NATIONAL TOXICOLOGY PROGRAM

Technical Report Series

No. 332

TOXICOLOGY AND CARCINOGENESIS

STUDIES OF

2-MERCAPTOBENZOTHIAZOLE

(CAS NO. 149-30-4)

IN F344/N RATS AND B6C3F1 MICE

(GAVAGE STUDIES)

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

Public Health Service
National Institutes of Health
NATIONAL TOXICOLOGY PROGRAM

The National Toxicology Program (NTP), established in 1978, develops and

evaluates scientific information about potentially toxic and hazardous
chemicals. This knowledge can be used for protecting the health of the
American people and for the primary prevention of disease. By bringing to-
gether the relevant programs, staff, and resources from the U S . Public
Health Service, DHHS, the National Toxicology Program has centralized
and strengthened activities relating to toxicology research, testing and test
developmenthalidation efforts, and the dissemination of toxicological in-
formation to the public and scientific communities and to the research and
regulatory agencies.

The NTP is made up of four charter DHHS agencies: the National Cancer
Institute (NCI), National Institutes of Health; the National Institute of En-
vironmental Health Sciences (NIEHS), National Institutes of Health; the
National Center for Toxicological Research (NCTR), Food and Drug Ad-
ministration; and the National Institute for Occupational Safety and
Health (NIOSH), Centers for Disease Control. In July 1981, the Carcino-
genesis Bioassay Testing Program, NCI, was transferred to the NIEHS.

2-Mercaptobenzothiazole,NTP TR 332
NTP TECHNICAL REPORT

ON THE

TOXICOLOGY AND CARCINOGENESIS

STUDIES OF 2-MERCAPTOBENZOTHIAZOLE

(CAS NO. 149-30-4)

IN F344/N RATS AND B6C3F1 MICE

(GAVAGE STUDIES)

Michael P. Dieter, Ph.D., Chemical Manager

NATIONAL TOXICOLOGY PROGRAM

P.O. Box S2233

Research Triangle Park, NC 27709

May 1988

NTP TR 332

NIH Publication No. 88-2588

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

Public Health Service

National Institutes of Health

NOTETOTHEREADER

This study was performed under the direction of the National Institute of Environmental Health Sci-
ences as a function of the National Toxicology Program. The studies described in this Technical Re-
port have been conducted in compliance with NTP chemical health and safety requirements and must
meet or exceed all applicable Federal, state, and local health and safety regulations. Animal care and
use were in accordance with the U.S.Public Health Service Policy on Humane Care and Use of Ani-
mals. All NTP toxicology and carcinogenesis studies are subjected to a data audit before being pre-
sented for public peer review.

Although every effort is made to prepare the Technical Reports as accurately as possible, mistakes
may occur. Readers are requested to identify any mistakes so that corrective action may be taken.
Further, anyone who is aware of related ongoing or published studies not mentioned in this report is
encouraged to make this information known to the NTP. Comments and questions about the
National Toxicology Program Technical Reports on Toxicology and Carcinogenesis Studies should be
directed to Dr. J.E. Huff, National Toxicology Program, P.O. Box 12233, Research Triangle Park, NC
27709 (919-541-3780).

These NTP Technical Reports are available for sale from the National Technical Information Service,
U.S.Department of Commerce, 5285 Port Royal Road, S p r i n ~ i e l dVA
, 22161 (703-487-4650). Single
copies of this Technical Report are available without charge (and while supplies last) from the NTP
Public Information Office, National Toxicology Program, P.O.Box 12233, Research Triangle Park,
NC 27709.

2-Mercaptobenzothiazole, NTP TR 332 2

CONTENTS

PAGE

NOTE TO THE READER ................................................................... 2

ABSTRACT ............................................................................... 5

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY ....................... 8

CONTRIBUTORS .......................................................................... 9

P E E R REVIEW PANEL ................................................................... io

SUMMARY OF PEER REVIEW COMMENTS .................................................. ii

I. INTRODUCTION ................................................................... 13

II. MATERIALS AND METHODS ........................................................ 19

PROCUREMENT AND CHARACTERIZATION OF 2-MERCAPTOBENZOTHIAZOLE ........20

PREPARATION AND CHARACTERIZATION OF DOSE MIXTURES ...................... 25

FIRST SIXTEEN-DAY STUDIES .................................................... 27

SECOND SIXTEEN-DAY STUDIES .................................................. 27

THIRTEEN-WEEK STUDIES ....................................................... 27

TWO-YEAR STUDIES ............................................................. 27

STUDY DESIGN ............................................................... 27

SOURCE AND SPECIFICATIONS OF ANIMALS .................................... 27

ANIMAL MAINTENANCE ....................................................... 30

CLINICAL EXAMINATIONS AND PATHOLOGY .................................... 30

STATISTICAL METHODS ....................................................... 31

III . RESULTS ......................................................................... 33

RATS .......................................................................... 34

SIXTEEN-DAY STUDIES ........................................................ 34

THIRTEEN-WEEK SI"NJIES ..................................................... 34

TWO-YEAR STUDIES .......................................................... 35

BODY WEIGHTS AND CLINICAL SIGNS ........................................ 35

SURVIVAL ................................................................. 38

PATHOLOGY AND STATISTICAL ANALYSES OF RESULTS ....................... 38

MICE .......................................................................... 46

SIXTEEN-DAY STUDIES ....................................................... -46

THIRTEEN-WEEK STUDIES .................................................... -47

TWO-YEAR STUDIES ........................................................... 48

BODY WEIGHTS AND CLINICAL SIGNS ........................................ 48

3 2.Mercaptobenzothiazole. NTP TR 332

CONTENTS (Continued)
PAGE

SURVIVAL . . . e. 51
PATHOLOGY AND STATISTICAL ANALYSES O F RESULTS , .....,.,.,........,..
.51

IV. DISCUSSION AND CONCLUSIONS ........,.......................................... .55

V. REFERENCES ................................................................... 0.59

APPENDIXES

APPENDIX A SUMMARY O F LESIONS IN MALE RATS IN THE TWO-YEAR GAVAGE

STUDY O F 2-MERCAPTOBENZOTHIAZOLE ..................................65
APPENDIX B SUMMARY O F LESIONS IN FEMALE RATS IN THE TWO-YEAR GAVAGE
STUDY O F 2-MERCAPTOBENZOTHIAZOLE ...,..............................89
APPENDIX C SUMMARY O F LESIONS IN MALE MICE IN THE TWO-YEAR GAVAGE
STUDY O F 2-MERCAPTOBENZOTHIAZOLE . .................................I11
APPENDIX D SUMMARY O F LESIONS IN FEMALE MICE IN THE TWO-YEAR GAVAGE
STUDY O F 2-MERCAPTOBENZOTHIAZOLE ................. . ....... .. .. I 3 1
APPENDIX E GENETIC TOXICOLOGY O F 2-MERCAPTOBENZOTHIAZOLE ,...........,....,. I @

APPENDIX F SENTINEL ANIMAL PROGRAM ,..........................................,161

APPENDIX G INGREDIENTS, NUTRIENT COMPOSITION, AND CONTAMINANT LEVELS IN

NIH 07 RAT AND MOUSE RAT'ION ........................................ .165

APPENDIX H AUDIT SUMMARY ...................................................... ,171

2-Mercaptobenzothiazole,NTP TR 332 4
2-MERCAPTOBENZOTHIAZOLE

CAS NO.149-30-4

C7H5NSZ Molecular weight 167.25

Synonyms and trade names: Captax, Dermacid, Mertax, Thiotax, 2(3H)-Benzothiazolethione,

2-Benzothiazolyl mercaptan

ABSTRACT

Toxicology and carcinogenesis studies of technical-grade 2-mercaptobenzothiazole (96%-97% pure), a

rubber accelerant and preservative, were conducted by administering the chemical by gavage in a
corn oil vehicle to groups of F344/N rats and B6C3F1 mice of each sex for 16 days, 13 weeks, or 2
years. 2-Mercaptobenzothiazole was nominated for study by the National Institute of Environmental
Health Sciences and the National Institute for Occupational Safety and Health.

