephone:  (310)825-9376
Fax:  (310)206-8107
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
                                      Columbia University
Patrick Kinney
Columbia Center for Children s
  Environmental Health
Columbia University School
  of Public Health
Division of Environmental
  Health Sciences
60 Haven Avenue, B-112
New York, NY 10032
Telephone:  (212)304-7275
Fax: (212)544-1943
E-mail: plk3@columbia.edu

Frederica Perera
Principal Investigator
Columbia Center for Children s
  Environmental Health
Columbia University School
  of Public Health
Division of Environmental
  Health Sciences
60 Haven Avenue, B-l 12
New York, NY 10032
Telephone:  (212)304-7275
Fax: (212)544-1943
E-mail: fppl@columbia.edu
                               Robin Whyatt
                               Columbia Center for Children s
                                Environmental Health
                               Columbia University School
                                of Public Health
                               Division of Environmental
                                Health Sciences
                               60 Haven Avenue, B-l 12
                               New York, NY 10032
                               Telephone:  (212)304-7275
                               Fax:  (212)544-1943
                               E-mail: rmw5@columbia.edu
Mary Wolff
Principal Investigator
Mount Sinai Center for Children s
  Environmental Health and Disease Prevention
Mount Sinai School of Medicine
One Gustave L. Levy Place, Box 1057
New York, NY 10029-6574
Telephone:  (212)241-6183
Fax:  (212)241-0407
E-mail: mary.wolff@mssm.edu
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                                    University of Washington
Gloria Cornado
Center of Child Environmental
  Health Risks Research
University of Washington, Box 354695
Department of Environmental Health
Seattle, WA 98195
Telephone: (206)685-2269
Fax: (206)685-4696

Elaine Faustman
Principal Investigator
Center of Child Environmental
  Health Risks Research
University of Washington, Box 354695
Department of Environmental Health'
Seattle, WA 98195
Telephone: (206)685-2269
Fax: (206)685-4696
E-mail:  faustman@u.washington.edu

Clement Furlong
Genetics Box 357360
University of Washington
Seattle, WA 98195-7360
Telephone: (206)543-1193
Fax: (206)543-0745
E-mail:  clem@u.washington.edu
Ruth Woods
Center of Child Environmental
  Health Risks Research
University of Washington, Box 354695
Department of Environmental Health
Seattle, WA 98195
Telephone: (206) 685-2269
Fax: (206)685-4696
E-mail:  rwoods@u.washington.edu

Alex Lee
University of Washington
E-mail:  calu@u.washington.edu

Kristina Dam
University of Washington
E-mail:  kdam@u.washington.edu
Robin Baker
Center for the Health Assessment
  of the Mothers and Children
  of Salinas
University of California
School of Public Health
312 Warren Hall
Berkeley, CA 94720-7360
Telephone: (510)642-9544
Fax: (510)642-5815
Debbie Bennett
Center for the Health Assessment
  of the Mothers and Children
  of Salinas
University of California
School of Public Health
312 Warren Hall
Berkeley, CA 94720-7360
E-mail: dhbennett@lbl.gov
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review1 Workshop
Kelly Birch
Center for the Health Assessment
  of the Mothers and Children
  of Salinas
University of California
School of Public Health
312 Warren Hall
Berkeley, CA 94720-7360
E-mail: kbirch@chori.org

Asa Bradman
Center for the Health Assessment
  of the Mothers and Children
  of Salinas
University of California
School of Public Health
312 Warren Hall
Berkeley, CA 94720-7360
Telephone:  (510)642-9544
Fax: (510)642-5815
E-mail: abradman@socrates.berkeley.edu

Rosemary Castorina
Center for the Health Assessment
  of the Mothers and Children
  of Salinas
University of California
School of Public Health
312 Warren Hall
Berkeley, CA 94720-7360
E-mail: rcastori@uclink4.berkeley.edu

Brenda Eskenazi
Principal Investigator
Center for the Health Assessment
  of the Mothers and Children
  of Salinas
University of California
School of Public Health
312 Warren  Hall
Berkeley, CA 94720-7360
Telephone:  (510)642-9544
Fax: (510)642-5815
E-mail: eskenazi@uclink4.berkeley.edu
                               David Gutzman
                               Center for the Health Assessment
                                 of the Mothers and Children
                                 of Salinas
                               University of California
                               School of Public Health
                               312 Warren Hall
                               Berkeley, CA 94720-7360
                               Telephone: (510)642-9544
                               Fax: (510)642-5815

