National Toxicology Program

U.S. Department of Health and Human Services

Annual Report 2010
National Toxicology Program

ANNUAL REPORT

For

Fiscal Year 2010

National Institute of Environmental Health Sciences 

National Institutes of Health

National Center for Toxicological Research

Food and Drug Administration

National Institute for Occupational Safety and Health

Centers for Disease Control and Prevention

July 2011

Department of Health and Human Services

National Toxicology Program

NIH Publication No. 11-5972
Table of Contents

Letter from the NIEHS/NTP Director.....................................................................................................................................1

NTP Highlights .................................................................................................................................................................................2

Overview of the NTP .....................................................................................................................................................................3

Resources and Planning..............................................................................................................................................................7

NIOSH/NTP ......................................................................................................................................................................................14

NCTR/NTP ..................................................................................................................................................................................... 18

NIEHS/NTP ..................................................................................................................................................................................... 30

Highlighted Activities ...................................................................................................................................................... 30

 Continued Oil Spill Initiatives.................................................................................................................................. 30

 NTP Laboratories Branch .......................................................................................................................................... 31

 Chemical Effects in Biological Systems (CEBS) ................................................................................................ 32

 Cellular Phone Radiation Emissions...................................................................................................................... 32

 Nanotechnology Safety Initiative.......................................................................................................................... 33

 Herbal Medicines and Dietary Supplements.................................................................................................... 34

Report on Carcinogens.................................................................................................................................................... 38

Center for the Evaluation of Risks to Human Reproduction ....................................................................... 41

NTP Interagency Center for the Evaluation of Alternative Toxicological Methods......................... 44

NTP Research and Testing Program.......................................................................................................................... 48 

 Nomination, Selection, Evaluation, and Review................................................................................................48

 Chronic Toxicity/Carcinogenicity Studies........................................................................................................... 52

 General Toxicology Studies...................................................................................................................................... 62

 Mutagenesis and Genetic Toxicity......................................................................................................................... 65

 Organ System Toxicity................................................................................................................................................ 67

 Disposition, Metabolism, and Toxicokinetic Studies...................................................................................... 70

 Genetic and Alternative Test Systems.................................................................................................................. 72

Cellular and Molecular Pathology ...................................................................................................................................... 74

NTP Postdoctoral Training Programs................................................................................................................................ 77

NTP Interagency Agreements

NIEHS/NCTR............................................................................................................................................................................. 78

NIEHS/NIOSH .......................................................................................................................................................................... 81

NTP/NHGRI/EPA ..................................................................................................................................................................... 84

NIEHS/EPA................................................................................................................................................................................. 87

Appendix 1

Agency Staff and Contact Information......................................................................................................................... 88

Appendix II

FY 2010 Bibliography........................................................................................................................................................... 90
Frequently Used Abbreviations

3TC lamivudine

AA aristolochic acid

ACB allele-specific competitive blocker 

ACD allergic contact dermatitis 

ADME absorption, distribution, metabolism, and 

excretion

Ag silver

AMPH amphetamine 

ATSDR Agency for Toxic Substances and Disease 

Registry

AZT zidovudine, 3’-Azido-3’-Deoxythymidine

BCOP bovine corneal opacity and permeability

BPA bisphenol A

BrdU bromodeoxyuridine

BSC Board of Scientific Counselors

CASRN CAS Registry Number

CDC Centers for Disease Control and Prevention

CDMA code division multiple access 

CEBS Chemical Effects in Biological Systems 

CERHR Center for the Evaluation of Risks to Human 

Reproduction

CNS central nervous system 

CNT carbon nanotube

CPSC U.S. Consumer Product Safey Commission

CYP cytochrome P450

DA Daicel adenosine triphosphate

DEHP di(2-ethylhexyl) phthalate

DEP diesel exhaust particles

DHHS Department of Health and Human Services

DILI drug-induced liver injury 

DTBBA dithiobisbenzanilide

ECVAM European Centre for the Validation of 

Alternative Methods

EGEHE ethylene glycol 2-ethylhexyl ether

ELISA enzyme-linked immunosorbent assay

ENU N-ethyl-N’-nitrosourea 

EPA Environmental Protection Agency

ESR electron spin resonance 

FDA U.S. Food and Drug Administration

FFPE formalin-fixed, paraffin-embedded 

FY fiscal year

GABA gamma-aminobutyric acid

GC/MS gas chromatography/mass spectroscopy

GMM Genetically Modified Model

GSM global system for mobile communication

HHE Health Hazard Evaluations

HPLC high performance liquid chromatography

HTS high throughput screening

IAG interagency agreement

IARC International Agency for Research on 

Cancer

ICATM International Cooperation on Alternative 

Test Methods

ICD irritant contact dermatitis 

ICE isolated chicken eye

ICCVAM Interagency Coordinating Committee on 

the Validation of Alternative Methods

IgE immunoglobulin E

JaCVAM Japanese Center for the Validation of 

Alternative Methods

KoCVAM Korean Center for the Validation of 

Alternative Methods 

LC/MS/MS liquid chromatography/tandem mass 

spectrometry

LLNA Local Lymph Node Assay

LTKB liver toxicity knowledgebase

mAb monoclonal antibody

MDIG mineral dust-induced gene

MLA Mouse Lymphoma Assay

Mn manganese

MOC Memorandum of Cooperation

MOU Memorandum of Understanding

MPTP 1-methyl-4-phenyl-1,2,3,6-

tetrahydropyridine

MRI magnetic resonance imaging 

MS mass spectrometry

NASH nonalcoholic steatohepatitis

NCGC NIH Chemical Genomics Center

NCI National Cancer Institute
NCTR National Center for Toxicological Research RACB reproductive assessment by continuous 

breeding NICEATM NTP Interagency Center for the Evaluation of 

Alternative Toxicological Methods RBC/RET red blood cell/reticulocyte 

NFV nelfinavir misylate RFR ardiofrequency radiation 

NHGRI National Human Genome Research Institute RoC Report on Carcinogens

NIEHS National Institute of Environmental Health ROS reactive oxygen species

Sciences RP retinyl palmitate

NIH National Institutes of Health RPT rabbit pyrogen test

NIOSH National Institute for Occupational Safety SACATM Scientific Advisory Committee on Alternative 

and Health Toxicological Methods

NMDA N-methyl-D-aspartic acid SAR structure-activity relationship

NMR Nuclear magnetic resonance SD Sprague-Dawley

nNOS neuronal nitric oxide synthase SDAR spectrometric data-activity-relationship 

NOAEL no-observed-adverse-effect level SOT Society of Toxicology

NTP National Toxicology Program SSL simulated solar light

NVP Nevirapine STP Society of Toxicologic Pathologists

OECD Organisation for Economic Co-operation and SULT1A1 sulfotransferase 1A1

Development TDI toluene diisocyanate

OPA orthophthalaldehyde TiO2 titanium dioxide

OSHA Occupational Safety and Health TOX NTP Toxicity Report Administration

TR NTP Technical Report PAH polycyclic aromatic hydrocarbon

TRRS Technical Reports Review Subcommittee PBPK physiologically based pharmacokinetic

TZD glitazones PBPK/PD PBPK/pharmacodynamic

UV ultraviolet PCBTF chloro-4-(trifluoromethyl) benzene

U.S. United States PCR polymerase chain reaction

VOC volatile organic chemical PET Positron Emission Tomography 

WC-Co tungsten carbide-cobalt PFAA perfluoralkyl acid

PFOA perfluorooctanoic acid 

PIG-A phosphatidylinositol glycan - 

complementation group A 

PPAR peroxisome proliferator-activated receptor

PWG pathology working group

QC quality control

qHCS quantitative high-content screens

qHTS quantitative high throughput screens

QSAR quantitative SAR

RA retinoic acid 

NTP Annual Report 2010 1

Letter from the NIEHS/NTP Director

As I look back on NTP for FY 2010, I am pleased to let you know that 

our prominence within the toxicology community continues both 

nationally and internationally. It was a busy year, and coordination of 

NTP’s activities with our federal partners enhanced our ability to address 

areas important to public health. The NTP responded immediately 

with coordinated efforts to address health issues associated with the 

Deepwater Horizon explosion and massive oil spill in the Gulf of Mexico. 

We initiated activities with other federal agencies to limit the adverse 

impacts of the oil and oil dispersants on human health, ecological 

health, and food sources. Early efforts by NTP focused on characterizing 

the toxicity of the oil and dispersants. While much progress has been 

made, we are continuing to tackle concerns regarding the safety of Gulf seafood, hazards to offshore 

and onshore cleanup workers, and potential long-term health impacts of residual oil in 

the environment. 

In 2010 we addressed important priorities in the President’s budget, including studies of nanomaterials 

and alternative test evaluations through recommendations of new in vitro ocular safety testing methods 

and new contact dermatitis test methods, and a workshop on alternative methods in 

vaccine potency and safety testing. We also directed efforts toward NIH Director Collins’ priorities 

in high throughput approaches, small molecule screening, and translational research. Cross-agency 

integration was enhanced by FDA joining the Tox21 collaboration, which merges federal research, 

funding, and testing tools, to develop ways to predict more effectively how chemicals affect human 

health and the environment. Additionally, we published ten NTP Technical Reports with the findings 

and conclusions from our cancer studies and introduced a more streamlined process for carrying out 

literature-based assessments of the potential for substances in our environment to cause adverse 

effects on human health.

The NTP is moving forward with new and renewed emphasis, cognizant that “Health and Environment” 

is a national priority. We are improving integration across research disciplines with all of our federal 

partners as new issues and technologies emerge. Now, and in the future, the NTP will continue to 

assemble the best individual and team science to address complex diseases and environmental impacts 

with the goal of improving our translation and communication of basic 

scientific findings into human health protection.

Linda S. Birnbaum, Ph.D., D.A.B.T., A.T.S.
FY 2010 Highlights, Public Meetings, and Events
NTP Annual Report 2010 3

Overview of the National Toxicology Program

Mission and Goals

Currently, the Toxic Substances Control Act Chemical Substance Inventory 

(http://www.epa.gov/oppt/existingchemicals/pubs/tscainventory/index.html),

first published in 1979, lists more than 80,000 chemicals as being available 

for sale and use in the United States. Approximately 850 active pesticide 

ingredients are formulated into approximately 17,000 pesticide products. 

An estimated 500 to 600 new industrial chemicals are introduced annually 

into U.S. commerce. The effects of many of these substances on human 

health are unknown, yet people and our environment may be exposed to 

them during their manufacture, distribution, use, and disposal or as pollutants 

in our air, water, or soil. While relatively few substances are thought to pose 

a significant risk to human health, safeguarding the public depends upon 

identifying the effects of these agents, and of certain naturally occurring 

chemicals, and determining the levels of exposure at which they may 

become potentially hazardous to humans.

The Department of Health, Education, and Welfare (now the Department of Health and Human Services, DHHS) 

established the National Toxicology Program (NTP) in 1978 as a focal point to coordinate toxicology testing in the 

Federal government. In carrying out its mission, the NTP has several goals:

• To provide evaluations of substances of public health concern

• To develop and validate improved (sensitive, specific, rapid) testing methods

• To develop approaches and generate data to strengthen the science base 

for risk assessment

• To communicate with all stakeholders including government, industry, academia, 

the environmental community, and the public

Organizational Structure and Oversight

Three agencies, the National Institute of Environmental Health Sciences (NIEHS) of the National Institutes of 

Health, the National Institute for Occupational Safety and Health (NIOSH) of the Centers for Disease Control and 

Prevention, and the National Center for Toxicological Research (NCTR) of the Food and Drug Administration (FDA),

form the core for the NTP (Figure 1). The NTP is located at the NIEHS, and the Director of the NIEHS serves as 

the NTP Director. Questions and inquiries about the NTP can be directed to the NTP Office of Liaison, Policy and 

Review (919-541-7539) or CDM@niehs.nih.gov.
4 National Toxicology Program

NTP Management

Dr. Linda Birnbaum, Director of NIEHS and NTP

Agency Program Management

NCTR: Dr. Paul Howard, Associate Director, Office of Scientific Coordination

NIEHS: Dr. John Bucher, Associate Director, NTP

NIOSH: Dr. Mark Toraason, Senior Fellow, Division of Applied Research and Technology

A list of the staff of the agencies involved with the program and their contact information are provided in Appendix 1.

Addressing Scientific, Regulatory, and Public Needs

The NTP is flexible and innovative in its approach toward addressing public health concerns related to exposures 

to chemical and physical agents at home, in the workplace, and in the environment. Over the years since its 

inception to take over the National Cancer Institute’s (NCI’s) cancer bioassy program, the NTP has expanded its 

scope beyond cancer to include examining the impact of substances on non-cancer outcomes, such as those 

affecting reproduction and development and the immune, respiratory, nervous, cardiovascular, and 

endocrine systems.

The NTP recognizes that initiatives addressing critical gaps in knowledge needed to evaluate environmental 

toxicants offer the best opportunities for preventing environmentally mediated diseases. Therefore, the NTP’s 

testing of substances continues to evolve to include more mechanism-based toxicology studies that focus on 

understanding the modes of action of agents under study. In recent years, the NTP has placed a greater emphasis 

on providing human relevance in interpreting and understanding toxicological information created from animal 

or in vitro cell models. This is important if we are to be at the forefront in research efforts to improve methods 

to assess risk that account for the entire sequence of events from initial chemical exposure to ultimate toxicity. 
Examples of activities the NTP covers include:

• Increased use of both information on mechanisms of action and scientific judgment in deliberations 

 for listings in the Report on Carcinogens

• Increased efforts to examine alternative testing methods that may provide better information than 

 current models while using fewer animals or causing less pain or distress, and may provide better data 

 for risk assessments

• Increased efforts to collect information on (1) a broader variety of both environmental and occupational 

 exposures, (2) potentially toxic mixtures of compounds, and (3) susceptibilities based on life stages 

 (e.g., neonatal, elderly)

Internationally, the NTP rodent bioassay is recognized as the standard for identifying carcinogenic agents; 

however, the NTP continues to work to reduce, refine, and replace the use of experimental animals and to 

develop and validate alternative testing methods. This effort led to the creation of the NTP Interagency Center 

for the Evaluation of Alternative Toxicological Methods (NICEATM) in 1998. The NTP will continue to work with 

the Interagency Coordinating Committee for the Validation of Alternative Methods (ICCVAM) through NICEATM 

in promoting the development, validation, and regulatory acceptance of new and revised alternative 

toxicological methods.

Strengthening existing partnerships and forging new ones are important to achieve NTP goals. Partnerships 

with other sister Federal agencies are increasing (interagency agreements [IAGs] are presented on page 78). The 

NTP continues to support an effort to evaluate the phototoxicity of various compounds through the NTP Center 

for Phototoxicology at the NCTR. The NTP is also contributing to toxicological assessments of emerging issues 

such as nanotechnology, radiofrequency radiation emissions from cellular phones, herbal medicines/dietary 

supplements, water disinfection by-products, and phthalates, and will provide this information to other agencies.

Regulatory agencies make decisions to protect public health based on scientific information from several 

sources (e.g., toxicology, human studies, and basic research). The NTP plays a critical role in providing needed 

scientific data, interpretation, and guidance in the appropriate uses of these data, to regulatory agencies as well 

as other groups involved in health-related research. The NTP is committed to using the best science available in 

setting priorities for future studies and in designing, conducting, and interpreting the findings of those studies. 

The American people and government agencies at state and Federal levels rely on the NTP to provide a strong 

scientific basis for making credible decisions that will protect public health. The NTP maintains an objective, 

science-based approach in dealing with critical issues in toxicology and is recognized by many groups for its 

scientific rigor, objectivity, and open approach in the continuing dialogue on appropriately applying scientific 

advances to applied toxicology research and testing.

Communication and Public Outreach

Maintaining open communications and ensuring dialogue with Federal and state agencies, industry, 

nongovernment groups, academia, and the public are goals of the NTP. NTP advisory groups (see page 8) 

provide regular scientific and public peer review and input. NTP conferences and workshops remain a priority 

and are designed to bring researchers, regulators, policy makers, and the public together to examine issues and 

achieve consensus on future directions in toxicology and risk assessment.

Distribution of NTP study results, program plans, initiatives, announcements, press advisories, and publications 

is accomplished in several ways to communicate as much as possible with the public. Information is routinely 

distributed to interested parties through Federal Register announcements on the NTP website (http://ntp.niehs.

nih.gov). The website offers access to information about the program that details and highlights ongoing and 
future initiatives, announcements, NTP centers, NTP publications, and study data. The public can subscribe to the 

NTP ListServ on the website to receive news and updates. Currently the ListServ has more than 4,000 subscribers. 

The NTP publishes the quarterly newsletter, NTP Update, which can be downloaded from the NTP website. NTP 

actively participates in the annual Society of Toxicology (SOT) meeting. At the 2010 SOT meeting in Salt Lake City, 

Utah, NTP staff participated in ten workshop/symposium/platform sessions, six education/information sessions, 

and presented more than 75 posters. 

The NIEHS/NTP Central Data Management Office oversees distribution (upon request) of specific chemical study 

information and printed NTP documents – NTP study status reports, final and draft copies of NTP Technical 

Report Series, and background documents for substances nominated to the NTP for study. On-line, searchable 

access is available for the Report on Carcinogens (RoC) and the NTP Technical, Toxicity, and Genetically Modified 

Models series reports (http://ntp.niehs.nih.gov). In FY 2010, 823,914 reports and other PDF documents were 

downloaded from the NTP website. Approximately one third of the requests were for documents related 

to the RoC.

The NTP is interested in and welcomes stakeholder input into its programs and priorities. Nominations, inquiries, 

and comments from the public and other interested parties are encouraged at any time. The NTP Office of 

Liaison, Policy and Review at the NIEHS under the direction of Dr. Mary S. Wolfe serves as the focal point for 

receiving input to the program and for overseeing the distribution of information about programs, workshops, 

initiatives, and other NTP projects. 

NTP Office of Liaison, Policy, and Review Central Data Management

NIEHS/NIH NIEHS/NIH

MD K2-03 MD K2-05

111 TW Alexander Drive, RTP, NC 27709 111 TW Alexander Drive, RTP, NC 27709

Phone: (919) 541-7539 Phone: (919) 541-3419

wolfe@niehs.nih.gov cdm@niehs.nih.gov
NTP Annual Report 2010 7

Resources and Planning

Current and Projected Research Capacity

The NTP relies on voluntary allocations from the program’s three core agencies (NIEHS, NCTR, and NIOSH) 

to support its various programs and initiatives. These allocations are specified after yearly appropriations are 

determined. As shown in Figure 2, the actual total allocations from the principals toward the NTP dropped from 

FY 2009 to FY 2010. For FY 2011, a 2% decrease in funding from FY 2010 levels is projected. FY 2011 funding 

is estimated to be 92% of FY 2008 funding. The NTP conducts its research studies mainly through contract 

laboratories or in-house at the core agencies, but also supports IAGs with other Federal agencies. Funds are 

used to sponsor workshops and conferences and to produce and distribute printed programmatic materials. 

In FY 2010, the NIEHS funded 46 contracts and held one workshop and two expert-panel meetings for the NTP. 

The NIEHS also funded IAGs with NIOSH, NCTR, EPA, and the NIH Chemical Genomics Center (NCGC) and held 

five scientific advisory meetings.

The NTP maintains an objective, science-based approach in dealing with critical issues in toxicology and 

continually sets priorities to improve the nation’s ability to evaluate the human health effects caused by 

environmental exposures.

In summary, the NTP is a comprehensive research program spanning several agencies committed to providing 

resources to support the NTP’s research and to communicating the knowledge learned to all stakeholders, 

public and private. The NTP’s efforts in testing, research, and assessing health hazards work to obtain the best 

scientifically valid data for health regulatory and research agencies to use to make appropriate decisions about 

potential human risks from exposure to environmental toxicants. Toward that end, the NTP is continually evolving 

to remain at the cutting edge of scientific research and development and application of technology.
8 National Toxicology Program

Advisory Boards and Committees

As shown in Figure 1 (page 4), the NTP relys on a number of external boards and committees for science and 

policy oversight and peer review. As needed, the program convenes Special Emphasis Panels and Working 

Groups to address specific topics.

NTP Board of Scientific Counselors 

The NTP Board of Scientific Counselors (BSC), a Federally chartered advisory group, provides scientific oversight 

to the NTP, including the RoC Center and the Center for the Evaluation of Risks to Human Reproduction (CERHR). 

The Secretary of DHHS appoints members to the BSC. The BSC can consist of up to 35 scientists, primarily from 

the public and private sectors, with scientific expertise relevant to the NTP’s activities. The BSC members serve 

rotating terms of up to four years and the BSC typically meets two to three times per year. Opportunities for 

public comment are scheduled at all meetings. The BSC’s Technical Reports Review Subcommittee (TRRS), which 

provides peer review of draft NTP long-term toxicology and carcinogenesis technical reports, met once during 

FY 2010. Following that meeting, the subcommittee was discontinued and beginning in FY2011, the peer review 

of the draft NTP Technical Reports will be done through expert peer review panel meetings. Dr. Lori White, 

the NTP Designated Federal Officer, manages the BSC. A list of members during FY 2010 is provided in Table 1.

BSC meetings in FY 2010

The BSC met three times in FY 2010 at NIEHS in Research Triangle Park, NC. Complete information about the 

meetings can be obtained at http://ntp.niehs.nih.gov/go/9741.

On December 9 – 10, 2009, the BSC reviewed the NTP Host Susceptibility Branch research and testing program, 

two contract concepts (analytical chemistry support and laboratory studies to evaluate toxicity following early life 

exposure to chemicals), CERHR’s revised evaluation process and proposed concept for a workshop on the effect 

of environmental exposures on diabetes and obesity, concepts for five testing program nominations (butterbur, 

evening primrose oil, hydroquinone, silica flour, and valerian extracts and oil), and NTP’s Dietary Supplement and 

Herbal Medicine Initiative.

BSC members and NTP staff at the June 2010 BSC meeting. 
NTP Annual Report 2010 9

The BSC met on May 10, 2010, to peer review the draft NTP Brief on Soy Infant Formula. The BSC also reviewed 

two CERHR proposals at that meeting, a research concept for assessing isoflavones in soy infant formula, and an 

approach for evaluating health effects of low-level lead exposure. The BSC also approved the TRRS report from 

the November 19, 2009 meeting.

At the June 21 – 22, 2010 meeting, the BSC peer reviewed draft substance profiles for three substances proposed 

for inclusion in the 12th Report on Carcinogens, glass wool fibers, formaldehyde, and cobalt-tungsten carbide: 

powders and hard metals. The BSC also reviewed a CERHR proposal for an evaluation of cancer chemotherapy 

during pregnancy, the NTP Chemical Effects in Biological Systems (CEBS) Data Management System, and a 

research concept for hydroxyurea.

TRRS meeting in FY 2010

The BSC TRRS met in public forum on November 19, 2009, at the NIEHS. The subcommittee peer reviewed the 

findings and conclusions of six draft NTP Technical Reports (1-bromopropane, ginseng, pulegone, milk thistle 

extract, bis(2-chloroethoxy)methane, diethylamine) of studies that used conventional rodent models. The 

subcommittee’s recommendations were reported to the BSC at the May 10, 2010 meeting.

Table 1. NTP Board of Scientific Counselors Membership Roster FY 2010

Name and Title Affiliation Term Ends BSC Service

Tracie E. Bunton, DVM, PhD, DACVP Eicarte LLC 12/27/10 BSC and TRRS

Toxicology Consultant Fairfield, PA

Edward W. Carney, PhD The Dow Chemical Company 12/27/10 BSC

Technical Leader, Developmental, Midland, MI

Reproductive and General Toxicology

Russell C. Cattley, VMD, PhD Amgen 12/27/10 BSC and TRRS

Executive Director Pathology Thousand Oaks, CA

David A. Eastmond, PhD University of California 06/30/12 BSC and TRRS

Professor and Chair, Department of Cell Riverside, CA

Biology and Neuroscience

Janan T. Eppig, PhD The Jackson Laboratory 06/30/12 BSC

Senior Staff Scientist Bar Harbor, ME

Elaine M. Faustman, PhD University of Washington 06/30/12 BSC

Professor and Director, Institute for Risk Analysis Seattle, WA

and Risk Communication; Department of 

Environmental and Occupational Health Sciences 

George Friedman-Jiménez, MD New York University School of Medicine 12/27/09 BSC

Assistant Professor, Departments of New York, NY

Environmental Medicine and Medicine 

William P. Janzen University of North Carolina 06/30/10 BSC

Professor, Division of Medicinal Chemistry and at Chapel Hill, NC

Natural Products; Director, Assay Development 

and Compound Profiling

Stephen W. Looney, PhD Medical College of Georgia 06/30/12 BSC and TRRS

Professor, Department of Biostatistics, Augusta, GA

Department of Oral Health and 

Diagnostic Science

Mitzi Nagarkatti, PhD University of South Carolina School 12/27/11 BSC and TRRS

Professor and Chair, Department of Pathology, of Medicine

Microbiology and Immunology Columbia, SC

Raymond F. Novak, PhD (chair 1/10 – 12/10) Wayne State University 12/27/10 BSC and TRRS

Director, Institute of Environmental Detroit, MI

Health Sciences 
10 National Toxicology Program

Name and Title Affiliation Term Ends BSC Service

Michael V. Pino, DVM, PhD Sanofi-Aventis 12/27/09 BSC and TRRS

Director of Pathology Bridgewater, NJ

Kenneth M. Portier, PhD (chair 1/09 – 12/09) American Cancer Society 12/27/09 BSC and TRRS

Director of Statistics Atlanta, GA

Jim E. Riviere, DVM, PhD, ATS North Carolina State University 12/27/09 BSC and TRRS

Burroughs Wellcome Fund Distinguished Raleigh, NC

Professor of Pharmacology

Diane M. Robins, PhD University of Michigan Medical School 12/27/09 BSC

Professor, Department of Human Genetics Ann Arbor, MI

Ruthann A. Rudel, MS Silent Spring Institute 12/27/11 BSC

Senior Scientist, Toxicology and Newton, MA

Environmental Health

Risk Assessment

James L. Sherley, MD, PhD Boston Biomedical Research Institute 06/30/11 BSC and TRRS

Senior Scientist Watertown, MA

Gina M. Solomon, MD, MPH Natural Resources Defense Council 12/27/11 BSC 

Senior Scientist San Francisco, CA

Justin G. Teeguarden, PhD Pacific Northwest National Laboratory 12/27/11 BSC and TRR

Senior Scientist, Fundamental and Richland, WA

Computational Sciences Directorate

NTP Board of Scientific Counselors Pending Members, FY 2010

Name and Title Affiliation Term Ends BSC Service

Miguel C. Fernández, MD, FACEP, FAAEM, University of Texas Health Science Center 06/30/13 BSC

FACMT, FAACT San Antonio TX

Professor of Surgery, Division 

of Emergency Medicine

Nicholas P. Jewell, PhD University of California 06/30/13 BSC

Professor of Biostatistics and Statistics Berkeley, CA

Dana Loomis, PhD University of Nebraska Medical Center 06/30/12 BSC

Professor and Chair, Department of Epidemiology Omaha, NE

Melissa A. McDiarmid, MD, MPH University of Maryland School of Medicine 06/30/13 BSC

Prof. of Epidemiology and Preventive Medicine Baltimore, MD

Director, Occupational Health Program

Lisa Minor, PhD In Vitro Strategies, LLC 06/30/13 BSC

Flemington, NJ

Richard Miller, DVM, PhD GlaxoSmithKline 06/30/13 BSC

Vice President, Safety Assessment Research Triangle Park, NC

Judith Zelikoff, PhD New York University School of Medicine 06/30/12 BSC

Professor, Environmental Medicine Tuxedo, NY

Director, Community Outreach

Additional information about the BSC, including minutes from its meetings, is available on the NTP website http://ntp.niehs.nih.gov/

go/164 or from Dr. Lori White, BSC Designated Federal Officer (whiteld@niehs.nih.gov).
NTP Annual Report 2010 11

Scientific Advisory Committee on Alternative Toxicological Methods

The Scientific Advisory Committee on Alternative Toxicological Methods (SACATM) is a federally chartered 

advisory committee established on January 9, 2002, in response to the ICCVAM Authorization Act of 2000 

(42 U.S.C. 285l-3(d)). SACATM advises ICCVAM, NICEATM, and the Director of the NIEHS regarding statutorily 

mandated duties of ICCVAM and activities of NICEATM (see page 44). SACATM provides advice on priorities and 

activities related to the development, validation, scientific review, regulatory acceptance, implementation, and 

national and international harmonization of new, revised, and alternative toxicological test methods. Alternative 

methods are those that reduce, refine (lessen or avoid pain and/or distress), or replace the use of animals in 

testing. SACATM also provides input on ways to foster partnerships and communication with interested parties. 

The NIEHS Director appoints 15 voting members to the SACATM, and membership includes representatives 

drawn from academia, state government, industry, and animal protection organizations. A list of members during 

FY 2010 is provided in Table 2. SACATM typically meets once a year and members serve rotating terms of up to 

four years. Dr. Lori White, NTP Designated Federal Officer, manages SACATM.

SACATM met on June 17 – 18, 2010, at the U.S. Environmental Protection Agency (EPA) in Research Triangle Park, 

NC. SACATM members discussed a number of important issues including the validation of alternative methods 

for assessing chemically induced eye injuries, assessment of acute and chronic pain in laboratory animals, and 

alternative methods of vaccine potency testing. Representatives from the FDA National Center for Toxicological 

Research and NIEHS/NTP presented updates on their current research activities including high throughput 

screening, validation of endocrine disruptor test methods, new model systems, approaches to identify biomarkers 

of disease and toxicity, and bio-imaging. Liaisons from Health Canada, the European Centre for the Validation 

of Alternative Methods (ECVAM), and the Korean Center for the Validation of Alternative Methods (KoCVAM) 

presented updates on the activities of their groups.

Members of SACATM and ICCVAM, international liaisons, and NIEHS/NTP staff at the June 2010 SACATM meeting.
12 National Toxicology Program

Table 2. Scientific Advisory Committee on Alternative Toxicological Methods Roster, FY 2010 

Name and Title Affiliation Term Ends

Laura Andrews, PhD, DABT Genzyme Corporation 06/30/12

Vice President, Pharmacology and Toxicology Framingham, MA

Karen K. Brown, PhD Pair O’ Docs Enterprises 06/30/11

President Parkville, MO

George B. Corcoran, PhD, ATS Wayne State University 06/30/11

Professor and Chairman Detroit, MI 

Department of Pharmaceutical Sciences

Helen E. Diggs, DVM, DACLAM College of Veterinary Medicine, Oregon State University 06/30/10

Associate Dean, Hospital Program Director Corvallis, OR

Veterinary Teaching Hospital

Marion F. Ehrich, PhD VA-MD Regional College of Veterinary Medicine 06/30/10

Professor, Biomedical Sciences and Blacksburg, VA

Pathology/Laboratory for Neurotoxicity Studies

Eugene L. Elmore, PhD University of California 06/30/12

Senior Project Scientist Department Irvine, CA

of Radiation Oncology

James Freeman, PhD (chair) ExxonMobil Biomedical Sciences, Inc. 06/30/10

Distinguished Toxicology Associate Annandale, NJ

Steven R. Hansen, DVM, MS, MBA, DABT, ABVT American Society for the Prevention of Cruelty to Animals 06/30/12

Chief Operating Officer Urbana, IL

Gwendolyn Y. McCormick, DVM, MS, DACLAM Boehringer Ingelheim Pharmaceuticals, Inc 06/30/12

Attending Veterinarian and Distinguished Ridgefield, CT

Research Fellow

Animal Resources Department

Sharon A. Meyer, PhD The University of Louisiana at Monroe 06/30/11

Associate Professor, Department of Toxicology Monroe, LA

Steven M. Niemi, DVM Massachusetts General Hospital 06/30/11

Director, Center for Comparative Medicine Charlestown, MA

Michael J. Olson, PhD, ATS GlaxoSmithKline 06/30/13

Director, Occupational Toxicology Corporate Research Triangle Park, NC

Environment, Health, Safety and Sustainability

Annie (Peiyong) Qu, PhD University of Illinois at Urbana-Champaign 06/30/10

Associate Professor, Department of Statistics Champaign, IL

Linda A. Toth, DVM, PhD Southern Illinois University School of Medicine 06/30/13

Associate Dean for Research Springfield, IL

and Faculty Affairs Professor

Department of Pharmacology

Gary Wnorowski, MBA, LAT Eurofins/Product Safety Laboratories 6/30/11

President Dayton, NJ

Additional information about SACATM, including minutes from its meetings, is available on the NTP website http://ntp.niehs.nih.gov/go/167 

(select “Advisory Board and Committees”) or from Dr. Lori White, Designated Federal Officer, NIEHS (whiteld@niehs.nih.gov).
NTP Annual Report 2010 13

NTP Executive Committee

The NTP Executive Committee provides programmatic and policy oversight to the NTP Director. The Executive 

Committee meets once or twice a year in closed forum. Members of this committee include the heads (or their 

designees) of the following Federal agencies:

• U.S. Agency for Toxic Substances and Disease Registry/National Center for Environmental Health

• U.S. Consumer Product Safety Commission

• U.S. Environmental Protection Agency

• U.S. Food and Drug Administration

• National Cancer Institute

• National Institute of Environmental Health Sciences

• National Institute for Occupational Safety and Health

• Occupational Safety and Health Administration

To enhance agency interactions, in FY 2010 the NTP began using agency Points of Contact (POCs) in lieu of 

formal committees to streamline communication and better utilize agency staff. Agency POCs have a dedicated 

responsibility and time commitment, are knowledgeable about the NTP mission and programs and their agency’s 

resources, and allow the most relevant agency expertise to be brought to bear on NTP issues. The responsibilities 

of the Interagency Committee for Chemical Evaluation and Coordination were assumed by the agency POCs.

Interagency Coordinating Committee on the Validation of Alternative Methods

ICCVAM is a permanent interagency committee of the NIEHS under NICEATM. The committee was formally 

established by the ICCVAM Authorization Act of 2000 (42 U.S.C. 285l-3). The purpose of ICCVAM is to establish, 

wherever feasible, guidelines, recommendations, and regulations that promote the regulatory acceptance of new 

or revised scientifically valid toxicological tests that protect human and animal health and the environment while 

reducing, refining, or replacing animal tests and ensuring human safety and product effectiveness (see http://

iccvam.niehs.nih.gov/about/process.htm). ICCVAM meets several times per year and consists of representatives 

from 15 Federal agencies that generate or use toxicological data to carry out their responsibilities to protect and 

improve the health and safety of people, animals, and the environment:

• U.S. Agency for Toxic Substances and Disease Registry/National Center for Environmental Health

• U.S. Consumer Product Safety Commission

• U.S. Department of Defense

• U.S. Department of Agriculture

• U.S. Department of Energy

• U.S. Department of the Interior

• U.S. Department of Transportation

• U.S. Environmental Protection Agency

• U.S. Food and Drug Administration

• National Cancer Institute

• National Institute of Environmental Health Sciences

• National Institutes of Health

• National Institute for Occupational Safety and Health

• National Library of Medicine

• Occupational Safety and Health Administration
14 National Toxicology Program

NIOSH/NTP

The National Institute for Occupational Safety and 

Health (NIOSH) is the Federal agency responsible for 

conducting research and making recommendations 

for the prevention of work-related injury and illness. 

The mission of NIOSH is to generate new knowledge 

in the field of occupational safety and health and 

to transfer that knowledge into practice for the 

betterment of workers. To accomplish this mission, 

NIOSH conducts scientific research, develops 

guidance and authoritative recommendations, 

distributes information, and responds to requests 

for workplace Health Hazard Evaluations (HHEs). 

NIOSH’s participation in the NTP is consistent with 

its mandate to protect workers’ health and safety 

under the Occupational Safety and Health Act and 

the Federal Mine Safety and Health Act. Setting 

priorities in occupational toxicological research 

is based upon several sources of information 

that are developed and maintained by NIOSH, 

including HHEs, industry-wide studies, gaps in 

knowledge identified while developing criteria for 

recommended standards or Criteria Documents, 

Current Intelligence Bulletins, hazard reviews or 

alerts, other technical reports, and information 

profiles on chemical hazards. Toxicological research 

on important occupational chemicals is conducted 

in genetic toxicology, carcinogenesis, toxicological 

characterization, chemical disposition, biological 

monitoring, reproductive and developmental 

toxicology, dermal toxicology, and exposure 

assessment. NIOSH research projects are 

conducted by the:

• Division of Applied Research and 

Technology – Cincinnati, Ohio 

• Division of Surveillance, Hazard Evaluations, 

and Field Studies – Cincinnati, Ohio 

• Education and Information Division – 

Cincinnati, Ohio

• Health Effects Laboratory Division – Morgantown, 

West Virginia 

NIOSH/NTP studies funded by NIOSH voluntary 

contributions are listed in Table 3.

NIOSH/NTP: Division of Applied Research and Technology

NIOSH/NTP: Division of Surveillance, Hazard Evaluations, 

and Field Studies 

NIOSH/NTP: Health Effects Laboratory Division
NTP Annual Report 2010 15

Table 3: NIOSH/NTP Projects FY 2010

NIOSH/NTP Project [Project Officer] Objective and/or Project Summary

Reproductive Health Assessment To evaluate reproductive health hazards using a health profile consisting of 

of Male Workers biomarkers for assessing male fecundity. Current efforts will focus on completing 

[Schrader] the Longitudinal Investigation of Fertility and the Environment (LIFE) project, which 

is a collaborative effort between NIOSH and the NICHD/NIH. This work includes 

development of new biomarkers to include in the male reproductive health profile.

Immunochemical Biological Monitoring To evaluate industrial and agricultural chemicals with known acute and chronic 

for Occupational Exposure and Disease toxicities which present a significant exposure risk for workers. Biological monitoring

[Striley] can assess exposure by analyzing acute and latent metabolites in various biological 

media. The goal of this project is to develop low-cost, rapid immunochemical and 

analytical chemistry biomonitoring methods that will be used to identify exposures

and evaluate potential interventions. Concurrent with development of exposure 

assessment methods, this project will identify and develop new multiplex 

immunochemical methods to evaluate biomarkers of occupational illness 

or subclinical signs of occupational illness. 

Orthophthalaldehyde (OPA) Hazard Assessment To conduct an assessment of occupational exposures to OPA and to determine if

[Toraason] healthcare workers are experiencing adverse effects associated with exposure. To 

assess exposure, this study will also develop analytical methods for environmental 

monitoring of OPA and determine the feasibility of an OPA biomarker. Because 

of the absence of published toxicological data on OPA, testing will be conducted 

in experimental animals. The toxicological testing will focus on dermal and 

respiratory irritation and sensitization. Dose-response data will be obtained for 

hazard identification risk assessment, which, along with health assessments, 

will serve as the basis for establishing exposure limits.

UV Native Fluorescence-Based Monitor To develop and evaluate a readily adaptable, next generation, direct reading,

for Workplace Exposures personal monitor for use in measuring worker exposure to a wide variety 

[Snawder] of chemicals including naphthalene and components of asphalt fume. The 

development of a monitor for volatile and semi-volatile workplace chemicals will 

be helpful in rapidly assessing chemical exposure and will result in more realistic 

occupational exposure assessments and allow for rapid interventions leading to 

reduced worker exposures and thus preventing occupational illness and disease.