Sixteen-Day and Thirteen-Week Studies: In 16-day studies, mean body weight gains of rats receiving
2,500 m g k g were 6-7 g lower than those of vehicle controls; 415 male and 515 female mice dosed with
3,000 mg/kg and 415 female mice dosed with 1,500 mg/kg died; lethargy and prostration occurred in
most of these animals after gavage. Based on these results, doses selected for both species in the 13-
week studies were 0,94 (mice only), 188,375,750, and 1,500 mg/kg.

In 13-week studies, no chemical-related deaths occurred in rats, but body weight gains in males dosed
with 1,500 mg/kg and in females dosed with 750 or 1,500 mg/kg were lower than those in the vehicle
control groups. Hepatomegaly occurred a t the two highest doses in males and at all doses in females;
however, no microscopic pathologic changes were noted in any tissue. More than half the mice dosed
with 1,500 mg/kg died, but no compound-related body weight changes occurred. Clinical signs in
mice were dose related and included lethargy in animals dosed with 375 mg/kg and lacrimation, sali-
vation, and clonic seizure in some dosed with 750 or 1,500 mg/kg. No association between these clini-
cal signs of toxicity and gross or microscopic pathologic effects was observed. Doses selected for the 2-
year studies were 0, 375, and 750 mg/kg for male rats and for mice of each sex and 0, 188, or 375
mg/kg for female rats.

Body Weight and Survival in the Two-year Studies: Fifty animals of each species and sex were ad-
ministered 2-mercaptobenzothiazole in corn oil by gavage 5 days per week for 103 weeks. Adminis-
tration of 2-mercaptobenzothiazole resulted in decreased survival in dosed male rats (vehicle control,
42/50; low dose, 22/50; high dose, 20/50) and in the high dose group of female mice (37150; 39/50;
22/50) but not in female rats (28/50; 31/50; 25/50) or in male mice (38150; 33/50; 30/50). No effect on
body weight gain in dosed rats was observed; in dosed mice, minor reductions occurred between weeks
3 and 64, with recovery thereafter. Postgavage lethargy and prostration occurred frequently in dosed
rats and mice.

Nonneoplastic and Neoplastic Effects in the Two-year Studies: The severity of nephropathy was in-
creased in dosed male rats. Ulcers and inflammation of the forestomach were prevalent in dosed rats,
as were increased incidences of epithelial hyperplasia and hyperkeratosis in male rats, but no neo-
plasms of the forestomach were observed. There were no increases of nonneoplastic lesions in mice
which were considered to be compound related.

5 2-Mercaptobenzothiazole,NTP TR 332
The incidences of a variety of tumors were increased in rats dosed with 2-mercaptobenzothiazole;
some of the increased incidences were not dose related. In low dose male rats, increased incidences
(P<O.Ol) were observed for mononuclear cell leukemia (7150; 16/50; 3/50) and pancreatic acinar cell
adenomas (2/50; 13/50; 6/49). Increased tumor incidences with dose-related trends (P< 0.05) included
pituitary gland adenomas in females (15/49; 24/50; 25/50), preputial gland adenomas or carcinomas
(combined) in males (160; 6/50; 5/50), adrenal gland pheochromocytomas or malignant pheochromo-
cytomas (combined) in males (18/50; 27/50; 24/49), and pheochromocytomas in females (1/50; 5/50;
6/50). These tumors were observed a t significantly greater incidences (PS0.05) in the high dose
groups than in the vehicle controls.

An increased incidence (P = 0.028) of hepatocellular adenomas or carcinomas (combined) was ob-

served only in low dose female mice (4/50; 12/49; 4/50). No significant increases in tumor incidences
were seen in male mice.

Genetic Toxicology: 2-Mercaptobenzothiazole was not mutagenic in Salmonella typhimurium strains

TA98, TA100, TA1535, or TA1537 with or without metabolic activation. In the presence of rat liver
S9,2-mercaptobenzothiazoleincreased the frequency of chromosomal aberrations and sister chroma-
tid exchanges (SCEs) in Chinese hamster ovary (CHO) cells, as well as mutations at the TK locus of
mouse L5178Y lymphoma cells.

Audit: The data, documents, and pathology materials from the 2-year studies of 2-mercaptobenzo-
thiazole were audited at the NTP Archives. The audit findings show that the conduct of the studies is
documented adequately and support the data and results given in this Technical Report.

Conclusions: Under the conditions of these 2-year gavage studies, there was some evidence of carcino-
genic activity* of 2-mercaptobenzothiazole for male F344/N rats, indicated by increased incidences of
mononuclear cell leukemia, pancreatic acinar cell adenomas, adrenal gland pheochromocytomas, and
preputial gland adenomas or carcinomas (combined). There was some evidence of carcinogenic activity
for female F344/N rats, indicated by increased incidences of adrenal gland pheochromocytomas and
pituitary gland adenomas. There was no evidence of carcinogenic activity of 2-mercaptobenzothiazole
for male B6C3F1 mice dosed with 375 or 750 mg/kg. There was equivocal evidence ofcarcinogenic ac-
tivity for female B6C3F1 mice, indicated by increased incidences of hepatocellular adenomas or carci-
nomas (combined).

*Explanation of Levels of Evidence of Carcinogenic Activity is on page 8.

A summary of the Peer Review comments and the public discussion on this Technical Report appears on pages 11-12.

2-Mercaptobenzothiazole, NTP TR 332 6

SUMMARY OF THE TWO-YEAR GAVAGE AND GENETIC TOXICOLOGY STUDIES O F
2-MERCAPTOBENZOTHIAZOLE

Male F344/N Rats Female F344/N Rats Male B6C3F1 Mice Female B6C3F1 Mice

Doses
375 or 750 mg/kg 2-mercapto- 188or 375 mg/kg 2-mercapto- 375 or 750 mg/kg 2-mercapto- 375 or 750 mg/kg
benzothiazole in corn oil, benzothiazole in corn oil, benzothiazole in corn oil, 2-mercaptobenzothiazole
5dhvk 5 d/wk 5 d/wk in corn oil, 5 d/wk

Survival rates in 2-year study

42/50; 22/50; 20150 28/50; 31/50; 25/50 38/50; 33/50; 30150 37/50; 39/50; 22/50
Nonneoplastic effects
Forestomach lesions; Forestomach lesions None None
nephropathy
Neoplastic effects
Mononuclear cell leukemia Adrenal gland pheochromo- None Hepatocellular adenomas
and pancreatic acinar cell cytomas and pituitary gland or carcinomas (combined)--
adenomas--low dose only; adenomas--trend and high dose low dose only
adrenal gland pheochromo-
cytomas and malignant
pheochromocytomas-trend
and high dose; preputial gland
adenomas or carcinomas
(combinedktrend and dosed

Level of evidence of carcinogenic activity

Some evidence Some evidence No evidence Equivocal evidence

Genetic toxicology
Not mutagenic in S. typhimurium strains TA98, TA100, TA1535, or TA1537 with or without metabolic activation; significant
increases in chromosomal aberrations and SCEs in CHO cells with S9; mutagenic at TK locus of mouse L5178Y lymphoma cells
with S9.