                               Rosana Hernandez
                               Center for the Health Assessment
                                 of the Mothers and Children
                                 of Salinas
                               University of California
                               School of Public Health
                               312 Warren Hall
                               Berkeley, CA 94720-7360
                               E-mail: sanie@uclink4.berkeley.edu

                               Nina Holland
                               Center for the Health Assessment
                                 of the Mothers and Children
                                 of Salinas
                               University of California
                               School of Public Health
                               312 Warren Hall
                               Berkeley, CA 94720-7360
                               Telephone: (510)642-8781
                               Fax: (510)642-0427
                               E-mail: ninah @ uclink4.berkeley.edu

                               Susan Neal
                               Center for the Health Assessment
                                 of the Mothers and Children
                                 of Salinas
                               University of California
                               School of Public Health
                               312 Warren Hall
                               Berkeley, CA 94720-7360
                               Telephone: (510)642-9544
                               Fax: (510)642-5815
                               E-mail: sneal@uclink4.berkeley.edu
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
                                 The Johns Hopkins University
Patrick Breysse
Center for Childhood Asthma
  in the Urban Environment
Department of Environmental
  Health Sciences
School of Public Health
615 N Wolfe Street
Baltimore, MD 21287
Telephone: (410)955-5883
Fax: (410)955-0229
E-mail: pbreysse@jhsph.edu

Peyton Eggleston
Principal Investigator
Center for Childhood Asthma
  in the Urban Environment
Johns Hopkins Hospital
600 N Wolfe Street, CMSC 1102
Baltimore, MD 21287
Telephone: (410)955-5883
Fax: (410)955-0229
E-mail: pegglest@jhmi.edu
Cynthia Rand
Center for Childhood Asthma
  in the Urban Environment
Johns Hopkins Hospital
600 N Wolfe Street, CMSC 1102
Baltimore, MD  21287
Telephone: (410)955-5883
Fax: (410)955-0229

Marsha Wills-Karp
Center for Childhood Asthma
  in the Urban Environment
Johns Hopkins Hospital
600 N Wolfe Street, CMSC 1102
Baltimore, MD  21287
Telephone: (410)955-5883
Fax: (410)955-0229
Gary Hunninghake
Principal Investigator
Children s Environmental Airway
  Disease Center
University of Iowa
Department of Internal Medicine
200 Hawkins Drive, C33-GH
Iowa City, IA 52242
Telephone:  (319)356-4187
Fax: (319)356-8101
E-mail: gary-hunninghake@uiowa.edu

James Merchant
Children s Environmental Airway
  Disease Center
University of Iowa
College of Public Health, 2707 SB
Iowa City, IA 52242
Telephone:  (319)355-9833
Fax: (319)355-9777
E-mail: james-merchant@uiowa.edu
Martha Monick
Children s Environmental Airway
  Disease Center
University of Iowa
Department of Internal Medicine, 100EMRB
Iowa City, IA 52242
Telephone:  (319)355-7590
Fax: (319)355-6530
E-mail: martha-monick@uiowa.edu

David Schwartz
Duke University Medical Center 2629
Room 275, Research Drive
Durham, NC 27710
Telephone:  (919)668-0380
Fax: (919)688-0494
E-mail: david.schwartz@duke.edu
The Office of Research and Development s National Center for Environmental Research
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Jerry Keeler
Michigan Center for the Environment
  and Children s Health
University of Michigan
School of Public Health
Department of Environmental Health
  Sciences
109 Observatory
Ann Arbor,  MI 48109-2029
Telephone:  (734) 936-1836
Fax: (734)764-9494
E-mail: jkeeler@umich.edu

Jiyoun Kim
Michigan Center for the Environment
  and Children s Health
University of Michigan
School of Medicine
Department of Pathology
Med.Sci4241
Ann Arbor,  MI 48109-2029
Telephone:  (734) 763-6454
E-mail: jiyoukim@umich.edu
                              Toby Lewis
                              Michigan Center for the Environment
                                and Children s Health
                              University of Michigan
                              School of Medicine
                              c/o Department of Health Behavior
                                and Health Education
                               1420 Washington Heights
                              Ann Arbor, MI 48109-2029
                              Telephone:  (734)615-2455
                              Fax: (734)763-7379
                              E-mail:  tobyl@med.umich.edu
                                            Agencies
Raymond Chavira
U.S. Environmental Protection Agency,
  Region 9
CMD-4-3
75 Hawthorne Street
San Francisco, CA 94105
Telephone:  (415)744-1926
E-mail: chavira.raymond@epa.gov