Analytical Research & To provide for the administrative needs and analytical instrumentation repair and

Development Infrastructure maintenance in support of Chemical Exposure and Monitoring Branch chemists

[Streicher] conducting research on sampling and analytical methods development for 

workplace chemicals. New methods needed to assess chemicals being investigated 

as part of the NIOSH/NTP exposure assessment IAG are developed in this project.

Diacetyl Exposure Assessment To develop and evaluate sampling and analytical methods for diacetyl and other

[Streicher] flavoring compounds to enable accurate exposure assessment and evaluation of

the effectiveness of control technology. Two sampling and analytical methods 

are being investigated for measurement of specific flavoring compounds, most 

notably diacetyl and 2,3-pentanedione in airborne particles and bulk powders. 

Finally, broader GC/MS method(s) will be developed for a range of compounds 

present in flavorings.

Chemical Exposure Monitoring with To investigate a direct reading exposure method that uses a personal photo

Indoor Positioning ionization detector chemical monitor with telemetry and an indoor positioning

[Brown] system to provide remote monitoring of a worker’s exposure to volatile organic 

chemicals (VOCs) with position and time. The personal monitor continuously 

samples and analyzes the workers breathing zone air for VOCs while recording 

their position and time of exposure. Indoor positioning is accomplished using a 

radio transmitter attached to the personal monitor and receivers place in the 

ceiling corners of the room. The positioning receivers communicate with each 

other and a remote laptop using wireless local area network (WLAN) technology. 

The remote laptop calculates and visualizes the worker position and exposure level.

Once developed, this technology will be applied to analyze workplace exposures 

to diacetyl.
16 National Toxicology Program

NIOSH/NTP Project [Project Officer] Objective and/or Project Summary

Titanium Dioxide (TiO2) To collect occupational exposure information for workers exposed to ultrafine and 

Nanoparticle Exposure Study fine TiO2. The data will be used by the Education and Information Division to

[Curwin] provide information for the Current Intelligence Bulletin on TiO2. The study 

objectives are (1) to develop a strategy to measure exposure to ultrafine particles, 

(2) to characterize exposure to ultrafine and fine TiO2 for various jobs and tasks at 

various facilities manufacturing and using TiO2, and (3) to evaluate a strategy for 

measuring workplace exposure to fine and ultrafine TiO2.

Exposure Assessment for To characterize workplace exposures to welding fumes with emphasis on 

Toxicologically Important Chemicals manganese, indium and indium compounds, diacetyl; 2-methoxy-4-nitroaniline, 

[Estill] and 2’, 2’’’-dithiobisbenzanilide, all of which have been nominated by various 

groups to the NTP. NIOSH will identify possible candidate industries, labor unions, 

workplaces, and uses and users; will determine if there is relevance for occupational

health; estimate the number of workers exposed; and perform limited workplace 

exposure sampling.

Exposures and Engineering Controls To conduct a complete exposure assessment, evaluate potential engineering 

in the Flavoring Industry controls within the flavoring industry, and create appropriate work practice advice 

[Curwin] to reduce occupational exposures in these industries. Although a dose response 

curve for various flavoring compounds and associated health effects has not been 

established, improved work practices and engineering control advice can minimize

occupational exposures. Data from initial research efforts have been very useful in 

preliminary rulemaking efforts for both OSHA and Cal-OSHA.

Development of New Immunodiagnostic and To address problems associated with measuring personal exposure to 

Detection Techniques for Indoor Fungi Paecilomyces variotii fungus in occupational settings by identifying the major 

[Green] allergens and to produce species-specific monoclonal antibodies towards these 

allergens. These antibodies will then be used to create practical immunoassays to 

detect this fungus in clinical and environmental samples.

Cutaneous Bioactivation of Xenobiotics: To develop an in vivo model of allergic skin sensitization that can discriminate 

Hapten vs. Prohapten between chemicals requiring metabolic activation for sensitization (prohaptens) 

[Siegel] and those that can sensitize without biological activation (haptens). The model 

will involve the dermal application of various pharmacological inhibitors of the 

cytochrome P450 (CYP) pathway prior to performing either the local lymph node 

assay (LLNA) or/and mouse ear swelling test. Selective inhibition of the CYP 

pathway should distinguish between direct acting haptens and metabolically 

activated pro-haptens. Validation of the models will be done using known direct 

acting haptens and prohaptens. Successful development of these models will 

produce data that strengthens in silico hazard predictive models and allows for 

substitution or modification of allergenic chemicals and drugs.

Immunotoxicological Evaluation of To identify occupational and environmental chemical hazards and evaluate immune 

Occupational Chemicals function and mechanism associated with exposure. This research will contribute to

[Anderson] increased identification of immunological hazards encountered in the workplace. 

Further evaluation of these compounds will allow for better risk assessment, which 

will ultimately establish occupational exposure limits.

Evaluation of Perfluoralkyl Acids To investigate perfluorooctanoic acid (PFOA), which is no longer used in

(PFAAs) Immunotoxicity manufacturing, but still persistent in the environment, and which has been shown 

[Franko] in a murine model to be both immunosuppressive and to have a potential role 

in asthma and allergy. Due to the potential health effects linked to PFOA exposure, 

replacement PFAAs are now being used in the manufacturing process but little 

is known about what effects these compounds will have on immune function. 

This project will evaluate the immunotoxic effects associated with individual 

PFAAs that are still in use, and will investigate the mechanism mediating the 

identified immunological alterations associated with PFOA exposure.
NTP Annual Report 2010 17

NIOSH/NTP Project [Project Officer] Objective and/or Project Summary

Airway Fungal Exposure and To compare the immunological health effects of lung exposure to fungal spores or

Allergic Sensitization in Mice hyphal fragment preparations. Agricultural, as well as construction and remediation 

[Templeton] workers, are exposed to elevated levels of fungi and can experience rhinitis, 

respiratory allergic symptoms, and/or asthma as a result of their exposure. This 

project will have two major areas of study: (1) determination of the health effects 

following aspiration of hyphal fragments from Stachybotrys chartarum and 

Alternaria alternata in the absence of intact spores in mice, and (2) comparison 

of the ability of aspirated spores or hyphal suspensions from Aspergillus spp., 

S. chartarum, and A. alternata to exacerbate respiratory allergy to ovalbumin.

Transient Dermal Exposure: To enhance knowledge and understanding of how industrial chemicals penetrate 

Model and Experiments the skin after the types of exposure that occur in occupational settings. Data from 

[Frasch] in vitro skin penetration experiments will be compared with predictions of 

computer models to enhance their predictive value. A final product of this 

research will be a user-friendly, interactive, web-based calculator that can estimate 

the amount of chemical that penetrates the skin during workplace exposures.

Indoor Chemistry of Consumer To investigate indoor reactant/consumer product reactions to more clearly define

Product Mixtures indoor workplace exposure, provide insight into important chemical structure(s) 

[Wells] that influence indoor air quality, and highlight potential analytical/sampling needs. 

The research direction will be influenced by indoor environment research, such as 

indoor pollutant characterization and measurements. The research results will yield 

more accurate exposure assessments, better analytical tools for HHE sampling, and 

improved engineering control methods to reduce chemical contaminants.

Immune and Inflammatory Aspects To understand the mechanisms of occupational rhinitis in occupational safety,

of Occupational Rhinitis health, and medicine. A combined study design using human and animal research 

[Johnson] will be employed to identify the orthologously conserved pathways and gene 

networks that characterize the pathobiology of occupational rhinitis induced by 

diisocyanates. The outcomes of this research will benefit occupational safety and 

health through improved diagnosis and prevention of allergic airways disease 

caused by diisocyanates.

Genetics in Occupational Diseases To investigate susceptibility gene variants that contribute to the development and

[Yucesoy] severity of occupational irritant contact dermatitis (ICD) and asthma using high 

density and high throughput genotyping platforms. Previous and on going studies 

showed that cytokine polymorphisms have a major influence on silicosis, dementia, 

accelerated decline in lung function, and vaccine efficacy. Understanding the 

genetic contribution to the development, progression, and outcomes of complex 

occupational diseases will help improve the accuracy of risk assessment and 

improve safe exposure levels for genetically susceptible groups in the workforce.

Investigation of the Genetic Fingerprint of To determine if there are different carcinogen-specific chromosomal (genetic)

Chemically-induced and Spontaneously- markers in spontaneously-occurring and chemically-induced mouse lung

occurring Lung Cancer Using a Mouse Model adenocarcinomas. Mice were exposed by inhalation to vanadium pentoxide, nickel

[Reynolds] oxide, or cumene (a benzene derivative), three chemicals to which workers in the 

construction and manufacturing sectors are exposed. NIOSH will analyze mouse 

lung tumors induced by single-wall carbon nanotubes (SWCNTs). Results from 

these studies will be used to establish biomarkers for early detection and 

therapeutic intervention of lung cancer in workers.

Workplace Exposure, Inflammation, To investigate particle-respiratory tract exposure that has been related to increased 

and Cardiovascular Toxicity mortality from cardiovascular diseases. This laboratory-based research will evaluate 

[Simeonova/Erdely] novel molecular mechanisms involved in the link between occupational exposures 

to ultrafine/nanosize particulates and the development of cardiovascular diseases. 

Planned studies will help to identify potential risk factors, biomarkers, and specific 

targets for prevention and therapeutic intervention of occupationally related 

cardiovascular diseases.
18 National Toxicology Program

NCTR/NTP

The National Center for Toxicological Research (NCTR), the FDA’s research center, plays a critical role carrying 

out the agency’s mission. NCTR, in partnership with researchers from elsewhere in FDA, other government 

agencies, academia, and industry, provides innovative technology, methods development, vital scientific training, 

and technical expertise. The unique scientific expertise of NCTR is critical in supporting FDA product centers and 

their regulatory roles. NCTR conducts an array of studies that reflect the NTP mission statement. 

These NCTR/NTP studies, funded by NCTR voluntary allocations, are listed in Table 4.

Table 4: NCTR/NTP Projects in FY 2010* 

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

DNA Adduct Formation, Mutations, and Patterns (1) To treat Big Blue rats subchronically with riddelliine, AA, and comfrey using

of Gene Expression in Big Blue Rats Treated with procedures appropriate for tumor induction, (2) to analyze DNA adduct formation

the Botanical Carcinogens Riddelliine, in the target tissues for carcinogenesis and in spleen lymphocytes, (3) to determine

Aristolochic Acid (AA), and Comfrey the cIITM mutant frequencies and the types of cIITM mutations in the target tissues

[Chen] of treated rats, (4) to determine global gene expression patterns in the target and 

surrogate tissues of treated rats, and (5) to correlate gene expression patterns with 

DNA adduct formation and mutation induction in treated rats.

Further Evaluation of the Types of Genetic (1) To determine if the L5178Y/TK+/- MLA adequately detects both aneuploidy and

Events Detected by the Mouse Lymphoma mitotic recombination, (2) to determine if L5178Y mouse lymphoma cells have active

Assay (MLA) and the Role of the Assay in recombinase functions which lead to a large proportion of mutants that result from

Mechanistically-based Risk Assessment recombinase-mediated rearrangements, and (3) to determine the fundamental

[Moore] genetic mechanism(s) causing the small and large colony thymidine kinase 

mutant phenotypes.

NCTR/NTP
NTP Annual Report 2010 19

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Evaluation of Novel Genetic Changes and (1) To determine the post-translational protein modifications in the protein 

Post-translational Modification in the Protein extracts of nigral and striatal tissues in substituted amphetamines and

Products of Specific Genes in Parkinson’s 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice, (2) to

Disease and in Substituted Amphetamine evalulate the effect of various nNOS inhibitors and peroxynitrite decomposition 

Neurotoxicity Using Quantitative Proteome catalysts on the post-translational protein modifications in the protein extracts 

Analysis in Mice Models and Human Subjects of nigral and striatal tissues in mice treated with substituted amphetamines and 

[Ali] (3) to determine protein-DNA interactions in nuclear extracts from nigral and 

striatal tissues in mice treated with substituted amphetamines and MPTP for the 

evaluation of novel post-translational changes in the proteins mediated by various 

transcription factor, (4) to determine the effect of various nNOS inhibitors on 

substituted amphetamine and MPTP-induced free radical production and 

monoamine concentrations in mouse brains, (5) to determine the nitrated protein 

on tyrosine hydroxylase by immunoprecipitation of tyrosine hydroxylase and 

co-localization of 3-nitrotyrosine in the presence and absence of nNOS inhibitors 

to correlate physiological effects with protein changes from objectives 1, 2 & 3, and 

(6) to determine the post-translational protein modifications in protein extracts 

and protein-DNA interactions in nuclear extracts of nigral and striatal tissues 

obtained from human subjects with Parkinson’s Disease.

Effect of p53 Genotype on Gene-Expression (1) To determine the effect of mutation in the p53 tumor suppressor gene on

Profiles in Mice Exposed to the Model Mutagen, gene-expression profiles in young and aged mice and (2) to determine the effect

N-ethyl-N’-nitrosourea (ENU) of mutation in the p53 tumor suppressor gene on gene expression profiles in

[Morris] young and aged mice exposed to the model mutagen ENU.

Methods for Support of a Functional (1) To establish and standardize for routine use procedures for whole cell and 

Proteomics Facility at NCTR subcellular organellar isolation for a variety of tissues; (2) to develop and standardize

[Yu] specific and sensitive markers of cell type and organellar purity and yield; (3) to 

identify, adapt, develop, and standardize appropriate 2-D protein separation 

techniques; and (4) to integrate results of objectives 1-3 to provide “front-end” 

components of a functional proteomics facility.

Dietary Modulation of the Renal (1) To demonstrate that the cystic kidney disease previously shown to be induced 

Toxicity of p-Nonylphenol and by p-nonylphenol in developing NCTR CD rats fed a soy-free diet is decreased in 

Di(2-ethylhexyl)phthalate (DEHP) incidence and/or severity in rats fed soy-containing diets; (2) to evaluate the 

[Delclos] renal toxicity of dietary DEHP in developing rats maintained on a soy-free diet; 

(3) to evaluate potential early markers of renal cystogenesis in p-nonylphenoland DEHP-treated rats and their modulation by soy-containing diets; (4) to evaluate 

the roles of modulation of antioxidant defenses and cyclooxygenase activities in 

the protective effect of soy against p-nonylphenol and, if demonstrated, 

DEHP-induced renal toxicity; and (5) as secondary objectives in the above 

studies to assess the effect of diet on hepatic, testicular, and lung toxicity of DEHP.

Allele-specific Competitive Blocker (ACB) - (1) To use newly established PCR-based methods to quantify the rat K-ras codon 

Polymerase Chain Reaction (PCR) Measurement 12 GGT ‘ GAT and GGT ‘ GTT mutant fractions in rat colonic mucosa, aberrant crypt 

of Azoxymethane-induced Rat K-ras codon foci, and tumors at specified times after colon tumor initiation by azoxymethane 

12 GGT-->GAT and GTT-->GTT Mutations in treatment and (2) to use these data in conjunction with K-ras mutant fraction data 

Colonic Aberrant Crypt Foci Isolated using generated from studies of human colon to determine how rodent data can be 

Laser Capture Microdissection [McKinzie] extrapolated to human disease.

Analysis of p53 Codon 270 CGT to TGT Mutation (1) To develop the ACB-PCR detection of mouse p53 codon 270 CGT >TGT mutation,

in Simulated Solar Light (SSL)-induced Skin (2) to measure the frequency of detection and levels of this mutation in mouse 

Tumors and Exposed Mouse Skin skin tumors, (3) to measure the frequency of this mutation in skin tissue from 

[Parsons] tumor-bearing animals, and (4) to measure the frequency of this mutation 

in skin exposed to decreasing levels of SSL.

Measurement of Cancer-associated Gene (1) To determine K-ras codon 12 GGT to GAT and GGT to GTT mutant frequencies 

Mutation in Colon Tumor and Nontumor Tissue in colonic aberrant crypt foci, adenomas, and carcinomas; first by DNA sequencing 

[Parsons] and, if mutation is not detected, then by ACB-PCR; (2) to determine K-ras codon 

12 GGT to GAT and GGT to GTT mutant frequencies in tumor-margin samples and 

tumor-distant, normal-appearing colonic epithelium from colon cancer patients; 

and (3) to determine K-ras codon 12 GGT to GAT and GGT to GTT mutant frequencies

in autopsy samples of colonic epithelium from colon disease-free individuals.
20 National Toxicology Program

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Assessment of Interindividual Variability in 1) To determine levels of expression of DNMT3a and DNMT3b in liver samples

Expression of DNA Methyltransferases (DNMT), from a pool of donors selected according to smoking status, gender, and age;

DNMT3a, and DNMT3b, in Liver and Identification (2) to determine the effect of tobacco smoke on DNMT1, 3a, and 3b expression

of Factors Influencing Expressions in liver cell systems; and (3) to assess the polymorphism frequency identified in

[Hammons] DNMT3b in the sample pool included in the study and assess whether it is 

correlated with expression.

Can Polymorphism in Human DNMT3b (1) To develop plasmid constructs with CC wildtype and TT homogenous variant of

Potentially Affect Individual Responses to DNMT3b to examine transcriptional activity, (2) to use activity assays to compare 

Xenobiotics and Chemopreventive Agents? transcriptional activity of gene constructs transfected into liver cells, and (3) to 

[Hammons] determine whether xenobiotics or dietary agents affect transcriptional activity 

differently in CC, CT or IT transfected cells.

Global and Locus-specific DNA (1) To determine if the temporal alterations in genomic methylation profile in 

Hypomethylation: A Common Mechanism preneoplastic liver tissue observed in the folate/methyl-deficient model of rat 

Involved in Genotoxic and Non-genotoxic endogenous hepatocarcinogenesis also occur in other carcinogenesis models; 

Rat Hepatocarcinogenesis (2) to identify genes that are consistently up-regulated or down-regulated in target 

[Pogribny] tissue during the promotion stage of carcinogenesis; and (3) to evaluate whether 

the global and locus-specific DNA hypomethylation, along with aberrant expression 

of related genes and changes in chromatin conformation, is specific only to target 

tissues and if may be used for early detection of chemicals with carcinogenic potential.

Carcinogenicity of Acrylamide and its Metabolite To compare the carcinogenicity of acrylamide and its metabolite glycidamide in

Glycidamide in Rodents: Neonatal Mouse Bioassay B6C3F1 mice treated neonatally.

[Beland]

Development of MitoChip, a Glass-based (1) To develop a MitoChip, a GeneChip Mitochondrial Resequencing Array, containing 

Oligonucleotide Microarray Containing genes associated with mitochondrial function such as oxidative phosphorylation, 

Mitochondrial and Nuclear Genes B-oxidation of free fatty acids, tricarboxylic acid cycle, apoptosis, as well as genes 

Associated with Mitochondrial Function involved in the replication, transcription, translation of mitochondrial DNA, DNA repair, 

[Desai] and regulation of DNA copy number; (2) to validate the MitoChip by evaluating gene 

expression profiles of AZT, an anti-HIV drug, and 3-nitropropionic acid, neurotoxins 

known to alter mitochondrial function; and (3) to verify the relative expression levels 

of differentially expressed genes by real-time quantitative PCR.

Assessment of Ketamine in the (1) To determine, using neurohistochemical approaches, if, and at what developmental

Developing Nonhuman Primate stages, ketamine exposure increases neuronal apoptosis/proliferation; (2) to determine, 

[Wang] using neurohistochemical approaches, the dose-response for ketamine to produce 

apoptosis at the most sensitive developmental stage; (3) to determine the 

reversibility or permanence of the response using behavioral, imaging, and 

neurohistochemical approaches; and (4) to determine, at the most sensitive 

stage and dose, genomic and proteomic responses to ketamine treatment.

Preclinical Metabonomic Biomarkers To examine the utility of metabonomics as an approach to produce predictive 

of Toxicity and Disease models of cardiovascular, renal, neural, and hepatic toxicity. The models will be 

[Beger] built using a variety of pattern-recognition technologies to determine how temporal 

endogenous metabolic changes found in nuclear magnetic resonance (NMR) and/or 

mass spectrometry (MS) spectra of urine, serum, and tissue are related to toxicity 

and disease state.

Phosphatidylinositol Glycan - Complementation (1) To develop flow-cytometric methods for the detection of cells with mutations 

Group A (PIG-A) Mutagenesis: Development of in the PIG-A gene using wild-type and mutant human lymphoblastoid cells, TK6,

Methods for the Identification and Molecular and WTK1, as a model and (2) to develop flow cytometric methods for the detection

Characterization of Mutations in the PIG-A Gene of hematopoietic cells with mutations in the PIG-A gene in C57Bl/6 mice.

in Human Lymphoblastoid Cells and C57Bl/6 Mice 

[Dobrovolsky]

PIG-A Mutagenesis: Development of (1) To develop methods for measuring PIG-A mutant frequencies in lymphocytes 

Methods for the Identification and Molecular cultured from rats, (2) to develop flow cytometric methods for measuring PIG-A 

Characterization of Mutations in the PIG-A mutant/variant frequencies in both the lymphocytes and red blood cells of 

Gene of F344 Rats rats, (3) to treat rats with either ENU or 7,12-dimethylbenz[α]anthracene and 

[Heflich] measure PIG-A mutant manifestation in lymphoctyes and compare results with 

PIG-A variant manifestation in red blood cells and Hprt mutant manifestation in 

lymphocytes, and (4) to determine the spectra of the PIG-A and Hprt mutations 

induced in the mutant lymphocytes detected in Objectives 1, 2 and 3.
NTP Annual Report 2010 21

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Phytoestrogens and Aging: Dose, (1) To evaluate the potential benefits or detrimental effects of dietary 

Timing, and Tissue phytoestrogens on breast cancer progression, adipose tissue, and the brain, 

[Doerge] using well-established laboratory animal models and (2) to complete additional 

studies to determine bioavailability of soy isoflavones in neonatal and adult male 

non-human primates.

Development of a Physiologically-based (1) To develop a PBPK/PD model for acrylamide and glycidamide, (2) to determine 

Pharmacokinetic/Pharmacodynamic (PBPK/PD) the mutagenicity of acrylamide and its metabolite glycidamide in Big Blue rats, 

Model for Acrylamide and (3) to determine the DNA adduct levels and the extent of mutagenicity of furan

[Doerge] and its metabolite cis-2-buten-4-dial in neonatal B6C3F1/Tk+/- mice.

Development of Liver Toxicity Knowledge To develop a Liver Ontology that characterizes liver pathology and toxicity. 

Base (LTKB) to Empower the FDA The ontology will be used as guidance for data collection/curation, classification 

Review Process and analysis with the following specific aims: (1) gene expression data – the primary 

[Tong] data collected for this project will be existing gene expression data. Other types 

of data such as data from proteomics, metabonomics, and genotyping studies 

(including genome-wide association studies) will be considered as the project 

progresses. (2) text mining – NCTR will conduct text mining on >13 million abstracts 

in PubMed and other public resources with an emphasis on liver-related data. 

The association between the liver-specific entities (i.e., genes/proteins, pathways, 

drugs, tissues and toxicity) will be established. (3) known data – there is substantial 

knowledge available in public domains on liver toxicity, including genes/proteins 

(e.g., signatures and biomarkers), pathways/networks, and chemicals/drugs. 

NCTR will assemble this knowledge in such a way that it can be integrated with 

other information in LTKB and effectively mined. (4) experiment – NCTR will 

conduct gene expression studies on well understood and characterized hepatic 

and non-hepatic compounds. The dataset will be used to validate the LTKB. Data 

from 50 compounds will be generated in the first year of this project and will serve 

as proof of principle. Data for 150 additional compounds will be collected in the 

following two years, assuming the first set of data supports the LTKB approach. 

(5) LTKB - the data/information generated from the specific aims will be analyzed 

(combined and correlated) to establish liver toxicity-related regulatory networks 

and genes/proteins-pathways-chemicals-disease associations. A set of training 

rules will be identified to minimize the false positives in LTKB.

Evaluation of the Genetic Toxicity and (1) To determine the baseline frequency of measures of genetic damage in a 

Behavioral Effects of Chronic Methylphenidate population of juvenile rhesus monkeys, (2) to determine the frequency of these 

Exposure in Juvenile Male Rhesus Monkeys measures of genetic damage in a population of juvenile rhesus monkeys at defined 

(Macaca mulatta) intervals during a chronic exposure to methylphenidate, (3) to determine if chronic 

[Morris] exposure to methylphenidate results in measurable effects on the behavior of 

juvenile rhesus monkeys utilizing the NCTR Operant Test Battery, and (4) to 

determine the plasma concentration of methylphenidate and its major metabolite 

ritalinic acid during the chronic exposure of juvenile rhesus monkeys to the drug.

Detection of DNA Adducts in Mice Treated To define dose-response curves for benzo[a]pyrene DNA adducts in the

with Benzo[a]pyrene at Low Exposure Levels A/J mouse lung, utilizing the application of HPLC-ES-MS/MS methodologies

[Fu] developed at NCTR.

Biotransformation of Isoflavonoid To use fecal samples of monkeys and rodents to determine if the metabolites 

Phytoestrogens by Colonic Microfloras of produced by intestinal microfloras of experimental animals exposed to 

Experimental Animals phytoestrogens are the same as those of humans or whether the animal-colonic 

[Rafii] bacteria metabolize them to different compounds. This information is necessary 

for extrapolation to humans of the data obtained from treatment of animals 

with phytoestrogens.

Molecular Mechanisms Underlying To elucidate molecular mechanisms of mitochondrial dysfunction that will address 

Gender-associated Differences in the Adverse gender-based differences in adverse effects of antiretroviral drugs such as AZT. 

Reactions to the Antiretroviral Agent, This will provide critical information to the FDA for development of guidelines 

Zidovudine (AZT): Role of Mitochondrial Toxicity to plan new treatment strategies to reduce the frequency and severity of 

[Desai] antiretroviral-related toxic effects in women, particularly in pregnant women.
22 National Toxicology Program

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Neurotoxicity Assessment of Manganese (Mn) (1) To evaluate the neurotoxicity of different size Mn nanoparticles using PC 12

Nanoparticles in PC12 Cells and in Mice cultured cells; (2) to determine if in vitro exposure to Mn nanoparticles selectively 

[Ali] induces specific genomic changes in PC12 cultured cells using oligonucleotide 

microarrays; (3) to determine if multiple doses of Mn nanoparticles produce ROS, 

alterations in lipid peroxidation and/or changes in antioxidant enzymes (catalase,

superoxide dismutase, glutathione peroxidase), and levels of glutathione in various

regions of the mouse brain; (4) to determine if single or multiple doses of Mn 

nanoparticles induce specific genomic changes in various regions of the mouse 

brain using oligonucleotide microarrays; (5) to determine if single or multiple 

doses of Mn nanoparticles produce significant changes in neurotransmitter 

concentrations in various regions of the mouse brain; and (6) to determine if 

single or multiple doses of Mn nanoparticles produce significant changes in the 

formation of 3-nitrotyrosine, an in vivo biomarker for oxidative stress, in various 

regions of the mouse brain.

Methods Development for High-resolution Advances in pediatric and obstetric surgery have resulted in an increase in 

Dedicated Positron Emission Tomography complexity, duration, and number of anesthetic procedures. To understand the 

(microPET) to Rodent Neuroplasticity and effects of anesthetic drugs on the developing nervous system and to minimize 

Toxicity During Development risks to children resulting from the use of anesthesia, this study will utilize microPET 

[Wang] to screen and evaluate in vitro and in vivo measurements from a broad range 

of pathophysiological or pharmacological parameters using specific tracers in the 

developing rat. Three different age groups of developing rats will be used: pregnant 

day 18 female rats, postnatal day 7 rat pups, and postnatal day 35 rats. This study will 

also attempt to elucidate the relationship between apoptosis-identifying ligands 

(specific tracers) and subsequent behavioral deficits.

Cancer Mutations as Biomarkers of (1) To develop the information necessary for the rational use of oncogene mutations 

Cancer Risk: Human Studies with as quantitative biomarkers of cancer risk using ACP-PCR to determine normal and 

Implications for Personalized Medicine pathological levels of relevant oncogene mutations in multiple human tissues 

[Parsons] and tumors; (2) to compare the information derived from human tissues with 

data generated in a parallel rodent protocol as an approach for incorporating 

carcinogenesis-relevant data into the rodent to human extrapolation necessary 

in cancer risk assessment; (3) to validate a streamlined ACP-PCR methodology and 

develop the methodology necessary to measure oncogene mutation frequency 

in cell-free DNA isolated from plasma; and (4) to convey to the regulatory riskassessment community, through a series of publications, the regulatory 

significance of the data regarding tumor-associated mutations which have 

and will be generated.

Liver Toxicity Biomarkers Study: To establish liver toxicity biomarkers and associated algorithms for use in preclinical

Phase 1, Entacapone and Tolcapone drug development that will predict the probability of occurrence of hepatocellular

[Beland] injury at any subsequent phase of drug development or following approval of the

drug for marketing. Emphasis will be placed upon drugs that do not demonstrate

“classical” signs of liver toxicity during preclinical stages of drug development.

Development of a New Safety Evaluation (1) To determine miRNA expression profiles of the tumor target tissues of rats 

Method Using MicroRNA (miRNA) and mice treated with genotoxic carcinogens AA, riddelliine, and comfrey; 

Expression Analysis as a Biomarker and non-genotoxic carcinogens, propiconazole, and triadimeforn, as well as 

for Detecting Carcinogens non-carcinogen myclobutanil using microarray technologies; (2) to develop a PCR 

[Chen] array containing the primers that are specifically used to amplify carcinogensis-related

miRNAs and use the PCR array to conduct time-course and dose-response studies 

for miRNA expression alterations in tissues of rats treated with carcinogens; and 

(3) to define the miRNA biomarker genes that are associated with carcinogen 

exposure by prediction of their target genes and determination of their 

biological functions.
NTP Annual Report 2010 23

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Neurotoxicity Assessment of Silver (Ag) (1) To evaluate the neurotoxicity of different sizes of Ag nanoparticles using cultured

Nanoparticles in PC-12 Cells and in Rats PC12 cells; (2) to determine if in vitro exposure to Ag nanoparticles selectively 

[Ali] induces specific genomic changes in cultured PC12 cells using microarrays; (3) to 

determine if single or multiple doses of Ag nanoparticles produce ROS, alterations 

in lipid peroxidation, and/or changes in antioxidant enzymes (catalase, superoxide 

dismutase, glutathione peroxidase) and glutathione levels in the rat brain; 

(4) to determine if single or multiple doses of Ag nanoparticles induce specific 

genomic changes in the rat brain as indicated with microarrays; (5) to determine 

if single or multiple doses of Ag nanoparticles produce significant changes in 

neurotransmitter concentrations in the brain in rat; (6) to determine if single or 

multiple doses of Ag nanoparticles produce significant changes in the formation 

of 3-nitrotyrosine, an in vivo biomarker for oxidative stress, in the rat brain; and 

(7) to determine if multiple doses of Ag nanoparticles produce morphological 

alterations in the blood-brain barrier, brain, or other visceral organs of the rat.

Development of High Throughput Methodology (1) To design high throughput methods for detecting PIG-A mutant human red 

for Detection of In Vivo Mutation and white blood cells by flow cytometric detection of cells lacking cell surface 

in the Endogenous PIG-A Gene protein markers anchored by glycosyl phosphatidyl inositol (e.g., CD59, CD48); 

of Human Blood Cells Using Flow Cytometry (2) to use the methods developed in Objective 1 to establish a normal range of 

[Dobrovolsky] PIG-A mutant frequencies in red and white blood cells and compare these ranges 

with those of different groups of human subjects hypothesized to have increased 

mutational loads. The groups will include, but will not be limited to (a) patients with 

the disease paroxysmal nocturnal hemoglobinurea, (b) patients undergoing 

radiation treatment or chemotherapy with DNA reactive drugs, and (c) patients 

predisposed to cancer due to inherited deficiencies in endogenous pathways; and 

(3) to compare, when the volumes of blood samples permit, red blood cell PIG-A 

mutant frequencies determined in Objective 2 with PIG-A mutant frequencies in 

white blood cells from these samples determined by limiting-dilution cloning, and 

to determine the PIG-A DNA sequence changes reponsible for the white blood 

cell mutants.

Evaluation of the Ability of Both the Agar To develop science-based best practice standard and tools to incorporate

and Microwell Versions of the MLA to Optimally translational and applied toxicological advancements into the regulatory

Detect the Mutagenic Potential and Potency of science process to create a seamless bench-to-bedside continuum.

Complex Chemical Mixtures 

[Moore]

Assessment of Gaseous Anesthetics in the (1) To evaluate dose-response effects of gaseous anesthetics to determine 

Developing Nonhuman Primate if prolonged exposure to nitrous oxide or isoflurane alone will result in an increase 

[Wang] in neuronal cell death and to determine if combinations of nitrous oxide and 

isoflurane will prevent or enhance each other’s effects on the developing nonhuman 

primate; (2) to determine if a relative high dose or prolonged exposure of 

developing nonhuman primates to nitrous oxide or isoflurane alone or in 

combination will induce long-term behavioral deficits, as well as long-lasting 

pathological changes; (3) to determine, using noninvasive imaging techniques 

[microPET and magnetic resonance imaging (MRI)], if a high dose or prolonged 

exposure of developing nonhuman primates to nitrous oxide or isoflurane alone 

or in combination will induce long-lasting pathological changes. MRI will be used 

to verify pathological evidence and look for volume changes. MicroPET will be 

used to examine the sensitivity for tracing low picomolar concentrations of 

radiolabeled molecules, which is useful for studying dynamic imaging in animal 

models of human diseases; and (4) to identify potential underlying mechanisms 

that could link alteration of mitochondrial function and elevation of ROS to gaseous 

anesthetic-induced neuronal cell death. L-carnitine will be used to attenuate 

neurological brain injury associated with mitochondria-related degenerative effects

induced by gaseous anesthetics in the developing nonhuman primate.

Evaluation of Growth and Pubertal To evaluate changes in pharmacokinetics and operant behavior testing in male 

Development In Male Rhesus Monkeys monkeys chronically exposed to MPH. The initial experiment was designed to 

(Macaca mulatta) Chronically Exposed to examine the genetic toxicity associated with chronic MPH treatment. This project 

Methylphenidate Hydrochloride (MPH) will begin after the completion of the genetic toxicity experiments, to avoid 

[Salminen] procedures that could compromise those genetic toxicity data, specifically 

anesthesia and x-ray radiation.
24 National Toxicology Program

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Development of Methods for Evaluating DNA To evaluate and establish methods and conditions that enhance the sensitivity and 

Damage using Single Cell Gel Electrophoresis reproducibility of the in vivo alkaline-comet assay for use in preclinical hazard 

(Comet Assay) in Rodents identification and genotoxicity testing of food ingredients and chemicals

[Aidoo] for regulatory purposes.

Laboratory Studies in Melamine and To determine chemical and biochemical properties of melamine and cyanuric acid 

Cyanuric Acid Biochemical Toxicology that may influence their toxicity and retention as tissue residues.

[Tolleson]

Characterizing the Amphetamine-Induced (1) To evaluate the effects of both acute and chronic amphetamine (AMPH) 

Changes in Vascular Tone, Vasotrauma exposure on the vasculature of the rat brain; (2) to examine vasculature within the 

and Alterations in Angiogenesis parenchyma of three brain regions: the striatum, parietal cortex, and the combined 

in Rodent Brain piriform and amygdaloid nuclear cortices where AMPH-induced neurodegeneration

[Bowyer] can occur; (3) to observe the effects of AMPH on the vasculature associated with pial

and arachnoid membranes (part of the meninges) and vasculature of the choroid 

plexus; and (4) to determine the alterations in AMPH-induced vascular gene 

expression after (a) 1-day, 4-dose exposure that produces hyperthermia [full time 

course of effects, not just first day], (b) a single exposure to a very high dose, 

(c) 9-day exposure that does not produce significant hyperthermia, and 

(d) 1-month exposure to AMPH included in the drinking water.

Chemical Inactivation of Protein Toxins (1) To identify cleaning/sanitizing treatments that result in elimination and/or 

on Food Contact Surfaces inactivation of protein toxins (abrin and ricin) on food contact surfaces, (2) to identify

[Tolleson] surrogate(s) that can be used to study chemical inactivation of abrin or ricin, and 

(3) to measure the loss of ricin and abrin biological and biochemical activities in the 

presence of cleaning/sanitizing solutions using RAW264.7 macrophage cytotoxicity 

assays and 28S rRNA adenosine N-glycosidase RTqPCR-based enzyme assays.

Method Development for Study of Microsomal Metabolism-Mediated Studies: (1) To determine whether or not the 

Antioxidant Properties in Dietary Supplements studied herbal dietary supplements can enhance or inhibit free radical formation, 

[Fu] mediated by microsomal metabolism, in a dose-dependent manner and 

(2) to determine whether or not the studied herbal dietary supplements can 

enhance or inhibit microsomal metabolism-mediated lipid peroxidation in a 

dose dependent manner. Cell Culture Studies: (1) To determine the toxic effects, 

including mitochondrial dehydrogenase activity, intracellular ROS concentration, 

and mitochondrial membrane potential, of the studied herbal dietary supplements 

in cells, including A549 human lung carcinoma cells and rabbit brain rBCECs cells 

(a normal cell line to assay the toxic effect on the CNS) and (2) to determine, by use

of the ESR oximetry technique, the inhibition/induction of lipid peroxidation by the 

studied herbal dietary supplements in A549 human lung carcinoma cells and 

rabbit brain rBCECs cells.

Use of Electron Spin Resonance (ESR) Chemical Reactions: (1) To determine whether or not nanomaterials can catalyze 

Spectroscopy to Characterize the a Fenton reaction to initiate hydroxyl radical formation in a nanoparticle size 

Interactions Between Nanoscale Materials dependent manner and (2) to determine whether or not nanomaterials and/

and Model Biological Systems or their cations can be reduced by natural reducing agents, such as ascorbic acid 

[Fu] and glutathione, leading to the formation of ROS. Microsomal MetabolismMediated Studies: (1) To determine whether or not nanomaterials enhance 

or inhibit free radical formation, mediated by microsomal metabolism, in a 

nanoparticle size-dependent manner and (2) to determine whether or not 

nanomaterials and/or their cations can enhance or inhibit microsomal metabolismmediated lipid peroxidation in a nanoparticle size-dependent manner. Cell Culture

Studies: (1) To determine the toxic effects, including mitochondrial dehydrogenase

activity, intracellular ROS concentration, and mitochondrial membrane potential, 

of nanomaterials of different particle size in cells, including A549 human lung 

carcinoma cells and rabbit brain rBCECs cells (a normal cell line to assay the 

toxic effect on CNS) and (2) to determine, by use of the ESR oximetry technique, 

the inhibition/induction of lipid peroxidation by nanomaterials of different 

particle size in A549 human lung carcinoma cells and rabbit brain rBCECs cells.
NTP Annual Report 2010 25

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Evaluation of the Applicability of Standard (1) To assess the genotoxicity of four types of nanoscale materials, carbon nanotubes, 

Assays for Assessing Genotoxicity of nanoscale titanium dioxide, nanoscale gold, and nanoscale silver in three standard 

Engineered Nanomaterials tests suggested by the FDA, Salmonella Ames test, mouse lymphoma assay, and 

[Chen] in vivo mouse micronucleus assay and (2) to evaluate the possible mechanisms of 

nanomaterial-induced genotoxicity using a transgenic mutation system, comet 

assay, and genomic analysis.