7 2-Mercaptobenzothiazole, NTP TR 332

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY
These studies are designed and conducted to characterize and evaluate the toxicologic potential, including carcinogenic activity, of
selected chemicals in laboratory animals (usually two species, rats and mice). Chemicals selected for NTP toxicology and carcinogenesis
studies are chosen primarily on the bases of human exposure, level of production, and chemical structure. Selection per se is not a n
indicator of a chemical’s carcinogenic potential.
Negative results, in which the study animals do not have a greater incidence of neoplasia than control animals, do not necessarily mean
that a chemical is not a carcinogen, inasmuch as the experimenta are conducted under a limited set of conditions. Positive results dem-
onstrate that a chemical is carcinogenic for laboratory animals under the conditions of the study and indicate that exposure to the
chemical has the potential for hazard to humans.
The National Toxicology Program describes the results of individual experiments on a chemical agent and notes the strength of the evi-
dence for conclusions regarding each study. Other organizations, such as the International Agency for Research on Cancer, assign a
strength of evidence for conclusions based on an examination of all available evidence including: animal studies such as those conducted
by the NTP, epidemiologic studies, and estimates of exposure. Thus, the actual determination of risk to humans from chemicals found to
be carcinogenic in laboratory animals requires a wider analysis that extends beyond the purview ofthese studies.
Five categories of evidence of carcinogenic activity are used in the Technical Report series to summarize the strength of the evidence ob-
served in each experiment: two categories for positive results (“Clear Evidence” and “Some Evidence”); one category for uncertain find-
ings (“Equivocal Evidence”); one category for no observable effects (“No Evidence”); and one category for experimenta that because of ma-
jor flaws cannot be evaluated (“Inadequate Study”). These categories of interpretative conclusions were first adopted in June 1983 and
then revised in March 1986for use in the Technical Reports series to incorporate more specifically the concept of actual weight of evidence
of carcinogenic activity. For each separate experiment (male rata, female rata, male mice, female mice), one ofthe following quintet is se-
lected to describe the findings. These categories refer to the strength of the experimental evidence and not to either potency or
mechanism.
0 Clear Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing a dose-related (i) in-
crease of malignant neoplasms, (ii) increase of a combination of malignant and benign neoplasms, or (iii) marked increase of be-
nign neoplasms if there is an indication from this or other studies of the ability of such tumors to progress to malignancy.
0 Some Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing a chemically related in-
creased incidence of neoplasms (malignant, benign, or combined) in which the strength of the response is less than that required
for clear evidence.
Equivocal Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing a marginal in-
crease of neoplasms that may be chemically related.
0 No Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing no chemically related in-
creases in malignant or benign neoplasms.
Inadequate Study of Carcinogenic Activity is demonstrated by studies that because of major qualitative or quantitative
limitations cannot be interpreted as valid for showing either the presence or absence of carcinogenic activity.
When a conclusion statement for a particular experiment is selected, consideration must be given to key factors that would extend the ac-
tual boundary of an individual category of evidence. This should allow for incorporation of scientific experience and current under-
standing of long-term carcinogenesis studies in laboratory animals, especially for those evaluations that may be on the borderline be-
tween two adjacent levels. These considerations should include:
0 The adequacy of the experimental design and conduct;
0 Occurrence of common versus uncommon neoplasia;
0 Progression (or lack thereon from benign to malignant neoplasia as well as from preneoplastic lesions;
0 Some benign neoplasms have the capacity to regress but others (of the same morphologic type) progress. At present, it is im-
possible to identlfy the difference. Therefore, where progression is known to be a possibility, the most prudent course is to assume
that benign neoplasms ofthose types have the potential to become malignant;
Combining benign and malignant tumor incidences known or thought to represent stages of progression in the same organ or
tissue;

0 Latency in tumor induction;

0 Multiplicity in site-specXc neoplasia;

Metastases;

Supporting information from proliferative lesions (hyperplasia) in the same site of neoplasia or in other experiments (same lesion

in another sex or species);

0 The presence or absence ofdose relationships;

The statistical significance ofthe observed tumor increase;

The concurrent control tumor incidence as well as the historical control rate and variability for a specific neoplasm;

0 Survival-adjusted analyses and false positive or false negative concerns;

0 Structure-activity correlations; and

0 In some cases,genetic toxicology.

These considerations together with the definitions as written should be used as composite guidelines for selecting one of the five cate-
gories. Additionally, the following concepts (as patterned from the International Agency for Research on Cancer Monographs) have been
adopted by the NTPto give further clarification of these issues:
The term chemical carcinogenesis generally means the induction by chemicals of neoplasms not usually observed, the induction by
chemicals of more neoplasms than are generally found, or the earlier induction by chemicals of neoplasms that are commonly ob-
served. Different mechanisms may be involved in these situations. Etymologically, the term carcinogenesis means induction of
cancer, that is, of malignant neoplasms; however, the commonly accepted meaning is the induction of various types of neoplasms or
of a combination of malignant and benign neoplasms. In the Technical Reports, the words tumor and neoplasm a r e used
interchangeably.

2-Mercaptobenzothiazole, NTP TR 332 8

CONTRIBUTORS

The NTP Technical Report on the Toxicology and Carcinogenesis Studies of 2-Mercaptobenzothiazole
is based on the 13-week studies that began in November 1980 (rats) or August 1980 (mice) and ended
in February 1981 (rats) or November 1980 (mice) and on the 2-year studies that began in July 1981
and ended in July 1983 at Physiological Research Laboratories (Minneapolis, Minnesota).
National Toxicology P r o g r a m

(Evaluated Experiment, Interpreted Results, a n d Reported Findings)

Michael P. Dieter, Ph.D., Chemical Manager

John Bucher, Ph.D. James Huff, Ph.D.
ScotL.Eustis,D.V.M.,Ph.D.
(Discipline Leaders and Principal Contributors)
Jack Bishop, Ph.D. Walter W. Piegorsch, Ph.D.
C.W. Jameson, Ph.D. G.N. Rao, D.V.M.,Ph.D.
E.E. McConnell, D.V.M. B.A. Schwetz, D.V.M., Ph.D.
John Mennear, Ph.D. James K. Selkirk, Ph.D.
N T P Pathology Working G r o u p

(Evaluated Slides and Prepared Pathology Report for R a t s on 4/8/86)

Steven Stefanski, D.V.M.,M.S. (Chair) (NTP) James Maclachlan, B.V.Sc., Ph.D.

Michael Elwell, D.V.M.,Ph.D. (NTP) North Carolina State University

Scot L. Eustis, D.V.M.,Ph.D. (NTP) Kunitoshi Mitsumori, D.V.M., Ph.D. (NTP)

Kiyoshi Imai, D.V.M., Ph.D. Linda Uraih, D.V.M. (NTP)

Hatano Research Institute

(Evaluated Slides and Prepared Pathology Report for Mice o n 4/10/86)
Linda Uraih, D.V.M. (Chair) (NTP) Kiyoshi Imai, D.V.M., Ph.D.
Ken Ayers, D.V.M. Hatano Research Institute
Burroughs Wellcome Laboratories Kunitoshi Mitsumori, D.V.M., Ph.D. (NTP)
Michael Elwell, D.V.M., Ph.D. (NTP) Steven Stefanski, D.V.M. (NTP)
Scot L. Eustis, D.V.M., Ph.D. (NTP)
Principal Contributors at Physiological Research Laboratories
(Conducted Studies a n d Evaluated Tissues)
D. EIsberry, Ph.D. M. Cowan, M.S.
A. Hall, D.V.M. J. Sagartz, D.V.M.
Principal Contributors at Experimental Pathology Laboratories, Inc.
(Provided Pathology Quality Assurance)
J. Gauchat Jerry Hardisty, D.V.M.