EdChu
U.S. Environmental Protection Agency
Ariel Rios Building, MC (1107A)
1200 Pennsylvania Avenue, NW
Washington, DC 20460
Telephone:  (202)564-2196
E-mail:  chu.edu@epa.gov
                               David J. Dix
                               U.S. Environmental Protection Agency
                               Mailroom
                               MD-72
                               Research Triangle Park, NC 27711
                               Telephone:  (919)541-2701
                               Fax:  (919)541-5138
                               E-mail: dix.david@epa.gov

                               Nigel Fields
                               U.S. Environmental Protection Agency
                               Ariel Rios Building, MC (8723R)
                               1200 Pennsylvania Avenue, NW
                               Washington, DC 20460
                               Telephone:  (202) 564-6936
                               Fax:  (202)565-2448
                               E-mail: fields.nigel@epa.gov
82
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  2001 EPA/NIEHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
Christopher S. Lau
U.S. Environmental Protection Agency
Mailroom
MD-67
Research Triangle Park, NC 27711
Telephone: (919)541-5097
Fax: (919)541-4017
E-mail:  lau.christopher@epa.gov

Boh Menzer
U.S. Environmental Protection Agency
Ariel Rios Building, MC (8701R)
1200 Pennsylvania Avenue, NW
Washington, DC  20460
Telephone: (202) 564-6949
Fax: (202)565-2444
E-mail:  menzer.robert@epa.gov
Brenda Perkovich
U.S. Environmental Protection Agency
Ariel Rios Building, MC (1107A)
1200 Pennsylvania Avenue, NW
Washington, DC 20460
Telephone: (202) 564-2707
E-mai 1: perkovich.brenda@epa.gov
Linda Bass
NEHS
79 T. Alexander Drive
P.O.Box 12233(EC-30)
Research Triangle Park, NC 27709
Telephone: (919)541-1307
Fax: (919)541-2503
E-mail:  bass@niehs.nih.gov

Gwen Collman
NEHS
79 T. Alexander Drive
P.O.Box 12233(EC-21)
Research Triangle Park, NC 27709
Telephone: (919) 541 -4980
Fax: (919)541-4937
E-mail:  collman@niehs.nih.gov

Allen Dearry
NffiHS
79 T. Alexander Drive
P.O. Box 12233 (EC-21)
Research Triangle Park, NC 27709
Telephone: (919)541-4943
Fax: (919)541-4397
E-mail: dearry@niehs.nih.gov
Ethel Jackson
NIEHS
79 T. Alexander Drive
P.O.Box 12233(EC-30)
Research Triangle Park, NC 27709
Telephone: (919)541-7846
Fax: (919)541-2503
E-mail: jackson4@niehs.nih.gov

Liam O Fallen
NffiHS
79 T. Alexander Drive
P.O.Box 12233 (EC-21)
Research Triangle Park, NC 27709
Telephone: (919)541-7733
Fax: (919)316-4606
E-mail: ofallon@niehs.nih.gov

Laura Williams-Boyd
NffiHS
79 T. Alexander Drive
P.O.Box 12233(EC-30)
Research Triangle Park, NC 27709
The Office of Research and Development s National Center for Environmental Research
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  2001 EPA/N1EHS/CDC Centers for Children s Environmental Health and Disease Prevention Research Progress Review Workshop
                          Centers for Disease Control and Prevention
Stephen Redd
National Center for Environmental Health
CDC
4770 Butbrd Highway, NE
Atlanta, GA 30341-3724
Telephone:  (770)488-7581
Fax: (770)488-3507
E-mail: scrl@cdc.gov
Lauren Fenster
Telephone:  (510)540-2001
E-mail: lfenster@dhs.ca.gov
     California Department of Health Services

                              Martha Harnly
                              E-mail:  mharnly@dhs.ca.gov
84
The Office of Research and Development s National Center for Environmental Research

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