Neurotoxicity Assessment of Carbon Nanotubes (1) To evaluate the neurotoxicity of single and multiple-wall and different sizes of 

(CNTs) and Gold Nanoparticles using a Brain CNTs using brain microvascular endothelial and PC12 cultured cells; (2) to determine, 

Microvascular Endothelial Cell System, PC12 using microarrays, if the in vitro exposure to these CNTs (single and multiple walls) 

Cultured Cells, and a Whole Animal Model would selectively induce specific genomic/proteomic changes in brain microvascular

(Rats or Mice) endothelial and PC-12 cultured cells; (3) to determine if acute or chronic exposure 

[Ali] of CNTs would produce ROS, alterations in lipid peroxidation and/or changes in 

antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase) and 

glutathione levels in mouse brain; (4) to determine if acute or chronic exposure of 

CNTs would induce specific genomic/proteomic changes in mouse brain as 

indicated with microarrays; (5) to determine if acute or chronic exposure of 

CNTs would produce significant changes in neurotransmitter concentrations, 

formation of 3-nitrotyrosine, an in vivo biomarker for oxidative stress, in mouse 

brain; and (6) to determine if acute or chronic exposure of CNTs would produce 

morphological alterations in the brain or visceral organs of the mice.

Effect of Urinary pH upon the Nephrotoxicity To determine the effect of urinary pH upon the renal toxicities elicited by a combined

of a Combined Exposure to Melamine and exposure of melamine and cyanuric acid.

Cyanuric Acid 

[Beland]

Assessment of the Nephrotoxicity of a To investigate the nephrotoxic effect of a seven-day co-exposure to melamine

Seven-day Combined Exposure to Melamine and cyanuric acid in Fischer 344 rats.

and Cyanuric Acid 

[Gamboa da Costa] 

Methylphenidate (Ritalin) Exposure during (1) To quantify the neurobehavioral toxicity associated with pre- and early postnatal 

Pregnancy: Assessment of Neurotoxicity treatment with methylphenidate in rats; (2) to evaluate a wide range of behaviors 

in Offspring at preweaning, adolescent, and adult ages to fully describe the potential effects; and

[Ferguson] (3) to evaluate the hypothesis that, at human-relevant serum levels of methylphenidate, 

alterations in baseline locomotor activity as well as locomotor response to a later 

methylphenidate challenge may be produced, and that higher doses will produce 

more significant effects.

Biomarkers of Liver Toxicity (1) To discover biomarkers of hepatotoxicity in preclinical studies that are more 

[Salminen] predictive of adverse effects in humans [these biomarkers may or may not be 

directly applicable to the clinic, but they must be predictive of human responses 

so that they can be used to extrapolate preclinical data to humans in safety 

assessment] and (2) to publish and make the biomarkers publicly available to 

fulfill FDA’s goal of developing science-based best practice standards, guidance, 

and tools to improve the regulatory processes. Farther-reaching goals include the 

qualification of such biomarkers (e.g., via the FDA/ European Medicines Agency 

qualification process) and potential translation for clinical use.

Development of Predictive Mitochondrial (1) To perform non-invasive measurements of heart rate, heart rate variability, and 

Biomarkers for Drug-induced Cardiotoxicity electrocardiogram using ECGenieTM; (2) to measure cardiac troponin T, creatine 

Using a System Biology Approach kinase MB, and cardiolipin levels in plasma as indicators of doxorubicin-induced 

[Desai] cardiac tissue damage; (3) to identify morphological changes in cardiac mitochondria 

in the left ventricular region by electron microscopy; (4) to use “omics” technologies

for analyte profiling in the heart, transcriptional profiling of approximately 906 

mitochondria-related genes using MitoChip, protein profiling by 2D-HPLC/MS/

MS, and measurement of endogenous metabolites by nuclear magnetic resonance 

(NMR) and MS; (5) to measure expression levels of 906 mitochondria-related genes 

in whole blood using MitoChip; (6) to measure levels of creatinine, creatine, lactate, 

Krebs cycle intermediates, and small ketone bodies in plasma using metabolomics; 

and (7) to integrate genomic, proteomic, and metabolomic endpoints in the heart 

tissue to define the molecular basis of doxorubicin-induced cardiac toxicity and 

to correlate “omics” data to genomic findings obtained in whole blood.
26 National Toxicology Program

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Development of an FDA Resource and (1) To develop a knowledge base for the sex differences in DILI through analyzing , 

Knowledge Base for Sex Difference in and modeling the molecular data in public domain; (2) to further augment the 

Drug-Induced Liver Injury (DILI) collection of the genomic data from public resources and through collaborations; 

[Tong] (3) to develop a standard data curation model for the sex-biased DILI in ArrayTrack 

to manage the collected data [the data curation model will be developed in 

accordance with the data standard developed in FDA for electronic data 

submission]; (4) to conduct the meta-analysis, text mining, and network analysis 

to develop a relationship between drugs, molecular signatures, liver-specific 

biomarkers, genes/proteins functions, pathways and sex-biased liver toxicity; 

(5) to gain understanding, at molecular level, of sex-biased differences in DILI; 

and (6) to provide knowledge and resources from this project for the FDA to utilize 

and reference when sex-related DILI issues arise during the various stages of the 

regulatory review process, thus impacting women’s health.

Collaborative Inter-agency Development (1) To develop in silico computational models to predict chemical interactions that 

of In Silico Computational Toxicology can inhibit CYP3A4 and CYP2D6 enzymes, (2) to have each government agency 

Modeling to Predict Adverse Drug/Chemical build and test a different type of in silico computational model to predict chemical 

Interactions with Cytochrome p450 Enzymes interactions that can inhibit these enzymes, and (3) to use several different 

[Beger] modeling methods to model compound-compound-enzyme interactions because 

no modeling technique is perfect and each technique has strengths and weaknesses

compared to other modeling techniques. NCTR will build structure-activity 

relationship (SAR) and spectrometric data-activity-relationship (SDAR) models 

of chemical interactions that inhibit CYP3A4 and CYP2D6 enzymes. CDER will 

build predictive quantitative SAR (QSAR) models with a training set of drugs and 

chemicals that inhibit CYP3A4 and CYP2D6 reversibly and irreversibly. ATSDR will 

build docking models of chemicals that contaminate drinking water and interact 

with CYP3A4 and CYP2D6 enzymes. These in silico models will be built with 

chemicals that have been reported to inhibit these P450 enzymes. Some of the 

chemicals used to form these models have been found in drinking water so these 

interactions could happen without polypharmacy. Cytochrome CYP3A4 and 

CYP2D6 are two of the major phase I drug metabolizing enzymes and drugs 

or chemicals that inhibit them could alter drug pharmacokinetics and associated 

drug toxicity or drug efficacy endpoints. Understanding these drug-enzyme 

inhibitory interactions and having the ability to accurately predict these 

interactions should drastically improve drug safety evaluation and increase 

protection of public health.

3D- and 4D-QSDAR Modeling Applied (1) To develop 3D- and 4D-QSDAR models for endocrine disruptors, lowestto Various Toxicological Endpoints observed-adverse-effects level (LOAEL), and no observed-adverse-effects level 

[Beger] (NOAEL), and other relevant toxicological endpoints; (2) to test the training models 

with external test sets and compare the training and testing results to previous 

QSDAR, quantitative structure-activity relationship (QSAR), and SAR models; and 

(3) to determine how the technique can be used to predict how 13C or 15N NMR 

spectra affects 3D-QSDAR modeling.

Relationship between Liver Epigenetic (1) To determine the role of epigenetic dysregulation in the etiology and 

Phenotype and Susceptibility to Nonalcoholic pathogenesis of dietary NASH-induced hepatocarcinogenesis in mice, 

Steatohepatitis (NASH)-induced 2) to determine whether or not interstrain-specific susceptibility of mice to 

Hepatocarcinogenesis in Mice NASH-induced hepatocarcinogenesis is associated with differences in individual 

[Pogribny] hepatic epigenetic phenotypes, (3) to determine the role of epigenetic dysregulation

in the etiology and pathogenesis of NASH-induced hepatocarcinogenesis in mice 

induced by tamoxifen administration, and (4) to determine whether or not aberrant 

epigenetic markers can be used as targets for prevention of NASH-induced 

hepatocarcinogenesis in mice.
NTP Annual Report 2010 27

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Methods Development for Toxicity Assays (1) To establish a high throughput assay system using zebrafish embryos for toxicity

using the Zebrafish Embryo as a Model System: assessments of FDA relevant compounds; (2) to monitor both traditional morphological

Whole Animal High Throughput Assays and behavioral endpoints of toxicity and the newer, more subtle organ-specific

for Chemical Testing toxicities; (3) to study the effect of methamphetamine on zebrafish embryos, 

[Kanungo] especially relating to sensory and motor neuron development; 4) to determine if

carbon nanotubes pass through the blood brain barrier in zebrafish embryos and 

have any toxic effects on early development; (5) to determine whether these 

nanomaterials generate ROS, cause the depletion of dopamine and its metabolites, 

dihydroxyphenylacetic acid and homovanillic acid, and alter markers of oxidative 

stress; and (6) to study the effect of nicotine on zebrafish embryos, especially 

relating to sensory and motor neuron development and the mechanism of action.

Long-term Consequences of Neonatal (1) To continue monitoring the cognitive capabilities of rhesus monkey subjects 

Ketamine Anesthesia in Rhesus Monkeys: exposed to a single, 24-hour bout of ketamine-induced anesthesia during the first 

Extended Congnitive Assessments week of life and (2) to extend the functional domains that are being assessed to 

[Kanungo] include performance of a temporal discrimination task (timing task), a counting 

task, and reversal learning tasks (cognitive flexibility). Data to date indicate that, 

compared to control animals, ketamine-exposed subjects exhibit significant deficits

in several aspects of brain function including learning, the ability to perform 

simple visual discriminations, motivation and speed of psychomotor processing. 

Continuing these observations will provide valuable information on the ultimate 

time course and severity of the observed deficits.

Pharmacokinetics of Aurimune (NCL74) (1) To conduct pharmacokinetic studies for various Nanotechnology 

and Docetaxel (Taxotere) in Characterization Laboratory-supplied nanomaterials in nonhuman primates and 

Nonhuman Primates (2) to perform measures of toxicity, specifically immunotoxicity, that are amenable 

[Matson] to blood analyses (e.g., clinical chemistries, hematology, complement activation, 

and cytokine profiles).

Assessing Acetaminophen-induced Liver Injury (1) To identify acetaminophen-dietary supplement interactions to allow warnings 

and the Influence of Dietary Supplements – to be provided to women and better assessment of the safety of the current 

Potential Synergistic Interactions therapeutic uses of acetaminophen and the protection afforded by the U.S. dietary

[Salminen] supplement regulations, (2) to identify differential sex-sensitivity to acetaminophen 

in combination with dietary supplements, and (3) to analyze biological pathways 

that are affected by the dietary supplements and acetaminphen individually and 

when combined to provide insight into the mechanism(s) of the interaction 

and why females are more sensitive to acetaminphen DILI.

Quality Control (QC) for Focused and (1) To develop and test a QC protocol on existing preclinical hepatotoxicity protocols 

Unfocused LC-MS-based Metabolomic (acetaminophen-induced injury and the influence of dietary supplements; liver 

Profiling of Blood Samples toxicity biomarkers) that can potentially be translated to clinical metabolomics 

[Bhattacharyya] experiments that are planned and (2) to test the hypothesis that adding QC will 

improve both intra-lab and inter-lab reproducibility of LC-MS-based metabolomics 

of blood samples.

Assessment of Iron Oxide Nanoparticle-induced (1) To determine if acute or chronic exposure of different sizes to iron oxide 

Neurotoxicity in Cell Cultures and Whole nanoparticles produces specific changes in the mitochondrial function, cell death, 

Animal Models and generation of ROS in different regions of rat and mice brain using in vivo

[Binienda] microdialysis, (2) to determine if acute or chronic exposure to iron oxide 

nanoparticles produces significant changes in neurotransmitter concentrations 

in different regions of mice/rat brains using microdialysis, (3) to determine if acute 

or chronic exposure to different sizes of iron oxide nanoparticles produce alterations 

in the brain free fatty acid levels, (4) to determine if acute or chronic exposure to 

different sizes of iron oxide nanoparticles produces alterations in lipid peroxidation 

and/or changes in antioxidant enzyme activity (catalase, superoxide dismutase, 

glutathione peroxidase) and glutathione levels in mice and rat brains, and (5) to 

determine if acute or chronic exposure to different sizes of iron oxide nanoparticles 

produces selective patterns of deposition and damage in different regions of rat 

and mice brain using in vivo MRI.
28 National Toxicology Program

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

Development and Application of a (1) To develop MitoChips for various mammalian species, including rat, non-human 

Mitochondria-specific Gene Array (MitoChip) primate, and human; (2) to perform transcriptional profiling of mitochondria-related 

for the Investigation of Clinical and Non-clinical genes using mitochondria-specific gene arrays to investigate the mechanisms 

Predictive Biomarkers of Toxicity of drug toxicities and degenerative diseases associated with mitochondrial 

[Desai] dysfunction in different mammalian species; and (3) to characterize species-specific 

transcriptional profiles to predict risk of drug toxicity or disease onset in different 

mammalian species.

Evaluation of Transgenic gpt-delta Mice (1) To dose by gavage daily for 28 days groups of gpt-delta transgenic 

developed at NCTR Breeding Colony for mice (8B-NCTR Breeding colony) with low, intermediate, or high doses of 

Detecting Point Mutations and Large Deletions cyclophosphamide, bleomycin, and 100 mg/kg ENU [as a positive control for 

[Manjanatha] induction of mutations in the transgenes gpt, red, and gam (spi- selection)]; 

(2) to characterize recovered mutants from target tissues for generation of gpt 

and spi mutational spectra; and (3) to analyze red blood cells for PIG-A mutant 

frequencies and erythrocytes for micronucleus frequencies, respectively.

In Vitro Assay to Predict Developmental (1) To use rodent in vitro organotypic and primary culture models to examine 

Neurotoxicity of Pediatric Anesthetics anesthetics including propofol (gamma-aminobutyric acid [GABA] A agonist), 

[Wang] baclofen (GABA B agonist), diazepam (GABA A agonist), pentobarbital (GABA 

A agonist & 2-amino-3-(5-methyl-3-oxo-1,2- oxazol-4-yl)propanoic acid) antagonist), 

etomidate (GABA A agonist), sevoflurane (N-methyl-D-aspartic acid [NMDA] 

antagonist & GABA agonist), fentanyl (opiate agonist), and anesthetic combinations 

commonly used in pediatric surgical procedures; (2) to determine the utility of in vitro

culture systems to predict in vivo outcomes in subsequent studies; (3) to determine 

the dose and time-course over which the potential neurotoxic effects of anesthetics 

are expressed in the developing brain; (4) to determine effective ways to protect 

against anesthetic-induced developmental neurotoxicity that have potential 

clinical utility; (5) to identify mechanisms that link altered NMDA receptor function 

and/or elevation of ROS to anesthetic-induced neuroapoptosis; and (6) to identify 

biomarkers such as genomic pathway signatures and determine their validity for 

predicting in vitro outcomes of pediatric anesthetic exposure.

PIG-A mutagenesis: an International (1) To generate data using a standardized protocol that, in combination with results 

Validation Study Comparing PIG-A from other investigators, will be used to determine the sensitivity, specificity, and 

Mutation in Rats with Other Biomarkers portability of the rat red blood cell/reticulocyte (RBC/RET) PIG-A gene mutation assay

of Genetic Toxicity and (2) to perform the in vivo Comet, MN, and PIG-A and Hprt lymphocyte gene 

[Heflich] mutation assays in conjunction with the RBC/RET PIG-A assay to determine how 

the RBC/RET PIG-A assay compares in terms of sensitivity and specificity with these 

other in vivo assays that have been used or considered for use as regulatory assays.

Development of a High Throughput Assay (1) To develop a high throughput in vivo mutation model that detects mutations 

for Measuring In Vivo Mutation in an induced by a range of mechanisms, including gene mutation, large deletions, and 

Autosomal Gene loss of heterozygosity, and (2) to evalute the basic properties and sensitivity of the 

[Bhalli] model in experiments employing well-characterized mutagens.

Effect of Soy-containing Diets on Ammonium To determine the effect of dietary whole soy and purified genistein, the principal soy 

Perchlorate-induced Thyroid Toxicity isoflavone, on the dose-response characteristics for perchlorate-induced thyroid 

in Sprague-Dawley (SD) Rats - II toxicity in male SD rats. The results from the previous study, while incomplete and 

[Doerge] therefore insufficient to inform regulatory policy, largely substantiate the original 

hypothesis that soy diets can adversely affect thyroid function in the presence of 

additional risk factors.
NTP Annual Report 2010 29

NCTR/NTP Project [Project Officer] Objective and/or Project Summary

PBPK Models for Bisphenol A (BPA) (1) To create PBPK models for BPA in three species of adult, neonatal, and pregnant 

[Fisher] (mother and fetus) and lactating (mother and neonate) laboratory animals (mouse, 

rat, and rhesus monkey); (2) to use these PBPK models to calculate internal measures 

of dose for both aglycone (i.e., active) and conjugated (i.e., inactive) forms of BPA; 

(3) to create human PBPK models for BPA (adult, child, and pregnant mother and fetus, 

and lactating mom and infant) using information obtained from the monkey, mouse, 

and rat, and limited information from the human published in the literature; (4) to use 

the human suite of PBPK models to extrapolate the internal doses of BPA associated 

with toxicity in laboratory animals to humans and to extrapolate dosimetry from 

regions of observation to low levels of exposure to BPA for which no experimental 

data exist; (5) to use this simulation protocol to help reduce the uncertainty in the 

assessment of health risks posed by BPA to human populations; (6) to use the human 

BPA PBPK model to help interpret biomonitoring data for BPA in urine and blood [the 

type of modeling is called reverse dosimetry]; (7) to consider a biologically-based dose 

response (BBDR) model for BPA, depending on the emerging knowledge about 

mode of action research for relevant adverse health effects; and (8) to propose for 

follow-on modeling beyond 3 years of this work to describe the interaction of BPA 

at target organs.

BPA Neurobehavioral Studies To establish the appropriate skills and techniques, which include complex behavioral 

[Ferguson] assessments and quantitative volumetric analysis of sexually dimorphic brain regions, 

for the conduct of developmental toxicity studies with BPA. 

DNA Methylation is Modulated by Lifestyle (1) To determine the effect of cigarette smoke condensate on DNA methylation of 

Factors and Environmental Agents several genes in lung cells, and (2) to assess the ability of other agents to modulate 

[Word] the effect of class-specific correlations on gene DNA methylation, either singularly 

or in various combinations.

Impact of Melamine on Human Intestinal (1) To determine if melamine impacts the population dynamics of the human intestinal

Microbiota: Does the Human Intestinal microbiota, and (2) to determine if the human intestinal microbiota metabolize

Microbiota have the Enzymatic Capacity melamine to cyanuric acid.

to Metabolize Melamine to Cyanuric Acid? 

[Cerniglia]

NCTR/ Arkansas Regional Laboratory/ Office (1) To support the needs of NCTR to characterize nanoscale materials used in 

of Regulatory Affairs Nanotechnology toxicology tests and to detect these materials in biological samples and (2) to 

Core Facility support the needs of Arkansas Regional Laboratory/Office of Regulatory Affairs 

to detect and characterize nanoscale materials in FDA-regulated products.

Analytical Assay for Photochemical (1) To provide support for analysis of the photoactivation of nanomaterials using the 

Generation of Hydroxyl Radical OH/coumarin-3-carboxylic acid assay, (2) to provide particle-size analysis for all 

[Howard] materials being analyzed by the OH method and other nanomaterials used in studies 

at NCTR and Arkansas Regional Laboratory/Office of Regulatory Affairs, and (3) to 

improve the assay using ultraviolet light diode laser (on hand) as a replacement 

for the existing broad band ultraviolet light A source.

*Funded by NCTR voluntary allocations
30 National Toxicology Program

NIEHS/NTP

Highlighted Activities

Gulf Oil Spill Response

Immediately following the Deepwater Horizon explosion on April 20, 2010, and the ensuing massive oil spill in 

the Gulf of Mexico, NTP and NIEHS began coordinated efforts with other Federal agencies (including FDA, OSHA, 

NHGRI, NIOSH, ATSDR, EPA, National Oceanographic and Atmospheric Administration, U.S. Fish and Wildlife 

Service, U.S. Geological Survey, and U.S. Coast Guard) to limit the adverse impacts of the oil and dispersants on 

human health, ecological health, and food sources. 

Early efforts by NTP focused upon assembly and distribution of information on characteristics and toxicity of oil 

and oil dispersants. The NTP is pursuing several lines of research to address concerns regarding the safety of Gulf 

seafood, hazards to offshore and onshore cleanup workers, and potential long-term health impacts of residual oil 

in the environment. NIEHS/NTP obtained samples of source oil, tar balls, and the dispersant, COREXIT® EC9500A. 

The NTP is conducting analytical chemistry studies to better understand the composition of the source oil, tar 

balls, and oiled sediment collected along the shoreline. The focus of these analyses is on components that are 

more likely to persist in the environment and potentially lead to residual human exposures. We will use the 

information from these analytical chemistry efforts to help understand exposures to response workers and to 

inform exposure assessments in the Gulf Long-term Follow-up Study (GuLF STUDY, see below). The NTP will also 

NIEHS/NTP staff at the NIEHS building, Research Triangle Park, NC Photo courtesy of Steve McCaw
NTP Annual Report 2010 31

use this information to develop a toxicology research program 

on polycyclic aromatic hydrocarbons (PAHs). The aim of this 

program is to further characterize the hazard of PAHs present 

in oil to determine if routinely monitoring PAHs in environmental 

media adequately captures the hazard of crude oil exposures, 

and to better understand whether there are combinations of 

different PAHs that should be considered in aggregate when 

establishing safe exposure levels.

The NTP’s toxicology assessments will inform the ongoing 

GuLF Study, which will assess the possible health effects of the 

oil spill on 55,000 cleanup workers and volunteers in coastal 

towns across Louisiana, Mississippi, Alabama, and Florida. This 

longitudinal cohort study, coordinated by NIEHS, is the largest 

health study of its kind ever conducted, and is an important 

component of the comprehensive federal response to the 

oil spill. Information about GuLF STUDY is available at 

http://nihgulfstudy.org/. NIEHS also issued a Request for 

Applications to fund university and community research 

in the Gulf on oil spill-related issues such as mental health, 

respiratory health, and chromosomal damage. 

Information about NIEHS oil spill response efforts is available 

at http://www.niehs.nih.gov/about/od/programs/gulfspill and 

information about the overall federal oil spill response efforts is available at 

http://www.restorethegulf.gov/ and http://cgvi.uscg.mil/media/main.php?g2_itemId=841814&g2_page=6.

Contact Information: Dr. Scott Masten, masten@niehs.nih.gov

NTP Laboratories Branch 

The NTP Laboratories Branch was initiated in FY 2010 to provide high quality laboratory capabilities and support 

for the performance of agent-specific, targeted research directly related to developing and applying tools of 

modern toxicology and molecular biology to the evaluation of specific substances of concern to the NTP, issues of 

central importance to programs of the NTP, or the development of new methods to advance the scientific mission 

of the NTP. Another focus of the branch is to develop better methods to study the developmental origins of adult 

diseases. The study design, review, and evaluation processes for the NTP Laboratories will be similar to those for 

other NTP projects to ensure the highest scientific quality and relevance to the NTP mission. 

Dr. Michael Waalkes is chief of the Branch and NIEHS scientists Drs. Susanne Fenton, Darlene Dixon, Jean Harry, 

and Daniel Morgan are all involved in Branch activities, while other NTP scientists are enlisted on an as-needed 

basis. One of the first studies initiated in the NTP Laboratories is to address data gaps identified in the CERHR soy 

infant formula evaluation, by conducting a series of lactation-only exposure studies in laboratory animals, with 

the goal of reducing areas of uncertainty for reaching conclusions on human infants fed soy infant formula. 

Contact Information: Dr. Michael Waalkes, waalkes@niehs.nih.gov
32 National Toxicology Program

Chemical Effects in Biological Systems (CEBS): 

An Integrated Data Management System for the NTP

CEBS is a data management tool that moved to the NTP in 2009. 

While it was part of the NIEHS Division of Intramural Research, 

CEBS was designed to house biological data from a variety of 

studies of different study designs and purposes deposited by 

NIEHS scientists and members of the public. The unifying theme 

of CEBS is that the data contained therein are of interest to 

toxicologists and environmental health scientists. Because 

CEBS was designed as a public reference repository it can faithfully 

house “any” type of biological measurements from “any” type of 

study design. This feature was designed into CEBS to allow data from different sources to be integrated in one 

place, to permit data mining and analysis. In FY 2010, the NTP was in the process of loading the NTP legacy data 

into CEBS so that users can mine data from NTP studies as well as from studies submitted from other sources.

CEBS consists of a database, a user interface, and tools for loading data and 

managing it for display. We have tested the capabilities of CEBS to house NTP 

data by loading data from legacy genetic toxicology studies, immunotoxicology 

studies, and current high-throughput assays into the CEBS database. The NTP is 

modifying the CEBS user interface to permit proper display of the conclusions 

and raw data from these studies so that users can access them. When this project 

is completed all the public NTP data will be accessible in CEBS for reference and 

for data mining.

CEBS captures the details of the study design and execution plus the biological responses of subjects in a way 

that permits searching, filtering, and sub-setting of the data. The user can view the details of each study, search 

for particular studies or study subjects of interest based on treatment, response, or other characteristics, and 

then either analyze the data within CEBS or download for import into other tools. CEBS can be accessed at 

http://cebs.niehs.nih.gov.

Contact Information: Dr. Jennifer Fostel, fostel@niehs.nih.gov

Cellular Phone Radiation Emissions

The NTP is conducting a multi-year study to address the potential health-related adverse 

effects of radiation emitted by cellular phones and cell towers. More than 270 million 

Americans use wireless communication devices; however, the potential health effects of 

long-term exposure to radiation emissions from these devices are unknown. The overall 

goal of the NTP’s studies is to determine the potential toxic and/or carcinogenic effects 

of exposure to cellular phone radiofrequency emissions in laboratory animals. The animal 

studies are being conducted in three phases: (1) pilot studies to determine appropriate field 

strengths of emissions, (2) subchronic toxicology using exposures of up to two months, and 

(3) chronic toxicology and carcinogenicity studies. In FY 2010, NTP was nearing completion 

of the second phase of the studies and expects to begin the third and final phase in 2011.

These NTP studies will provide information regarding the safety of exposure to radiofrequency radiation and 

strengthen the science base for determining any potential health effects in humans. These data could contribute 

to information used by the Federal government, including the FDA, in making decisions about radiofrequency 

radiation health issues to protect public health and safety.

Contact Information: Dr. Michael Wyde, wyde@niehs.nih.gov
NTP Annual Report 2010 33

Nanotechnology Safety Initiative

Nanoscale materials are materials that have at least one 

dimension in the size range of approximately 1-100 nm 

in size. While they are already appearing in commerce 

as industrial and consumer products and as novel 

drug delivery formulations, little research has focused 

on the potential toxicity of manufactured nanoscale 

materials. Also, the unique and diverse physicochemical 

properties of nanoscale materials suggest that their 

toxicological properties may differ from those of 

materials of similar composition but larger size.

The NTP is currently engaged in a broad-based 

research program to address potential human 

health hazards associated with the manufacture 

and use of nanoscale materials. This initiative is 

driven by the current and anticipated future focus 

on nanotechnology research and development. 

Ongoing research activities are initially focusing 

on several classes of materials, including nanoscale 

metal oxide powders (titanium dioxide and cerium 

oxide), cadmium-based fluorescent nanocrystalline 

semiconductors (quantum dots), carbon fullerenes, 

carbon nanotubes, and metal-based nanoparticles 

(nanosilver and nanogold).

The ultimate goal of this research program is to 

evaluate the toxicological properties of several 

nanoscale material classes that represent a cross 

section of composition, size, surface coatings, and 

physicochemical properties, and to use these properties 

to investigate fundamental questions concerning if 

and how nanoscale materials interact with biological 

systems and potentially cause adverse effects in 

humans. 

In FY 2010, the NTP was invited to participate in a 

number of working and advisory groups including the Scientific Oversight Committee for the National Cancer 

Institute’s Nanotechnology Characterization Laboratory and the Interagency Working Group on Nanotechnology 

Environmental and Health Implications (NEHI) of the Nanoscale Science, Engineering, and Technology 

Subcommittee of the President’s National Science and Technology Council. 

Current NTP projects include evaluating the pharmacokinetics of nanoscale silver in rodents. The NTP has 

completed two subchronic, 90-day inhalation studies of fullerene C60 (exposures to each of two particle sizes, 

50 nm and 1 μm, each in both rats and mice via nose-only inhalation). The NTP has completed physicochemical 

characterization of 24 multi-walled nanotubes of varying sizes, lengths, and vendors. These data are being 

used to develop subchronic inhalation studies of selected multi-walled nanotubes. Dr. Walker gave six invited 

presentations on NTP’s nanomaterials program in FY 2010.

Contact Information: Dr. Nigel Walker, walker3@niehs.nih.gov.

NIOSH field investigator taking instrument readings during a 

filter collector drum change by a technician at a nano metal 

oxide plant.

Scanning electron micrograph of multiwalled carbon nanotubes.

Titanium dioxide
34 National Toxicology Program

Herbal Medicines and Dietary Supplements

Herbal and dietary supplements are a major focus area for the NTP. The NTP is currently studying many of the 

25 top-selling herbal and dietary supplements. Table 5 lists the substances for which studies are completed, 

ongoing, or planned. The substances can be loosely classified as “women’s health” (black cohosh, gum guggul, 

dong quai), “cancer chemopreventives” (e.g., green tea extract, indole carbinol, resveratrol, melatonin), 

“anti-aging” (e.g., ginseng, glucosamine/chondroitin sulfate, Ginkgo biloba, vincamine), weight loss/sports 

aids (e.g., Usnea lichen/usinic acid, chitosan, Garcinia cambogia, bitter orange extract), and “multipurpose” 

(e.g., Aloe vera, Echinacea, kava, milk thistle, pulegone/pennyroyal, and senna laxative).

Contact Information: Dr. Nigel Walker, walker3@niehs.nih.gov.

Table 5: Herbal Ingredients and Dietary Supplements for Which Studies Are Completed, Ongoing, or Planned 

Substance [CASRN] Project Leader Use and/or Rationale

Aloe vera Boudreau Humans have widespread oral and dermal exposure to Aloe vera which lacks 

(oral) [8001-97-6] adequate toxicity information; there is a suspicion of carcinogenicity. The 1998 FDA 

Special Nutritional Adverse Event Monitoring System lists Aloe vera exposure as 

associated with numerous adverse effects in humans.

Arbutin Chan Consumer exposure occurs through food, cosmetics, and dietary supplements. 

[497-76-7] There is a lack of adequate toxicological data and suspicion of toxicity based on 

chemical structure. 

Bitter orange extract Hansen Consumer exposure is through increasing dietary supplement use (it is the most 

common replacement for ephedra, banned from use by the FDA in 2003, in dietary 

supplements for weight loss). There is suspicion of toxicity and a lack of adequate 

toxicity data.

Butterbur Howard Used as a dietary supplement. There is suspicion of toxicity based on 

(Petasites hybridus) extract pharmacological activity of constituents and the potential presence of hepatotoxic

[90082-63-6] pyrrolizidine alkaloids.

Chitosan Chhabra Chitosan is a popular dietary supplement used for weight loss. Several subacute

[9012-76-4] studies in animals show that it has hypercholesterolemic properties and may 

influence weight gain; it may also cause vitamin and mineral deficiencies. There is 

a potential for vitamin E depletion and osteoporosis from ingestion.

Chondroitin sulfate/ Leakey Chondroitin sulfate is a dietary supplement widely used alone and in combination 

Glucosamine with glucosamine to alleviate pain and inflammation from osteoarthritis. No data are

[9007-28-7] available on the possible adverse or toxic effects from long-term exposure.

Dong quai Wyde Dong quai has widespread use as a dietary supplement and in Chinese herbal 

(Angelica sinensis root) medicine as an antispasmodic or blood purifier and for reducing pain, dilating

and extract blood vessels, and stimulation, as well as for relaxing uterine muscles. There is 

suspicion of toxicity based on estrogenic activity and chemical structure, and 

there is a lack of adequate toxicity data.

Echinacea purpurea, Irwin Echinacea is the most popular herbal supplement in the United States, creating

extract widespread human exposure. It is used to stimulate the immune system, and there 

[90028-20-9] is a lack of scientific literature supporting its safety or efficacy.
NTP Annual Report 2010 35

Substance [CASRN] Project Leader Use and/or Rationale

Epigallocatechin Chan Epigallocatechin is a potential cancer chemopreventive agent. It is an antioxidant 

(Green tea extract) thought to prevent tumorigenesis by protecting cellular components from oxidative

[989-51-5] damage via free radical scavenging. It is a major component of the polyphenolic 

fraction of green tea. It requires evaluation regarding its toxicity.

Evening primrose oil Germolec Evening primrose oil is used as a dietary supplement, particularly

(Oenothera biennis extract) for autoimmune conditions. There is a lack of adequate toxicological data.

[90028-66-3]

Garcinia cambogia extract Wyde Garcinia cambogia is marketed as an ephedra-free diet aid. There is consumer

[90045-23-1] exposure through increasing dietary supplement use and a lack of adequate 

toxicity data.

Ginkgo biloba extract Chan There is potential for widespread exposure through use as a dietary supplement 

[90045-36-3] for “improving brain functioning” or “promoting radical scavenging activity.” Ginkgo 

biloba clearly demonstrates biological activity and may be consumed in rather large 

doses for an extended period of time. Some ingredients are known mutagens or 

suspected carcinogens.

Ginseng Chan Ginseng has widespread use as a dietary supplement. There is a possibility 

[50647-08-0] that ginseng and ginsenosides may have anticarcinogenic activity, and a lack 

of toxicity information.

Glucosamine Leakey Glucosamine is a widely used dietary supplement, both alone and in combination 

[3416-24-8] with chondroitin sulfate, to alleviate pain and inflammation from osteoarthritis. 

There are no data on the possible adverse or toxic effects from long-term exposure.

Gum guggul extract Wyde Gum guggul has expanding use as a dietary supplement and has demonstrated 

biological effects on lipid metabolism, thyroid hormone homeostasis, female 

reproductive tissues, and endogenous nuclear hormone receptors, as well as the 

potential for serious drug interactions. There is a lack of available information to 

adequately assess safe use in humans.

Indole-3-carbinol Wyde Indole-3-carbinol is marketed as a dietary supplement with projected rapid 

[700-06-1] growth in sales. It is found in cruciferous vegetables and is under review at NCI 

as a chemopreventive agent for breast cancer. Substantial evidence exists that 

indole-3-carbinol can reduce the risk of cancers induced by several carcinogens 

when administered to animals. 

Kava kava extract Chan Kava kava is a dietary supplement with widespread use. It has also been promoted 

[9000-3-8] as a substitute for Ritalin (methylphenidate) in children. Insufficient toxicity data are 

available. NCI recommended testing kava extract standardized to 30% kavalactones.

Melatonin Travlos Melatonin, a hormone produced by the pineal gland, has become very popular 

[73-31-4] as an over-the-counter hormone supplement as well as being used as a 

chemotherapeutic agent in cancer. There is a lack of toxicity information and 

a suggestion that melatonin may have the potential to cause ocular toxicity.

Milk thistle extract Dunnick Milk thistle extract is a popular dietary supplement thought to have beneficial 

[84604-20-6] effects on the liver; however, there is limited information on its safety. Metabolism 

Silymarin [65666-07-1] studies are needed to resolve questions regarding bioavailability of orally 

Silybin [22888-70-6] administered milk thistle extract. Milk thistle fruits contain silymarin, the active 

flavonoid constituent; one of silymarin’s principal components is silybin.

Pulegone Chan The nomination of pulegone and menthofuran for testing is based on 

(Pennyroyal) the potential for human exposure and the absence of carcinogenicity data. 

[89-82-7] Pulegone is a major constituent of pennyroyal, and menthofuran is one of the 

metabolites of pulegone.

trans-Resveratrol Germolec trans-Resveratrol is found in grapes and wine and is currently marketed in pure 

[501-36-0] or extract form as a dietary supplement. It has numerous reported beneficial 

effects but toxicity is poorly characterized.
36 National Toxicology Program

Substance [CASRN] Project Leader Use and/or Rationale

Retinyl palmitate Howard Retinyl palmitate was nominated for phototoxicity and photocarcinogenicity 

testing based on the increasing widespread use of this compound in cosmetic retail 

products for sun-exposed skin. There is a need to investigate the biochemical and 

histological changes in skin caused by retinyl palmitate and the association between 

topical application of retinoids and enhancement of photocarcinogenesis.

Senna (powdered) Dunnick The safety of laxatives is currently being reassessed by the FDA as a result 

[8013-11-4] of the testing of phenolphthalein for carcinogenicity in rodents. Senna has been 

reported as positive in the Ames test and a preliminary 2-year rat study showed 

an increase in lymph node hyperplasia. The FDA Center for Drug Evaluation and 

Research is requesting a p53 hemizygous study to complement a 2-year rat study 

sponsored by the manufacturer.

alpha/beta Thujone mixture Hooth Thujone was identified through a review of direct food additives given “generally 

[546-80-5] recognized as safe” status by the FDA. It has known toxicity that has caused it 

[471-15-8] to be banned from some products. Twenty-four direct food additives in the FDA 

Priority-Based Assessment of Food Additives contain thujone. There is a potential 

for widespread consumer and worker exposure.

Usnea barbata, extract Leakey Usnea barbata is used as a dietary supplement for weight loss. 

[84696-53-7] Insufficient toxicity data are available. 

Usnic acid and Usnea herb Leakey Usnic acid and Usnea herb have widespread use in dietary supplements and personal 

[125-46-2] care products. Adequate toxicological data are lacking, and there are numerous 

human adverse event reports.

Valerian DeVito toxicological data and concern for adverse developmental and reproductive effects.

Valeriana officinalis L. 

root extract

[8057-49-6]

Valerian oil

[8008-88-6]

Vincamine Chan Consumer exposure to vincamine occurs through dietary supplement use. 

[1617-90-9] There is a suspicion of toxicity and a lack of adequate toxicological data.

1

Testing Status and study results for completed studies can be found at http://ntp.niehs.nih.gov/ – select 

“Testing Status of Agents at NTP” and “Study Results and Research Projects.”
NTP Annual Report 2010 37

Table 6. Use of NTP Study Data or Recommendations by Federal and State Regulatory Agencies in FY 2010 

Agency, Title, Additional Information NTP Information Cited

California Environmental Protection Agency • TR 201 - Bioassay of 2,3,7,8-tetrachlorodibenzo-p-dioxin [1746-01-6]

Office of Environmental Health for possible carcinogenicity (gavage study)

Hazard Assessment (OEHHA)

• TR 109 - Carcinogenesis bioassay of 2,3,7,8-tetrachlorodibenzo-p-dioxin

Announcement of Publication in Osborne-Mendel rat and B6C3F1 mice (gavage study)

of the Final Public Health Goals for

Benzo(A)Pyrene, Methoxychlor, and • Report of the NTP ad hoc panel on chemical carcinogenesis testing and

TCDD (Dioxin) In Drinking Water evaluation. Board of Scientific Counselors. 1984.