Roger Brown, D.V.M.

Principal Contributors at Carltech Associates, Inc.

(Contractor for Technical Report Preparation)
William D. Theriault, Ph.D. John Warner, M.S.
Abigail C. Jacobs, Ph.D.

9 2-Mercaptobenzothiazole, NTP TR 332

PEER REVIEW PANEL

The members of the Peer Review Panel who evaluated the draft Technical Report on 2-mercaptoben-
zothiazole on March 4,1987, are listed below. Panel members serve as independent scientists, not as
representatives of any institution, company, or governmental agency. In this capacity, Panel mem-
bers have five major responsibilities: (a)to ascertain that all relevant literature data have been ade-
quately cited and interpreted, (b) to determine if the design and conditions of the NTP studies were
appropriate, (c) to ensure that the Technical Report presents the experimental results and conclu-
sions fully and clearly, (d) to judge the significance of the experimental results by scientific criteria,
and (e) to assess the evaluation of the evidence of carcinogenicity and other observed toxic responses.
National Toxicology Program Board of Scientific Counselors

Technical Reports Review Subcommittee

Robert A. Scala, Ph.D. (Chair)

Senior Scientific Advisor, Medicine and Environmental Health Department

Research and Environmental Health Division, Exxon Corporation

East Millstone, New Jersey

Michael A. Gallo, Ph.D. Frederica Perera, Dr. P.H.*

Associate Professor, Director of Toxicology Division of Environmental Sciences
Department of Environmental and Community School of Public Health
Medicine, UMDNJ - Rutgers Medical School Columbia University
Piscataway, New Jersey New York, New York
Ad Hoc Subcommittee Panel of Experts
Charles C. Capen, D.V.M., Ph.D. Franklin E. Mirer, Ph.D.*
Department of Veterinary Pathobiology Director, Health and Safety Department
Ohio State University International Union, United Auto
Columbus, Ohio Workers, Detroit, Michigan
Vernon M. Chinchilli, Ph.D. (Principal Reviewer) James A. Popp, D.V.M., Ph.D. (Principal
Department of Biostatistics Reviewer) Head, Department of
Medical College of Virginia Experimental Pathology and Toxicology
Virginia Commonwealth University Chemical Industry Institute of Toxicology
Richmond, Virginia Research Triangle Park, North Carolina
John J. Crowley, Ph.D.* I.F.H. Purchase,B.V.Sc.,Ph.D.,F.R.C. Path.*
Division of Public Health Science Director, Central Toxicology Laboratory
The Fred Hutchinson Cancer Research Center Imperial Chemical Industries, PLC
Seattle, Washington Alderley Park, England
Kim Hooper, Ph.D. (Principal Reviewer) Andrew Sivak, Ph.D.
Hazard Evaluation System and Vice President, Biomedical Science
Information Services
Arthur D. Little, Inc.
Department of Health Services
Cambridge, Massachusetts
State of California

Berkeley, California

Donald H. Hughes, Ph.D.

Scientific Coordinator, Regulatory Services
Division, The Procter and Gamble Company
Cincinnati, Ohio

+Unableto attend meeting

2-Mercaptobenzothiazole,NTP TR 332 10
SUMMARY OF PEER REVIEW COMMENTS

ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF

2-MERCAPTOBENZOTHIAZOLE

On March 4,1987, the draft Technical Report on the toxicology and carcinogenesis studies of 2-mer-
captobenzothiazole received peer review by the National Toxicology Program Board of Scientific
Counselors’ Technical Reports Review Subcommittee and associated Panel of Experts. The review
meeting was held at the National Institute of Environmental Health Sciences, Research Triangle
Park, North Carolina.

Dr. M. Dieter, NTP, began the discussion by reviewing the experimental design, results, and proposed
conclusions (some evidence of carcinogenic activity for male and female rats, no evidence of carcino-
genic activity for male mice, equivocal evidence of carcinogenic activity for female mice).

Dr. Hooper, a principal reviewer, agreed with the conclusions as written. However, he argued that in-
creased incidences of preputial gland adenomas or carcinomas (combined) should be included in sup-
port of the conclusion for male rats. Dr. Dieter agreed that it was valid to include the preputial gland
tumors along with the mononuclear cell leukemia as some evidence of carcinogenic activity and that
the conclusion and other appropriate sections of the Technical Report could be revised to reflect this
change. Dr. S. Eustis, NIEHS, commented that this tumor was not originally included in the list of
evidence because although the incidence of preputial gland tumors in this study was twice the histori-
cal mean, the incidence also fell within the historical range. Dr. Hooper noted the lack of tumors in
high dose male rats compared with a n elevated tumor incidence in low dose male rats for several neo-
plasms, including mononuclear cell leukemia. Dr. Dieter said that there was just one other tumor be-
sides mononuclear cell leukemia, pancreatic acinar cell adenomas in male rats, for which there was
an effect only a t the low dose. Dose-related increases occurred in two tumor types, including adrenal
gland tumors in male and female rats and pituitary gland tumors in female rats.

As a second principal reviewer, Dr. Popp agreed in principle with the conclusions. He said that the
issue for decision was whether the conclusions for rats should remain as written or be lowered to
equivocal evidence of carcinogenic activity.

As a third principal reviewer, Dr. Chinchilli agreed with the conclusions as written. He asked that an
incidence table for mononuclear cell leukemia in female rats be added to the Results section.

Dr. Harold Grice, Cantox, Inc. Canada, representing the Chemical Manufacturers Association, men-
tioned several factors that he felt made interpretation of the increased tumor rates in male rats diffi-
cult. These factors included reduced survival in both dose groups, compound-induced kidney toxicity,
gavage stress, and postgavage lethargy. Dr. Grice thought that the conclusion for male rats should be
lowered to equivocal evidence of carcinogenic activity.

Since the low dose animals were placed in the cage racks nearest the room fluorescent lights and
because cages were not rotated in these studies, there was speculation as to whether photoactivation
of the chemical might have been a factor in toxicitylcarcinogenicity. Although the incidence of eye
lesions (retinopathy and cataracts) could be correlated with cage position, there was no consensus
that increased tumor rates in low dose rats could be associated with exposure to light.

11 2-Mercaptobenzothiazole, NTP TR 332

SUMMARY OF PEER REVIEW COMMENTS (Continued)

In other discussion, Dr. Hooper thought that the small but significant increase in renal neoplasms in
male rats (tubular cell adenomas and transitional cell papillomas/carcinomas) might have been
chemically associated. Dr. Eustis said that the renal tumors were not considered chemically related
because the two cell types are generally not combined and the tumors were split between dose groups.

Dr. Hooper moved that the Technical Report on 2-mercaptobenzothiazole be accepted with the revi-
sions discussed and the conclusions as written for male and female rats, some evidence of carcinogenic
activity, for male mice, no evidence of carcinogenic activity, and for female mice, equivocal evidence
of carcinogenic activity. He asked that the increased incidences of preputial gland adenomas or car-
cinomas (combined) in male rats be cited. Dr. Gallo seconded the motion, which was approved unani-
mously with seven votes.