• TR 521 - Toxicology and carcinogenesis studies of 

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in female Harlan SD rats 

(gavage study)

• Symposium on Significance of Foci of Cellular Alteration in the Rat Liver. 

Toxicol Pathol 17:557-735. 1989.

U. S. EPA NTP 11th Report on Carcinogens classified the following as reasonably 

Community Right-to-Know Toxic anticipated to be a human carcinogen:

Chemical Release Reporting 1-Amino-2,4- Dibromoanthraquinone [81-49-2] 

2,2-bis(Bromomethyl)-1,3- propanediol [3296-90-0] 

EPA is proposing to add 16 chemicals to the list Furan [110-00-9] 

of toxic chemicals subject to reporting under Glycidol [556-52-5] 

section 313 of the Emergency Planning and Isoprene [78-79-5] 

Community Right-to-Know Act (EPCRA) of 1986 Methyleugenol [93-15-2]

and section 6607 of the Pollution Prevention 1,6-Dinitropyrene [42397-64-8] 

Act of 1990 1,8-Dinitropyrene [42397-65-9] 

April 6, 2010 – 75 FR 17333 6-Nitrochrysene [7496-02-8] 

4-Nitropyrene [57835-92-4] 

o-Nitroanisole [91-23-6] 

Nitromethane [75-52-5] 

Phenolphthalein [77-09-8]

Tetrafluoroethylene [116-14-3] 

Tetranitromethane [509-14-8] 

Vinyl Fluoride [75-02-5]

Use of NTP Products by Other Agencies

Federal and state regulatory agencies use NTP study data and recommendations in considering the need to 

regulate and test specific chemicals to protect human health. Table 6 lists NTP data and recommendations used 

in FY 2010.
38 National Toxicology Program

Report on Carcinogens

The Report on Carcinogens (RoC) is a congressionally mandated listing [Section 301(b)(4) of the Public Health 

Services Act, 42 U.S.C. 241(b)(4)] of substances (1) that either are known to be human carcinogens or may 

reasonably be anticipated to be human carcinogens and (2) to which a significant number of persons residing 

in the United States are exposed. Each substance listed in the RoC has a profile, which contains the listing status, 

a summary of the cancer studies supporting the listing status, information on human exposure, and Federal 

regulations to reduce exposure. The RoC is a cumulative report and consists of substances newly reviewed 

in addition to those listed in previous editions. The Secretary of Health and Human Services has delegated 

preparation of the RoC to the NTP, with assistance from other Federal health and regulatory agencies. 

Dr. Ruth Lunn is the Director of the RoC Center. SRA International, Inc., provided contract support for 

preparation of the RoC in FY 2010.

Figure 3: NTP Report on Carcinogens Review Process 
NTP Annual Report 2010 39

Reviewing nominations to the RoC is a multi-step, formal, and open process, as shown in Figure 3 and on the 

RoC website (http://ntp.niehs.nih.gov/go/15208). The nomination of substances for listing in or removal from the 

RoC is open to all interested individuals and groups. The RoC Center prepares a draft background document on 

each substance under review that is peer-reviewed by an external scientific ad hoc panel at a public meeting. 

Three independent scientific groups make recommendations to the NTP on the listing status of candidate 

substances using established listing criteria. Taking into account recommendations from the three review 

groups and the public comments, the NTP drafts a substance profile, which contains its preliminary listing 

status recommendation, the science that supports the recommendation, and information on use, production, 

exposure, and current regulations. The NTP BSC meets publicly to peer review the draft substance profile. 

Public comments are solicited several times during the review process and are provided to each review group 

as available. The NTP prepares the draft RoC (which contains substance profiles for newly proposed and listed 

substances and substances listed in previous editions of the RoC) for review and comment by the NTP Executive 

Committee. The NTP Director receives the input from all reviews plus the public comments and submits the 

final draft RoC to the Secretary, DHHS, for review and approval. 

Candidate substances for the 12th RoC are listed in Table 7. Scientific review of candidate substances for the 12th

edition of the RoC, which was carried out using a formal process (see Figure 3), was completed in FY 2010 with the 

review of formaldehyde. Formaldehyde is currently listed in the 11th RoC as reasonably anticipated to be a human 

carcinogen; it was reviewed for the 12th RoC for a possible change in listing status. The RoC Center convened 

an expert panel to peer review the draft background document for formaldehyde on November 2-4, 2009, in 

Research Triangle Park, NC. The expert panel recommended listing formaldehyde as a known human carcinogen 

based on sufficient evidence of carcinogenicity from studies in humans and supporting studies on mechanisms 

of carcinogenesis. 

The NTP BSC peer reviewed the draft substance profiles for cobalt-tungsten carbide: hard metals and powders, 

glass wool fibers, and formaldehyde at the June 21-22, 2010 meeting.

Table 7: Candidate Substances for the 12th Report on Carcinogens in FY2010 

Candidate Substance Primary Uses/Exposures Basis of Nomination [CASRN] Nominator

Aristolochic Acids A family of nitrophenanthrene carboxylic acids that IARC finding of sufficient evidence of

NIEHS occurs naturally in plants in the Aristolochiaceae carcinogenicity in animals and limited

family, primarily of the genera Aristolochia and evidence in humans.

Asarum. Botanical products from plants containing (IARC Monograph Vol. 82, 2002).

aristolochic acids are used in traditional folk 

medicines, particularly in Chinese herbal medicine, 

and have been used inadvertently as part of a 

weight-loss regimen.

Captafol A broad-spectrum fungicide that has been widely IARC finding of sufficient evidence of carcinogenicity

[2425-06-01] used since 1961 for the control of fungal diseases in animals (IARC Monograph Vol. 53, 1991). 

NIEHS in fruits, vegetables, and some other plants. Use IARC also noted that captafol gives positive results

of captafol in the United States was banned in 1999. in many genetic assays, including the in vivo assay

for dominant lethal mutation.

Cobalt–Tungsten Composites of tungsten-carbides with a metallic Recent human cancer studies on the hard metal

Carbide: Powders cobalt, used to make cutting and grinding tools, manufacturing industry showing an association 

Hard Metals dies, and wear products for a broad spectrum of between exposure to hard metals (cobalt tungstenNIEHS industries, including mining and oil and gas drilling. carbide) and lung cancer.

Formaldehyde Primarily used to produce resins for the production Nominated for reconsideration based on the 2004

[50-00-0] of many different products, including plastics, IARC review, which concluded that there was

NIEHS – for adhesives and binders for wood products, pulp sufficient evidence for the carcinogenicity of

reclassification and paper, and synthetic fibers, and in textile formaldehyde in humans (IARC Monograph

It is also used as a disinfectant and preservative finishing. Vol. 88, 2004).

and as an intermediate for many industrial chemicals.
40 National Toxicology Program

Candidate Substance Primary Uses/Exposures Basis of Nomination [CASRN] Nominator

Glass Wool Fibers Glass wool fibers, which are a type of synthetic Insulation glass wool: IARC finding of limited evidence

North American vitreous fibers, are an inorganic fibrous material of carcinogenicity in animals and evaluation as not 

Insulation manufactured primarily from glass and processed classifiable as to its carcinogenicity to humans

Manufacturers inorganic oxides. The major uses of glass wool are (Group 3; IARC Monograph Vol. 81, 2002).

Association nominated in thermal, electrical, and acoustical insulation, Special-purpose glass fibers: IARC finding of 

glass wool (respirable weatherproofing, and filtration media. Some sufficient evidence of carcinogenicity in animals

size) for delisting special-purpose glass wool fibers are used for (IARC Monograph Vol. 81, 2002).

high-efficiency air filtration media and acid 

battery separators.

ortho-Nitrotoluene A chemical intermediate used in the synthesis Results of an NTP bioassay (NTP Technical Report

[88-72-2] of azo dyes. It is also used (either directly or as 504, 2002), which reported clear evidence of

NIEHS an intermediate) in the production of other dyes, carcinogenic activity in rats and mice.

agricultural chemicals, rubber chemicals, pesticides, 

petrochemicals, pharmaceuticals, and explosives.

Riddelliine Found in a class of plants growing in the western Results of an NTP bioassay (NTP Technical Report

[2346-96-0] United States. Cattle, horses, and sheep ingest 508, 2003), which reported clear evidence of

NIEHS these toxic plants. carcinogenic activity in male and female rats 

and mice.

Styrene Used worldwide in the production of polymers, IARC finding of limited evidence of carcinogenicity

[100-42-5] which are incorporated into products such as in animals and limited evidence of carcinogenicity

Private Individual rubber, plastic, insulation, fiberglass, pipes, in humans (IARC Monograph Vol. 82, 2002). 

automobile parts, food containers, and 

carpet backing.

International Agency for Research on Cancer (IARC). IARC Monographs are available from http://monographs.iarc.fr/.

NTP Technical Reports are available at http://ntp.niehs.nih.gov/ see “NTP Study Reports.”

Contact Information: Report on Carcinogen Center, Dr. Ruth Lunn, lunn@niehs.nih.gov. 

RoC website: http://ntp.niehs.nih.gov select “Report on Carcinogens.”
NTP Annual Report 2010 41

Center For the Evaluation of Risks to Human Reproduction

The Center for the Evaluation of Risks to Human Reproduction (CERHR) was established in 1998 to serve as an 

environmental health resource to the public and regulatory and health agencies. CERHR publishes monographs 

that assess the potential for substances to cause adverse effects on reproduction and development in humans. 

CERHR selects substances for evaluation based on several factors, including production volume, extent of 

human exposures, public concern about the chemical hazard, and the extent of published data from studies 

of reproductive or developmental toxicity. Dr. Kristina Thayer is the Director of CERHR. 

Revision of the CERHR Evaluation Process

The NTP revised the CERHR evaluation process (Figure 4) for preparation of NTP monographs on environmental 

substances to (1) enhance scientific development of CERHR products and to maximize efficiency and utilization 

of resources and (2) broaden the scope of analysis topics beyond those traditionally associated strictly with 

reproduction and development. The process was changed to allow flexibility to better tailor an evaluation 

to meet programmatic needs depending upon the topic, scope, nature and extent of the literature, scientific 

complexity, and time considerations. The new process retains elements of the past, including opportunity 

for external scientific input, public input at multiple points, interagency input, and external peer review, and 

addresses the weakness of limited flexibility. NTP presented this change in process to the BSC in December 

2009 (http://ntp.niehs.nih.gov/go/9741). Overall, the BSC supported the change in the evaluation process to 

allow flexibility in the approach used to gain input.
42 National Toxicology Program

Evaluation of Soy Infant Formula

CERHR convened a public, expert panel meeting on December 16-18, 2009, in Alexandria, Virginia to evaluate soy 

infant formula. Soy formula is an infant food made using soy protein and other components. It is fed to infants 

as a supplement or replacement for human milk or cow milk formula. Soy formula contains isoflavones, naturally 

occurring compounds found primarily in beans and other legumes including soybeans, peanuts, and chickpeas. 

The 14-member ad hoc scientific panel reviewed and evaluated the available scientific data on soy infant formula. 

In their deliberations, the expert panel considered the quality and strength of the scientific evidence that soy 

formula or its isoflavone constituents might cause adverse effects on human development. The expert panel also 

identified gaps in the available scientific data on the possible effects of soy formula and suggested areas where 

additional research is needed. The expert panel expressed minimal concern for adverse developmental effects in 

infants fed soy infant formula. 

Based upon information in the expert panel report, public comments, and other literature since the expert panel 

met, the NTP prepared its NTP Brief with its preliminary conclusion regarding soy formula and potential effects on 

human reproduction and development. The BSC peer reviewed the draft NTP Brief at its meeting on May 10, 2010. 

The BSC supported the NTP’s conclusion of minimal concern for adverse effects on development in infants who 

consume soy infant formula. This level of concern represents a “2” on the five-level scale of concern used by the 

NTP that ranges from negligible concern (“1”) to serious concern (“5”). The NTP released the final NTP Brief on Soy 

Infant Formula on September 16, 2010 (http://cerhr.niehs.nih.gov/evals/genistein-soy/soyformula/soyformula.html).

The NTP BSC further supported a research program proposed by CERHR to address some key data gaps

 identified in the soy infant formula evaluation. CERHR initiated this research program, which includes a series 

of lactation-only exposure studies with the goal of reducing areas of uncertainty for reaching conclusions on 

human infants fed soy infant formula from the laboratory animal data. These studies will be carried out in the 

new NTP Laboratory Branch (see page 31).

Evaluation of Low-Level Lead

CERHR began an evaluation of the adverse effects of low-level lead exposure, which will focus on epidemiological 

data on blood lead levels <10 μg/dL, because the health effects at higher blood lead levels are well established. 

The current occupational exposure limit is 40 μg/dL in workers (including women of childbearing age), although 

epidemiological data show adverse effects on reproduction and development at lower blood lead levels. The 

scope of the evaluation will include epidemiological findings on health effects other than reproduction and 

development (e.g., cardiovascular effects in adults) in order to maximize the utility of the evaluation. The evaluation 

is designed to address (1) the weight of the evidence for adverse health effects associated with blood lead levels 

of <10 μg/dL, (2) the health effect(s) associated with blood lead levels <10 μg/dL, (3) the life stages (prenatal, 

childhood, adolescence, or adulthood) in which lead’s effects are identified, (4) the blood lead level associated 

with the health effect(s), and (5) additional biomarkers of exposure associated with the effect (e.g., bone lead) 

and the relationship of this biomarker to the blood lead level. The NTP BSC reviewed the proposed approach for 

the evaluation at the May 10, 2010 meeting and gave its support. Information is available at http://cerhr.niehs.nih.

gov/evals/Lead/lead.html.
NTP Annual Report 2010 43

Evaluation of the Role of Environmental Chemicals in the Development of Diabetes and Obesity

CERHR presented a concept proposal at the December 9-10, 2010 BSC meeting for a State of the Science 

Workshop on environmental exposures and diabetes/obesity. The justification for this assessment is that 

diabetes and obesity are major threats to human health. Forty percent of people over the age of 20 have diabetes 

or pre-diabetes and ~70% of type 2 diabetes risk can be attributed to overweight/obesity. Diabetes and obesity 

are often discussed in the context of developmental origins of adult disease or prenatal programming. There is 

growing scientific and public interest in this issue. The goals and expected outcomes of the assessment are 

to (1) complete a critical assessment of the current literature and (2) focus future research directions. CERHR 

scheduled the workshop for January 2011 (see http://cerhr.niehs.nih.gov/evals/diabetesobesity/index.html).

Evaluation of Cancer Chemotherapy during Pregnancy

CERHR is evaluating the evidence for developmental effects of exposure to cancer chemotherapy in utero, 

with the main focus on clinical data in humans, to be supplemented with biomedical and toxicological literature 

in animals. The goal of the monograph is to provide clinicians, patients, and researchers with a comprehensive 

review of the incidence and types of adverse effects seen in humans exposed in utero to cancer chemotherapeutic 

agents. The key objectives of the document are to (1) identify the complete published scientific literature on 

chemotherapy during pregnancy in humans, focusing on the most common cancers occurring during pregnancy; 

(2) critically evaluate the strength and consistency of the literature on embryo, fetal, and postnatal outcomes 

in humans by cancer type, chemotherapeutic agent, and trimester of exposure; (3) develop weight of evidence 

conclusions on the occurrence of adverse effects at different gestational stages, by agent; and (4) identify data 

gaps and research needs for evaluating the effects of exposure to cancer chemotheraputics in utero. The NTP BSC 

reviewed the proposal at the June 21-22, 2010 meeting and supported NTP moving forward (http://cerhr.niehs.nih.

gov/evals/pregnancychemo/index.html).

CERHR staff recognition

Dr. Michael Shelby, former CERHR Director, was recognized by his peers at the Environmental Mutagen Society 

meeting in October 24-28, 2009 for his 30 years of contribution to Mutation Research as an author, reviewer, and 

editor, and for his long career in mutagenesis research.

Contact Information: NTP Center for the Evaluation of Risks to Human Reproduction, Dr. Kristina Thayer, Director, thayer@niehs.nih.gov. 

CERHR website: http://cerhr.niehs.nih.gov
44 National Toxicology Program

NTP Interagency Center for the Evaluation

of Alternative Toxicological Methods

The development, validation, acceptance and harmonization of new, revised, and alternative toxicological 

test methods are coordinated in the Federal government through ICCVAM. NIEHS established ICCVAM in 1997 

to implement a directive in the 1993 NIH Revitalization Act to develop a process to achieve the regulatory 

acceptance of scientifically valid alternative testing methods. Alternative methods are those methods that 

reduce, refine, or replace the use of animals. NICEATM was established in 1998 to administer ICCVAM, provide 

scientific support for ICCVAM activities, and conduct independent validation studies on promising test methods. 

The ICCVAM Authorization Act of 200 (42 U.S.C. 285l-3) established ICCVAM as a permanent interagency committee 

under NICEATM, and specified the purposes and duties of the committee. Dr. William Stokes (Rear Admiral, 

U.S. Public Health Service) is the NICEATM Director and Executive Director of ICCVAM. Integrated Laboratory 

Systems, Inc., provided contract support for NICEATM in FY 2010.

ICCVAM and NICEATM work to promote the validation and regulatory acceptance of new, revised, and alternative 

toxicological test methods that are more predictive of human and ecological effects than those currently available 

and that refine, reduce, and replace animal use whenever possible. The desired outcomes from these new methods

are to improve agencies’ abilities to assess risk and make regulatory decisions, promote more humane animal use, 

and reduce and replace animal use. NICEATM, in conjunction with ICCVAM, convenes scientific peer review panels 

to evaluate the validation status of proposed alternative testing methods for which validation studies have been 

completed to assess their accuracy and reproducibility across multiple labs. ICCVAM then develops formal test 

method recommendations for acceptance consideration by agencies, and proposes recommended test methods 

for adoption by international organizations. NICEATM and ICCVAM also convene workshops and expert panels 

to evaluate the adequacy of existing methods, identify promising test methods for further development and 

validation, evaluate the interim validation status of methods, and evaluate proposed validation studies. 

NICEATM receives test method nominations and submissions for ICCVAM to consider and review (see 

http://iccvam.niehs.nih.gov). Test methods can be nominated for validation studies or technical reviews. The 

ICCVAM evaluation process involves an initial assessment by NICEATM of the adequacy and completeness of the 

test method nomination or submission, and a determination by ICCVAM of the priority of the proposed method 

for technical evaluation or validation studies. Once a proposed test method is accepted for evaluation or validation,

ICCVAM assembles an interagency working group with appropriate scientific and regulatory expertise to collaborate 

with NICEATM on the evaluation process. Depending on the validation status of the proposed test method, 

ICCVAM, in conjunction with NICEATM, develops recommendations and priorities for appropriate evaluation activities. Such efforts might include an expert workshop, an expert panel meeting, a peer review meeting, 

an expedited peer review process, or a validation study. Information on the status and results of NICEATM and 

ICCVAM activities, including meeting reports and background documents, is available on the NICEATM-ICCVAM 

website. ICCVAM has contributed to the approval or endorsement of 33 alternative safety-testing methods by 

Federal regulatory agencies since its establishment in 1997. Recent NICEATM publications, meetings, and test 

methods currently under review, and project status are presented in Tables 8, 9, 10, and 11 respectively. 

On June 11, 2010, NICEATM published the ICCVAM Biennial Progress Report, which detailed ICCVAM activities and 

progress from 2008-2009. Among the highlights included in the report are ICCVAM’s recommendations of two 

in vitro ocular safety test methods, two in vitro assays to assess the potential of chemicals and products to cause 

acute oral poisoning, and an updated test method protocol and procedures for the murine local lymph node 

assay. A pdf version of the report can be accessed on line at http://iccvam.niehs.nih.gov/about/ICCVAMrpts.htm. 
NTP Annual Report 2010 45

Dr. Stokes delivered the keynote address at an event held to recognize the establishment of KoCVAM, the inaugural 

KoCVAM International Symposium, which was held at Seoul National University in South Korea on November 3, 

2009. In his talk, “Validation and Regulatory Acceptance of Alternative Methods for Safety Testing: Recent Progress 

and Future Directions,” he stressed the importance of high quality validation studies to enable regulatory 

acceptance and use of new safety test methods. He also visited the Korean FDA and the National Institute 

of Food and Drug Safety, the parent agency of KoCVAM.

Dr. Stokes delivered the plenary lecture at the 22nd Annual Meeting of the Japanese Society for Alternatives to 

Animal Experiments (JSAAE), held November 13-15, 2009, at Osaka University. While in Japan, he also participated 

in an international workshop on dermal safety of cosmetics and chemicals and attended an advisory committee 

meeting of the Japanese Center for the Validation of Alternative Methods (JaCVAM). He also met with the 

JaCVAM director and staff members to discuss joint validation and evaluation activities with NICEATM and ICCVAM.

Alternative methods of vaccine safety testing were the focus of an 

international workshop held September 14-16, 2010, at NIH in Bethesda, 

MD. The workshop, which was organized by NICEATM and ICCVAM in 

partnership with ECVAM, JaCVAM, and Health Canada, hosted nearly 

200 scientists from 13 countries. Participants at the International Workshop on Alternative Methods to Reduce, Refine, and Replace the Use 

of Animals in Vaccine Potency and Safety Testing: State of the Science 

and Future Directions worked to advance the availability of alternative 

methods for vaccine potency and safety testing while ensuring continued protection of human and animal health. NICEATM and ICCVAM 

have included vaccine testing as one of four high priority areas in their 

2008-2012 Five-Year Plan.

In FY 2011, NICEATM-ICCVAM is planning to convene the Independent 

Peer Review Panel Meeting: Evaluation of an In Vitro Stably-Transfected 

Estrogen Receptor Transcriptional Activation Assay for Identification of 

Potential Endocrine Disruptor Activity, scheduled for March 29-30, 2011, at NIH in Bethesda, MD. NICEATM-ICCVAM

will also be holding a series of workshops in FY 2011, Best Practices for Regulatory Safety Testing.

Table 8: NICEATM-ICCVAM Publications in FY 2010 

Date Title

Mar 23, 2010 NICEATM-ICCVAM Brochure (revised March 2010)

Jun 10, 2010 ICCVAM 2008-2009 Biennial Report

Jun 29, 2010 ICCVAM Test Method Evaluation Report on the Murine Local Lymph Node Assay: DA A Nonradioactive 

Alternative Test Method to Assess the Allergic Contact Dermatitis Potential of Chemicals and Products

Jun 29, 2010 ICCVAM Test Method Evaluation Report on the Murine Local Lymph Node Assay: BrdU-ELISA A 

Nonradioactive Alternative Test Method to Assess the Allergic Contact Dermatitis Potential of Chemicals 

and Products

Sep 13, 2010 ICCVAM Test Method Evaluation Report: Recommendations for Routine Use of Topical Anesthetics, 

Systemic Analgesics, and Humane Endpoints to Avoid or Minimize Pain and Distress in Ocular Safety Testing

Sep 13, 2010 ICCVAM Test Method Evaluation Report on a Proposed In Vitro Testing Strategy for U.S. Environmental 

Protection Agency Ocular Hazard Classification and Labeling of Antimicrobial Cleaning Products

Sep 13, 2010 ICCVAM Test Method Evaluation Report: Current Validation Status of In Vitro Test Methods Proposed for 

Identifying Eye Injury Hazard Potential of Chemicals and Products

Sep 13, 2010 ICCVAM Test Method Evaluation Report: Recommendation to Discontinue Use of the Low Volume 

Eye Test for Ocular Safety Testing
46 National Toxicology Program

Table 9: NICEATM-ICCVAM Workshops and Peer Review Meetings in FY 2010 

Date Meeting Topics

Jun 17-18, 2010 Scientific Advisory Committee on • Regulatory acceptance and availability of ICCVAM-recommended

Alternative Toxicological Methods alternative test methods

(U.S. EPA, Research Triangle Park, NC) • Assessment of acute and chronic pain in animals

• Federal agency research, development, translation and validation

activities relevant to the NICEATM-ICCVAM Five-Year Plan

• Current issues in the validation of alternative methods for 

assessing chemically-induced eye injuries

• Updates on international collaborations

• Alternative methods of vaccine potency testing

Sep 14-16, 2010 International Workshop on Alternative • Review of the state of the science of alternative methods 

Methods to Reduce, Refine, and Replace in vaccine potency and safety testing, and discussion of ways 

the Use of Animals in Vaccine Potency to promote their implementation

and Safety Testing: State of the Science • Identification of knowledge and data gaps that must be addressed 

and Future Directions to develop alternative methods to further reduce, refine, and

(Natcher Conference Center, replace the use of animals in vaccine potency and safety testing

NIH, Bethesda, MD) • Identification and prioritization of research, development, and 

validation efforts needed to address these knowledge and data 

gaps in order to advance alternative methods for vaccine potency

and safety testing, while ensuring continued protection of human

and animal health

Table10: Actions on Nominations or Submissions by NICEATM-ICCVAM in FY 2010 

Test Method Nomination or Submission Nominator or Sponsor/Activity Status

Request for assessment of the validation of the LLNA for U. S.CPSC/ICCVAM has recommended that the LLNA may be used 

classification of sensitizers to test any chemical or product for allergic contact dermatitis 

hazard potential unless the chemical or product to be tested has

properties that may interfere with the ability of the LLNA to 

detect sensitizing substances.

MCF-7 Estrogenic Activity Cell Proliferation Assay CertiChem, Inc./Validation study ongoing, completion expected 

Spring 2011. 

BG1Luc ER TA test method (also known as the LUMI-CELL® Xenobiotic Detection Systems, Inc./Validation study complete. 

test method) NICEATM staff and international collaborators presented an update

on the international validation study of the BG1Luc ER TA test 

method at the 2010 meeting of the Society of Toxicology. Expert 

panel to review background draft review document, March 2011.

In vitro test method for assessment of the eye irritation potential Alternative Testing Steering Committee: S.C. Johnson and Son, Inc.,

of antimicrobial cleaning products The Procter and Gamble Company, and The Accord Group/ 

(ACMPs) ICCVAM concluded that there are currently insufficient data with 

which to adequately demonstrate that the proposed strategy 

can classify test substances to all four EPA ocular hazard 

categories. ICCVAM also concluded that the data were insufficient

to support definitive recommendations on an alternate AMCP 

testing strategy to classify substances in all four EPA ocular 

hazard categories. ICCVAM recommended future studies that 

could further characterize the usefulness and limitations of a 

testing strategy for AMCPs. These recommendations were 

transmitted to U.S. Federal agencies, September 2010.
NTP Annual Report 2010 47

Table 11: NICEATM-ICCVAM Recommendations in FY 2010 

Test Method ICCVAM Recommendations/Agency Status

Routine Use of Topical Anesthetics, Recommended to U.S. Federal agencies, September 2010

Systemic Analgesics, and Humane 

Endpoints in Ocular Safety Testing

Cytosensor microphysiometer (CM) Recommended to U.S. Federal agencies, September 2010

ocular test method

Proposed in vitro testing strategy that uses Further studies recommended, September 2010

three in vitro test methods to assess the 

eye irritation potential of antimicrobial 

cleaning products

OECD Test Guideline (TG) 487, In Vitro November 2009: ICCVAM and its Genetic Toxicity Working Group (GTWG) 

Micronucleus Assay recommended approval of TG 487 based on the results of the cytotoxicity study data 

provided by European Union and U.S. laboratories from November 2008 to July 2009

Expanded applicability domain for the LLNA ICCVAM recommended to U.S. Federal agencies in 2010

LLNA performance standards; updated LLNA U.S. acceptance in 2010

test method protocol (20% animal reduction) OECD adoption (TG 429), July 2010

Reduced LLNA method U.S. acceptance in 2010

(40% animal reduction) OECD adoption (TG 429), July 2010

Nonradioactive LLNA versions ICCVAM recommended to U.S. Federal agencies, June 2010

(BrdU-ELISA and DA) OECD adopted test guidelines (TG 442A, TG 442B), July 2010

Contact information: NTP Interagency Center for the Evaluation of Alternative Toxicological Methods, Dr. William Stokes, Director. 

niceatm@niehs.nih.gov or NICEATM/ICCVAM website http://iccvam.niehs.nih.gov
48 National Toxicology Program

NTP Research and Testing Program

Nomination, Selection, Evaluation, and Review

Nominations for Study

The NTP maintains a balanced research and testing program that provides data addressing a wide variety of 

issues important to public health. The NTP actively seeks to identify and select for study chemicals and other 

substances for which sufficient information is not available to adequately evaluate potential human health 

hazards. The NTP accomplishes this goal through a formal open nomination and selection process. Substances 

considered appropriate for study generally fall into two broad, yet overlapping, categories:

 1. Substances judged to have high concern as a possible public health hazard based on the extent of human 

exposure and/or suspicion of toxicity.

 2. Substances for which toxicological data gaps exist and additional studies would help assess potential human 

health risks, e.g., by extrapolating data across species or by evaluating dose-response relationships.

Input is also solicited regarding the nomination of studies that test hypotheses to enhance the predictive ability 

of future NTP studies, address mechanisms of toxicity, or fill significant gaps in the knowledge of the toxicity of 

classes of chemical, biological, or physical substances. Increased efforts continue to focus on:

• Improving the quality of the nominations of chemicals, environmental agents, or issues for study 

so that public health and regulatory needs are addressed.

• Broadening the base and diversity of nominating organizations and individuals.

• Increasing nominations for studying non-cancer toxicological end points.

The nomination process is open to the public. The NTP routinely solicits nominations at conferences and 

workshops; through the NTP newsletter, Federal Register notices, the NTP website (http://ntp.niehs.nih.gov), 

and from interested individuals and groups. Also, NCI, FDA, NIOSH, and NIEHS routinely identify and forward 

nominations to the NTP. The NTP also reviews environmental occurrence and human exposure databases 

and the scientific literature to identify substances of potential interest.

Contact Information: Office of Nomination and Selection, Dr. Scott Masten, masten@niehs.nih.gov. 

Nomination website: http://ntp.niehs.nih.gov/ select “Nominations to the Testing Program.”

Review and Selection Process

Reviewing and selecting substances and issues nominated for study is a multi-step process (see figure 5 and 

http://ntp.niehs.nih.gov/go/156) that addresses a broad range of concerns from participating representatives 

from the NIEHS, other Federal agencies, the NTP Board of Scientific Counselors (see page 8), the NTP Executive 

Committee (see page 13), and the public. This multi-step evaluative process provides the NTP with direction 

and guidance to ensure that its testing program addresses toxicological concerns in all areas of public health 

and that there is balance among the types of substances selected for study (e.g., industrial chemicals, consumer 

products, therapeutic agents). Figure 5 summarizes the study nomination review process, and Table 12 lists the 

nominations reviewed in FY 2010.
NTP Annual Report 2010 49

NTP Study Nomination Review Process

Table 12: Nominations Reviewed in FY 2010 

Substance [CASRN] Nominator Nomination Rationale Study Recommendation

Butterbur NIEHS • Use as a dietary supplement • Chemical characterization and

(Petasites hybridus) extract • Lack of toxicological data in vitro toxicity screening assays

[90082-63-6] • Suspicion of toxicity based on pharmacological for multiple butterbur preparations

activity of constituents • Subchronic toxicity studies

• Potential presence of hepatotoxic • Multigeneration developmental

pyrrolizidine alkaloids and reproductive toxicity studies

 • Chronic toxicity and 

 carcinogenicity studies 

Evening primrose oil NIEHS • Use as a dietary supplement, particularly for • Subchronic toxicity studies

(Oenothera biennis extract) autoimmune conditions • Immunotoxicity studies

[90028-66-3] • Lack of adequate toxicological data • Multigeneration reproductive 

 toxicity studies

Hydroquinone FDA • Use in drugs and cosmetics • Comparative metabolism 

[123-31-9] • Evidence of carcinogenicity from oral exposures and disposition studies in mice

in prior NTP studies and rats by oral and dermal routes

• Insufficient toxicological data for regulatory of administration

hazard determination • Subchronic toxicity studies and 

 carcinogenicity studies (dermal 

 route of administration)

 • Multigeneration reproductive 

 toxicity studies (oral route 

 of administration)
50 National Toxicology Program

Substance [CASRN] Nominator Nomination Rationale Study Recommendation

Hydroxyurea NIEHS • Long-term safety concern when used as therapy for • Multigenerational developmental 

[127-07-1] Private sickle cell anemia; and reproductive toxicity studies

Individual • Critical data need, identified by CERHR, for including neurotoxicity and

multi-generation experimental animal studies to immunotoxicity assessments

assess the long-term effects of prenatal and postnatal • Carcinogenicity studies

exposures on postnatal development including 

developmental neurotoxicity, reproductive function, 

and carcinogenicity

Silica flour Private • Use in skin care and pharmaceutical products • Chemical characterization of silica 

[14808-60-7] Individual • Inhalation exposures associated with flour-containing products

autoimmune disease • Absorption and tissue disposition

• Lack of toxicity data for oral and dermal exposures following oral administration

• Insufficient data to evaluate dose-response for renal • Immunotoxicity studies

and autoimmune effects by any route of exposure (oral route of exposure)

Isoflavones in NIEHS • Widespread exposure • Studies to determine the feasibility 

soy infant formula • Concern for potential adverse effects soy of direct oral administration 

on human development; of infant formula to rodents (rats 

• Address certain critical data gaps and research and/or mice) during the period 

needs identified in the NTP CERHR Expert Panel of lactation

Report on Soy Infant Formula • Reproductive development 

 and fertility study in rodents with 

 soy infant formula and a mixture 

 of isoflavones at the ratio found 

 in soy infant formula (oral 

 administration to pups during 

 period of lactation)

 • Uterotrophic assay to assess 

 interactions among the individual

 isoflavones in soy infant formula 

 • Pharmacokinetic studies to assess

 metabolism of daidzin to equol 

 during the period of lactation

Valerian NIEHS • Use as a dietary supplement • Chemical characterization 

(Valeriana officinalis L.) • Lack of toxicological data short-term in vivo toxicity 

root extract • Concern for adverse developmental and screening assays for multiple 

[8057-49-6], reproductive effects valerian preparations 

Valerian oil • Subchronic toxicity and 

[8008-88-6] developmental and reproductive 

 toxicity studies including 

 neurotoxicity assessments

 • Carcinogenicity studies

Evaluation 

In carrying out its mission, the NTP provides toxicological evaluations on substances of public health concern. 

The NTP can initiate bioassays to characterize potential carcinogenicity of only a small fraction of the thousands 

of substances for which there is little or no information. Many more substances are also studied to assess a 

variety of non-cancer health-related effects including, but not limited to, reproductive and developmental 

toxicities, immunotoxicity, neurotoxicity, and genotoxicity. Other biological parameters are often assessed, such 

as quantifying the disposition and excretion of substances, identifying and correlating biochemical markers with 

exposure and metabolism, and examining genetic polymorphisms in human drug metabolizing enzymes to 

understand the susceptibility of individuals and populations to xenobiotic-induced toxicity.
NTP Annual Report 2010 51

An NTP project review committee evaluates a study’s project plan (e.g., design, methods, hypothesis, etc.). 

The toxicological evaluation for carcinogenicity generally involves repeated administration of a substance to 

groups of laboratory animals for up to two years. Many short-term, subchronic studies are designed to provide 

dose-setting information for longer, chronic exposure studies and to address specific gaps in the toxicology 

database. The adverse health effects from short- or long-term exposures to different dose levels of a substance 

are examined by observation, histopathology, and several toxicology endpoints, comparing them with control 

groups of animals not exposed to the substance. Many substances are also studied using protocols specifically 

designed to address how the substance causes particular toxic effects. The NTP has specific requirements that 

testing laboratories comply with the Animal Welfare Act of 1966 and adhere to the principles in the “Guide for 

the Care and Use of Laboratory Animals” (NRC, 1996). General information about the objectives and procedures 

of NTP study protocols is available on the NTP website (http://ntp.niehs.nih.gov/go/type). Current testing status 

can be found at http://ntpsearch.niehs.nih.gov/index.html?col=010stat.

The NTP carries out toxicology and carcinogenesis testing through two primary mechanisms: laboratory studies 

conducted in contract laboratories (see table 13) and studies conducted via IAGs at agencies (see page 78). In 

addition to toxicology research on compounds and exposures, the NTP supports developing new techniques 

and methods to improve the ability to identify and assess potential environmental toxicants and to develop 

and validate novel and alternative testing methods that will reduce, replace, or refine animal use. The NTP also 

supports developing improved statistical methods for designing and evaluating the results of toxicology studies.

Table 13: NTP Contracts That Supported NTP Testing Activities in FY 2010 

Description Contractor

Archives and Specimen Repository Experimental Pathology Labs

Cell Phone Radiation IIT Research Institute

Chemistry Support Midwest Research Institute

Chemistry Support Research Triangle Institute

Chemistry Support (2) Battelle Memorial

Effects of Chemicals in Developing Animals Research Triangle Institute

Evaluation of Test Agents in Laboratory Animals Battelle Memorial

Evaluation of Test Agents via Inhalation Exposure Battelle Memorial

Evaluate Toxicological Potential of Test Agents Battelle Memorial

Inhalation Toxicology Alion Science & Technology

Investigative Research Support Contract Integrated Laboratory Systems

NTP Computer and Statistical Support Constella

NTP Computing and CEBS Support Vistronix, Inc.

NTP Information Systems Support Z-Tech Corporation

NTP Information Systems Support SciMetrika

NTP Technical Reports Preparation Service Biotechnical Services

Preparation of Background and Nomination Information Integrated Laboratory Systems

Perform Cell Dose Response Standard Curves Hemogenix, Inc

Provantis Software for NTP INSTEM LSS

Quality Assessment/Pathology Support Experimental Pathology Labs

Quality Assessment Reports for Audits and Inspections Dynamac

Statistical Support Services SRA International, Inc.

Therapeutic Agents to Induce Immunotoxicity Virginia Commonwealth University
52 National Toxicology Program

Review and Dissemination 

The results of toxicology and carcinogenesis studies undergo rigorous peer review and are published 

in several NTP report series:

• Technical Reports (TR). This series presents the results of long-term, generally 2-year, toxicology and 

carcinogenicity studies, typically conducted in rats and mice. Results of genetic toxicology, absorption, 

distribution, metabolism, and excretion (ADME), and toxicokinetic studies are often included in the reports. 

The Technical Reports Review Subcommittee of the NTP BSC (see Advisory Boards and Committees, page 8) 

evaluates the draft reports at a public meeting. Starting in 2011, the draft reports will be reviewed by ad hoc

expert panels.

• NTP Toxicity (TOX) Reports. TOX reports are prepared for studies where the substance exposure period is 

short term, generally up to 13 weeks. Draft reports are typically peer reviewed by letter review.

• Genetically Modified Models (GMM) Reports. NTP began the GMM report series in May 2003. 

This report series presents the results of substances evaluated by NTP in transgenic mouse strains 

(e.g., p53+/- heterozygous and Tg.AC mice). Starting in 2011, the draft reports will be reviewed by 

ad hoc expert panels.

Abstracts of the TR, TOX, and GMM series are posted on the NTP website, and PDF files of completed reports are 

available at the NTP website (http://ntp.niehs.nih.gov/go/reports) and are also catalogued in PubMed. Pathology 

data from the NTP rodent studies included in these reports undergo several reviews. The final review is by a 

pathology working group (PWG), a panel of experts convened by the NTP to review the microscopic evaluations. 

After the NTP PWG review, pathology tables and body weight and survival graphs for the completed studies 

are made publicly available (http://ntp.niehs.nih.gov/go/peerreview) until draft study reports are completed 

for peer review. 

When the draft reports are completed for peer-review, the abstracts are put on the NTP website with the list of 

reports. The pathology tables and the body weight and survival graphs are accessible at the end of the abstract 

text. Following peer review, the NTP finalizes the report, posts it on its website (http://ntp.niehs.nih.gov/go/reports) 

and provides electronic links to the final pathology tables and the curves for body weight and survival. Study 

summaries for other types of studies, such as immunotoxicity, developmental toxicity, and reproductive toxicity 

studies, are also available on the “NTP Study Reports” page on the website. 

Chronic Toxicity/Carcinogenicity Studies

In the area of general toxicology assessments, the scope and types of studies performed are dictated mainly 

by the data needs for the specific substance being studied. General toxicology studies usually fall into two 

categories: subchronic or pre-chronic studies, and 2-year chronic toxicology and carcinogenicity studies. 

Two-year studies in rodents are a method by which chemicals or physical agents are identified as having 

the potential to be hazardous to humans.

The chronic toxicology and carcinogenicity studies in conventional rodent models generally use both male 

and female rats (Fischer 344/N, Harlan Sprague Dawley, or Wistar Han) and mice (B6C3F1 hybrid), with three 

exposure levels plus untreated controls, in groups of 50 animals, for two years; other rodent models (e.g., 

genetically modified mice) are used as needed. If adequate data exist in the literature for one rodent species 

(rats or mice), then typically only the remaining species is studied. The NTP interfaces its testing with regulatory 

agencies and the private sector to minimize duplication of effort. Studies ongoing, initiated, completed, and 

published in FY 2010 are listed in Tables 14, 15, 16, and 17, respectively. 
NTP Annual Report 2010 53

The NTP describes the results of individual experiments on a chemical agent and notes the strength of evidence 

for conclusions of each study. Negative results, in which the study animals do not have a greater incidence of 

neoplasia than do control animals, do not necessarily mean that a chemical is not a carcinogen, because the 

experiments are conducted under a limited set of conditions. Positive results demonstrate that a chemical 

is carcinogenic for laboratory animals under the conditions of the study and indicate that exposure to the 

chemical has the potential to be hazardous to humans. 

Table 14: Chronic Toxicity/Carcinogenicity Studies Ongoing During FY 2010 

Study [CASRN] Species/Strain Route Study Length Project Leader

Acrylamide [79-06-1] Rats: F344 (NCTR) Water 2 years Beland

Mice: B6C3F1/NCTR BR 

(C57BL/6N x C3H/HEN MTV-)

Aging cohort study - Mice: 129S1/SvImJ Not applicable 2 years French

129/SvlmJ mouse

Aging cohort study - Mice: B6C3F1 (Jackson) Not applicable 2 years French

B6C3F1J mouse

Aging cohort study - Mice: C3H/HeJ Not applicable 2 years French

C3H/HeJ mouse

Aging cohort study - Mice: C57BL/6J (Jackson) Not applicable 2 years French

C57/BL/6J mouse

Aging cohort study - Mice: CAST/EiJ (M. m. castaneus) Not applicable 2 years French

CAST/EiJ mouse

Aging cohort study - Mice: NZO/HiLtJ Not applicable 2 years French

NZO/HiLtJ mouse

Aging cohort study - Mice: PWK/PhJ Not applicable 2 years French

PWK/PhJ mouse

Aging cohort study - Mice: WSB/EiJ (M. m. domesticus) Not applicable 2 years French

WSB/EIJ mouse

Aging cohort study – Mice: A/J Not applicable 2 years French

A/J mouse

Aging cohort study - Mice: NOD. B10Sn-H2(b)/J Not applicable 2 years French

NOD. B10Sn-H2(b)/J 

Aloe vera whole leaf Rats: F344 (NCTR) Water 2 years Boudreau

extract (native) Mice: B6C3F1/NCTR BR

(C57BL/6N x C3H/HEN MTV-)

Antimony trioxide [1309-64-4] Rats: Wistar Han Inhalation 2 years Stout 

Mice: B6C3F1

3’-Azido-3’- [30516-87-1] Mice: B6C3F1/NCTR BR Gavage 2 years Beland

deoxythymidine (AZT) (C57BL/6N x C3H/HEN MTV-)

*3’-Azido-3’- [30516-87-1] Mice: B6C3F1/NCTR BR Gavage 9 months Leakey

deoxythymidine (AZT) (C57BL/6N x C3H/HEN MTV-)

AZT/drug combinations Mice: B6C3F1/NCTR BR Gavage 2 years Beland

transplacental/ neonatal (C57BL/6N x C3H/HEN MTV-)

study

AZT/drug combinations Mice: B6C3F1/NCTR BR In utero 2 years Beland

transplacental (C57BL/6N x C3H/HEN MTV-) 

carcinogenesis study

2,3-Butanedione (diacetyl) [431-03-8] Rats: Wistar Han Inhalation 2 years Morgan

Mice: B6C3F1
54 National Toxicology Program

Study [CASRN] Species/Strain Route Study Length Project Leader

Cobalt [7440-48-4] Rats: F344/NTac Inhalation 2 years Hooth

Mice: B6C3F1

Dibutyl phthalate [84-74-2] Rats: Harlan SD Feed 2 years Blystone

Mice: B6C3F1

N,N-Dimethyl-p-toluidine [99-97-8] Rats: F344/N Gavage 2 years Dunnick

Furan 110-00-9 Rats: F344 (NCTR) Gavage 2 years Beland

Mice: B6C3F1

Ginkgo biloba extract [90045-36-6] Rats: F344/N Gavage 2 years Chan

Mice: B6C3F1

Glycidamide [5694-00-8] Rats: F344 (NCTR) Water 2 years Beland

Mice: B6C3F1/NCTR BR

(C57BL/6N x C3H/HEN MTV-)

Green tea extract Rats: Wistar Han Gavage 2 years Chan

Mice: B6C3F1

2-Hydroxy-4- [131-57-7] Rats: Harlan SD Feed 2 years Auerbach

methoxybenzophenone Mice: B6C3F1

Indole-3-carbinol [700-06-1] Rats: Harlan Sprague Dawley Gavage 2 years Wyde

Mice: B6C3F1

Kava kava extract [9000-38-8] Rats: F344/N Gavage 2 years Behl/Chan

Mice: B6C3F1

Metalworking fluids Rats: Wistar Han Inhalation 2 years Morgan

(CIMSTAR 3800) Mice: B6C3F1

Metalworking fluids Rats: Wistar Han Inhalation 2 years Morgan

(Trim VX) Mice: B6C3F1

Methyl trans-styryl ketone [1896-62-4] Rats: F344/N Topical 2 years Cunningham

Mice: B6C3F1 application

Pentabromodiphenyl oxide [32534-81-9] Rats: Wistar Han Gavage 2 years Dunnick

(technical) (DE 71) Mice: B6C3F1

PFOA [335-67-1] Rats: Harlan SD Feed 2 years Blystone

PFOA [335-67-1] Rats: Harlan SD Feed 2 years Blystone

beta-Picoline [108-99-6] Rats: F344/N Water 2 years Wyde

Mice: B6C3F1

Pyrogallol [87-66-1] Rats: F344/N Topical 2 years MercadoMice: B6C3F1 application Feliciano

All-trans-retinyl palmitate [79-81-2] Mice: SKH-1 Hairless Topical 1 year Boudreau

application

Styrene-acrylonitrile trimer Rats: F344/N Feed 2 years Behl/Chhabra

Tetrabromobisphenol A [79-94-7] Rats: Wistar Han Gavage 2 years Dunnick

Mice: B6C3F1

alpha/beta-Thujone mixture [76231-76-0] Rats: F344/N Gavage 2 years Hooth

Mice: B6C3F1

Trimethylolpropane [15625-89-5] Rats: F344/N Topical 2 years Surh/Chhabra

triacrylate Mice: B6C3F1 application

Tripelennamine [154-69-8] Rats:F344/N Feed 11 months Jackson

Hydrochloride Mice: B6C3F1

Vinylidene chloride [75-35-4] Rats: F344/N Inhalation 2 years Wyde

Mice: B6C3F1

Bromodichloroacetic acid [71133-14-7] Rats: F344/NTac Water 2 years Hooth

(water disinfection byproduct) Mice: B6C3F1

Zinc carbonate, basic 5263-02-5 Rats: Harlan SD Feed 2 years Wyde
NTP Annual Report 2010 55

Table 15: Chronic Toxicity/Carcinogenicity Studies Initiated During FY 2010 

Study [CASRN] Species/Strain Route Study Length Project Leader

Aging Cohort Study – Mice: CAST/EiJ Not applicable 2 years French

CAST/EiJ mouse (M. m. castaneus)

Aging Cohort Study - Mice: NZO/HiLtJ Not applicable 2 years French

NZO/HiLtJ mouse

Aging Cohort Study - Mice: C3H/HeJ Not applicable 2 years French

PWK/PhJ mouse

Aging Cohort Study - Mice: PWK/PhJ Not applicable 2 years French

C57/BL/6J mouse

Aging Cohort Study - Mice: WSB/EIJ Not applicable 2 years French

WSB/EIJ mouse (M. m. domesticus)

Aging Cohort Study - Mice: NOD. B10Sn-H2(b)/J Not applicable 2 years French

NOD. B10Sn-H2(b)/J 

Dibutyl phthalate 84-74-2 Rats: Harlan SD, Mice:B6C3F1 Feed 2 years Blystone

2-Hydroxy-4- 131-57-7 Rats:Harlan Sprague-Dawley, Feed 2 years Auerbach

methoxybenzophenone Mice: B6C3F1
56 National Toxicology Program

Only summaries of carcinogenic activity conclusions are included in the tables. 

Complete information is available in the full study reports found at http://ntp.niehs.nih.gov/.
NTP Annual Report 2010 57
58 National Toxicology Program

Only summaries of carcinogenic activity conclusions are included in the tables. Complete information is available in the full study reports 

found at http://ntp.niehs.nih.gov/. 
NTP Annual Report 2010 59
60 National Toxicology Program

The NTP anticipates that six Technical Reports will undergo peer review in FY 2010, as shown in Table 18.

Table 18: Technical Reports expected to undergo peer review in FY 2011 

Technical 

Chemical [CASRN] Report Use

Number

Acrylamide [79-06-1] TR-575 Acrylamide, a water-soluble α,β-unsaturated amide, is a contaminant 

Inhalation in baked and fried starchy foods, including French fries, potato chips, 

and bread, as a result of Maillard reactions involving asparagine and 

reducing sugars. Additional sources of acrylamide exposure include 

cigarettes, laboratory procedures involving polyacrylamide gels, 

and various occupations (e.g, monomer production and 

polymerization processes).

AIDS Therapeutics: TR-569 With the increased administration of multidrug regimens to pregnant

3’-Azido-3’-Deoxythymidine [30516-87-1] women who are HIV-1 positive, along with the increased efficacy 

(AZT) of these combinations, determining the long-term consequences 

Lamivudine (3TC) [134678-17-4] of the antiretroviral agents in noninfected children becomes important. 

Nevirapine (NVP) [129617-40-2] The goal of the current study was to determine the carcinogenicity of 

Melfinavir Misylate (NFV) [159989-65-8] combinations of antiretroviral drugs in male and female B6C3F1 mouse 

pups exposed transplacentally and monitored for 2 years.

• AZT was synthesized initially for use as an anticancer agent and was later

 reported to block the infectivity and cytopathic effects, in vitro, of 

 human immunodeficiency virus type-1 (HIV-1), due to the inhibition 

 (by AZT 5′-triphosphate) of viral reverse transcriptase. Pregnant women 

 who are positive for HIV-1 are given AZT to manage the infection and to 

 prevent maternal-to-fetal transmission of the virus.

• 3TC was synthesized initially as a racemate and then in enantiomerically 

 pure forms. 3TC (as 3TC 5′-triphosphate) is thought to inhibit viral reverse

 transcriptase by competing with deoxycytidine 5′-triphosphate for 

 incorporation into HIV-1 DNA. When used for the management of HIV-1 

 infections, 3TC is always used in combination with another nucleoside 

 reverse transcriptase inhibitor (e.g., AZT) and either a protease inhibitor 

 (e.g., nelfinavir mesylate, NFV) or a non-nucleoside reverse transcriptase 

 inhibitor (e.g., nevirapine, NVP).

•NVP a non-nucleoside reverse transcriptase inhibitor, was first 

 synthesized in 1991. NVP inhibits HIV-1 reverse transcriptase 

 noncompetitively by binding to an allosteric site on the enzyme; this 

 action is specific for HIV-1 reverse transcriptase. NVP is usually given as 

 part of a three-drug regimen. Typical regimens in adults and adolescents

 include NVP and 3TC or emtricitabine and AZT or tenofovir.

•NFV synthesis was reported in 1997. NFV acts by inhibiting HIV-1 

 protease, the enzyme responsible for cleavage of the polyprotein 

 resulting from the gag and gag-pol genes of HIV-1. This inhibition 

 results in an immature, noninfectious virus. NFV is always used in 

 combination with other antiretroviral agents, typically two nucleoside 

 reverse transcriptase inhibitors (e.g., AZT and 3TC). 
NTP Annual Report 2010 61

Chemical [CASRN] Report Use

A Nondecolorized TR-577 Aloe barbadensis Miller (Aloe vera) has enjoyed a long history of lay

Whole Leaf Extract acceptance as an herbal remedy and is perhaps the most popular herbal

of Aloe barbadensis Miller remedy in use today. In recent times, the oral consumption of Aloe vera

(Aloe Vera) has been promoted as a prophylaxis and treatment to alleviate a variety 

of unrelated systemic conditions.

α,β-Thujone [76231-76-0] TR-570 Thujone is a monoterpene ketone that exists in two stereoisomeric forms

(-)-3-isothujone, or α-thujone, and (+)-3-thujone, or β-thujone. Thujone 

occurs in nature as a mixture of α- and β-isomers and is the primary 

constituent of essential oils derived from a variety of plants. Although 

thujone itself is banned as a food additive in the United States, it appears 

in many other natural flavoring substances that are approved food 

additives and flavorings.

Kava Kava Extract [9000-38-8] TR-571 Kava beverages, made from dried roots of the shrub Piper methysticum,

have been used ceremonially and socially in the South Pacific and in 

Europe since the 1700s. The drink is reported to have pleasant mild 

psychoactive effects, similar to alcoholic beverages. In the United States, 

kava kava is an herbal product used extensively as an alternative to 

anti-anxiety drugs such as Xanax® and Valium.®

Methyl trans-Styryl Ketone [1896-62-4] TR-572 Methyl trans-styryl ketone is used as a synthetic flavoring agent

and a fragrance additive in food and personal care products.

Retinoic Acid (RA) [302-79-4] TR-568 Topical retinoids, compounds that are metabolites, analogs, or derivatives

Retinyl Palmatate (RP) [79-81-2] of retinol and possess biological vitamin A activity, are among the most 

used adjunctive agents for the mitigation of fine wrinkles, mottled 

hyperpigmentation, and tactile roughness of photodamaged and 

chronically aged skin. RA is the most active biological form of vitamin A 

and remains the medical treatment of choice for photoaged skin. RA 

is the major storage form of vitamin A in the skin and, because RP is also 

the most stable of available vitamin A esters, it is readily incorporated into

the oil phase of cosmetic creams or lotions. Therefore, the topical 

application of RP is a practical strategy for increasing the levels of 

vitamin A in the skin.

Styrene-acrylonitrile (SAN) TR-573 Styrene-acrylonitrile trimer is a byproduct of the production

Trimer of acrylonitrile styrene plastics and is created in specific manufacturing 

processes for polymers of acrylonitrile and styrene.
62 National Toxicology Program

General Toxicology Studies 

The NTP performs prechronic toxicity studies to address specific deficiencies in toxicology databases for 

chemicals, such as an understanding of toxicity with repeated exposures; to identify target organs for more 

in-depth systems toxicology evaluations and mechanistic studies; and to provide dose-setting information 

for possible chronic studies.

Although designs are flexible, prechronic studies usually involve exposures of rats and mice of both sexes to 

substances for periods of 14-90 days. Assessments almost always include tissue histopathology, clinical pathology, 

and sperm motility or measurements of estrous cycle length. The frequency of malformed red blood cells 

(micronucleated erythrocytes, a measure of chromosomal damage) is determined as an in vivo measure 

of genotoxic potential.

The study protocol may include more detailed or focused studies when findings published in the existing 

scientific literature or identified in initial NTP studies suggest a target organ or system. The study protocol may 

include separate studies of reproductive, genetic, or immunological toxicity based on the outcome of the toxicity 

screens and may use additional endpoints to improve our understanding of the mechanisms and modes 

of action of a chemical.

In some cases, the NTP uses alternative models, including genetically modified mouse models and 

non-mammalian models, for prechronic studies. Such studies are presented in the section “Genetic 

and Alternative Test Systems” (page 72). Tables 19-21 list the toxicity studies that were ongoing, initiated, and 

completed, respectively, during FY 2010. Information is available at http://ntp.niehs.nih.gov/go/reports. 

Table 22 lists toxicity studies planned for FY 2011.
NTP Annual Report 2010 63

Table 19: Ongoing Prechronic Toxicity Studies During FY 2010

Study Route Project Study [CASRN] Species/Strain Length Leader

*3’-Azido-3’-deoxythymidine (AZT) [30516-87-1] Mice: P53 +/- (FVB/N) Gavage 9 months Leakey

Glucosamine hydrochloride [9007-28-7] Rats: Zucker – Obese Gavage 13 weeks Leakey

+ chondroitin sulfate [66-84-2] (HsdHlr: ZUCKER-Leprfa)

Rats: Zucker - Lean 

(HsdHlr: ZUCKER-Lepr+)

Microcystin-LR (TGMX) [101043-37-2] Rats: Wistar Han Intravenous 1/2/6/24 hours Walker 

Pregnancy rate comparison study Rats: Harlan SD Not applicable 16 weeks Vallant

(Indianapolis Facility),

Rats: Harlan SD 

(Dublin Facility)

QT drugs (bepridil hydrochloride) [74764-40-2] Dog: Beagles Oral (capsule) 1 day Hooth

QT drugs (diltiazem hydrochloride) [33286-22-5] Dog: Beagles Oral (capsule) 1 day Hooth

QT drugs (loratadine) [79794-75-5] Dog: Beagles Oral (capsule) 1 day Hooth

QT drugs (lovastatin) [75330-75-5] Dog: Beagles Oral (capsule) 1 day Hooth

QT drugs (sotalol hydrochloride) [959-24-0] Dog: Beagles Oral (capsule) 1 day Hooth

QT drugs (terfenadine) [50679-08-8] Dog: Beagles Oral (capsule) 1 day Hooth

Sodium tungstate, dihydrate 10213-10-2 Rats: Harlan SD Water 13 weeks Hooth

Mice: B6C3F1

tris(2-Chloroisopropyl)phosphate 13674-84-5 Rats: Harlan SD Feed 90 days Stout

Mice: B6C3F1

Usnea lichen Rats: F344 (NCTR) Feed 90 days Leakey

Mice:B6C3F1/NCTR BR 2 weeks

(C57BL/6N x C3H/HEN MTV-) 

(+)-Usnic acid [7562-61-0] Rats: F344 (NCTR) Feed 2 weeks Leakey

Mice: B6C3F1/NCTR BR 90 days

(C57BL/6N x C3H/HEN MTV-) 
64 National Toxicology Program

Table 20: Toxicity Studies Initiated during FY 2010

Chemical [CASRN] Species/Strain Route Study Length Project Leader

Bisphenol A [80-05-7] Rats: SD (NCTR) Gavage 90 days Delclos

Black cohosh [84776-26-1] Mice: B6C3F1 Gavage 90 days MercadoFeliciano

bis(2-Chloroethoxy)methane [111-91-1] Mice: C57BL/6J (Jackson) Gavage 10 days, Dunnick

 Mice: C3H/HeJ 3 days

Ephedrine + caffeine combination Mice: C57BL/6J (Jackson) Gavage 10 days, Dunnick

 Mice: C3H/HeJ, 3 days

 Mice:C3H/HeJ

Insertional mutagenesis Mice:B6.SJL-Ptprc[a] Whole-body 8 weeks Irwin

(Radiation Levels) Pepc[b]/BoyJ exposure

Insertional mutagenesis II (SIN vector) Mice: B6.SJL-Ptprc[a] Intravenous 8 plus Irwin

 Pepc[b]/BoyJ, 3 weeks

 Mice: B6.SJL-Ptprc[a] 

 Pepc[b]/BoyJ

2,3-Pentanedione [600-14-6] Rats: Wistar Han Inhalation 90 days Morgan

 Mice: B6C3F1

Table 21: Toxicity Studies Completed during FY 2010

Chemical [CASRN] Species/Strain Route Study Length Project Leader

Abrasive blasting agents Rats: Harlan SD Inhalation 39 weeks, Roycroft

(blasting sand) Rats:F344/NTac 14 days

Abrasive blasting agents Rats: Harlan SD Inhalation 39 weeks, Roycroft

(specular hematite) Rats: F344/NTac 14 days

Acetoin [513-86-0] Rats: Wistar Han Inhalation 90 days, Morgan

Mice:B6C3F1 14 days

Cell phone radiation Rats: Harlan SD Whole-body 5 days, Wyde

(code division multiple access [CDMA]) Mice: B6C3F1 49 days

Cell phone radiation Rats:Harlan Sprague-Dawley, Whole-body 49 days Wyde

(global systems mobile [GSM]) Mice: B6C3F1 exposure 5 days

p-Chloro-a,a,a-trifluorotoluene [98-56-6] Rats: Harlan SD Inhalation 90 days Stout

Mice: B6C3F1

Glucosamine [3416-24-8] Rats: Zucker - Obese Gavage 13 weeks Leakey

(HsdHlr:ZUCKER-Leprfa)

Rats: Zucker - Lean 

(HsdHlr:ZUCKER-Lepr+)

Gum guggul extract Rats: Harlan SD Gavage 13 weeks Wyde

Mice: B6C3F1

Insertional mutagenesis II (SIN vector) Mice: B6.SJL-Ptprc[a] Intravenous 8 plus Irwin

Pepc[b]/BoyJ, 3 weeks

Ionic liquid toxicity Rats: Harlan SD Water 14 days Hooth

Mice: B6C3F1

2-Methoxy-4-nitroaniline [97-52-9] Rats: Harlan SD Feed Range Surh

Mice: B6C3F1 finding, 

14 days

Myristicin [607-91-0] Rats: F344/NTac Gavage 13 weeks Wyde

Mice:B6C3F1

Resveratrol [501-36-0] Rats: Wistar Han, Gavage 2 weeks, Germolec

Rats: F344/NTac, 13 weeks

Mice: B6C3F1

Serotype 5 adeno-associated Mice: BALB/c Intraductal 13 weeks Irwin

viral vector (rAAV5SCTLA4:Ig) cannulation

Vincamine [1617-90-9] Mice B6C3F1 Gavage 2 weeks Chan
NTP Annual Report 2010 65

Table 22: Toxicity Studies Planned for FY 2011*

Chemical [CASRN] Species/Strain Route Study Length Project Leader

3-Aminopyridine [462-08-8] Rats: F344/NTac Gavage 14 days Dunnick

Mice: B6C3F1

2-Aminopyridine [504-29-0] Rats: F344/NTac Gavage 14 days Dunnick

Mice: B6C3F1

4-Aminopyridine [504-24-5] Rats: F344/NTac Gavage 14 days Dunnick

Mice: B6C3F1

Comparison study of aminopyridines/ Rats: F344/NTac Gavage 1 & 4 hours Dunnick

troponin levels Mice: B6C3F1

Microcystin mixture (TGMX) Rats: Wistar Han Intravenous 1/2/6/24 hours Walker

Pyridine [110-86-1] Mice: B6C3F1 Gavage 14 days Dunnick

Serotype 2 adeno-associated Mice: BALB/c Intraductal 13 weeks Irwin

viral vector hAQP1 (rAAV2hAQP1) Cannulation

* Known test articles as of 10/1/2010. Others may be scheduled as protocols are finalized.

Mutagenesis and Genetic Toxicity 

Genetic toxicity test results are used to help interpret toxicity, carcinogenicity, or other in vivo test results 

and to provide a database for use in structure-activity analyses. Analysis of the early, multi-test database 

showed that positive results for a chemical in the Salmonella gene mutation assay were sometimes correlated 

with carcinogenicity in several species/sexes of rodents and at several tissue sites. Subsequently, studies of the 

correlation between mutagenicity test data and rodent carcinogenicity showed a strong association between 

clearly positive results in long-term mouse peripheral blood micronucleus tests and rodent carcinogenicity. 

The importance of genetic toxicity test data in assessing exposure hazard for NTP chemicals is underscored 

by the fact that most organic chemicals (other than hormones) identified as human carcinogens by the 

International Agency for Research on Cancer (IARC) are genotoxic, and the vast majority of them are 

detected by both the Salmonella assay and rodent micronucleus tests. Additional assays may be conducted 

with certain chemicals to gain further insight into the types of DNA and chromosomal damage induced 

by a chemical. Substances tested for genetic toxicity during FY 2010 are listed in Table 23. Information is 

available at (http://ntp.niehs.nih.gov/go/reports).
66 National Toxicology Program

Table 23: Ongoing and Completed Genetic Toxicity Studies During FY 2010

Chemical [CASRN] Testing Battery

2-Aminoanthracene [613-13-8] Salmonella

3’-Azido-3’-deoxythymidine (AIDS) [30516-87-1] Salmonella

Acetoin [513-86-0] Micronucleus

Antimony trioxide [1309-64-4] Micronucleus

2,6-Diethylaniline (2,6-DEA) [579-66-8] Salmonella

Dimethylamine borane [74-94-2] Salmonella

2’,2’’’-Dithiobisbenzanilide [135-57-9] Salmonella

Efavirenz [154598-52-4] Salmonella

2-Ethylaniline [578-54-1] Salmonella

3-Ethylaniline [587-02-0] Salmonella

4-Ethylaniline [589-16-2] Salmonella

Gum guggul extract Micronucleus

2-metyl-6-ethylanline [24549-06-2] Salmonella

Milk thistle extract [84604-20-6] Salmonella

Nelfinavir [159989-64-7] Salmonella

Nevirapine [129618-40-2] Salmonella

ortho-Phthalaldehyde [643-79-8] Micronucleus

p-Chloro-a,a,a-trifluorotoluene [98-56-6] Micronucleus

p-Toluidine [106-49-0] Salmonella

Pulegone [89-82-7] Salmonella

Senna (powdered) [8013-11-4] Micronucleus

Senna (powdered) [8013-11-4] Salmonella

Sennoside B [128-57-4] Salmonella

Sodium tungstate, dihydrate [10213-10-2] Micronucleus

Sodium tungstate, dihydrate [10213-10-2] Salmonella

Styrene-acrylonitrile trimer SANTRIMER2 Micronucleus

3TC (AIDS initiative) [134678-17-4] Salmonella

Thiodiglycolic acid [123-93-3] Salmonella

m-Toluidine [108-44-1] Salmonella

tris(2-Chloroisopropyl)phosphate [13674-84-5] Micronucleus

4,7,10-Trioxatridecane-1,13-diamine [4246-51-9] Salmonella

tris(2-Chloroisopropyl)phosphate [13674-84-5] Micronucleus

Vincristine sulfate salt [2068-78-2] Micronucleus

Zinc carbonate, basic [5263-02-5] Micronucleus
NTP Annual Report 2010 67

Organ System Toxicity

Nervous System, Developmental, and Reproductive Toxicity

Behavioral and neurological alterations in response to deleterious environmental agents often represent the 

earliest evidence of toxicity. These testing batteries examine the sensory, motor, autonomic, and peripheral 

nervous systems. The Functional Observational Battery employs observational screening, while the NIEHS 

test battery uses automated test systems to evaluate the various nervous system components. 

As part of its charge to test chemicals of concern for potential toxicity, the NTP evaluates developmental and 

reproductive toxicity primarily by using teratology and Reproductive Assessment by Continuous Breeding (RACB) 

study designs (see http://ntp.niehs.nih.gov/go/33668). The RACB study design was developed by the NTP to 

identify potential hazards from toxic effects on male and/or female reproduction, to characterize that toxicity, 

and to define the dose-response relationships for each compound. The study design has evolved over the years: 

initially the studies mainly used mice as the test species; now, they use rats almost exclusively. As our knowledge 

has improved and use of sensitive end points has increased, these advances have been incorporated into 

revisions of the study design. 

Table 24 lists completed and ongoing nervous system, developmental, and reproductive studies 

during FY 2010, and Table 25 lists studies planned for FY 2011.

Table 24: Ongoing and Completed Organ Systems Toxicity Studies During FY 2010

Chemical [CASRN] Species/Strain Route Project Leader Testing Battery

Acrylamide [79-06-1] Rats: F344 (NCTR) Gavage Beland Neurotox assessment

Bitter orange Rats: SD Gavage Hansen Teratology

Bitter orange with caffeine Rats: SD Gavage Hansen Teratology

Black cohosh [84776-26-1] Rats: Harlan SD Gavage Mercado- RACB - range-finding

Feliciano

n-Butyl-p-hydroxybenzoate [94-26-8] Rats: Harlan SD Gavage Blystone RACB - range-finding

 Nonylphenol [84852-15-3] Rats: SD (NCTR) Feed Newbold, Multigenerational

Delclos

p-Synephrine [94-07-5] Rats: SD Gavage Hansen Teratology

Synephrine and caffeine [94-07-5] Rats: SD Gavage Hansen Teratology

[58-08-2]

3,3’,4,4’-Tetrachloroazobenzene [14047-09-7] Rats: Crl:CD (SD) Gavage Hooth Teratology - 

pre-implantation

3,3’,4,4’-Tetrachloroazobenzene [14047-09-7] Rats: Crl:CD (SD) Gavage Hooth Postnatal developmental 

toxicity

tris(2-Chloroisopropyl)phosphate [13674-84-5] Rats: Harlan SD Gavage Stout Prenatal developmental 

toxicity

Vinclozolin [50471-44-8] Rats: Wistar Han Gavage Bishop Modified RACB

Table 25: RACB Studies Planned for FY 2011

Chemical [CASRN] Species/Strain Route

Black cohosh [84776-26-1] Rats: Harlan SD Gavage

Butyl paraben [94-26-8] Rats: Harlan SD Feed

Diisobutyl phthalate [84-69-5] Rats: Harlan SD Feed

4-Methylimidazole [822-36-6] Rats: Harlan SD Feed

Sodium tungstate, dihydrate [10213-10-2] Rats: Harlan SD TBD
68 National Toxicology Program

Immunotoxicity

NTP immunotoxicity studies address adverse effects on the immune system that may result from exposure 

to environmental chemicals, biological materials, or therapeutic agents. The identification of substances that 

have potential to cause injury to the immune system is of considerable public health significance as alterations 

in immune function can lead to increased incidence of hypersensitivity disorders, autoimmune or infectious 

disease, or neoplasia. Immunotoxicity caused by exposure to chemicals can be divided into two broad research 

areas: (1) studies of altered hematopoietic (blood cell development) or other immunologic events associated 

with exposure of humans and animals to chemicals and (2) studies of immune-mediated hypersensitivity (allergy 

and autoimmunity) resulting from exposure to environmental chemicals or therapeutics. In the former case, 

the immune system acts as a passive target (nonspecific) for the foreign substance, and the result may be an 

increased incidence or severity of infectious disease or neoplasia because of the inability to respond adequately 

to the invading agent. In hypersensitivity (i.e., allergy), the immune system responds to small molecular weight 

compounds that bind to host tissue, recognizing the complex as foreign antigen. This immune response to the 

chemical-host tissue complex may lead to diseases, such as respiratory tract allergies (e.g., asthma, rhinitis) or 

allergic contact (skin) dermatitis. Autoimmunity, another form of immune-mediated disease, is characterized 

by an immune response against constituents of the body’s own tissues (autoantigens). Table 26 lists completed 

and ongoing immunotoxicity studies during FY 2010.

Table 26: Ongoing and Completed Immunotoxicity Studies During FY 2010

Chemical [CASRN] Species/Strain Route Project Leader Testing Battery

Abrasive blasting agents Rats: Harlan Inhalation Roycroft Immunosuppression

(blasting sand) Sprague Dawley

Abrasive blasting agents Rats: Harlan Inhalation Roycroft Immunosuppression

(specular hematite) Sprague Dawley 

Autumn Sunset True Color Mice: CBA/Ca Jackson Subcutaneous Howard Hypersensitivity

Concentrate injection

3’-Azido-3’-deoxythymidine [30516-87-1] Mice: B6C3F1 Gavage Irwin Immunosuppression-

(AZT) range finding,

developmental

Black cohosh [84776-26-1] Mice: B6C3F1 Gavage Mercado- Immunotoxicity 

Feliciano

2,3-Butanedione (diacetyl) [431-03-8] Mice: BALB/c Topical Morgan Hypersensitivity

application

2,3-Butanedione (diacetyl) [431-03-8] Mice: BALB/c Inhalation Morgan Immunosuppressionrange finding

tert-Butyl hydroperoxide [75-91-2] Mice: BALB/c Topical Chhabra Hypersensitivity

application

o-Cresol [95-48-7] Mice: BALB/c Topical Chhabra Hypersensitivity 

application 

Dibenz(a,h)anthracene [53-70-3] Mice: B6C3F1 Gavage Germolec Immunosuppressionrange-finding, 

developmental,

immunosuppressionfull protocol

1,3-Dichloropropene [542-75-6] Mice: B6C3F1 Water Yang Immunosuppression- 

(Telone II) full protocol

Dimethylamine borane [74-94-2] Mice: BALB/c Topical Germolec Hypersensitivity

application

Double dark fudge Mice: CBA/ Ca Jackson Subcutaneous Howard Hypersensitivity

true color concentrate range finding

Double fudge concentrate Mice: CBA/Ca Jackson Subcutaneous Howard Hypersensitivity 

injection 
NTP Annual Report 2010 69

Chemical [CASRN] Species/Strain Route Project Leader Testing Battery

Echinacea purpurea, extract [90028-20-9] Mice: B6C3F1 Gavage Irwin Immunosuppressionfull protocol 

Elmiron (sodium [37319-17-8] Mice: B6C3F1 Gavage Germolec Immunosuppressionpentosanpolysulfate) full protocol

Ethanone, 1-(1,2,3,4,5,6,7, [54464-57-2] Mice: BALB/c Topical Chan Hypersensitivity 

8-octahydro-2,3,8, application

8-tetramethyl2-naphthalenyl)- (Iso-E Super) 

Genistein [446-72-0] Mice: Gavage Germolec Autoimmunity

NOD/MrKTac

Gum guggul extract Mice: B6C3F1 Gavage Wyde Immunosuppressionrange-finding

Heptachlor [76-44-8] Mice: BALB/c Topical Germolec Hypersensitivity

application

Ionic liquid (1-butyl- [79917-90-1] Mice: BALB/c Topical Hooth Hypersensitivity

3-methylimidazolium chloride) application 

Ionic liquid (1-butyl-1- [479500-35-1] Mice: BALB/c Topical Hooth Hypersensitivity

methylimidazolium chloride) application

Ionic liquid [1124-64-7] Mice: BALB/c Topical Hooth Hypersensitivity

(n-butylpyridinium chloride) application

Ionic liquid [65039-09-0] Mice: BALB/c Topical Hooth Hypersensitivity

(1-ethyl-3-methylimidazolium application

chloride)

Lovastatin [75330-75-5] Mice: B6C3F1 Gavage Germolec Immunosuppressionrange finding

Monoclonal antibody protein Mice: B6C3F1 Intraperitoneal Germolec Immunosuppressiontherapeutics (CD-4) injection full protocol

Monoclonal antibody protein Mice: B6C3F1 Intraperitoneal Germolec Immunosuppressiontherapeutics (CD-8) injection full protocol

Fullerene-C60 (1 micron) [99685-96-8] Mice: B6C3F1 Inhalation Walker ImmunosuppressionWistar Han range finding

Fullerene-C60 (50 nm) [99685-96-8] Rats: Wistar Han Inhalation Walker ImmunosuppressionMice: B6C3F1 range finding

1,5-Naphthalene diisocyanate [3173-72-6] Mice: BALB/c Topical Germolec Hypersensitivity

application

Nelfinavir mesylate [159989-65-8] Mice: B6C3F1 Gavage Germolec Immunosuppressionfull protocol,

developmental

Nevirapine [129618-40-2] Mice: B6C3F1 Gavage Germolec Immunosuppressionfull protocol,

developmental

Perfluorodecanoic acid [335-76-2] Rats: Harlan SD Gavage Blystone Immunosuppression

Phenol [108-95-2] Mice: B6C3F1 Water Germolec Immunosuppressionfull protocol

Resveratrol [501-36-0] Mice: B6C3F1 Gavage Germolec Immunosuppressionrange finding

Rosewood True Mice: CBA/Ca Jackson Subcutaneous Howard Hypersensitivity

Color Concentrate injection

Sodium tungstate, dihydrate [10213-10-2] Mice: B6C3F1 Water Hooth Immunosuppressionfull protocol

3,3’,4,4’-Tetrachloroazobenzene [14047-09-7] Rats: Sprague Dawley Gavage Behl Immunosuppressionrange finding

3,3’,4,4’-Tetrachloroazobenzene [14047-09-7] Rats: Crl:CD (SD) Gavage Behl Developmental
70 National Toxicology Program

Disposition, Metabolism and Toxicokinetic Studies

Complete dosimetry of a chemical or physical agent describes its ADME in the body at differing levels of exposure, 

over all ages, via several routes of exposure, and under varying genetic backgrounds in humans and test animals. 

Data from NTP chemical disposition and toxicokinetic studies are used in these studies. Substances evaluated during 

FY 2010 are listed in Table 27, and studies planned for FY 2011 are listed in Table 28. Most studies are conducted in 

intact laboratory animals; some require incubating human and rodent tissues (liver slices) with the chemical. This 

information provides dosimetric data that can be combined with other anatomical, biochemical, and physiological 

information to develop models based on biochemistry and physiologically based pharmacokinetics. Such models 

are used increasingly in risk assessment to extrapolate between species, across dose ranges, and across different 

routes of exposure.