2-Mercaptobenzothiazole,NTP TR 332 12
I. INTRODUCTION

Production, Use, and Exposure

Acute Toxicity
Dermal Toxicity
Reproductive Toxicity
Biochemical Effects
Absorption, Distribution, and Metabolism
Genetic Toxicology
Carcinogenicity
Study Rationale

13 2-Mercaptobenzothiazole,NTP TR 332
I. INTRODUCTION

2-MERCAPTOBENZOTHIAZOLE

CAS No. 149-30-4

C7H5NS2 Molecular weight 167.25

Synonyms and trade names: Captax, Dermacid, Mertax, Thiotax, 2(3H)-Benzothiazolethione,

2-Benzothiazolyl mercaptan

2-Mercaptobenzothiazole forms pale, yellow, as 2-mercaptobenzothiazole (Stinson, 1983). 2-

nionoclinic needles or leaflets with a dis- Mercaptobenzothiazole, however, serves as a n
agreeable odor; it has a melting point of 180.2"- intermediate for other sulfenamide derivatives
181.7" C and a specific gravity of 1.42. The (Santadonato et al., 19761, so these production
chemical is insoluble in water but soluble in al- figures may be underestimated. 2-Mercaptoben-
cohol, acetone, benzene, and chloroform (Haw- zothiazole is used commercially as an accelerant
ley, 1981). The octano1:water partition coef- in the rubber vulcanization process and as a pre-
ficient is 41:l (Hansch and Leo, 1979). 2-Mer- servative for textile or cordage materials; the so-
captobenzothiazole exists in the thioketo form in dium salt is used as a corrosion inhibitor in pe-
the solid crystalline state but converts to the thi- troleum products.
oenol form upon reaction with metals (Santo-
donato et al., 1976). 2-Mercaptobenzothiazole is 2-Mercaptobenzothiazole was found to contami-
a weak acid and will form salts in basic solutions nate medicinal products that came in contact
with a wide variety of metal ions. In acid solu- with rubber stoppers made with this accelerator
tions in the presence of iron, 2-mercaptobenzo- (Petersen et al., 1981) and was found in aqueous
thiazole is reduced to benzothiazole, whereas in extracts of rubber baby bottle nipples (Blosczyk
the presence of ozone and potassium iodide, it and Doemling, 1982). Since manufacturing
dimerizes to 2-mercaptobenzothiazole disulfide. processes occur in closed, continuous systems
(Santadonato et al., 1976), employee exposure to
Production, Use, and Exposure 2-mercaptobenzothiazole would probably occur
through dermal contact or inhalation of dust
2-Mercaptobenzothiazole is produced by reacting during packaging, transport, or the use of rubber
aniline, carbon disulfide, and sulfur at elevated products. Consumer exposure occurs through di-
temperature and pressure; generally, the prod- rect contact with stretch garments (Bauer,
uct is then purified by dissolving it in a base to 1972), shoes (Fisher, 1977), rubber pharmaceu-
remove the dissolved organics. Reprecipitation tical products (Petersen et al., 19811, and baby
is accomplished by the addition of acid (Kirk- bottle nipples containing 2-mercaptobenzothia-
Othmer, 1982). zole (Blosczyk and Doemling, 1982).
Acute Toxicity
2-Mercaptobenzothiazole is produced in the
United States by two major tire companies The reported oral LD50 values in mice and rats
(Goodyear and Uniroyal) and by Monsanto Com- range between 2,000 a n d 3,000 mg/kg
pany. Production in the United States was (Vorob'eva and Mezentsera, 1968; Vanderbilt,
6,531,000 pounds in 1984 (USITC, 19851, and 1975; Guess and O'Leary, 1969; Monsanto, 1982;
198,414 pounds was imported in 1981 (USITC, Uniroyal, 19751, and intraperitoneal LD50 val-
1983). The use of smaller tires on cars and ues range between 100 and 400 mg/kg in mice
trends toward reduction in length of automobile (Guess and O'Leary, 1969; Doull et al., 1962). 2-
trips may result in a decline in future needs for Mercaptobenzothiazole (110 or 300 mg/kg ad-
production of rubber-processing chemicals such ministered by intraperitoneal injection) was

2-Mercaptobenzothiazole, NTP TR 332 14

I. INTRODUCTION

shown to exert neurotoxic and hepatotoxic ef- biosynthesis, was inhibited 40% below control
fects in mice after acute or short-term exposure values in brain tissue taken from mice 1 hour
(Johnson et al., 1970; Guess and O’Leary, 1969). after a 200 mg/kg intraperitoneal injection of 2-
mercaptobenzothiazole. In the same tissues
Dermal Toxicity used for in vitro studies, there was 47% inhibi-
tion after less than 7 pM 2-mercaptobenzothia-
2-Mercaptobenzothiazole was shown to be a very zole was added to t h e reaction m i x t u r e .
strong contact allergen in guinea pigs (Maurer Grassetti et al. (1970) showed that 1 mM 2-mer-
et al., 1979) but was judged a moderate contact captobenzothiazole added in vitro affected carbo-
sensitizer in humans (Goodwin et al., 1981). hydrate metabolism in Ehrlich ascites tumor
Rubber additives, such as salts of 2-mercapto- cells, causing a slight inhibition of the hexose
benzothiazole, have been reported to cause monophosphate shunt pathway and a moderate
dermatitis in humans (Bauer, 1972). 2-Mercap- stimulation of the tricarboxylic acid cycle. An
tobenzothiazole was more soluble in a salt solu- intraperitoneal injection of 100 mg/kg 2-mer-
tion approximating human perspiration than in captobenzothiazole lowered blood glucose con-
water (Ito et al., 1979). The sensitizing proper- centrations in rabbits 5 hours after administra-
ties of 2-mercaptobenzothiazole were reviewed tion (Chiba, 1969).
by Fisher (1973) and Santodonato et al. (19761,
who noted that allergic contact dermatitis in hu- Absorption, Distribution, a n d Metabolism
mans is often caused by rubber products.
Absorption, tissue distribution, and metabolism
Reproductive Toxicity studies of radiolabeled 2-mercaptobenzothiazole
in guinea pigs showed that the chemical was ab-
Embryotoxic effects of 2-mercaptobenzothiazole sorbed through the skin and that abrasion in-
in rats were reported (Aleksandrov, 19821, but creased this rate; initially, the kidney, liver, and
these results were not corroborated in more ex- thyroid gland were the principal organs of up-
tensive studies in rats administered 200 mg/kg take, with the thyroid gland ultimately attain-
2-mercaptobenzothiazole by intraperitoneal in- ing the highest concentration of 2-mercapto-
jection on days 1-15 of gestation (Hardin et al., benzothiazole 48 hours after subcutaneous
1981). There were no chemically related histo- injection; 90% of the compound was conjugated
pathologic effects in maternal tissues, and no with glucuronides and sulfates and excreted in
maternal toxicity, fetal toxicity, or teratogenesis the urine 6 hours after injection (Nagamatsu et
was observed. In a long-term study, no cumula- al., 1979). The urinary metabolites of [W-mer-
tive effects on reproduction or lactation were ob- captol2-mercaptobenzothiazole in rats dosed by
served in rats fed ad libitum 5,000 ppm of a for- intraperitoneal injection consisted of conjugates
mulation containing 2.4% 2-mercaptobenzothia- of glutathione, glucuronic acid, and inorganic
zole and 27.6% dimethyldithiocarbamate sulfate (Colucci and Buyske, 1965); these au-
through the second generation (Lehman, 1965). thors proposed three possible metabolic path-
ways for 2-mercaptobenzothiazole which started
Biochemical Effects with a benzothiazole-2-glutathionemetabolite
and proceeded either through benzothiazole-2-
Biochemical studies suggested that 2-mercapto- cysteine to benzothiazole-2-mercapturic acid
benzothiazole was capable of enzyme inhibition that was eliminated in the urine, or to benzo-
in vivo and in vitro (Johnson et al., 1970; thiazole-2-mercaptan that then was eliminated
Grassetti et al., 1970). Dopamine j3-hydroxylase, in the urine as either benzothiazole-2-mercapto-
an enzyme in the pathway for norepinephrine glucuronide or as inorganic sulfate.