 Table 27: Ongoing and Completed Disposition, Metabolism and Toxicokinetic Studies During FY 2010

Chemical [CASRN] Species/Strain Route Project Leader

Anatase (TiO2) [1317-70-0] Mice: Tg.AC (FVB/N) and FVB/N Topical application NCTR

Benzene [71-43-2] Mice: Gavage Cunningham

• BALB/cByJ

• WSB/EiJ (M. m. domesticus)

• CAST/EiJ (M. m. castaneus)

• C3H/HeJ

• DBA/2 Jackson

• BTBR T+ tf/J 2, C57BL/6

• FVB/NJ

• KK/HIJ

• AKR/J

• NZW/LacJ

• A/J

• NOD/LtJ

• B6C3F1

• PWD/PhJ (M. m. musculus)

• MOLF/EiJ (M. m. molossinus)

• 129S1/SvImJ

2,3-Butanedione [431-03-8] Rats: Harlan Sprague Dawley Oropharyngeal Waidyanatha

Mice: B6C3F1

2,3-Butanedione [431-03-8] In vitro Waidyanatha

2-Butene-1,4-diol [110-64-5] In vitro NCTR

n-Butyl-p-hydroxybenzoate [94-26-8] Rats: Harlan Sprague Dawley Intravenous Blystone

Cumene [98-82-8] Rats: Fischer 344 Gavage Chan

Mice: B6C3F1

1,3-Dichloro-2-propanol [96-23-1] Rats: Harlan Sprague Dawley Gavage Chan

Mice: B6C3F1

Di(2-ethylhexyl) phthalate 117-81-7 Monkey: Rhesus Gavage Delclos 

N,N-Dimethylacetoacetamide [2044-64-6] Rats: F344/N Charles River Gavage Waidyanatha

Dimethylamine borane [74-94-2] Human skin cells In vitro Germolec

Rats: Harlan Sprague Dawley

Dimethylethanolamine [108-01-0] Mice: B6C3F1 Gavage Irwin

Rats: Wistar

2’,2’’’-Dithiobisbenzanilide [135-57-9] Rats: Harlan Sprague Dawley Gavage Sanders

Mice: B6C3F1

Ephedrine + [299-42-3] Rats: F344 Gavage Dunnick

caffeine combination [58-08-2] 

Furan [110-00-9] Rats: F344 (NCTR) Gavage Irwin

Furan [110-00-9] Rats: Tg.Lac1/C57BL/6 (Big Blue) Gavage Irwin

Ginkgo biloba extract [90045-36-6] Human liver microsomes In vitro Chan

and hepatocytes

Gum guggul extract Human liver microsomes In-vitro Wyde 
NTP Annual Report 2010 71

Chemical [CASRN] Species/Strain Route Project Leader

2-Hydroxy- [131-57-7] Mice: B6C3F1 Gavage Auerbach

4-methoxybenzophenone Rats: Sprague Dawley

Ionic liquid (1-butyl- [479500-35-1] Mice: B6C3F1 Intravenous Hooth

1-methylpyrrolidinium chloride) Rats: F344/N

Ionic liquid [1124-64-7] Mice: B6C3F1 Topical application Hooth

(n-butylpyridinium chloride) Rats: F344/N

Isocyanuric acid [108-80-5] Rats: Fischer 344 Gavage NCTR

Isocyanuric acid [108-80-5] Rats: F344 (NCTR) Intravenous NCTR

Melamine [108-78-1] Rats: F344 (NCTR) Gavage Tolleson

Melamine cyanurate [37640-57-6] Rats: F344 (NCTR) Gavage Tolleson

Melamine + cyanuric Rats: F344 (NCTR) Feed Tolleson

acid combination

Melamine + cyanuric Rats: F344 (NCTR) Intravenous Tolleson

acid combination

2-Methoxy-4-nitroaniline [97-52-9] Rats: Harlan Sprague Dawley Gavage Auerbach

Mice: B6C3F1

L-beta-Methylaminoalanine [15920-93-1] Rats: Harlan SD Gavage Sanders

Mice: B6C3F1

Fullerene C60 [99685-96-8] Rats: F344/N Intratracheal Sanders

 Rutile TiO2 [1317-80-2] Mice: SKH-1 Hairless Topical application NCTR

Nanoscale silver [7440-22-4] Rats: SD Gavage Walker

Nanoscale silver [7440-22-4] Rats: SD Intravenous Walker

Nanoscale silver [7440-22-4] Rats: SD Gavage Walker

Silver acetate [563-63-3] Rats: SD Intravenous Boudreau

Silver acetate [563-63-3] Rats: SD Gavage Boudreau

Triclosan [3380-34-5] Mice: B6C3F1/NCTR BR Topical application Fang

(C57BL/6N x C3H/HEN MTV-)

Toxicogenomics study Human liver microsomes In vitro Waidyanatha

of allylbenzene and 

propenylbenzene class 

flavor constituents 

• Anethole [104-46-1]

• Estragole [140-67-0]

• Eugenol [97-53-0]

• Isoeugenol [97-54-1]

• Isosafrole [120-58-1]

• Methyleugenol [93-15-2]

• Myristicin [607-91-0]

• Safrole [94-59-7]

Table 28: Disposition, Metabolism and Toxicokinetic Studies Planned for FY 2011

Chemical [CASRN] Species/Strain Route

Bisphenol AF [1478-61-1] Rats: Harlan SD Gavage

Mice: B6C3F1

2-Ethylhexyl [5466-77-3] Mice: B6C3F1 Dermal

p-methoxycinnamate Rats: Harlan SD

Tris(4-chlorophenyl)methane [27575-78-6] Mice: B6C3F1 Intravenous

Rats: Harlan SD

Tris(4-chlorophenyl)methanol [3010-80-8] Mice: B6C3F1 Gavage

Rats: Harlan SD
72 National Toxicology Program

Genetic and Alternative Test Systems

Biomolecular Screening

In 2008, the NTP established a high throughput screening (HTS) initiative, representing a new paradigm in 

toxicological testing. During 2010 the NTP continued using this HTS approach to screen for mechanistic targets 

active within cellular pathways critical to carcinogenicity, reproductive and developmental toxicity, genotoxicity, 

neurotoxicity, and immunotoxicity. The NTP’s HTS program is administered through the Biomolecular Screening 

Branch (BSB).

The goals of the HTS Program are: 

• To prioritize substances for further in-depth toxicological evaluation (to judiciously allocate efforts 

and resources to maximize public health impact)

• To identify mechanisms of action for further investigation (e.g., disease-associated pathways)

• To develop predictive models for biological responses in humans and animals (predictive toxicology)

Much of the research conducted in support of the HTS program is coordinated with the EPA, the National Human 

Genome Research Institute (NHGRI), and the FDA through a Memorandum of Understanding (see page 84).

Contact Information: Dr. Raymond Tice, Chief BSB, tice@niehs.nih.gov. HTS website: http://ntp.niehs.nih.gov/go/28213

Non-mammalian models – C. elegans

The NTP is currently evaluating Caenorhabditis elegans ( C. elegans) as a study organism for assessing the effects of 

potential developmental and neurological toxicants on multi-cellular organisms. C. elegans is a roundworm about 

1 mm in length that lives freely in soil and feeds on bacteria. The use of C. elegans is consistent with NTP’s strategy 

to reduce the number of mammals used in testing. Several toxicology assays for feeding, growth, reproduction, 

and movement of C. elegans have been developed. Table 29 lists ongoing and completed C. elegans studies.

Table 29: Ongoing and Completed C. Elegans Studies During FY 2010 

Chemical [CASRN]

3-Aminopyridine [462-08-8]

2-Aminopyridine [504-29-0]

4-Aminopyridine [504-24-5]

Secondary screens using the C. elegans model (http://www.niehs.nih.gov/research/atniehs/labs/bmsb/index.

cfm) continue to be developed; the 309 compounds used in Phase I of EPA’s ToxCastTM Program were screened in 

C. elegans using a growth assay and the data compared with the results of a zebrafish embryo assay conducted 

by the EPA. The Tox21 partners continue to identify and use a full spectrum of secondary and tertiary screening 

assays to further define and characterize activities identified in initial quantitative high throughput screens (qHTS) 

and quantitative high content screens (qHCS).

Contact Information: Jonathan Freedman, freedma1@niehs.nih.gov. 

Toxicogenomics Studies

The NTP is working to bring the latest toxicogenomics technology into its testing program to help revolutionize 

the way NTP conducts its studies. Toxicogenomics examines how the entire genetic structure, or genome, is 

involved in an organism’s response to environmental toxicants. It applies gene and protein technologies to 

environmental medicine by studying the effect of toxicants on gene activity and specific proteins produced by 

genes. This information could be useful to identify biomarkers of disease and exposure to toxic substances and 

for understanding individual genetic susceptibilities. 
NTP Annual Report 2010 73

Preliminary toxicogenomic studies suggest that gene expression often is predictive for phenotypic alterations. 

The NTP is interested in determining if differential gene expression (DGE) analysis can provide indicators of 

toxicity at earlier time points and at lower doses than is possible with traditional toxicology parameters. DGE 

may provide more than a genotypic link to a morphology, because it is expected to provide insights into the 

pathogenesis of the disease and how different rodent models respond to toxicants. 

Perhaps the most exciting potential of toxicogenomics is the possibility to identify biomarkers of exposure 

or biomarkers of effect. Changes that can be found in easily obtainable samples (blood, urine) could then be 

monitored in clinical studies. When the technology is validated, it will allow repeated sampling during long-term 

NTP studies to determine whether chemical exposures can be detected or whether developing cancers will 

provide a genetic signature. 

The NTP is currently evaluating study conditions that may contribute to gene expression (e.g., animal and tissue 

variability), best methods of tissue sampling, and establishing standards for conducting toxicogenomic studies 

under laboratory conditions. However, a long-term goal of the NTP is to find better and more accurate methods 

of predicting potential carcinogenicity, because current NTP carcinogenicity studies take four to five years to 

complete and are costly. Planned or ongoing NTP toxicogenomic studies are listed in Table 30.

Table 30: Toxicogenomics Studies (Planned or Ongoing) in FY 2010 

Study [CASRN] Species/Strain Study Route Length Project Leader

Aflatoxin B1 [1162-65-8] Rats: F344/N Feed 90 days Irwin

Effect of the estrous cycle Rats: Wistar Han Not applicable 90 days Irwin

on hepatic transcriptome

Microcystin mixture [101043-37-2] Rats: F344/N Intravenous 1/2/6/24 hrs Walker

[96180-79-9]

Microcystin – LR [101043-37-2] Rats: Wistar Han Intravenous 1/2/6/24 hrs Walker

Microcystin – LA [96180-79-9] Rats: F344/N Intravenous 1/2/6/24 hrs Walker

Evaluation of rat liver carcinogens Rats: F344/N Tac Feed 8 weeks Irwin

and non-carcinogens administered Gavage 13 weeks

by feed in: Water

• Acetaminophen [103-90-2]

• Aflatoxin B1 [1162-65-8]

• 1-Amino-2,4-dibromoanthraquinone [81-49-2]

• Ascorbic acid [93-15-2]

• Methyleugenol [50-81-7]

• N-nitrosodimethylamine [62-75-9]

• 1-Tryptophan [73-22-3]

Toxicogenomics study of allylbenzene Rats: F344/N Tac Gavage 90 days Irwin

and propenylbenzene class

 flavor constituents 

• Anethole [104-46-1]

• Estragole [140-67-0]

• Eugenol [97-53-0]

• Isoeugenol [97-54-1]

• Isosafrole [120-58-1]

• Methyleugenol [93-15-2]

• Myristicin [607-91-0]

• Safrole [94-59-7]

Microarray analysis studies: [51974-40-4] DeVito

• 2,3,4,7,8-Pentabromodibenzofuran [13116-92-2] • 1,2,3,7,8-Pentabromodibenzofuran [107555-93-1]

• 2,3,7,8-Tetrabromodibenzofuran [67733-57-7]

• 2,3-Dibromo-

 7,8-dichlorodibenzo-p-dioxin [50585-40-5]

• 2,3,4,7,8-Pentachlorodibenzofuran [57117-31-4]

• 1,2,3,7,8-Pentachlorodibenzofuran [57117-41-6]

Pentabromodiphenyl oxide [32534-81-9] Rats: Wistar Han Gavage Gestation Dunnick

(technical) (DE 71) day 6 -

Postnatal 

day 21
74 National Toxicology Program

Cellular and Molecular Pathology

The Cellular and Molecular Pathology Branch (CMPB) is internationally recognized as a leader in toxicologic 

pathology. The CMPB has developed workshops, symposia, atlases, study sets, seminars, and other educational 

media for use by NTP pathologists (both CMPB staff and NTP contractors) as well as the global community. 

The NTP often sets standards that are highly valued by regulators at EPA, FDA, and NIOSH as well as other 

agencies and institutes around the world. 

Digital Pathology 

Digital photomicroscopy and slide scanning technologies are now routinely used for NIEHS/NTP studies and 

have led to the ability to publish and present stellar microscopic images, and through web-based telepathology, 

to conduct pathology peer reviews and seek instantaneous opinions from pathologists at remote locations 

around the world. During the evaluation of an NTP study, pathologists identify representative lesions for 

photomicroscopy or slide scanning and inclusion in the new pathology database. It is estimated that 70% of the 

NTP studies require photography, ranging from approximately 5-20 photographs per study. Routinely, the images 

are used for the PWG reviews. The NTP, in partnership with NCTR, is currently evaluating the diagnostic quality 

of digital histopathology by directly comparing the diagnoses given for a digital image to the corresponding 

glass slide. To date, ten PWGs have been evaluated and reveal promising results. During the past decade the 

NTP has established and maintained a state-of-the-art approach to capturing and archiving digital images of 

rodent pathology in a database that has grown to over 60,000 images. The images are primarily obtained by light 

microscopy but also include electron microscopy as well as MRI and microcomputed tomography (micro-CT). 

New types of imaging, such as MRI and micro-CT, for both in vivo and ex vivo studies, provide significant increases 

in data content and efficiency and allow a more complete examination of tissues and organs from animals 

involved in experimental studies. The images are categorized by species, sex, organ system, treatment, and dose. 

Atlas for Documenting Diagnostic Criteria for Non-neoplastic Lesions

In efforts to standardize and improve the diagnostic accuracy of non-neoplastic lesions in the NTP database, 

CMPB pathologists have been working to improve and revise the NTP’s Pathology Code Table and nomenclature 

for non-neoplastic lesions of the mouse and rat. Groups of two to three pathologists are generating terminology 

for specific organ systems and have been gathering or generating photomicrographic images of each lesion 

for the atlas. Lesion thresholds and severity scoring are also being addressed. Dr. Mark Cesta is leading the 

effort, which has completed drafts for six organ systems thus far. Additionally, NTP pathologists are in working 

groups for the Society of Toxicologic Pathology’s International Harmonization of Nomenclature and Diagnostic 

Criteria (INHAND) project, which is developing an internationally accepted nomenclature standardization of 

rodent neoplastic and non-neoplastic lesions. The project is relying on NTP expertise and using the NTP’s digital 

database of >60,000 pathology images for examples for the atlases. Approximately eight organ system chapters 

have been submitted for review. 

Hepatobiliary Atlas of the Embryonic Mouse

Dr. Susan Elmore, Julie Foley, and Dr. Laura Crawford (a veterinary student in the Summers of Discovery program 

at NIEHS) published Histology atlas of the developing mouse hepatobiliary system with emphasis on embryonic 

days 9.5-18.5. (Toxicol Pathol. 2010;38(6):872-906). This atlas provides phenotyping information to better address 

in utero exposure toxicity studies, and an investigation into causes of embryonic, fetal, or perinatal deaths.
NTP Annual Report 2010 75

Immunotoxicology Pathology Peer Review

Because of a constant effort to improve data quality, the NTP is applying the same level of rigorous pathology 

peer review to its immunotoxicology studies as it has traditionally applied to its subchronic and chronic 

carcinogenicity bioassays. This effort has included both the development of NTP specifications and pathology 

training for these studies. Six studies have been completed to date and have final pathology data.

Reproductive and Developmental Toxicology Pathology Peer Review

As with the immunotoxicology studies, the NTP has decided to apply the same rigorous pathology peer 

review process to its reproductive and developmental toxicity studies. These multigenerational RACB studies 

have been redesigned to provide a more complete picture of the potential for agents to cause reproductive 

and developmental toxicity. An NTP reproductive and developmental toxicity study pathology specifications 

document has been developed.

CL Davis Foundation Independent Study Center

CMPB is designated an Independent Study Center by the CL Davis Pathology Foundation and as such serves 

in-house and area trainees with study materials kept either at the NIEHS or NTP Archive in Research Triangle Park, 

NC. The study sets have been upgraded over the years with many new CDs comprising diseases of cattle, pigs, 

horses, primates, cat and dog, fish, wildlife, and ophthalmic pathology. These are in addition to rodent videos and 

other web-based teaching materials. 

RTP Rodent Pathology Courses

CMPB staff is involved in the organization and presentation of biennial Research Triangle Park Rodent Pathology 

Courses. These meetings are designed to showcase local pathology talent but also include international 

pathologists and basic and medical scientists. The September 26-28, 2010 meeting in Cary, NC was focused on 

hepatobiliary pathology. Drs. David Malarkey and Susan Elmore of CMPB were on the organizing committee. 

They and Dr. Mark Hoenerhoff and Julie Foley gave presentations. 

11th Annual NTP Satellite Symposium

The 11th Annual NTP Satellite Symposium was held on June 19, 2010, in advance of the jointly sponsored 29th 

Society of Toxicologic Pathology (STP) and International Federation of Societies of Toxicologic Pathologists (IFSTP) 

Annual Meeting in Chicago, Illinois. Chaired by Dr. Susan Elmore, this symposium was one of the most highly 

attended interactive sessions of the meeting. The theme for the 2010 symposium was Pathology Potpourri and 

included presentations of unusual lesions in the liver, pancreas, preputial gland, kidney, nasal septum, retina, skin, 

lung, ovary, and vascular system. Also included were example definitions and discussion of proposed INHAND 

nomenclature for proliferative and nonproliferative entities of the central and peripheral nervous system and 

cardiovascular system. Drs. David Malarkey and Deepa Rao also participated. 

Modified Sampling of the Nervous System in Routine NTP Studies

In the past, routine evaluation of the nervous system in short and long-term studies conducted in rodents by the 

NTP were limited to three histological sections of brain; in occasional circumstances, three sections of the spinal 

cord and a section of the sciatic nerve were also included. The increasing occurrence of human neurological 

diseases and subsequent cost burden on the economy, the role of unidentified environmental stressors in 

neurodegenerative disorders such as Parkinson’s disease and autism, multiple therapeutic drug-induced 

neuropathies observed in human clinical trials, and the exponential increase in use of chemicals with unknown 

neurotoxic potential within the environment or workplace, necessitate a more extensive evaluation of the 
76 National Toxicology Program

nervous system during routine screening for toxicity and carcinogenicity of chemicals, drugs, and biological agents 

by the NTP. Hence, the NTP has modified its protocol toward a more comprehensive histopathological evaluation 

of the nervous system that will allow a more complete examination of key anatomic subsites not previously 

examined. It is expected that this modified approach will increase the sensitivity of detecting and achieving 

mechanistic understanding of neurotoxicants and neurocarcinogens important to human neurologic and 

neurodegenerative disorders. 

Molecular Mechanisms of Chemically Induced Neoplasia 

NTP studies have identified several genetic alterations in chemically induced rodent neoplasms that have 

potential mechanistic implications for human cancer. Identifying such genetic alterations in NTP rodent models 

exposed to environmental contaminants, occupational chemicals, and other compounds demonstrates the 

importance of these studies in evaluating potential human health risks. These studies provide convincing 

molecular data that serve to complement the traditional gold standard histopathology data. NTP staff has 

initiated several studies investigating the molecular mechanisms of tumorigenesis in NTP studies. Studies 

included investigation of genetic and epigenetic alterations in liver, lung, mesothelioma, and colon tumors 

induced by compounds in NTP studies. Protocols to further standardize frozen tissue collection from NTP 

chronic studies for molecular biology analysis, including gene expression and protein analysis, have been 

initiated. Using the high quality samples obtained from the NTP chronic studies in conjunction with 

high throughput global gene expression profiling, important similarities were discovered between mouse 

and human tumors at the molecular level. Additionally, this technology provided important information on 

chemical-specific events leading to tumorigenesis. It is becoming increasingly important in NTP studies to 

identify pre-disease early genetic and epigenetic events in the process of toxicity and carcinogenicity in order 

to establish predictive as well as preventative measures to minimize human health hazards. Evaluation of 

alterations in gene and protein expression in tissues during the pre-neoplastic stages will provide important 

data about the initiating events that will provide critical information on predictive events in the pathogenesis 

of chemically induced neoplasms in NTP studies. NTP contributors were Drs. Mark Hoenerhoff, Arun Pandiri, 

Stephanie Lahousse, and Robert Sills, Ms. Hue Hua Hong, and Mr. Tai-Vu Ton. 

Contact Information: Cellular and Molecular Pathology Branch, Dr. Robert C. Sills, Chief, sills@niehs.nih.gov.
NTP Annual Report 2010 77

Laboratory Animal Medicine Training Program

This four-year training program, under the direction of Dr. Angela King-Herbert, includes clinical and surgical 

responsibilities, management of animal care facilities, participation in research projects, and laboratory animal 

pathology. The training program is a collaborative effort between NIEHS and the University of North Carolina 

at Chapel Hill. Fellows interact with laboratory animal veterinarians at NIEHS and at local area academic,

 industrial, and government facilities to receive didactic and hands-on training in laboratory animal medicine. 

Two postdoctoral fellows are currently in the program, Drs. Jacquelyn Tubbs and Dr. Coralie Zegre-Cannon. Both 

are scheduled to take the American College of Laboratory Animal Medicine certifying examination in June 2011.

Contact Information: Dr. Angela King-Herbert, kingher1@niehs.nih.gov.

Toxicological Pathology Training Program

Since formalizing a training program in toxicological pathology in 2003, Dr. David Malarkey, training coordinator, 

and other CMPB staff have mentored 11 post-doctoral fellows and over 30 veterinary student externs. Three 

post-doctoral fellows, Drs. Michael Boyle, Torrie Crabbs, and Deepa Roa, and five veterinary students participated 

in the program during 2010. The program is designed to introduce students to the field and career opportunities 

in veterinary and toxicological pathology while also providing hands-on projects often leading to abstracts 

and publications. Post-doctoral fellows learn rodent and toxicological pathology, participate in NTP and other 

DIR research projects, work to achieve accuracy of NTP pathology data by assisting the NTP pathologist on 

NTP studies, and continue education towards achievement of board certification by the American College 

of Veterinary Pathologists.

Contact Information: Dr. David Malarkey, malarkey@niehs.nih.gov.

Toxicology and Carcinogenesis Training Program

Trainees in this program, under the direction of Dr. Paul Foster, learn to perform all aspects of contracted 

toxicology studies for carcinogenic or non-carcinogenic endpoints (e.g., reproductive and developmental effects, 

immune system function). They learn about NTP efforts in molecular toxicology and HTS and receive training 

applicable to regulatory or industrial toxicology. By serving as study scientists in non-laboratory positions, they 

evaluate the toxicity of substances of interest to the NTP. They actively participate in designing, conducting, and 

evaluating studies and interact extensively with chemistry, pathology, toxicokinetics, toxicogenomics, genetics, 

epidemiology, statistics, and molecular biology staff. Four postdoctoral fellows are currently in the program, 

Drs. Minerva Mercado-Feliciano, In Ok Surh, Mamta Behl, and Sang-Hyun Kim.

Contact Information: Dr. Paul Foster, foster2@niehs.nih.gov.

NTP Postdoctoral Training Programs
78 National Toxicology Program

NIEHS/NCTR Interagency Agreement

In 1992, the FDA entered into an IAG with NIEHS. The interagency agreement (IAG) is an instrument that allows 

chemicals nominated to the NTP to be studied for toxicity using the unique resources and facilities at NCTR. The 

research conducted under the IAG allows the FDA to better assess study design input and initial data on the safety 

of FDA-regulated products. It has allowed continued collaborative toxicity testing on compounds of interest 

to the FDA and NTP and has led to investigations of mechanisms of action and assessments of toxicity for many 

classes of chemicals including cosmetics, endocrine-disrupting compounds, food contaminants, food cooking 

byproducts, dietary supplements, drugs, and anesthetics. The IAG supports the Phototoxicity Research and 

Testing Laboratory at the NTP Center for Phototoxicology and the Nanotechnology Core Facility at the NCTR/

Office of Regulatory Affairs. All toxicology studies conducted under the IAG are designed with input from 

FDA regulatory scientists, NCTR and NIEHS scientists, scientists from other agencies, and invited subject 

matter experts. The IAG uses resources from public funds and exceptional scientific expertise to provide the 

best possible assessment of product safety through toxicological studies. Table 31 lists projects completed or 

ongoing in FY 2010.

Interagency Agreements

Table 31: NIEHS/NCTR Interagency Agreement Projects in FY 2010 

Study [CASRN] [Principal Investigator] Objective and/or Rationale

para-Nonylphenol: (1) To determine the effects of p-nonyphenol, an intermediate in the production 

Evaluation of Reproductive of surfactants and other industrial products, on reproduction and on the 

Effects over Multiple Generations development of reproductive and other hormone-sensitive organs when administered

[84852-15-3] [Delclos] to CD rats over five generations; (2) to determine if subtle effects observed in the

dose-range-finding study are magnified through multiple generations; and (3) to

evaluate the reversibility of any observed effects. 

Perinatal Carcinogenicity of Drug To determine the carcinogenicity, genotoxicity, and metabolism of antiretroviral 

Combinations Used to Prevent drug combinations administered to mice transplacentally, perinatally, or neonatally. 

Mother-to-Child Transmissions of HIV The studies include 14-day range-finding as well as 2-year chronic studies in

[30616-87-1], [134678-17-4] [Beland] both groups.

Effect of Topically Applied Skin Creams (1) To determine if retinyl palmitate applied to the skin of SKH-1 hairless mice alters

Containing Retinyl Palmitate on the the incidence of tumors produced by simulated solar and/or UV light, and

Photocarcinogenicity of Simulated Solar (2) to determine the mechanisms of tumor promotion by retinyl palmitate.

Light in SKH-1 Mice [79-81-2] [Boudreau]

Bioassays in the F-344 Rat and B6C3F1 Mouse (1) To use 14-day studies determine dose ranges for future toxicology studies, 

Administered Aloe vera Plant Constituents in (2) to use 90-day studies to determine the toxic effects and highest tolerated doses

Drinking Water [8001-97-6] [Boudreau] of the different Aloe vera components, including the gel and whole leaf extracts 

[filtered and non-filtered], and (3) to use 2-year chronic studies to determine the 

chronic toxicity and carcinogenic potential of Aloe vera whole leaf exctract in 

the rat and mouse by oral exposure administered in the drinking water.

Toxicity Studies of a Combination of To evaluate the potential toxicity and carcinogenicity of perinatal and chronic

AIDS Drugs in p53 (+/-) Transgenic Mice exposures to the AIDS drugs AZT and 3TC in C3B6F1trp53(+/-) haplodeficient F1 

[30616-87-1; 134678-17-4] [Leakey] transgenic mice. The study includes both a range-finding and a 6-9-month 

chronic phase.

Genotoxicity and Carcinogenicity of To compare the carcinogenicity of acrylamide and its metabolite glycidamide in

Acrylamide and Its Metabolite Glycidamide B6C3F1 mice and F344 rats treated chronically for two years.

in Rodents [79-06-1; 5694-00-8] [Beland]

Developmental Neurotoxicity Assessment of To determine the consequences of long-term exposure to acrylamide on a variety of

Acrylamide in Rats: Long-term Studies developmental milestones and measures of nervous system integrity throughout life.

[79-06-01] [Paule] 
NTP Annual Report 2010 79

Study [CASRN] [Principal Investigator] Objective and/or Rationale

Genotoxicity and Carcinogenicity of Acrylamide To complete 13-week studies, 90-day studies, and 2-year bioassays for

and its Metabolite Glycidamide in Rodents — acrylamide and glycidamide.

Range-finding/Subchronic/2-year Chronic 

Carcinogenicity Studies 

[79-06-1; 5694-00-8] [Beland] 

Subchronic Toxicity Studies of Chondroitin (1) To investigate the potential toxicity of chondroitin sulfate and glucosamine,

Sulfate and Glucosamine Combinations in administered by oral gavage in male rats, and (2) to determine whether subchronic

Fischer 344 and Diabetic Goto-Kakizaki Rats exposure of glucosamine or chondroitin sulfate potentiates the pathological effects

[3416-24-8, 9007-28-7] [Leakey] of noninsulin-dependent diabetes in obese diabetic rats.

Toxicity Studies of Glucosamine and To investigate the potential toxicity of glucosamine and glucosamine/chondroitin

Glucosamine/Chondroitin Sulfate sulfate combinations, administered by oral gavage in male rats.

Combinations in Obese and Lean Zucker Rats 

[3416-24-8; 9007-28-7] [Leakey]

DEHP Toxicokinetics in Neonatal Male (1) To quantify the metabolism and disposition of multiple, single intravenous doses 

Rhesus Monkeys after Intravenous and of DEHP administered to male rhesus monkeys during the first 12 postnatal weeks, 

Oral Dosing [117-81-7] [Delclos] (2) to quantify the metabolism and disposition of multiple, single oral doses of DEHP 

administered to male rhesus monkeys during the first 12 postnatal weeks, (3) to use 

the results of this work to evaluate the feasibility and utility of a subchronic toxicity 

study of DEHP, and (4) to utilize blood and testicular tissue from the infant monkeys 

to establish methods to be utilized in the subchronic study and/or estimate variability 

in the endpoints to aid in determining the number of animals that will be required in 

each dose group for a subchronic study. 

The Immunogenicity of Permanent To investigate the possibility of immunostimulation by a select group of permanent

Makeup Inks and Their Components makeup inks that were associated with adverse event reports to the FDA.

[Howard]

Effects of Sedatives on the Metabolism of (1) To determine if the metabolic profile of DEHP is affected by sedatives potentially 

DEHP Administered by Intravenous Injection useful for intravenous injection studies of DEHP in neonatal rhesus monkeys and/or

and the Relationship of DEHP Metabolism to in common use in neonatal intensive care units and (2) to examine DEHP metabolism 

Biological Effects in Neonatal Rats in neonatal rodents dosed intravenously and orally and relate this metabolism to 

[117-81-7] [Delclos] biological effects.

Maintenance of the Transgenic p16/p19(-/-) To maintain the NCTR p16/p19 breeding colony for the development of a strain of

Haplodeficient [NCTR strain code 7V] haplodeficient mice that could be used in GMM 9-month studies for rapid drug 

Breeding Colony [Leakey] evaluation for carcinogenesis.

Determination of Carcinogenic Mechanisms To determine the pharmacokinetic mechanisms, mutagenesis, and hepatotoxicity of 

for Furan in Male Fischer 344 Rats low doses of the carcinogen furan in rodents. 

[110-00-9] [Doerge]

Subchronic Studies of Usnic Acid in To evaluate the subchronic toxicity of usnic acid in male and female

Fischer 344 Rats and B6C3F1 Mice Fischer 344 rats and B6C3F1 mice.

[125-56-2] [Leakey]

Subchronic Studies of Usnea Lichen To evaluate the subchronic hepatotoxicity of Usnea lichen in male and

in Fischer 344 Rats and B6C3F1 Mice female Fischer 344 rats and B6C3F mice.

[84696-53-7] [Leakey]

Physiological Effects of Bitter Orange in Rats To determine potential physiological effects of synthetic synephrine as well as an 

[94-07-5] [Hansen] extract from the botanical Citrus aurantium alone and combined with caffeine in rats 

with and without exercise.

Developmental Toxicity of Bitter Orange To determine potential developmental toxicity of synthetic synephrine and Citrus 

in Rats [94-07-5] [Hansen] aurantium extract in rats. 

Determine Potential Developmental To determine the potential developmental toxicity of synthetic synephrine, 

Toxicity of Synthetic Synephrine and Citrus aurantium, and possible potentiation of the toxicity by caffeine.

Citrus aurantium Extract in Rats 

[CAS 94-07-5] [Hansen] 

Mechanisms of Nevirapine Carcinogenicity To determine the mechanism by which nevirapine induces liver tumors in rats.

[129618-40-2] [Beland]
80 National Toxicology Program

Study [CASRN] [Principal Investigator] Objective and/or Rationale

A Toxicological Evaluation of Nanoscale (1) To determine if reducing the size of silver particles to nanoscale changes the 

Silver Particles in Rodents absorption, biodistribution, and excretion as well as the toxicity, and (2) to determine

[7440-22-4] [Boudreau] the effects of reducing the particle size to nanoscale of a compound that has been 

regarded as nontoxic in micron and above sizes.

Evaluation of the Toxicity of Bisphenol A (BPA) (1) To assess the toxicity of BPA in perinatally dosed rats study, and (2) to evaluate

in Male and Female Sprague Dawley Rats estrogenic endpoints from the F1 generation dosed perinatally with BPA, via gavage.

Exposed Orally from Gestation Day 6 

through Postnatal Day 90 [80-05-7] [Delclos]

Two-Year Carcinogenicity Bioassay of Furan To determine the dose-response relationship for the carcinogenicity

in F344 Rats [110-00-9] [Beland] of furan in F344 rats.

The Role of Perinatal Development To develop additional data from rat and monkey exposures that will be used in the 

on Toxicokinetics of BPA [80-05-7] [Doerge] creation and validation of a PBPK model to predict internal exposures to free BPA 

in the appropriate target tissues of fetuses and babies that are derived from food 

contact and medical device exposures.

Assessment of the Nephrotoxic Effect To investigate the toxic effects noted in cats and small children who were exposed to 

of a Combined Exposure to Melamine melamine and cyanuric acid via adulterated food supplies. The toxicity was noted 

and Cyanuric Acid [108-78-1; 64-18-6] in the kidneys of the pets and children. The study will include a 28-day NOAEL study. 

[Gamboa Da Costa]

Neurotoxicity Assessment of Cell Phone (1) To determine whether power levels of RFR that are emitted from mobile phones

Radiofrequency Radiation (RFR) using Rat produce any changes in the CNS of mice and rats, and (2) to determine if there

and Bovine Brain Microvascular Endothelial are disruptions in the blood brain barrier after being subjected to RFR radiation.

Cells as Model Blood Brain Barrier Systems, 

PC-12 Cultured Cells, and Whole Animal 

Models (mice and rats) [Ali]

Range-finding, Mechanistic and Toxicokinetic To complete a standard 14-day range-finding study with both compounds

Studies of Usnic Acid and Usnea barbata Herb administered in feed.

in Fischer 344 Rats and B6C3F1 Mice 

[125-46-2, 84696-53] [Leakey]

Vehicle Selection for Triclosan Dermal To determine ADME toxicokinetics after an appropriate solvent is determined

Toxicity Studies in B6C3F1 Mice for dermal application.

[3380-34-5] [Fang]

Contact Information: Dr. Paul Howard, paul.howard@fda.hhs.gov and Dr. Nigel Walker, walker3@niehs.nih.gov.
NTP Annual Report 2010 81

NIEHS/NIOSH Interagency Agreement – Comprehensive Assessment 

of Occupationally-Relevant Exposures

The NTP is coordinating an effort with NIOSH to better understand worker exposures, identify occupational 

health research gaps, and educate workers. Current efforts listed in Table 32 address worker exposures to welding 

fumes, nano-sized materials, food flavorings, and indium compounds. 

Table 32: NIEHS/NIOSH Interagency Agreement on Occupationally Relevant Exposures 

Study [Principal Investigator] Objective and/or Rationale

Administrative Support To enable NIOSH scientists to (1) participate in review and oversight of NTP activities 

[Toraason] and (2) attend NTP-related meetings in Research Triangle Park, NC and Washington, DC.

Assess the Feasibility of (1) To identify industries (e.g., construction, shipbuilding, railroad, and manufacturing), companies, 

an Occupational Exposure and/or unions involved in welding operations where the potential for substantial manganese exposure 

Assessment of Welding exists for exposure assessments; (2) to develop methods to identify specific manganese compounds, 

Fumes with Emphasis on different valence states, and potential solubility contained within various welding fumes matrices; and 

Manganese Compounds (3) to characterize welding fume exposures based on welding-associated jobs, tasks, and processes.

[Hanley]

Exposure Assessment of (1) To characterize workplace inhalation exposures to diacetyl in food production industries that use 

Diacetyl and Other Flavorings food flavorings, (2) to document high-exposure activities and processes in flavored food production 

in Food Production Industries industries, (3) to identify work practices and procedures that affect exposure, (4) to document 

[Curwin] engineering controls, and (5) to field test novel techniques for both gravimetric and volatile sampling.

Exposure Assessment of (1) To identify worker populations at increased risk of inhalation and surface exposure to DTBBA during 

Dithiobisbenzanilide (DTBBA) a manufacturing process, (2) to develop a NIOSH analytical method for quantitatively assessing DTBBA 

in a Manufacturing Setting airborne particulate and surface exposures, and (3) to characterize industry-wide occupational 

[Wurzelbacher] exposures, including total number of workers, and evaluate patterns of exposure to DTBBA.

Assessment of Use of Indium (1) To contact and visit companies to determine indium materials being used, jobs and processes with 

and Indium Compounds potential indium exposure, exposure controls, and indium use trends; (2) to attend the Semiconductor 

in the Workplace Environmental Safety and Health Association meeting to establish contacts in industry and other 

[Hines] organizations involved in indium-related applications; and (3) to conduct preliminary sampling 

for indium, if possible.

Exposure Assessment of (1) To identify workplaces engaged in the synthesis, manufacture, and use of engineered nanomaterials,

Engineered Nanoparticles and (2) to characterize workplace exposure to selected engineered nanoparticles.

[Geraci]

Exposure Assessment of (1) To identify worker populations at elevated risk of inhalation and surface exposure to PCBTF during 

1-Chloro-4-(trifluoromethyl) manufacturing processes, (2) to update a previously published analytical method for quantitatively 

benzene (PCBTF) assessing PCBTF airborne vapors and surface exposures to allow the use of capillary column 

[Harper] chromatography, and (3) to characterize industry-wide occupational exposures, including 

total number of workers, and evaluate patterns of exposure to PCBTF.

Cardiovascular Toxicity (1) To evaluate potential cardiovascular toxicity of fullerene C60 in animal models using molecular and 

Assessment of Subchronic biochemical analysis of cardiovascular tissue and blood samples from NTP inhalation studies, and 

Inhalation Exposure to (2) to correlate the findings with the results from histopathology, particle distribution, and blood 

Fullerene C60 [Erdely] chemistry studies.

Contact Information: Dr. Dori Germolec, germolec@niehs.nih.gov and Dr. Mark Toraason, mht1@cdc.gov.
82 National Toxicology Program

NIEHS/NIOSH Interagency Agreement – Immunotoxicology 

The goal of this IAG is to provide support of NTP hazard identification activities aimed at preventing diseases 

or adverse effects caused by environmental exposure to chemical or physical agents. These cooperative studies 

continue to improve risk assessment by measuring what constitutes an adverse health effect on the immune system 

in humans. The studies, listed in Table 33, evaluate unique cohorts of individuals from professions associated with 

immune-mediated occupational diseases, including asthma, contact dermatitis, allergy to mold spores, chronic 

beryllium disease, allergic rhinitis, and silicosis. These cohorts are being studied for a number of endpoints including 

impact of genetic polymorphisms on development of inflammatory disease and clinical outcomes, and identification 

of unique immunological biomarkers for disease. The NIOSH Laboratory for Occupational Genomics serves as a 

resource for obtaining samples from individuals with occupational and occupationally related diseases.

Table 33: NIEHS/NIOSH Interagency Agreement on Immunotoxicology Studies, FY 2010 

Study [Principal Investigator] Objective and/or Rationale

Chronic Sinusitis and Mold Exposure To investigate the role of fungi in chronic sinusitis and to determine if the 

[Beezhold] prevalence of fungal sensitization is different to other seasonal allergens in chronic 

rhinosinusitis patients. To date, the chronic rhinosinusitis and control patients have 

been recruited and tested. The dataset has been collected, statistically analyzed, 

and a manuscript is currently being prepared. 

Heading off Environmental To evaluate the effectiveness of a novel asthma case management intervention 

Asthma in Louisiana among children with asthma after Hurricane Katrina in New Orleans and to assess

[Beezhold] total immunoglobulin E (IgE) and mold-specific IgE in asthmatic children before 

and after intervention. This project is completed. All sera have been screened 

and the dataset has been statistically analyzed. 

NIEHS Agricultural Pesticide Study To evaluate allergic sensitization in a cohort of 677 farmers with or without 

[Beezhold] pesticide exposures. To date, 677 farmers have been recruited and the serum total 

IgE and specific IgE quantified using Phadia ImmunoCap. Serum concentrations 

of the fungal mycotoxin deoxynivalenol have also been quantified. Currently, 

a manuscript is being prepared by NIOSH and NIEHS collaborators. The second 

phase of the study is about to begin and will evaluate allergic sensitization in 

a cohort of >2000 farmers located in Iowa and North Carolina. 

A marker for Aspergillus terreus exposure To develop new and improved methods for detecting fungal exposure. In this 

[Beezhold] project the emerging opportunistic fungal pathogen A. terreus has been used

as a model fungal species. Terrelysin, a cytolysin and potential biomarker of fungal 

infection, was characterized and a recombinant protein produced. The recombinant

terrelysin was then used to immunize mice to produce terrelysin specific monoclonal

antibodies (mAbs). Seven specific mAbs were produced and used to characterize

the production of terrelysin. mAb binding sites are currently being determined

in epitope mapping experiments. NIOSH anticipates the development of a sensitive

immunoassay that may be used in the serological detection of this biomarker.

Animal model for airway exposure to To develop a murine model of dry fungal exposure to better mimic natural 

dry fungal aerosols human exposures to fungi. In collaboration with aerosol scientists at NIOSH, an 

[Beezhold] acoustical generation system has been developed. In preliminary experiments, 

histopathology and bronchoalveolar lavage fluid analysis demonstrated pulmonary

deposition of conidia following dry fungal bioaerosol exposure. Future experiments

are planned to further characterize the exposures and to characterize the immune 

responses associated with various fungal species that frequently colonize waterdamaged buildings or are occupationally relevant.

The Role of Genetic Variation in (1) To investigate whether the 24-hour irritant patch test is predictive of 

Environmental and Occupational occupational hand dermatitis caused by high exposure to hand washing in

Diseases - ICD health care workers, and (2) to investigate association between genetic variations 

[Yucesoy] in specific candidate genes (with emphasis on variants of cytokines, MHC region, 

antioxidant enzyme genes and genes related to skin barrier integrity) and irritation 

threshold levels of the subjects with development of ICD. This study is in 

collaboration with Case Western Reserve University and West Virginia University. 
NTP Annual Report 2010 83

Study [Principal Investigator] Objective and/or Rationale

The Role of Genetic Variation in Environmental (1) To investigate genetic factors in individuals predisposed to develop ACD, 

and Occupational Diseases - Allergic specifically induced by nickel and (2) to investigate genetic factors involved in 

Contact Dermatitis (ACD) the development of ACD in individuals sensitized to weak allergens, individuals 

[Yucesoy] sensitized to allergens that require metabolism in the skin, and individuals 

who react to more than three allergens of the standard screening series. 

This study is in collaboration with Case Western Reserve University 

and Dartmouth-Hitchcock Medical Center. 

Role of Genetic Variation in Environmental (1) To investigate whether genetic variations in specific candidate genes 

and Occupational Diseases – (e.g., cytokine, MHC region, antioxidant enzyme genes) are associated with

Occupational Asthma asthma induced by diisocyanates, and (2) to investigate potential associations 

[Yucesoy] between genetic variations in candidate genes and occupational asthma caused by 

low molecular weight agents. This project is in collaboration with the University of 

Montreal and the University of Cincinnati.

The Role of Genetic Variation in Environmental To investigate the contribution of genetic variations in the MHC region to the

and Occupational Diseases – Chronic Beryllium development of beryllium sensitization and CBD. This study is in collaboration

Disease (CBD) – [Yucesoy] with National Jewish Medical and Research Center.

Investigations into Health Effects Caused (1) To identify and measure the reaction products of gas-phase compounds 

by Exposure to Indoor Air Reaction Products present in the indoor environment, especially oxygenated organics. 

(supportive animal studies) (2) to further develop and validate a novel in vitro exposure method utilizing 

[Wells, Anderson] realistic indoor chemistry scenarios to expose cells and tissues to these indoor 

air reaction products, (3) to complete both in vitro and in vivo assays to assess 

adverse health effects caused by indoor air reaction products, and (4) to further 

investigate the role of structurally similar indoor air chemicals present in mixtures 

in the indoor environment.

Toluene Diisocyanate (TDI) (1) To produce and characterize 35 monoclonal antibodies against 2,4-TDI protein

Monoclonal Antibody Production and and 13 monoclonal antibodies against 2,6-TDI-protein; (2) to complete characterCharacterization (improved methods) ization of all monoclonal antibodies, especially with respect to minimal epitope

[Siegel] required for reactivity of the monoclonal antibodies to a TDI-reacted chemical; 

and (3) to examine the utility of select monoclonal antibodies for immunoassay, 

immunohistochemistry, and proteomic analyses of tissues after TDI exposure.

Immunological Mechanisms of Occupational (1) To use specific inhalation challenge with suspected diisocyanates to diagnose

Rhinitis Induced by Diisocyanate allergic rhinitis in workers from Canada and Spain who are exposed to diisocyanates

Exposure (supportive animal studies and then clinically phenotyped for upper and lower airway disease, (2) to collect

and field studies) nasal mucosal samples from workers with diisocyanate rhinitis and use whole

[Johnson] genome approaches to study mechanisms of disease, (3) to identify molecular 

targets in the human nasal mucosa for biomarker development and validation 

using the NIOSH TDI rhinitis mouse model, and (4) to develop the mucosal 

sampling technique and identified biomarker(s) into a simple and non-invasive 

tool for worker health surveillance and early diagnosis of sensitization/disease.

Assessing Exposures to Asthmagenic Cleaning (1) To investigate the use of cleaning and disinfectant products which have been

and Disinfecting Products Among identified as an important risk factor for asthma in healthcare workers; (2) to identify

Healthcare Workers and quantify exposures to cleaning and disinfectant product-associated asthmagens

[Virji] or their surrogates in six healthcare occupations; (3) to characterize job-tasks and

activities, work practices and workplace factors and evaluate their impact on

exposure levels; (4) to develop exposure assessment survey instruments and

questionnaires; and (5) to revise an existing asthma-specific JEM for an 

epidemiologic study of asthma in healthcare. Exposure assessment for selected 

VOCs was conducted in two Veterans Administration health care facilities in 2010. 

This work will ultimately lead to identification of those agents most associated 

with asthma in the healthcare environment, enabling prioritization for detailed

immunotoxicological evaluation as well as preventive intervention studies.

Determination of Total IgE, Antinuclear To assess members of the Upper Midwest Health Study (1) to determine

Antibodies and Atopy In Plasma From Members total IgE, antinuclear antibodies, and atopy in plasma, and (2) to test for

of Upper Midwest Health Study, A Case-Control common allergies.

Study of Intracranial Gliomas (field study) [Biagini]

Antibody Levels in Systemic Lupus To measure total serum IgA, IgM and IgG in sera from systemic lupus

Erythematosus (field study) erythematosus patients to investigate whether adjustment for total

[Biagini] immunoglobulin levels may unmask differences in serum IgE levels.

Contact Information: Dr. Dori Germolec, germolec@niehs.nih.gov and Dr. Mark Toraason, mht1@cdc.gov.
84 National Toxicology Program

NTP/NHGRI/EPA Interagency Agreement – High Throughput Screening, 

Toxicity Pathway Profiling, and Biological Interpretation of Findings

A five-year Memorandum of Understanding (MOU), High Throughput Screening, Toxicity Pathway Profiling and 

Biological Interpretation of Findings, was signed on February 14, 2008. With this MOU, NTP partnered with the 

NHGRI’s NIH Chemical Genomics Center (NCGC) and the U.S. EPA’s National Center for Computational Toxicology 

located within the Office of Research and Development. A central component of this MOU is exploration of 

qHTS and tests using phylogenetically lower animal species (e.g., fish, worms), as well as high–throughput, 

whole-genome analytical methods to evaluate mechanisms of toxicity. Ultimately, the data generated by these 

new tools will be provided to risk assessors to use to protect human health and the environment. On July 19, 

2010, an amended five-year MOU was announced that was signed by the original three partners plus the U.S. 

Food and Drug Administration (FDA). The FDA brings to the partnership experience in human diseases, animal 

models of human disease, and expertise in toxicity pathway analysis and computational toxicology. Their active 

participation is in recognition of the FDA’s commitment to developing new methods to evaluate the toxicity of 

the substances they regulate. 

The goals of this MOU are to investigate the use of these new tools to (1) identify mechanisms of chemically 

induced biological activity, (2) prioritize chemicals for more extensive toxicological evaluation, and (3) develop 

more predictive models of in vivo biological response. The results should yield test methods for toxicity testing 

that are more scientifically and economically efficient and models for risk assessment that are more biologically 

based. This should reduce or replace animals in regulatory testing and is anticipated to occur in parallel with 

an increase in our ability to evaluate the large numbers of chemicals that currently lack adequate toxicological 

evaluation. Through this partnership, it is possible to pool resources, overcome the resource limitations of a single 

agency, build on existing expertise, and avoid the need to create a new administrative and support structure.

By the end of FY 2010, a library of approximately 2,800 compounds provided to the NCGC for qHTS by NTP 

and EPA had been tested in approximately 100 different assays. These assays broadly evaluated the ability of 

compounds (1) to induce, in various cell lines, cytotoxicity, apoptosis, DNA damage, changes in methylation 

status, mitochondrial toxicity, and various stress response pathways (e.g., antioxidant, hypoxia, heat shock), or 

(2) to act as an agonist or antagonist in 12 different nuclear receptors including the estrogen and androgen 

receptors. Included among these studies were those to evaluate differential sensitivity among cells from humans 

by exposing ~80 CEPH panel HapMap cell lines to 240 toxic chemicals and evaluating the extent of cytotoxicity 

and caspase 3/7 induced by each chemical .

During FY 2010, the partners continued to work on establishing a library of ~10,000 compounds that broadly 

characterizes and defines the chemical-biological space occupied by chemicals of toxicological concern. This 

library, to be completed by mid-2011, will be tested at the NCGC with qHTS and qHCS that provide information 

on critical cellular pathways. Chemical analysis studies to determine the identity, purity, and stability of all 

compounds in this library were initiated and will continue through FY2011. Starting in FY2011, this library will 

be screened in triplicate against a battery of nuclear receptor and stress response pathway assays at the NCGC. 

Testing was started of a subset of these compounds (~700) in Phase II of EPA’s ToxCastTM Program (http://

www.epa.gov/ncct/toxcast/). The data from these assays, along with full chemical characterization and assay 

protocol details, are being deposited into publicly accessible, relational databases such as the National Library 

of Medicine’s PubChem (http://pubchem.ncbi.nlm.nih.gov/), EPA’s ACToR (http://www.epa.gov/ACToR/) and NTP’s 

CEBS (http://www.niehs.nih.gov/research/resources/databases/cebs/). 

In FY 2011, the MOU will also evaluate the differential sensitivity of cells from humans by screening >1000 

densely sequenced lymphoblastoid cell lines against 180 toxic chemicals and evaluating the resulting extent of 

cytotoxicity. To conduct the level of screening planned for FY 2011 and in subsequent years, the NCGC purchased 

a new robot in FY 2010 with operation scheduled for March 2011.
NTP Annual Report 2010 85

The NCGC BioPlanet

No one comprehensive and uniform 

resource covers all known annotations 

of pathways and no single platform allows 

integrated browsing, retrieval, and analysis 

of information from the many existing 

individual pathway resources; therefore, 

the NCGC (with support from the NTP) 

built an integrated pathway resource 

that hosts information on ~1100 human 

pathways from manually curated and 

publicly available resources. The NCGC 

BioPlanet complements this pathway 

warehouse by allowing easy browsing, 

visualization, and analysis of the universe 

of pathways. The main view of the BioPlanet 

shows the mapping of all known human 

pathways on a 3D globe, where each spot represents a pathway. Selecting a pathway on the globe will place 

all components of the selected pathway in the detailed view window. Detailed descriptions of all genes in the 

selected pathway are shown below the 3D graphics. When multiple pathways are selected at the same time, the 

view shows all unique gene components within selected pathways. Since the coordinates of genes are generated 

globally and are not tied to any specific pathway, this allows the visualization of multiple, predefined pathways as 

one extended pathway that better shows the interaction between different biological processes. The BioPlanet is 

searchable by any gene or pathway identifier, and also by disease relevance (prevalence of disease genes), toxicity 

relevance (occurrence of genes in toxicology literature), and availability of probing assays in PubChem. 

NTP Archives

NTP archival tissues represent a significant and underutilized resource, so in FY 2010 NTP initiated pilot studies 

to evaluate the extent to which gene expression signatures can be reliably derived from the molecular analysis 

of tissue samples collected from the laboratory animals used in NTP’s toxicological studies and stored as 

formalin- fixed, paraffin-embedded (FFPE) tissues in the NTP archives. Signature expression profiles are 

critical sets of altered transcripts or proteins that distinguish toxicity and disease from a comparable normal 

state. Development of such signatures would further our understanding of pathological changes, mechanisms, 

and critical pathways in agent-induced toxicity. Further, such signatures could contribute to the identification 

of useful targets for in vitro assays, to an evaluation of the correlation between in vitro test results and in vivo

toxicological outcomes, and to the development of predictive models of toxicity. 

DrugMatrix®

Related to the goal of developing analysis tools and approaches to allow an integrated assessment of HTS 

endpoints and associations with findings from traditional toxicology and cancer models, the NTP acquired 

DrugMatrix, a toxicogenomics reference database, the accompanying extensive frozen tissue archives, and 

the informatics system. The NTP acquired this resource to expand the ability to develop predictive models for 

toxicological effects based on gene signatures, to provide additional tools for linking in vitro data to in vivo gene 

signatures and disease outcomes, and to provide additional tissue samples for NextGen-based investigations. It is 

anticipated this database will be made publicly accessible to the international scientific community through CEBS.
86 National Toxicology Program

Mouse Methylome Project

An individual’s response to exposure-related toxicity and concomitant disease is influenced at the genome 

level by genetic, epigenetic, gene-gene interactions (intrinsic factors) and interaction with the environment 

(extrinsic factors). Individual DNA sequence variation does not account for all of the heritability for susceptibility 

to toxicity and diseases such as asthma, cancer, diabetes, etc. An intrinsic factor that quantitative and molecular 

geneticists believe is the basis for the observed “missing heritability” is the methylome, an individual’s genomewide methylated CpG sequence pattern. The methylome (a component of the epigenome) may be the major 

epigenetic modifier of susceptibility to cancer and other chemical exposure-related diseases. Presently, 

there is no mouse reference database for the methylome. The DNA sequence data has significantly increased 

our knowledge of the genomic structure of the inbred mouse and provided the basis for imputation of the 

haplotype structure of more than 90 inbred strains used in biological research. The absence of a methylome 

reference database for the mouse significantly handicaps understanding of the mouse model in toxicology 

and environmentally related diseases and designing and conducting hypothesis based genetic and epigenetic 

research studies to understand the associated mechanisms. Two high content technologies have recently been 

developed that (1) determine genome-wide methylated CpG sites by deep sequencing of bisulfite treated 

genomic DNA to determine sequence context and cytosine methylation variation (BIS-Seq) and RNA (RNA-Seq) 

and (2) fractionate DNA sequences using differential restriction and/or affinity capture (MMDE-seq) to enrich for 

methylated DNA sequences. Together, these tools allow targeted interrogation of CpG regions of interest using 

bioinformatic data mining tools. These technologies will allow creation of a definitive map of the mouse liver 

methylome from the two parental strains (C57BL/6N and C3H/HeN) and their F1 hybrid (B6C3F1/N) offspring 

that exhibit dramatically different rates of interstrain and sex-dependent spontaneous liver cancers.

Contact Information: Dr. Raymond Tice, BSB Chief, tice@niehs.nih.gov. HTS website: http://ntp.niehs.nih.gov/go/28213
NTP Annual Report 2010 87

NIEHS/EPA Interagency Agreement – The Phthalate Initiative

Di(2-ethyl)hexyl phthalate (DEHP) and other phthalates have been nominated to the NTP for testing. To address 

these nominations, the NIEHS and EPA signed The Phthalate Initiative IAG in June 2008, which was renewed in July 

2009 and 2010. Many aspects of this IAG fall under nominations previously approved by the BSC for the study of 

peroxisome proliferators (begun in the 1990s), the nomination of DEHP by the FDA in 2004, and the critical data 

need highlighted in the NTP Monograph on DEHP issued in 2006. 

These studies will clarify how the peroxisome proliferator-activated receptor alpha (PPARα) develops in the rat 

and its relationship to DEHP-related cancer and other developmental toxicities. The studies will also provide 

critical data for future mixture studies using various models to inform on potential risks for toxicity over time 

and during development. Recent data have indicated that because phthalate esters have similar modes of 

action in utero, they show dose-addition when administered in combination; therefore, it would be appropriate 

to consider cumulative risk for the class, since human subjects (including fetuses) are typically exposed to multiple 

phthalates.

The initiative has two specific aims:

(1) Undertake an ontogeny study of PPARα in the rat to determine when the receptor is first expressed 

 in target tissues. This study will test the hypothesis that PPARα is developmentally regulated in the rat 

 and unlikely to contribute to toxicity initiated in utero after exposure to DEHP.

(2)Undertake perinatal phthalate mixture studies. These studies will test the hypothesis that exposures 

 to mixtures of phthalates, based on their individual potencies, would result in dose-addition for cancer 

 (and potentially other) outcomes.

Based on data generated in 2010, using a specific short-term in utero screen developed in the initiative to 

evaluate individual phthalates and mixtures in the Harlan SD rat, data have been developed for approximately 

11 different phthalates and derivatives. This model is also appropriate for studying developmental effects of 

various phthalates. Phthalates’ ability to alter fetal testicular testosterone production, induce changes in specific 

fetal testicular genes (especially related to steroidogenesis and testicular descent), and induce specific male 

reproductive tract malformations has been termed the “phthalate syndrome.”

Thus far, data indicating a positive response has been obtained from butylbenzyl, di-n-butyl, di-iso-butyl, 

di-ethylhexyl, di-pentyl di-n-hexyl, di-iso heptyl, di-n-heptyl and di-cyclohexyl phthalate and three different 

preparations of di-iso-nonyl phthalate (a complex mixture). Brominated DEHP, diethyl phthalate, and di-octyl 

terephthalate were all negative in this fetal phthalate screen. Specific potency factors have been developed based 

on this screening information. 

These data should guide selection of individual studies to investigate potential short-term biomarkers for toxicity, 

which in turn should support future perinatal bioassays and phthalate mixture work. The NTP has now designed 

both perinatal and standard bioassays to evaluate developmental and cancer potential from different exposure 

paradigms for DEHP, which should begin in early 2011. This information will be extremely valuable in assessing 

potential cumulative risk of phthalates, as has been advocated by a recent National Academy of Sciences 

committee report and efforts by EPA’s Integrated Risk Information System (IRIS) program.

In the ontogeny study, the NTP is evaluating the potential role of activation of PPAR pathways in the disruption 

of male sexual differentiation and testis development using custom-designed PCR plates. These plates allow the 

determination of relative mRNA expression of PPARa, PPARδ, and PPARγ and several genes activated by PPARa

including CYP4a1 on a single sample simultaneously. For six positive phthalates examined so far, none of the PPAR 

genes, CYP4a1, or any other genes “downstream” of PPARa are activated by any of the phthalates at a single high 

dosage level.

Contact Information: Dr. Paul Foster, foster2@niehs.nih.gov
88 National Toxicology Program

Appendix I

NTP staff are listed alphabetically by agency

NCTR/NTP

Beland, Frederick (870) 543-7205 frederick.beland@fda.hhs.gov

Boudreau, Mary (870) 543-7526 mary.boudreau@fda.hhs.gov

Brakebill, Nancy (870) 543-7785 nancy.brakebill@fda.hhs.gov

Chidambaram, Mani (870) 543-7368 mani.chidambaram@fda.hhs.gov

Churchwell, Mona (870) 543-7698 mona.churchwell@fda.hhs.gov

Corlett, Angela (870) 543-7403 angela.corlett@fda.hhs.gov

Couch, Letha (870) 543-7137 letha.couch@fda.hhs.gov

Cozart, Christy (870) 543-7602 christy.cozart@fda.hhs.gov

Delclos, Barry (870) 543-7372 barry.delclos@fda.hhs.gov

Doerge, Daniel (870) 543-7943 daniel.doerge@fda.hhs.gov

Duffy, Peter (870) 543-7054 peter.duffy@fda.hhs.gov

Fang, Jia-Long (870) 543-7612 jia-long.fang@fda.hhs.gov

Felton, Robert (870) 543-7504 robert.felton@fda.hhs.gov

Ferguson, Sherry (870) 543-7589 sherry.ferguson@fda.hhs.gov

Fisher, Jeffrey (870) 543-7703 jeffrey.fisher@fda.hhs.gov

Gamboa, Goncalo (870) 543-7400 goncalo.gamboa@fda.hhs.gov

George, Nysia (870) 543-7047 nysia.george@fda.hhs.gov

Guo, Lei (870) 543-7048 lei.guo@fda.hhs.gov

Hansen, Deborah (870) 543-7480 deborah.hansen@fda.hhs.gov

Heflich, Robert (870) 543-7493 robert.heflich@fda.hhs.gov

Heinze, Thomas (870) 543-7468 thomas.henize@fda.hhs.gov

Howard, Paul C. (870) 543-7672 paul.howard@fda.hhs.gov

Law, Charles (870) 543-7013 charles.law@fda.hhs.gov

Leakey, Julian (870) 543-7916 julian.leakey@fda.hhs.gov

Lewis, Sherry (870) 543-7627 sherrym.lewis@fda.hhs.gov

McDaniel, Lea (870) 543-7213 lea.mcdaniel@fda.hhs.gov

Mendoza, Maria (870) 543-7844 maria.mendoza@fda.hhs.gov

Mittelstaedt, Roberta (870) 543-7190 roberta.mittelstaedt@fda.hhs.gov

Moon, Steven (870) 543-7124 steven.moon@fda.hhs.gov

Nichols, Jasyl (870) 543-7210 jasyl.nichols@fda.hhs.gov

Nysia, George 870-543-7047 nysia.george@fda.hhs.gov

Paule, Merle (870) 543-7147 merle.paule@fda.hhs.gov

Pogribna, Marta (870) 543-7681 marta.pogribna@fda.hhs.gov

Pogribny, Igor (870) 543-7096 igor.pogribny@fda.hhs.gov

Siitonen, Paul (870) 543-7656 paul.siitonen@fda.hhs.gov

Shaddock, Joseph (870) 543-7280 joseph.shaddock@fda.hhs.gov

Stingley, Robin (870) 543-7350 robin.stingley@fda.hhs.gov

Thorn, Brett (870) 543-7338 brett.thorn@fda.hhs.gov

Trbojevich, Raul (870) 543-7490 raul.trbojevich@fda.hhs.gov 

Twaddle, Nathan (870) 543-7178 nathan.twaddle@fda.hhs.gov

Vanlandingham, Michelle (870) 543-7458 michelle.vanlandingham@fda.hhs.gov

VonTungeln, Linda (870) 543-7620 linda.vontungeln@fda.hhs.gov

Wang, Cheng (870) 543-7259 cheng.wang@fda.hhs.gov

Weis, Connie (870) 543-7378 connie.weis@fda.hhs.gov

Willingham, Evelyn (870) 543-7938 evelyn.willingham@fda.hhs.gov

Woodling, Kellie (870) 543-7491 lkellie.woodling@fda.hhs.gov

NIEHS/NTP

Alden, Charles (919) 541-3687 alden@niehs.nih.gov 

Allen, Bonnie (919) 541-3449 allenb@niehs.nih.gov

Alpers, Scott (919) 541-4377 alpers@niehs.nih.gov

Andrews, Danica (919) 541-2595 andrewsda@niehs.nih.gov

Behl, Mamta (919) 541-2784 behlmv@niehs.nih.gov

Auerbach, Scott (919) 541-4505 auerbachs@niehs.nih.gov

Birnbaum, Linda (919) 541-3201 birnbaumls@niehs.nih.gov

Bishop, Jack (919) 541-1876 bishop@niehs.nih.gov

Blystone, Chad (919) 541-2741 blystonecr@niehs.nih.gov

Bowden, Beth (919) 541-3355 bowden1@niehs.nih.gov

Boyd, Windy (919) 541-9810 boydw@niehs.nih.gov

Boyle, Michael (919) 541-1676 boylemc@niehs.nih.gov

Brown, Quashana (919) 541-4069 brownqd@niehs.nih.gov

Bucher, John (919) 541-4532 bucher@niehs.nih.gov

Casey, Warren (919) 316-4729 warren.casey@nih.gov

Caspary, William (919) 541-2150 caspary@niehs.nih.gov

Castro, Lysandra (919) 541-3373 castro@niehs.nih.gov

Cesta, Mark (919) 541-3229 cesta@niehs.nih.gov

Chan, Po-Chuen (919) 541-7561 chanp@niehs.nih.gov

Chhabra, Rajendra (919) 541-3386 chhabrar@niehs.nih.gov

Clayton, Natasha (919) 541-7843 clayton2@niehs.nih.gov

Collins, Bradley (919) 541-4666 collin10@niehs.nih.gov

Cora, Michelle (1919) 316-4717 coramc@niehs.nih.gov 

Corniffe, Glenda (919) 541-4880 corniffe@niehs.nih.gov

Cunningham, Michael (919) 541-3799 cunning1@niehs.nih.gov

Cunny, Helen (919) 541-5717 cunnyh@niehs.nih.gov

de Serres, Frederick (919) 541-0718 deserres@niehs.nih.gov

DeVito, Michael (919) 541-4142 devitom@niehs.nih.gov

Di, Xudong (919) 316-4611 dix3@niehs.nih.gov

Dixon, Darlene (919) 541-3814 dixon@niehs.nih.gov

Dunlap, Paul (919) 541-2679 dunlapp@niehs.nih.gov

Dunnick, June (919) 541-4811 dunnickj@niehs.nih.gov

Elmore, Susan (919) 541-3474 elmore@niehs.nih.gov

Fenton, Suzanne (919) 541-4141 fentonse@niehs.nih.gov

Fields, Sally (919) 541-3971 fields3@niehs.nih.gov

Fisher, Kristen (919) 541-7739 fisherk2@niehs.nih.gov

Flagler, Norris (919) 541-2397 flagler@niehs.nih.gov

Flake, Gordon (919) 541-0037 flake@niehs.nih.gov

Foley, Julie (919) 541-3772 foley1@niehs.nih.gov

Fostel, Jennifer (919) 541-5055 fostel@niehs.nih.gov

Foster, Paul (919) 541-2513 foster2@niehs.nih.gov

Frawley, Rachel (919) 541-2151 frawleyr@niehs.nih.gov

Freedman, Jonathan (919) 541-7899 freedma1@niehs.nih.gov

French, Jef (919) 541-2569 french@niehs.nih.gov

Gao, Xiaohua (919) 316-4611 gaox3@niehs.nih.gov

Germolec, Dori (919) 541-3230 germolec@niehs.nih.gov

Guy, Robbin (919) 541-4363 guyr2@niehs.nih.gov

Hall, Carolyn (919) 541-4995 hall7@niehs.nih.gov

Herbert, Ronald (919) 541-4613 herbert1@niehs.nih.gov

Hoenerhoff, Mark (919) 541-3440 hoenerhm@niehs.nih.gov

Hong, Hue-Hua (919) 541-2141 hong5@niehs.nih.gov

Hooth, Michelle (919) 316-4643 hooth@niehs.nih.gov

Howdeshell, Kembra (919) 316-4708 howdeshellkl@niehs.nih.gov

Irwin, Richard (919) 541-3340 irwin@niehs.nih.gov

Jahnke, Gloria (919) 541-3376 jahnke@niehs.nih.gov

Jayaram, Beby (919) 541-1480 jayaram1@niehs.nih.gov

Jensen, Heather (919) 541-4075 jensen1@niehs.nih.gov

Jeter, Shawn (919) 541-3476 jeter@niehs.nih.gov

Johnson, Frank (919) 541-3503 johnso54@niehs.nih.gov

Jones, Tina (919) 541-2836 jones30@niehs.nih.gov

Kamel, Freya (919) 541-1581 kamel@niehs.nih.gov

Kim, Yongbaek (919) 316-4559 Kim16@niehs.nih.gov

Kim, Sang-Hyun (919) 541-9468 sang-hyum.kim@nih.gov
NTP Annual Report 2010 89

King, Debra (919) 541-0698 kingdh@niehs.nih.gov

King-Herbert, Angela (919) 541-3464 kingher1@niehs.nih.gov

Kissling, Grace (919) 541-1756 kissling@niehs.nih.gov

Koivisto, Christopher (919) 541-3241 koivist@niehs.nih.gov

Lahousse, Stephanie (919) 541-3069 lahousses@niehs.nih.gov

Lasko, Denise (919) 541-0255 lasko@niehs.nih.gov

Litton, Dan (919) 541-2100 litton@niehs.nih.gov

Lunn, Ruth (919) 316-4637 lunn@niehs.nih.gov

Malarkey, David (919) 541-1745 malarkey@niehs.nih.gov

Malling, Heinrich (919) 541-3378 malling@niehs.nih.gov

Masinde, Tiwanda (919) 541-4069 marsh2@niehs.nih.gov

Masten, Scott (919) 541-5710 masten@niehs.nih.gov

McCarley, Deborah (919) 541-2384 mccarley@niehs.nih.gov

McIntyre, Barry (919) 316-4747 mcintyrebs@niehs.nih.gov

Mercado-Feliciano, Minerva (919) 541-2756 mercadofelicim@niehs.nih.gov

Merrick, Alex (919) 541-1531 merrick1@niehs.nih.gov

Moore, Alicia (919) 541-7914 moore5@niehs.nih.gov

Morgan, Daniel (919) 541-2264 morgan3@niehs.nih.gov

Ney, Elizabeth (919) 541-5182 ney@niehs.nih.gov

Newbold, Retha (919) 541-0738 newbold1@niehs.nih.gov

Nichols, Sharon (919) 541-7992 nicholss2@niehs.nih.gov

Orihuela Garcia, Ruben (919) 541-0540 ruben.orihuela@nih.gov 

Ovwigho, Pamela (919) 541-3278 ovwighope@niehs.nih.gov

Person, Rachel 919) 541-0685 rachel.person@nih.gov

Qu, Wei 919) 541-0540 qu@niehs.nih.gov

Rowley, Michael (919) 541-3436 rowley@niehs.nih.gov

Rao, Deepa (919) 541-0036 raod2@niehs.nih.gov

Rebolloso, Yvette (919) 541-4209 rebollo1@niehs.nih.gov

Rice, Julie (919) 541-2533 ricej2@niehs.nih.gov

Rider, Cynthia (919) 541-7638 ridercv@niehs.nih.gov

Roach, Donna (919) 541-3023 roachd@od.nih.gov

Robinson, Veronica (919) 541-2238 godfrey@niehs.nih.gov

Rooney, Andrew (919) 316-4704 rooneyaa@niehs.nih.gov

Roycroft, Joseph (919) 541-3627 roycroft@niehs.nih.gov

Sanders, Michael (919) 541-1872 sander10@niehs.nih.gov

Shelby, Michael (919) 541-3455 shelby@niehs.nih.gov

Shockley, Keith (919) 541-3033 shockleykr@niehs.nih.gov

Sills, Robert (919) 541-0180 sills@niehs.nih.gov

Smith, Cynthia (919) 541-3473 smith19@niehs.nih.gov

Soward, Sharon (919) 541-5132 soward@niehs.nih.gov

Spencer, Diane (919) 541-2759 spencer2@niehs.nih.gov

Stanko, Jason (919) 541-0461 stankojp@niehs.nih.gov

Stockton, Patricia (919) 541-4471 stockton@niehs.nih.gov

Stokes, William (919) 541-7997 stokes@niehs.nih.gov

Stout, Matthew (919) 541-3489 stoutm@niehs.nih.gov

Sun, Yang (919) 541-3024 suny2@mail.nih.gov

Surh, In (919) 541-3862 surhi@niehs.nih.gov

Sutton, Deloris (919) 541-5112 sutton2@niehs.nih.gov

Taylor, Kyla (919) 316-4707 taylorkw@niehs.nih.gov

Teng, Christina (919) 541-0344 teng1@niehs.nih.gov

Thayer, Kristina (919) 541-5021 thayer@niehs.nih.gov

Tice, Raymond (919) 541-4482 tice@niehs.nih.gov

Tokar, Erik (919) 541-3951 tokare@niehs.nih.gov

Ton, Thai-Vu (919) 541-2118 ton@niehs.nih.gov

Travlos, Gregory (919) 541-0653 travlos@niehs.nih.gov

Tubbs, Jacquelyn (919) 541-2919 tubbsj@niehs.nih.gov

Vallant, Molly (919) 541-5234 vallant@niehs.nih.gov

Waalkes, Michael (919) 541-2328 waalkes@niehs.nih.gov

Waidyanatha, Suramya (919) 541-2144 waidyanathas@niehs.nih.gov

Walker, Nigel (919) 541-4893 walker3@niehs.nih.gov

White, Lori (919) 541-9834 whiteld@niehs.nih.gov

Wilson, Ralph (919) 541-4140 wilson23@niehs.nih.gov

Withers, Sheila (919) 541-5066 withersg@niehs.nih.gov

Witt, Kristine (919) 541-2761 witt@niehs.nih.gov

Wolfe, Mary (919) 541-7539 wolfe@niehs.nih.gov

Wyde, Michael (919) 316-4640 wyde@niehs.nih.gov

Yu, Xianhong (919) 316-4599 yu1@niehs.nih.gov

Yu, Yuanyuan (919) 541-3024 xuy5@niehs.nih.gov

Zegre-Cannon, Coralie (919) 316-4559 cannonc@niehs.nih.gov

NIOSH/NTP

Anderson, Stacy (304) 285-6174, X2 dbx7@cdc.gov

Beezhold, Don (304) 285-5963 zec1@cdc.gov

Biagini, Raymond (513) 533-8196 reb4@cdc.gov 

Castranova, Vincent (304) 285-6032 vic1@cdc.gov

Curwin, Brian (513) 841-4432 bic4@cdc.gov

DeBord, D. Gayle (513) 533-8212 ded4@cdc.gov 

Erdely, Aaron (304) 285-5903 efi4@cdc.gov 

Estill, Cherie (513) 841-4476 clf4@cdc.gov

Franko, Jennifer (304) 285-6174, x8 hfy0@cdc.gov 

Frasch, H. Frederick (304) 285-5755 hbf9@cdc.gov 

Geraci, Chuck (513) 533-8339 ciu9@cdc.gov 

Green, Brett (304) 285-5884 dox6@cdc.gov

Hanley, Kevin (513) 841-4113 kwh0@cdc.gov 

Harper, Martin (304) 285-6043 zzg7@cdc.gov

Hines, Cynthia (513) 841-4453 cjh8@cdc.gov

Johnson, Victor (304) 285-6249 vcj6@cdc.gov 

Joseph, Pius (304) 285-6240 pcj5@cdc.gov

Leonard, Steve (304) 285-5831 stl0@cdc.gov 

Luster, Michael (304) 285-5940 myl6@cdc.gov 

Lynch, Dennis (513) 533-8213 dwl1@cdc.gov 

Munson, Albert E. (304) 285-6121 akm5@cdc.gov 

Reynolds, Steve (304) 285-5806 sir6@cdc.gov 

Schrader, Steven (513) 533-8210 sms4@cdc.gov 

Siegel, Paul (304) 285-5855 pds3@cdc.gov 

Snawder, John (513) 533-8496 jts5@cdc.gov 

Streicher, Robert (513) 841-4296 rps3@cdc.gov 

Striley, Cynthia (513) 533-8123 chs3@cdc.gov 

Templeton, Steven (304) 285-5721 goy6@cdc.gov

Toraason, Mark (513) 533-8207 mht1@cdc.gov 

Virji, Mohammed (304) 285-5797 fnb8@cdc.gov

Wells, Ray (304) 285-6341 ozw0@cdc.gov

Wurzelbacher, Steve (513) 841-4322 srw3@cdc.gov

Yucesoy, Berran (304) 285-5993 yab7@cdc.gov
90 National Toxicology Program

NTP FY 2010 Publication List 

Ali R, Mittelstaedt RA, Shaddock JG, Bhalli JA and Heflich RH. 2010. Micronucleus induction by Ochratoxin A 

in CHO and TK6 cells. Environ Mol Mutagen 51(7):728-728.

Ali SF, Bondy SC. 2010. Red wine but not ethanol at low doses can protect against the toxicity of 

methamphetamine. Brain Res 1346:247-250. 

Anderson SE, Jackson LG, Franko J, Wells, JR. 2010. Evaluation of dicarbonyls generated in a simulated indoor 

air environment using an in vitro exposure system. Toxicol Sci 115(2):453-461. 

Anderson SE, Munson AE, Butterworth LF, Germolec D, Morgan DL, Roycroft JA, Dill J, Meade BJ. 2010. 

Whole-body inhalation exposure to 1-bromopropane suppresses the IgM response to sheep red blood cells 

in female B6C3F1 mice and Fisher 344/N rats. Inhal Toxicol 22(2):125-132.

Anderson SE, Umbright C, Sellamuthu R, Fluharty K, Kashon M, Franko J, Jackson LG, Johnson VJ, Joseph, P. 2010. 

Irritancy and allergic responses induced by topical application of ortho-phthalaldehyde. Toxicol Sci 115(2):435-443. 

Antonini JM, Roberts JR, Chapman RS, Soukup JM, Ghio AJ, Sriram K. 2010. Pulmonary toxicity and extrapulmonary

tissue distribution of metals after repeated exposure to different welding fumes. Inhal Toxicol 22(10):805-816. 

Antunes AMM, Godinho ALA, Martins IL, Justino GC, Beland FA, Marques, MM. 2010. Amino acid adduct formation 

by the nevirapine metabolite, 12-Hydroxynevirapine – a possible factor in nevirapine toxicity. Chem Res Toxicol

23(5):888-899. 

Arias-Loza PA, Muehlfelder M, Elmore SA, Maronpot R, Hu K, Blode H, Hegele-Hartung C, Fritzemeier KH, Ertl G, 

Pelzer T. 2009. Differential effects of 17 beta-estradiol and of synthetic progestins on aldosterone-salt-induced 

kidney disease. Toxicol Pathol 37(7):969-982.

Auerbach SS, Bristol DW, Peckham JC, Travlos GS, Hébert CD, Chhabra RS. Toxicity and carcinogenicity studies 

of methylene blue trihydrate in F344N rats and B6C3F1 mice. Food Chem Toxicol 48(1):169-177.

Auerbach SS, Shah RR, Mav D, Smith CS, Walker NJ, Vallant MK, Boorman GA, Irwin RD. Predicting the 

hepatocarcinogenic potential of alkenylbenzene flavoring agents using toxicogenomics and machine learning. 

Toxicol Appl Pharmacol 243(3):300-314. 

Auerbach SS, Thomas R, Shah R, Xu H, Vallant MK, Nyska A, Dunnick JK. 2010. Comparative phenotypic assessment 

of cardiac pathology, physiology, and gene expression in C3H/HeJ, C57BL/6J, and B6C3F1/J mice. Toxicol Pathol

38:923-942.