15 2-Mercaptobenzothiazole, NTP TR 332

I. INTRODUCTION

Genetic Toxicology In NTP cytogenetic assays, significant increases

in chromosomal aberrations and sister chroma-
2-Mercaptobenzothiazole demonstrated no mu- tid exchanges (SCEs) were observed in cultured
tagenic activity in bacteria, but it is clearly clas- Chinese hamster ovary (CHO) cells after expo-
togenic as well as genotoxic to mammalian cells sure to 2-mercaptobenzothiazole a t 351-451
in culture. Donner et al. (1983) found no in- pg/ml in the presence of Aroclor 1254-induced
crease in Salmonella typhimurium his' revert- male Sprague Dawley rat liver S9; no significant
ant colonies after exposure to 2-mercaptobenzo- induction of chromosomal aberrations or SCEs
thiazole; a n early study by Szybalski (1958) was observed without S9 (Tables E3 and E4).
showed no induction of mutations in Escherichia Although the in vitro cytogenetic data indicate
coli strain SD-4-73 after exposure to 2-mercapto- that the chemical is a clastogen, intraperitoneal
benzothiazole. Neither the doses used nor the injection of 300 mg/kg 2-mercaptobenzothiazole
source and purity of the 2-mercaptobenzothia- dissolved in corn oil did not produce a significant
zole were provided by the authors. Two labora- increase in the number of micronucleated poly-
tories investigated the mutagenicity of 2-mer- chromatic erythrocytes in the bone marrow of
captobenzothiazole for NTP in the S. typhimuri- CD-1 mice (Pharmakon, 1984).
umlmicrosome assay with a preincubation proto-
col with strains TA98, TA100, TA1535, and One published study presents data from a series
TA1537 with and without Aroclor 1254-induced of short-term tests designed to evaluate the
male Sprague Dawley r a t or Syrian hamster genotoxic activity of four rubber accelerators, in-
liver S9. In the study conducted a t EG&G Ma- cluding 2-mercaptobenzothiazole disulfide, a
son Research Institute, questionable mutagenic structural analog resulting from the dimeriza-
activity was noted only in strain TA98 in the tion of 2-mercaptobenzothiazole (Hinderer et al.,
presence, but not in the absence, of S9 from 1983). Results showed that 2-mercaptobenzo-
either species. The Case Western Reserve Uni- thiazole disulfide (80% pure and containing 30
versity study detected no mutagenic activity in ppm morpholine, a nonmutagen in NTP Sal-
any of the four strains of S. typhimurium tested monella studies; Haworth et al., 1983) did not in-
under any conditions (Zeiger et al., 1987; Appen- duce gene reversion in Salmonella and E. coli
dix E, Table El). WP2 uvrA- with or without metabolic activa-
tion, was negative in the BALB/3T3 transforma-
Exposure of V79 cells to doses of 50-300 pg/ml2- tion assay in the absence of S9, and did not in-
mercaptobenzothiazole for 4 hours resulted in no duce chromosomal aberrations in cultured CHO
increase in 6-thioguanine resistant mutants cells with or without S9. The maximum concen-
(Donner et al., 1983). Results from a CHO/ tration of 2-mercaptobenzothiazole disulfide
HGPRT forward mutation assay conducted both tested in the chromosomal aberration assay was
with and without exogenous metabolic activa- 10.0 pg/ml, whereas the NTP cytogenetic tests
tion, at 2-mercaptobenzothiazole doses of up to used 2-mercaptobenzothiazole a t concentrations
300 pg/ml, were negative (Pharmakon, 1984). in excess of 350 pg/ml. Exposure of mouse
The results of a mouse lymphoma forward muta- L5178Y lymphoma cells t o 2-mercaptobenzo-
tion assay showed mutagenic activity for 2-mer- thiazole disulfide in the absence of exogenous
captobenzothiazole a t the highest doses tested metabolic activation resulted in no increase in
(100 and 150 pg/ml) in the absence of exogenous forward mutations at the TK+'- locus; in the
metabolic activation with concomitant extreme presence of activation, the two highest doses (15
toxicity (Litton, 1985). With S9 activation, tox- and 30 pg/ml) did produce a significant increase
icity was reduced, and a significant increase in over background rates in the number of mutant
mutations was again noted a t the highest doses colonies.
tested (80 and 100 pg/ml). 2-Mercaptobenzo- Carcinogenicity
thiazole induced forward mutations in mouse
L5178Y lymphoma cells only in the presence of 2-Mercaptobenzothiazole did not cause increased
Aroclor 1254-induced male F344 rat liver S9 tumor incidences in two hybrid mouse strains
(Table E2). (C57BL/6 X C3H/Anf and C57BL/6 X AKR)

2-Mercaptobenzothiazole, NTP TR 332 16

I. INTRODUCTION

after 18 months of chemical administration Study Rationale

(Innes et al., 1969). The F1 generation of hybrids
was administered 100 mg/kg 2-mercaptobenzo- 2-Mercaptobenzothiazole was nominated for
thiazole in 0.5% gelatin by gavage from 7 to 28 study by the National Institute of Environ-
days of age and then was fed 323 ppm 2-mercap- mental Health Sciences and the National Insti-
tobenzothiazole ad libitum for the remainder of tute for Occupational Safety and Health because
the study. There were 18 mice of each sex and of potential widespread human exposure and to
strain per dose group and four untreated control determine structure-activity relationships with
groups containing 12-18 mice of each sex and other sulfur-containing compounds. Since the
strain. Lehman (1965) also reported no increase salts of 2-mercaptobenzothiazole are hydrolyzed
in tumor incidence in 10 rats (unspecified strain) to the parent compound in vivo and these salts
of each sex fed a mixture of 5,000 ppm of a for- a r e marketed as fungicides and bactericides
mulation containing 2.4% 2-mercaptobenzothia- (Foltinova and Bloeckinger, 1970), the genotoxic
zole and 27.6% dimethyldithiocarbamate (a di- effects of noncytocidal concentrations of 2-mer-
etary 2-mercaptobenzothiazole concentration of captobenzothiazole were also examined.
120 ppm) for 2 years.

17 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole,NTP TR 332 18
II. MATERIALS AND METHODS

PROCUREMENT AND CHARACTERIZATION OF

2-MERCAPTOBENZOTHIAZOLE
PREPARATION AND CHARACTERIZATION OF
DOSE MIXTURES
FIRST SIXTEEN-DAY STUDIES
SECOND SIXTEEN-DAY STUDIES
THIRTEEN-WEEK STUDIES
TWO-YEAR STUDIES
Study Design
Source and Specifications of Animals
Animal Maintenance
Clinical Examinations and Pathology
Statistical Methods

19 2-Mercaptobenzothiazole, NTP TR 332

II. MATERIALS AND METHODS

PROCUREMENT AND plates and a ch1oroform:methanol (96:4) solvent

CHARACTERIZATION OF system; a major spot, a minor spot, a trace im-
2-MERCAPTOBENZOTHIAZOLE purity, and a slight trace impurity in lot no. 39-
7-D were detected by ultraviolet (254 and 366
2-Mercaptobenzothiazole (Captax) was obtained nm) light and a n iodoplatinate spray. Thin-
i n two lots from R.T. Vanderbilt Co., Inc. layer chromatography with a hexanes:diethyl-
(Norwalk, Connecticut) (Table 1). Purity and ether (40:60) solvent system detected a major
identity analyses of both lots were conducted a t spot, three trace impurities, and one slight trace
Midwest Research Institute (MRI) (Kansas City, impurity in lot no. V10479 and a major spot, a
Missouri). MRI reports on the analyses per- trace impurity, and a slight trace impurity in lot
formed in support of the 2-mercaptobenzothia- no. 39-7-D. High-performance liquid chromatog-
zole studies are on file at NIEHS. Chemical raphy on a pBondapak C l 8 column with a
identity was confirmed by infrared, ultraviolet/ water/l% acetic acid:acetonitrile/l% acetic acid
visible, and nuclear magnetic resonance spec- (49:51) mobile phase at a flow rate of 1 ml/min-
troscopy (Figures 1 to 4). Ute and detection a t 313 nm indicated six im-
Lot no. V10479 was obtained as a light green purities with peak areas greater than 0.1% that
powder with a melting point of 175"-178" C;lot of the major peak and a relative combined area
no. 39-7-D was obtained as a light green-yellow of 2.2% (lot no. V10479) and five impurities with
powder. The purity of the two lots was deter- peak areas greater than 0.1% and a relative
mined by elemental analysis, water analysis, combined area of 1.7% (lot no. 39-7-D).
nonaqueous titration of the sulfhydryl group,
thin-layer chromatography, and high-perfor- Stability studies performed by the same high-
mance liquid chromatography. Cumulative data performance liquid chromatographic system
indicated that lot no. V10479 was approximately with a 5050 solvent ratio a t a flow rate of 1.5
96% pure and lot no. 39-7-D was approximately ml/minute and detection a t 254 nm indicated
97% pure. The water content of lot no. V10479 that 2-mercaptobenzothiazole was stable on stor-
by Karl Fischer titration was 1.358, and that of age for 2 weeks at 60" C. Further confirmation
lot no. 39-7-D was 0.25%. Titration of the sulf- of the stability of the bulk chemical during the
hydryl group with 0.1 N tetrabutylammonium toxicity studies (storage at 25" C) was obtained
hydroxide indicated that lot no. V10479 was by titration with 0.1 N tetrabutylammonium
96.3% pure and lot no. 39-7-D, 96.8% pure. A hydroxide and the same high-performance liquid
major spot, three trace impurities, and one slight chromatographic system that was used for the
trace impurity in lot no. V10479 were detected stability studies. No degradation was seen over
by thin-layer chromatography with silica gel the course of the studies.