Azad N, Iyer A, Vallyathan V, Wang L, Castranova V, Stehlik C, Rojanasakul Y. 2010. Role of oxidative/nitrosative 

stress-mediated Bcl-2 regulation in apoptosis and malignant transformation. Ann N Y Acad Sci 1203:(1-6). 

Azad N, Iyer AKV, Wang LY, Lu YJ, Medan D, Castranova V, Rojanasakul Y. 2010. Nitric oxide-mediated Bcl-2 

stabilization potentiates malignant transformation of human lung epithelial cells. Am J Respir Cell Mol Biol

42(5):578-585. 

Bach U, Hailey JR, Hill GD, Kaufmann W, Latimer KS, Malarkey DE, Maronpot RM, Miller RA, Moore RR, Morrison JP, 

Nolte T, Rinke M, Rittinghausen S, Suttie RW, Travlos GS, Vahle JL, Willson GA and Elmore SA. 2010. Proceedings of 

the 2009 National Toxicology Program satellite symposium. Toxicol Pathol 38(1):9-36. 
NTP Annual Report 2010 91

Barnard DE, Lewis SM, Teter BB, Thigpen JE. 2009. Open- and closed-formula laboratory animal diets and their 

importance to research. J Am Assoc Lab Anim Sci 48(6):709-713. 

Bauer AK, Fostel J, Degraff LM, Rondini EA, Walker C, Grissom SF, Foley J, Kleeberger SR. 2009. Transcriptomic 

analysis of pathways regulated by toll-like receptor 4 in a murine model of chronic pulmonary inflammation and 

carcinogenesis. Mol Cancer 8:107. 

Beezhold KJ, Castranova V, Chen F. 2010. Microprocessor of microRNAs: regulation and potential for therapeutic 

intervention. Mol Cancer 9:134. 

Beland FA. 2009. Abstracts of the 32nd Annual Meeting of the United Kingdom Environmental Mutagen Society, 

12-15 July 2009 at the University of Leeds, UK. Mutagenesis 24(6):523-523. 

Bell DR, Clode S, Fan MQ, Fernandes A, Foster PMD, Jiang T, Loizou G, MacNicoll A, Miller BG, Rose M, 

Tran L and White S. 2010. Interpretation of studies on the developmental reproductive toxicology of 

2,3,7,8-tetrachlorodibenzo-p-dioxin in male offspring. Food Chem Toxicol 48(6):1439-1447. 

Benbrahim-Tallaa L, Tokar EJ, Diwan BA, Coppin JF, Waalkes MP. 2009. Cadmium malignantly transforms normal 

human breast epithelial cells into a basal-like phenotype. Environ Health Perspect 117(12):1847-1852. 

Bhalli JA, Dobrovolsky VN, Heflich RH. 2010. Pig-a mutation manifestation in mice following single and split doses 

of N-ethyl-N-nitrosourea. Environ Mol Mutagen 51(7):722-722. 

B’Hymer C, Cheever KL. 2010. Evaluation of a procedure for the simultaneous quantification of 

4-ketocyclophosphamide, cyclophosphamide, and Ifosfamide in human urine. J Chromatogr Sci 48(5):328-333. 

Binienda ZK, Ross IA, Gough B, Riccio T, Kim CS, Ali SF. Striatal dopamine in the response of brain and liver 

unsaturated and saturated free fatty acids following administration of C75 in CD-1 mice. Neurosci Lett 469(3):324-327. 

Binienda ZK, Beaudoin MA, Gough B, Ali SF, Virmani A. 2010. Assessment of 3-nitropropionic acid-evoked 

peripheral neuropathy in rats: neuroprotective effects of acetyl-l-carnitine and resveratrol. Neurosci Lett

480(2):117-121. 

Birnbaum LS, Zenick H, Branche CM. 2009. Environmental justice: a continuing commitment to an evolving 

concept. Am J Public Health Suppl 3:S487-9. Erratum in: Am J Public Health. 2010 Feb;100(2):199. 

Birnbaum LS. 2009. Applying research to public health questions: timing and the environmentally relevant dose. 

Environ Health Perspect 117(11):A478. 

Birnbaum LS. 2009. NIEHS supports partnerships in environmental public health. Prog Community Health Partnersh

3(3):195-196. 

Blanco I, Janciauskiene S, Nita I, Fernandez-Bustillo E, Carcaba V, Gallo C, Alvarez-Rico M, de Serres F, Beridze N. 

2010. Low plasma levels of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF), 

and vascular endothelial growth factor (VEGF) in patients with alpha1-antitrypsin deficiency-related fibromyalgia. 

Clin Rheumatol 29(2):189-197. 

Blanco IB, Astudillo A, Iglesias FD, Janciauskiene S, Fernandez VC, Alvaro CG, Argiz HC, de Serres FJ, Bustillo, EF. 

2010. Intravenous infusions of purified alpha-1 antitripsyn effectively controls symptoms and reverts muscle 

biopsy changes in an MZ alpha-1 antitripsyn deficiency and fibromyalgia syndrome Patient. J Musculoskelet Pain

18(2):167-172. 
92 National Toxicology Program

Blystone CR, Kissling GE, Bishop JB, Chapin RE, Wolfe GW, Foster PM. 2010. Determination of the di-(2-ethylhexyl) 

phthalate NOAEL for reproductive development in the rat: importance of the retention of extra animals to 

adulthood. Toxicol Sci 116(2):640-646. 

Borm P, Castranova V. 2009. Toxicology of nanomaterials: permanent interactive learning. Part Fibre Toxicol 6:28. 

Bowman JD, Miller CK, Krieg EF, Song RG. 2010. Analyzing digital vector waveforms of 0-3000 Hz magnetic fields 

for health studies. Bioelectromagnetics 31(5):391-405. 

Boyd WA, Crocker TL, Rodriguez AM, Leung MCK, Wade Lehmann D, Freedman JH, Van Houten B, Meyer JN. 2010. 

Nucleotide excision repair genes are expressed at low levels and are not detectably inducible in Caenorhabditis 

elegans somatic tissues, but their function is required for normal adult life after UVC exposure. Mutat Res 683

(1-2):57-67. 

Boyd WA, McBride SJ, Rice JR, Snyder DW, Freedman JH. 2010. A high-throughput method for assessing chemical 

toxicity using a Caenorhabditis elegans reproduction assay. Toxicol Appl Pharmacol 245(2):153-159. 

Burch JB, Svendsen E, Siegel PD, Wagner SE, Von Essen S, Keefe T, Mehaffy J, Martinez AS, Bradford M, Baker 

L, Cranmer B, Saito R, Tessari J, Linda P, Andersen C, Christensen O, Koehncke N, Reynolds SJ. 2010. Endotoxin 

exposure and inflammation markers among agricultural workers in Colorado and Nebraska. J Toxicol Environ 

Health A 73(1):5-22. 

Caldwell J, Lunn R, Ruder A. 2010. Trichloroethylene. In identification of research needs to resolve the 

carcinogenicity of high-priority IARC carcinogens. Views and expert opinions of an IARC/NORA expert group 

meeting, Lyon, France, 30 June – 2 July 2009. IARC Technical Publication No. 42. 

Caldwell J, Lunn R. 2010. Dichloromethane, methylene chloride (DCM). In identification of research needs to 

resolve the carcinogenicity of high-priority IARC carcinogens. Views and expert opinions of an IARC/NORA expert 

group meeting, Lyon, France, 30 June – 2 July 2009. IARC Technical Publication No. 42

Cannon CZ, Godfrey VL, King-Herbert A, Nielsen JN. 2009. Metastatic uterine adenocarcinoma in an 8-year-old 

gilt. J Am Assoc Lab Anim Sci 48(6):795-800. 

Cao LY, Taylor JS, Sood A, Murray D, Siegel PD. 2010. Allergic contact dermatitis to synthetic rubber gloves: 

Changing trends in patch test reactions to accelerators. Arch Dermatol 146(9):1001-1007. 

Card JW, Carey MA, Voltz JW, Bradbury JA, Ferguson CD, Cohen EA, Schwartz S, Flake GP, Morgan DL, Arbes Jr SJ, 

Barrow DA, Barros SP, Offenbacher S, Zeldin, DC. 2010. Modulation of allergic airway inflammation by the oral 

pathogen Porphyromonas gingivalis. Infect Immun 78(6):2488-2496. 

Carey MA, Bradbury JA, Rebolloso YD, Graves JP, Zeldin DC, Germolec DR. 2010. Pharmacologic inhibition of COX-1 

and COX-2 in influenza A viral infection in mice. PLoS One 5(7):e11610. 

Carreon T, Hein MJ, Viet S, Hanley KW, Ruder AM, Ward EM. 2009. Increased bladder cancer risk among workers 

exposed to o-toluidine and aniline: a reanalysis. Occup Environ Med 67:348-350. 

Castranova V, Fedan J. 2010. Dr. Val Vallyathan: In memoriam. Part Fibre Toxicol 7:24 

Cederroth CR, Zimmermann C, Beny JL, Schaad O, Combepine C, Descombes P, Doerge DR, Pralong FP, Vassalli JD, 

Nef S. 2010. Potential detrimental effects of a phytoestrogen-rich diet on male fertility in mice. Mol Cell Endocrinol

321(2):152-160. 
NTP Annual Report 2010 93

Cesta MF, Ryman-Rasmussen JP, Wallace DG, Masinde T, Hurlburt G, Taylor AJ, Bonner JC. 2010. Bacterial 

lipopolysaccharide enhances PDGF signaling and pulmonary fibrosis in rats exposed to carbon nanotubes. 

Am J Respir Cell Mol Biol 43(2):142-151. 

Chambliss KL, Wu QA, Oltmann S, Konaniah ES, Umetani M, Korach KS, Thomas GD, Mineo C, Yuhanna IS, Kim SH, 

Madak-Erdogan Z, Maggi A, Dineen SP, Roland CL, Hui DY, Brekken RA, Katzenellenbogen JA, Katzenellenbogen 

BS, Shaul, PW. 2010. Non-nuclear estrogen receptor alpha signaling promotes cardiovascular protection but not 

uterine or breast cancer growth in mice. J Clin Invest 120(7):2319-2330. 

Chan PC, IARC Working Group. 2010. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. 

Ingested nitrates and nitrites, and cyanobacterial peptide toxins. Lyon, France. Vol 4 http://monographs.iarc.fr/

ENG/Monographs/vol94/index.php

Chan PC, Ramot Y, Malarkey DE, Blackshear P, Kissling G, Travlos G, Nyska A. 2010. 14-Weeks toxicity study of green 

tea extract in rats and mice. Toxicol Pathol 38:1070-1084.

Chandra SA, Nolan MW, Malarkey DE. 2010. Chemical carcinogenesis of the gastrointestinal tract in rodents: 

An overview with emphasis on NTP carcinogenesis bioassays. Toxicol Pathol 38(1):188-197. 

Chang Q, Pan J, Wang X, Zhang Z, Chen F, Shi X. 2010. Reduced reactive oxygen species-generating capacity 

contributes to the enhanced cell growth of arsenic-transformed epithelial cells. Cancer Res 70(12):5127-5135. 

Chelonis J, Johnson T, Ferguson S, Kubacak B, Edwards M, Paule M. 2010. Effects of methylphenidate on 

motivation in children with attention-deficit/hyperactivity disorder. Neurotoxicol Teratol 32(4):504. 

Chen F, Li S, Castanova V. 2009. Signature genes for both hepatoblastoma and hepatocellular carcinoma. 

Eur J Gastroenterol Hepatol 21(10):1220-1221. 

Chen F, Li S, Castranova V. 2009. Overlapping signature genes between hepatocellular carcinoma and intrahepatic 

cholangiocarcinoma. Eur J Gastroenterol Hepatol 21(11):1320-1321.

Chen T, Mei N, Fu PP. 2010. Genotoxicity of pyrrolizidine alkaloids. J Appl Toxicol 30(3):183-96. 

Chiang HM, Yin JJ, Xia Q, Zhao Y, Fu PP, Wen KC, Yu H. 2010. Photoirradiation of azulene and guaiazulene-Formation 

of reactive oxygen species and induction of lipid peroxidation. J Photochem Photobiol A Chem 211(2-3):123-128. 

Chipinda I, Ajibola RO, Morakinyo MK, Ruwona TB, Simoyi RH, Siegel PD. 2010. Rapid and simple kinetics screening 

assay for electrophilic dermal sensitizers using nitrobenzenethiol. Chem Res Toxicol 23(5):918-25. 

Cimafranca MA, Davila J, Ekman GC, Andrews RN, Neese SL, Peretz J, Woodling KA, Helferich WG, Sarkar J, Flaws 

JA, Schantz SL, Doerge DR, Cooke PS. 2010. Acute and chronic effects of oral genistein administration in neonatal 

mice. Biol Reprod 83(1):14-121. 

Collins BJ, Stout MD, Levine KE, Kissling GE, Melnick RL, Fennell TR, Walden R, Pritchard JB, Fernando RA, Burka LT, 

and Hooth MJ. 2010. Exposure to hexavalent chromium resulted in significantly higher tissue chromium burden 

compared with trivalent chromium following similar oral doses to male F344/N rats and female B6C3F1 mice. 

Tox Sci 118(2):368-379.

Cowin PA, Gold E, Aleksova J, O’Bryan MK, Foster PM, Scott HS, Risbridger GP. 2010. Vinclozolin exposure in utero

induces postpubertal prostatitis and reduces sperm production via a reversible hormone-regulated mechanism. 

Endocrinology 151:783-792.
94 National Toxicology Program

Crawford LW, Foley JF, Elmore SA. 2010. Histology atlas of the developing mouse hepatobiliary system with 

emphasis on E9.5 to E18.5. Toxicol Pathol 38:872-906.

Cruz VPDL, Elinos-Calderón D, Carrillo-Mora P, Silva-Adaya D, Konigsberg M, Morán J, Ali SF, Chánez-Cárdenas ME, 

Pérez-De La Cruz G, Santamaría A. 2010. Time-course correlation of early toxic events in three models of striatal 

damage: modulation by proteases inhibition. Neurochem Int 56(6-7):834-842. 

Cuevas E, Lantz S, Newport G, Divine B, Wu Q, Paule MG, Tobón-Velasco JC, Ali SF, Santamaría A. 2010. On the early 

toxic effect of quinolinic acid: involvement of RAGE. Neurosci Lett 474(2):74-78. 

Cummings KJ, Gaughan DM, Kullman GJ, Beezhold DH, Green BJ, Blachere FM, Bledsoe T, Kreiss K, Cox-Ganser J. 

2010. Adverse respiratory outcomes associated with occupational exposures at a soy processing plant. Eur Respir J 

36(5)1007-1015.

Cunningham ML, Collins BJ, Hejtmancik MR, Herbert RA, Travlos GS, Vallant MK, Stout MD. 2010. Effects of 

the PPAR alpha agonist and widely used antihyperlipidemic drug gemfibrozil on hepatic toxicity and lipid 

metabolism. PPAR Res 2010:681963.

Curwin BD, Hein MJ, Barr DB, Striley C. 2010. Comparison of immunoassay and HPLC-MS/MS used to measure 

urinary metabolites of atrazine, metolachlor, and chlorpyrifos from farmers and non-farmers in Iowa. J Expo Sci 

Environ Epidemiol 20(2):205-212. 

Davis RR, Custer DA, Krieg E, Alagramam K. 2010. N-Acetyl L-cysteine does not protect mouse ears from the effects 

of noise. J Occup Med Toxicol 5:11. 

DeBord DG, Schulte P, Butler MA, McCanlies E, Reutman S, Ruder A, Schill A, Schubauer-Berigan M, Schuler C, 

Weston A. 2009. Genetics in the Workplace: Implications for Occupational Safety and Health. Washington, DC: 

U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, National Institute for 

Occupational Safety and Health, DHHS (NIOSH) Publication No. 2010-101.

De Serres FJ, Blanco I, Fernández-Bustillo E. 2010. Ethnic differences in alpha-1 antitrypsin deficiency in the United 

States of America. Ther Adv Respir Dis 4(2):63-70. 

Dertinger SD, Phonethepswath S, Weller P, Stankowski LF, Roberts DJ, Shi J, Krsmanovic L, Vohr HW, Custer L, 

Gleason C, Henwood A, Sweder K, Giddings A, Lynch AM, Gunther WC, Thiffeault CJ, Shutsky TJ, Fiedler RD, Bhalli JA,

Heflich RH. 2010. Pig-a Mutation assay: Evaluation of inter-laboratory transferability and reproducibility. 

Environ Mol Mutagen 51(7):P86. 

Dietert RR, DeWitt JC, Germolec DR, Zelikoff JT. 2010. Breaking patterns of environmentally influenced disease for 

health risk reduction: immune perspectives. Environ Health Perspect 118(8):1091-1099. 

Ding M, Zhao J, Bowman L, Lu Y, Shi X. 2010. Inhibition of AP-1 and MAPK signaling and activation of Nrf2/ARE 

pathway by quercitrin. Int J Oncol 36(1):59-67. 

Dinse GE, Peddada SD, Harris SF, Elmore SA. 2010. Comparison of NTP historical control tumor incidence rates in 

female Harlan Sprague Dawley and Fischer 344/N rats. Toxicol Pathol 38(5):765-775. 

Dobrovolsky VN, Shaddock JG, Pearce MG, Heflich RH. 2010. Differential Induction of Pig-a and Hprt mutation 

following exposure of rats to ionizing radiation. Environ Mol Mutagen 51(7): 724-724. 

Doerge DR, Twaddle NC, Vanlandingham M, Fisher JW. 2010. Pharmacokinetics of bisphenol A in neonatal and 

adult Sprague-Dawley rats. Toxicol Appl Pharmacol 247(2):158-65. 
NTP Annual Report 2010 95

Draper DW, Madenspacher JH, Dixon D, King DH, Remaley AT, Fessler MB. 2010. ATP-binding cassette transporter 

G1 deficiency dysregulates host defense in the lung. Am J Respir Crit Care Med 182(3):404-12. 

Elmore S A. 2010. Enhanced histopathology evaluation of lymphoid organs. In: Methods in Molecular Biology; 

Immunotoxicity Testing (Humana Press). Ed. Dietert R. Chapter 22:323-339.

Fan X, Lobenhofer EK, Chen M, Shi W, Huang J, Luo J, Zhang J, Walker SJ, Chu TM, Li L, Wolfinger R, Bao W, 

Paules RS, Bushel PR, Li J, Shi T, Nikolskaya T, Nikolsky Y, Hong H, Deng Y, Cheng Y, Fang H, Shi L. Tong W. 2010. 

Consistency of predictive signature genes and classifiers generated using different microarray platforms. 

Pharmacogenomics J 10(4):247-257. 

Fang F, Xu Q, Park Y, Huang XM, Hollenbeck A, Blair A, Schatzkin A, Kamel F, Chen HL. 2010. Depression and the 

subsequent risk of Parkinson’s disease in the NIH-AARP Diet and Health Study. Mov Disord 25(9):1157-1162. 

Fang JL, Stingley RL, Beland FA, Harrouk W, Lumpkins DL, Howard P. 2010. Occurrence, efficacy, metabolism, 

and toxicity of triclosan. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 28(3):147-171. 

Fedan JS, Piedimonte G. 2010. Dr. R Shiffmann’s Asthmador: relevant to our times, or a snake oil remedy? 

Curr Opin Pharmacol 10(3):213-217. 

Fenton SC, Rudel RA, Euling SY, Makris SL. 2010. Mammary gland development as a sensitive indicator of early life 

exposures: recommendations from an interdisciplinary workshop. Birth Defects Res A Clin Mol Teratol 88(5):359-359. 

Ferguson SA, Berry KJ. 2010. Chronic oral treatment with isotretinoin alters measures of activity but not anxiety in 

male and female rats. Neurotoxicol Teratol 32(5):573-578. 

Ferguson SA, Boctor SY. 2010. Cocaine responsiveness or anhedonia in rats treated with methylphenidate during 

adolescence. Neurotoxicol Teratol 32(4):432-442. 

Ferguson SA, Garey J, Smith ME, Twaddle NC, Doerge DR, Paule MG. 2010. Preweaning behaviors, developmental 

landmarks, and acrylamide and glycidamide levels after pre- and postnatal acrylamide treatment in rats. 

Neurotoxicol Teratol 32(3):373-382. 

Foster PMD. 2010. The cons of the extended one-generation study protocol. Birth Defects Res A Clin Mol Teratol

88(5):325. 

Fowler BA, Schubauer-Berigan M, Hines CJ. 2010. Indium phosphide and other indium compounds: includes 

indium phosphide, indium arsenide, indium tin oxide, CIS, CIGS. In Identification of Research Needs to Resolve the 

Carcinogenicity of High-priority IARC Carcinogens. Views and Expert Opinions of an IARC/NORA Expert Group 

Meeting, Lyon, France, 30 June – 2 July 2009. IARC Technical Publication No. 42. Lyon, France: International Agency 

for Research on Cancer 42:16-24.

French J E, Leblanc B, Long GG, Morton D, Storer R, Leighton J, Swenburg J, Tsuda H. Panel discussion: alternative 

mouse models for carcinogenicity assessment. Toxicol Pathol 38(1):72-5. 

Gamboa da Costa G, Singh R, Arit VM, Mirza A, Richards M, Takamura-Enya T, Schmeiser HH, Farmer PB, Phillips 

DH. 2009. Quantification of 3-nitrobenzanthrone-DNA adducts using online column-switching HPLC-electrospray 

tandem mass spectrometry. Chem Res Toxicol 22(11):1860-1868. 

Gao X, Yu L, Castro L, Moore AB, Hermon T, Bortner C, Sifre M, Dixon D. 2010. An endocrine-disrupting chemical, 

fenvalerate, induces cell cycle progression and collagen type I expression in human uterine leiomyoma and 

myometrial cells. Toxicol Lett 196(3):133-141. 
96 National Toxicology Program

Garey J, Binienda Z, Paule M. 2010. Effect of lifetime exposure to acrylamide on motor nerve conduction velocity 

in rats at one year of age. Neurotoxicol Teratol 32(4):504-505. 

Germolec DR, Frawley RP, Evans E. 2010. Markers of inflammation. Methods Mol Biol 598:53-73.

Ghosh S, Hoenerhoff MJ, Clayton N, Myers P, Stumpo DJ, Maronpot RR, Blackshear PJ. 2010. Left-sided cardiac 

valvulitis in tristetraprolin-deficient mice: the role of tumor necrosis factor alpha. Am J Pathol 176(3):1484-1493. 

Glass R, Rosenthal J, Jessup CM, Birnbaum L, Portier C. 2009 Tackling the research challenges of health and climate 

change. Environ Health Perspect 117(12):A534. 

Godin SJ, DeVito MJ, Hughes MF, Ross DG, Scollon EJ, Starr JM, Setzer RW, Conolly RB, Tornero-Velez R. 2010. 

Physiologically based pharmacokinetic modeling of deltamethrin: development of a rat and human diffusion-limited 

model. Toxicol Sci 115(2):330-343. 

Gohlke JM, Stockton PS, Sieber S, Foley J, Portier CJ. 2009. AhR-mediated gene expression in the developing mouse 

telencephalon. Reprod Toxicol 28(3):321-328. 

Goodman JI, Augustine KA, Cunnningham ML, Dixon D, Dragan YP, Falls JG, Rasoulpour RJ, Sills RC, Storer RD, Wolf 

DC, Pettit SD. 2010. What do we need to know prior to thinking about incorporating an epigenetic evaluation into 

safety assessments? Toxicol Sci 116(2):375-381. 

Grajewski B, Waters MA, Yong LC, Tseng CY, Zivkovich Z, Cassinelli RT. 2010. Airline pilot cosmic radiation and 

circadian disruption exposure assessment from logbooks and company records. Am J Epidemiol 171(S86-S86) 

Green BJ, Tovey ER, Beezhold DH, Perzanowski MS, Acosta LM, Divjan AI, Chew GL. 2009. Surveillance of fungal 

allergic sensitization using the fluorescent halogen immunoassay. J Mycol Med 19(4):253-261. 

Guo L, Dial S, Shi Q, Xia Q, Mei N, Li Q, Chan PC, Fu P. 2010. Gene expression profiling in male B6C3F1 mouse livers 

exposed to kava identifies - changes in drug metabolizing genes and potential mechanisms linked to kava toxicity. 

Food Chem Toxicol 48, 686-696.

Guo L, Mei N, Liao W, Chan PC, Fu P. 2010. Ginkgo biloba extract induces gene expression changes in xenobiotics 

metabolism and the Myc-centered network. OMICS J Int Biol 14:75-90.

Guo L, Mei N, Xia QS, Chen T, Chan PC, Fu PP. 2010. Gene expression profiling as an initial approach for mechanistic 

studies of toxicity and tumorigenicity of herbal plants and herbal dietary supplements. J Environ Sci Health C 

Environ Carcinog Ecotoxicol Rev 28(1):60-87. 

Guo, L, Shi Q, Dial S, Xia Q, Mei N, Li QZ, Chan PC, Fu P. Gene expression profiling in male B6C3F1 mouse livers 

exposed to kava identifies changes in drug metabolizing genes and potential mechanisms linked to kava toxicity. 

Food Chem Toxicol 48(2):686-696. 

Hakk H, Diliberto JJ, Birnbaum LS. 2009. The effect of dose on 2,3,7,8-TCDD tissue distribution, metabolism and 

elimination in CYP1A2 (-/-) knockout and C57BL/6N parental strains of mice. Toxicol Appl Pharmacol 241(1):119-126. 

Hamilton RF, Wu N, Porter D, Buford M, Wolfarth M, Holian A. 2009. Particle length-dependent titanium dioxide 

nanomaterials toxicity and bioactivity. Part Fibre Toxicol 6:35. 

Hanley KW, Petersen MR, Cheever KL, Luo L. 2010. Bromide and N-acetyl-S-(n-propyl)-L-cysteine in urine from 

workers exposed to 1-bromopropane solvents from vapor degreasing or adhesive manufacturing. Int Arch Occup 

Environ Health 83(5):571-84. 
NTP Annual Report 2010 97

Hard, GC, Flake GP, Sills RC. 2009. Re-evaluation of kidney histopathology from 13-week toxicity and two-year 

carcinogenicity studies of melamine in the F344 rat: Morphologic evidence of retrograde nephropathy. Vet Pathol

46(6):1248-1257. 

Harrison JC, Ham JE. 2009. β-Ionone reactions with the nitrate radical: Rate constant and gas-phase products.

Int J Chem Kinetics 41(10):629-641. 

Hein MJ, Waters MA, Ruder AM, Stenzel MR, Blair A, Stewart PA. 2010. Statistical modeling of occupational 

chlorinated solvent exposures for case-control studies using a literature-based database. Ann Occup Hyg

54(4):459-472. 

Hines CJ, Yau AY, Zuniga MM, Wells JR, Nilsen Hopf NB, Camann DE. Development of a personal dual-phase air 

sampling method for phthalate diesters. J Environ Monit 12(2):491-499. 

Hines, CJ, Hopf NB, Deddens JA, Silva MJ, Calafat AM. 2009. Estimated daily intake of phthalates in occupationally 

exposed groups. J Expo Sci Environ Epidemiol 21(2):133-141. 

Hladik WJ, Stover J, Esiru G, Harper M, Tappero J. 2009. The contribution of family planning towards the prevention 

of vertical HIV transmission in Uganda. PLoS One 4(11):e7691.

Hobbs CA, Recio L, Shepard K, Green A, Winters J, Baldetti C, Streicker M, Witt KL. 2010. Genotoxicity of 

hydroxyurea in mice measured at doses relevant to pediatric sickle cell anemia patients. Environ Mol Mutagen

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in axial skeleton. J Biol Chem 285(21):16206-16217. 

Shpyleva SI, Tryndyak VP, Kovalchuk O, Starlard-Davenport A, Chekhun VF, Beland FA, Pogribny IP. 2010. Role of 

ferritin alterations in human breast cancer cells. Breast Cancer Res Treat 126(1)63-71

Shvedova AA, Kagan VE. 2010. The role of nanotoxicology in realizing the ‘helping without harm’ paradigm 

of nanomedicine: lessons from studies of pulmonary effects of single-walled carbon nanotubes. J Intern Med

267(1):106-118. 

Shvedova AA, Kagan VE. Fadeel B. 2010. Close encounters of the small kind: adverse effects of man-made 

materials interfacing with the nano-cosmos of biological systems. Annu Rev Pharmacol Toxicol 50:63-88. 

Siegel PD, Fowler JF, Storrs FJ, Sasseville D, Pratt M, Bledsoe TA, Law BF, Beezhold D, Zug K, Fowler LM. 2010. 

Allergen content of patient problem and nonproblem gloves: relationship to allergen-specific patch-test findings. 

Dermatitis 21(2):77-83. 

Siegel PD, Law BF, Fowler JF Jr., Fowler LM. 2010. Disproportionated rosin dehydroabietic acid in neoprene surgical 

gloves. Dermatitis 21(3):157-159. 

Singh BP, Nyska A, Kissling GE, Lieuallen W, Johansson SL, Malarkey DE, Hooth MJ. 2010. Urethral carcinoma and 

hyperplasia in male and female B6C3F1 mice treated with 3,3 ‘,4,4 ‘-tetrachloroazobenzene (TCAB). Toxicol Pathol 

38(3):372-381. 

Slikker W, Liu F, Zhang X, Xou X, Patterson T, Paule M, Wang C. 2010. Genomic response to perinatal 

anesthetic-induced neuronal cell death. Birth Defects Res A Clin Mol Teratol 88(5):379-379. 
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Smith J, Sammons D, Robertson S, Biagini R, Snawder J. 2010. Measurement of multiple drugs in urine, water, and 

on surfaces using fluorescence covalent microbead immunosorbent assay. Toxicol Mech Methods 20:587-593.

Smoak KA, Aloor JJ, Madenspacher J, Merrick BA, Collins JB, Zhu X, Cavigiolio G, Oda MN, Parks JS, Fessler MB. 

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Dopaminergic neurotoxicity following pulmonary exposure to manganese-containing welding fumes. 

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Sriram K, Lin GX, Jefferson AM, Roberts JR, Wirth O, Hayashi Y, Krajnak KM, Soukup JM, Ghio AJ, Reynolds SH, 

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Starlard-Davenport A, Tryndyak V, Kosyk O, Ross SR, Rusyn I, Beland FA, Pogribny IP. 2010. Dietary methyl 

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Starlard-Davenport A, Tryndyak VP, James SR, Karpf AR, Latendresse JR, Beland FA, Pogribny IP. 2010. Mechanisms 

of epigenetic silencing of the Rassf1a gene during estrogen-induced breast carcinogenesis in ACI rats. 

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Stefaniak AB, Harvey CJ, Bukowski VC, Leonard SS. 2010. Comparison of free radical generation by pre- and 

post-sintered cemented carbide particles. J Occup Environ Hyg 7(1):23-24. 

Swartz CD, Lentz CA, Green AS, Prajapati MH, Recio L, Witt KL. 2010. Mutagenicity of three herbal products, 

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Templeton SP, Buskirk AD, Green BJ, Beezhold DH, Schmechel D. 2010. Murine models of airway fungal exposure 

and allergic sensitization. Med Mycol 48(2):217-228. 

Teng CT. 2010. Lactoferrin: the path from protein to gene. Biometals 23(3):359-364. 

Thigpen J, Kissling G, Collins B, Setchell K, Adsit F, Caviness G, Locklear J, Whiteside T, Grant M, Forsythe D. 2010. 

The impact of rodent diets and bedding in studies evaluating the estrogenic activity of endocrine disruptor 

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Tokar EJ, Benbrahim-Tallaa L, Ward JM, Lunn R, Sams RL 2nd, Waalkes MP. 2010. Cancer in experimental animals 

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Tokar EJ, Diwan BA and Waalkes MP. 2010. Arsenic exposure transforms human epithelial stem/progenitor cells 

into a cancer stem-like phenotype. Environ Health Perspect 118(1):108-115.

Tokar EJ, Diwan BA, Waalkes MP. 2010. Arsenic exposure in utero and nonepidermal proliferative response in 

adulthood in Tg.AC mice. Int J Toxicol 29(3):291-296. 
106 National Toxicology Program

Tokar EJ, Diwan BA, Waalkes MP. 2010. Early life inorganic lead exposure induces testicular teratoma and renal 

and urinary bladder preneoplasia in adult metallothionein-knockout mice but not in wild type mice. Toxicology

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Tokar EJ, Qu W, Liu J, Liu W, Webber MM, Phang JM, Waalkes MP. 2010. Arsenic-specific stem cell selection during 

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Trembath D, Rao D, Morrison J, Malarkey D. 2010. Comparative pathology of medulloblastoma in F-344 rats and 

humans. J Neuropath Exp Neurol 69(5):30. 

Tryndyak VP, Beland FA, Pogribny IP. 2010. E-cadherin transcriptional down-regulation by epigenetic and 

microRNA-200 family alterations is related to mesenchymal and drug-resistant phenotypes in human breast 

cancer cells. Int J Cancer 126(11):2575-2583.

Tubbs JT, Blankenship-Paris T, Travlos G, King-Herbert AP, Kissling G, Clark J, Goulding D. 2010. Buprenorphine, 

meloxicam, and flunixin meglumine postoperative analgesia in mice. J Am Assoc Lab Anim Sci 49(5):692-692. 

Turpin EA, Antao-Menezes A, Cesta MF, Mangum JB, Wallace DG, Bermudez E, Bonner JC. 2010. Respiratory syncytial

virus infection reduces lung inflammation and fibrosis in mice exposed to vanadium pentoxide. Respir Res 11:20. 

Tvermoes BE, Boyd WA, Freedman JH. 2010. Molecular characterization of numr-1 and numr-2: genes that increase 

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Tyl RW, Seely JC, Marr MC, Myers CB, Youn VY, Browning DB, Bishop JB. 2010. Gonadal responses to vinclozolin 

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Wan YW, Qian Y, Rathnagiriswaran S, Castranova V, Guo NL. 2010. A breast cancer prognostic signature predicts 

clinical outcomes in multiple tumor types. Oncol Rep 24(2):489-494. 

Wan YW, Sabbagh E, Raese R, Qian Y, Luo DJ, Denvir J, Vallyathan V, Castranova V, Guo NL. 2010. Hybrid models 

identified a 12-gene signature for lung cancer prognosis and chemoresponse prediction. PLoS One 5(8):e12222 

Wang C, Zhang XA, Liu F, Paule MG, Slikker W. 2010. Anesthetic-induced oxidative stress and potential protection. 

Sci World J 10:1473-1482. 

Wang J, Rahman MF, Duhart HM, Newport GD, Patterson TA, Murdock RC, Hussain SM, Schlager JJ, Ali SF. 2009. 

Expression changes of dopaminergic system-related genes in PC12 cells induced by manganese, silver, or copper 

nanoparticles. Neurotoxicology 30(6):926-933. 

Wang L, Mercer RR, Rojanasakul Y, Qiu A, Lu Y, Scabilloni JF, Wu N, Castranova V. 2010. Direct fibrogenic effects of 

dispersed single-walled carbon nanotubes on human lung fibroblasts. J Toxicol Environ Health A 73(5):410-422. 

Wang RS, McDaniel LP, Manjanatha MG, Shelton SD, Doerge DR, Mei N. 2010. Mutagenicity of acrylamide and 

glycidamide in the testes of big blue mice. Toxicol Sci 117(1):72-80. 

Ward EM, Schulte PA, Straif K, Hopf NB, Caldwell JC, Carreón T, DeMarini DM, Fowler BA, Goldstein BD, Hemminki 

K, Hines CJ, Pursiainen KH, Kuempel E, Lewtas J, Lunn RM, Lynge E, McElvenny DM, Muhle H, Nakajima T, 

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Steenland K, Stevens RG, Vineis P, Zahm SH, Zeise L, Cogliano VJ. 2010. Research recommendations for selected 

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Waters MA, Grajewski B, Pinkerton LE, Hein MJ, Zivkovich Z. 2009. Development of historical exposure estimates 

of cosmic radiation and circadian rhythm disruption for cohort studies of Pan Am flight attendants. Am J Ind Med

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Whaley MJ, Rose C, Martinez J, Laher G, Sammons DL, Smith JP, Snawder JE, Borrow R, Biagini RE, Plikaytis B, 

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for measuring serum antibodies to pneumococcal polysaccharides. Clin Vaccine Immunol 17(5):862-869. 

Witt KL, Malarkey DE, Hobbs CA, Davis JP, Kissling GE, Caspary W, Travlos G, Recio L. 2010. No increases in 

biomarkers of genetic damage or pathological changes in heart and brain tissues in male rats administered 

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Wu N, Wang J, Tafen DN, Wang H, Zheng JG, Lewis JP, Liu X, Leonard SS, Manivannan A. 2010. Shape-enhanced 

photocatalytic activity of single-crystalline anatase TiO2 (101) Nanobelts. J Am Chem Soc 132(19):6679-6685. 

Yoshizawa K, Brix AE, Sells DM, Jokinen MP, Wyde M, Orzech DP, Kissling GE, Walker NJ, Nyska A. 2009. 

Reproductive lesions in female Harlan Sprague-Dawley rats following two-year oral treatment with dioxin and 

dioxin-like compounds. Toxicol Pathol 37(7):921-937. 

Yu L, Moore AB, Dixon D. 2010. Receptor tyrosine kinases and their hormonal regulation in uterine leiomyoma. 

Semin Reprod Med 28(3):250-259.

Yuan BZ, Chapman J, Reynolds SH. 2009. Proteasome inhibitors induce apoptosis in human lung cancer cells 

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Yucesoy B, Johnson VJ, Fluharty K, Kashon ML, Slaven JE, Wilson NW, Weissman DN, Biagini RE, Germolec DR, 

Luster MI. 2009. Influence of cytokine gene variations on immunization to childhood vaccines. Vaccine

27(50):6991-6997. 

Zegre Cannon C, Kissling G, J Hoenerhoff M, King-Herbert A, Blankenship-Paris T. 2010. Evaluation of dosages and 

routes of administration of tramadol analgesia in rats using hot-plate and tail-flick tests. Lab Anim 39:342-351. 

Zeidler-Erdely PC, Kashon ML, Li SQ, Antonini JM. 2010. Response of the mouse lung transcriptome to welding 

fume: effects of stainless and mild steel fumes on lung gene expression in A/J and C57BL/6J mice. Respir Res 11:70. 

Zhang D, Hu X, Qian L, O’Callaghan JP, Hong JS. 2010. Astrogliosis in CNS pathologies: Is there a role for microglia? 

Mol Neurobiol 41(2-3):232-241. 

Zhang XD, Zhao JS, Bowman L, Shi XG, Castranova V, Ding M. 2010. Tungsten carbide-cobalt particles activate Nrf2 

and its downstream target genes in JB6 cells possibly by ROS generation. J Environ Pathol Toxicol Oncol 29(1):31-40. 

Zhang YB, Ali SF, Dervishi E, Xu Y, Li ZR, Casciano D, Biris AS. 2010. Cytotoxicity effects of graphene and single-wall 

carbon nanotubes in neural phaeochromocytoma-derived PC12 Cells. ACS Nano 4(6):3181-3186. 

Zhu H, O’Brien JJ, O’Callaghan JP, Miller DB, Zhang Q, Rana M, Tsui T, Peng Y, Tomesch J, Hendrick JP, Wennogle LP, 

Snyder GL. 2010. Nerve agent exposure elicits site-specific changes in protein phosphorylation in mouse brain. 

Brain Res 1342(C):11-23
National Toxicology Program

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www.ntp.niehs.nih.gov NIH Publication No. 11-5972