TABLE 1. IDENTITY AND SOURCE OF 2-MERCAPTOBENZOTHIAZOLEUSED IN THE GAVAGE

STUDIES
~~~~ ~

First Sixteen-Day Second Sixteen-Day Thirteen-Week Two-year

Studies Studies Studies Studies

Lot Numbers Used

V 10479 V10479 V10479 V10479,39-7-D

Date of Initial Use

2111/80 4/28/80 Rats--11/17/80; V10479--7/14/81 (rats),
mice--8/18/80 7/28/81 (mice);
39-7-D--1/21/83

Supplier
R.T. Vanderbilt Co., Same as first 16-d studies Same RS first 16-d studies Same as first 16-d studies
Inc. (Norwalk,CT)

2-Mercaptobenzothiazole,NTP TR 332 20
0
z

c4

0
21 2-Mercaptobenzothiazole, NTP TR 332
0
i
2

s
c!
i
0
0
I
i-
W

u
z
c

2-Mercaptobenzothiazole, NTP TR 332 22

23 2-Mercaptobenzothiazole, NTP' TR 332
Y

E
3

p:
E-c
W

z
It +

2-Mercaptobenzothiazole,NTP TR 332 24
II. MATERIALS AND METHODS

PREPARATION AND 5" C. Samples exposed to air and light for 3

CHARACTERIZATION OF hours at room temperature also showed no loss
DOSE MIXTURES of 2-mercaptobenzothiazole.
Weighed amounts of 2-mercaptobenzothiazole
and corn oil were mixed as described in Table 2. Analyses for 2-mercaptobenzothiazole in dose
Stability studies of dose mixtures were per- mixtures were performed by the study and ana-
formed with high-performance liquid chroma- lytical chemistry laboratories by extracting
tography on a yBondapak C l 8 column with a samples with methanol and determining the ab-
wateracetonitrile (65:35) mobile phase at a flow sorption a t 320 nm (study laboratory) or 322 nm
rate of 1 ml/minute and ultraviolet detection at (analytical chemistry laboratory). Dose mix-
313 nm after extraction with methanol; the stud- tures were analyzed three times during the 13-
ies indicated that 2-mercaptobenzothiazole (20 week studies; concentrations of 2-mercapto-
mg/ml) in corn oil was stable for a t least 14 days benzothiazole ranged from 91% to 109% of the
when stored in the dark a t room temperature or target concentration (Table 3).

TABLE 2. PREPARATION AND STORAGE OF DOSE MIXTURES IN THE GAVAGE STUDIES O F

2-MERCAPTOBENZOTHIAZOLE

First Sixteen-Day
Second Sixteen-Day
Thirteen-Week
Two-year
Studies
Studies
Studies
Studies

Preparation
A Polytron@homogenizer operated Same as first 16-d studies Similar to first 16-d studies Same a s first 16-d
a t low intensity for 2 min was used studies
to suspend 2-mercaptobenzothiazole
in corn oil

Maximum Storage Time

14 d 14d 14 d 14 d

Storage Conditions
Room temperature in the dark Same as first 16-d studies Same as first 16-d studies 25'C in the dark

TABLE 3. RESULTS O F ANALYSIS OF DOSE MIXTURES IN THE THIRTEEN-WEEK GAVAGE STUDIES

O F 2-MERCAPTOBENZOTHIAZOLE

Target Determined
Determined as

Date Mixed Concentration (mg/ml) Concentration (mg/ml) (a)

Percent of Target

09124180 9.4 8.5 91

18.8 17.5 93

37.5 37.1 99

75.0 73.7 98

150.0 158.1 105

11126180 37.5 37.7 101

75.0 73.7 98
150.0 146.3 98
300.0 290.5 97

0210418 1 37.5 40.8 109

75.0 75.8 101
150.0 152.5 102
300.0 289.0 96

(a) Results of duplicate analysis

25 2-Mercaptobenzothiazole, NTP TR 332

II. MATERIALS AND METHODS

During the 2-year studies, periodic analysis of mixtures were within specifications 100% of the
dose preparations indicated that concentrations time. Results of periodic referee analyses per-
varied from 93.3% to 108.0% of the target con- formed by the analytical chemistry laboratory
centration (Table 4). Because 42/42 dose mix- indicated generally good agreement with the re-
tures analyzed were within 10% of the target sults from the study laboratory (Table 5).
concentration, it is estimated t h a t the dose

TABLE 4. RESULTS OF ANALYSIS OF DOSE MIXTURES I N THE TWO-YEAR GAVAGE STUDIES

O F 2-MERCAPTOBENZOTHIAZOLE

Concentration of 2-Mercaptobenzothiazole in Corn Oil

for Target Concentration (mg/ml) (a)
Date Mixed 37.5 75.0 150.0

07/08/81
37.8 74.8 140.0
0811218 1
37.8 76.9 149.0
10/20/81
36.6 72.8 147.5
10/28/81
38.2 72.6 146.7
01/13/82
39.1 70.2 157.5
04/07/82
38.7 74.2 146.1
05/05/82
39.8 76.0 147.2
06/23/82
38.3 76.2 144.3
09/08/82
37.1 74.4 144.0
11/17/82
35.4 76.7 149.3
12/01/82
37.8 79.4 156.9
02/16/83
39.9 78.3 158.3
04/27/83
37.7 75.7 151.4
06/08/83
39.0 81.0 147.2

Mean (mg/ml)
38.1 75.7 149.0
Standard deviation
1.22 2.83 5.39
Coefficientof variation (percent)
3.2 3.7 3.6
Range (mg/ml)
35.4-39.9 70.2-81.0 144.0-158.3
Number of samples
14 14 14
~~

(a)Results of duplicate analysis

TABLE 5. RESULTS OF REFEREE ANALYSIS O F DOSE MIXTURES I N THE THIRTEEN-WEEK AND

TWO-YEAR GAVAGE STUDIES O F 2-MERCAPTOBENZOTHIAZOLE

Determined Concentration (mg/ml)

Target Concentration Study Referee
Date Mixed (mg/ml) Laboratory (a) Laboratory (b)

Thirteen-Week Studies
09/24/80 150.0 158.1 134.0
11/26/80 37.5 37.7 36.0

Two-year Studies
07/08/81 150.0 140.0 145.0
05/05/82 37.5 39.8 37.1
11/17/82 75.0 76.7 75.0
04/27/83 150.0 151.4 144.0

(a) Results of duplicate analysis

(b)Results of triplicate analysis

2-Mercaptobenzothiazole,NTP TR 332 26
II. MATERIALS AND METHODS

FIRST SIXTEEN-DAY STUDIES (The 13-week study in rats reported in this Tech-
nical Report was a second study. In the first
Male and female F344/N rats and B6C3F1 mice study in rats, 3,000 mg/kg groups all died during
were obtained from Charles River Breeding Lab- week 1.) Animals were housed five per cage.
oratories and held for 14 days before the studies Feed and water were available ad libitum. Fur-
began. Rats were 6 weeks old when placed on ther experimental details are summarized in Ta-
study, and mice were 6-8 weeks old. Groups of ble 6.
five males and five females were administered 0,
156, 313,625, 1,250, or 2,500 mg/kg 2-mercapto- Animals were checked two times per day; mori-
benzothiazole in corn oil by gavage (12 doses bund animals were killed. Individual animal
over 16 days). Rats and mice were observed weights were recorded weekly. At the end of the
twice per day and were weighed on days 1,8, and 13-week studies, survivors were killed. A ne-
15. A necropsy was performed on all animals. cropsy was performed on all animals except
Details of animal maintenance are presented in those excessively autolyzed or cannibalized. Tis-
Table 6. sues and groups examined are listed in Table 6.

SECOND SIXTEEN-DAY STUDIES TWO-YEAR STUDIES

Study Design
Male and female B6C3F1 mice were obtained
from Harlan Industries and held for 19 days be- Groups of 50 male rats and 50 male and 50 fe-
fore the studies began. Mice were 5-6 weeks old male mice were administered 0, 375, or 750
when placed on study. Groups of five males and mg/kg 2-mercaptobenzothiazole in corn oil by
five females were administered 0, 188, 375, 750, gavage, 5 days per week for 103 weeks. Groups
1,500, or 3,000 mg/kg 2-mercaptobenzothiazole of 50 female rats were administered 0, 188, or
in corn oil by gavage (12 doses over 16 days). 375 mg/kg 2-mercaptobenzothiazole in corn oil
Mice were observed twice per day and were by gavage on the same schedule.
weighed on days 1, 8, and 15. A necropsy was
performed on all animals. Details of animal Source and Specifications of Animals
maintenance are presented in Table 6.
The male and female F344/N rats and B6C3F1
THIRTEEN-WEEK STUDIES (C57BL/6N, female X C3H/HeN MTV-, male)
mice used in these studies were produced under
Thirteen-week studies were conducted to evalu- strict barrier conditions a t Charles River Breed-
ate the cumulative toxic effects of repeated ad- ing Laboratories under a contract to the Carci-
ministration of 2-mercaptobenzothiazole and to nogenesis Program. Breeding stock for the
determine the doses to be used in the 2-year foundation colonies a t the production facility
studies. originated at the National Institutes of Health
Repository. Animals shipped for study were
Four- to five-week-old male and female F344/N progeny of defined microflora-associated parents
rats and B6C3F1 mice were obtained from that were transferred from isolators to barrier-
Charles River Breeding Laboratories, observed maintained rooms. Rats were shipped to the
for 19 days (rats) or 18 days (mice), and distrib- study laboratory at 4-5 weeks of age and mice, a t
uted to weight classes and then to cages accord- 6 weeks of age. The animals were quarantined
ing to a table of random numbers. Cages were a t the study laboratory for 13 days. Thereafter,
assigned to dosed and vehicle control groups ac- a complete necropsy was performed on five ani-
cording to a table of random numbers. Groups of mals of each sex and species to assess their
10 rats of each sex were administered 0, 188, health status. The rats were 46 days old and the
375, 750, or 1,500 mg/kg 2-mercaptobenzo- mice, 56 days old when placed on study. The
thiazole in corn oil by gavage, 5 days per week health of the animals was monitored during the
for 13 weeks. Groups of 10 mice of each sex were course of the studies according to the protocols of
administered 0,94,188,375,750, or 1,500 mg/kg the NTP Sentinel Animal Program (Appen-
2-mercaptobenzothiazole on the same schedule. dix F).

27 2-Mercaptobenzothiazole, NTP TR 332

TABLE 6. EXPERIMENTAL DESIGN AND MATERIALS AND METHODS IN THE GAVAGE STUDIES
OF 2-MERCAPTOBENZOTHIAZOLE

First Sixteen-Day Second Sixteen-Day Thirteen-Week Two-year

Studies Studies Studies Studies

EXPERIMENTAL DESIGN

Size of S t u d y Groups
5 males and 5 females 5 male and 5 female 10 males and 10 females 50 males and 50 females
of each species mice of each species of each species

Doses
0,156,313,625,1,250, 0,188,375,750,1,500, Rats--O,188,375,750, or 1,500 Rats--male: 0,375,or 760 mg/kg
or 2,500 m g k g 2-mercapto- or 3,000 m g k g 2-mercapto- mg/kg 2-mercaptobenzothiazole 2-mercaptobenzothiazole in
benzothiazole in corn oil benzothiazole in corn oil in corn oil by gavage; dose VOL- corn oil by gavage; female:
by gavage; dose Vol--rats: by gavage; dose VOL- 5 mg/kg; mice--0,94,188,375, 0,188, or 375 mgkg; dose VOL-
5 ml/kg except 10 m l k g 10ml/kg 750, or 1,500 mg/kg; dose VOL- 5 mlkg; mice--O,375,or
for 2,500 m g k g groups; 10 mlkg 750 mgkg; dose vol--10 m l k g
mice: 10mVkg

Date of First Dose

211 1/80 4/28/60 Rats-1 1/17/80;mice--8/18/80 Rata--7/14/81; mice--7/28/81

Date of Last Dose

2/26/80 5/13/80 Rats--2/13/81;mice--11/16/80 Rats--7/4/83; mice--7/19/83

Duration of Dosing
5 d/wk, 12 doses over Same as first 16-d studies 5d/wkfor 13 wk 5 d/wk for 103 wk
16 d
Type and Frequency of Observation
Observed 2 X d; weighed Same as first 16-d studies Same as first 16-d studies Observed 2 X d; weighed
initially and 1 x wk 1 Xwkfor12wkandl x 4 w k
thereafter thereaffer

Necropsy and Histologic Examination

Necropsy performed on Necropsy performed on Necropsy performed on all ani- Necropsy and histologic exams
all animals; histologic all animals; histologic mals; histologic exams per- performed on all animals; tis-
exams performed on all exams not performed on formed on some animals from sues examined: same as for
vehicle control and rata or mice all groups. Tissues examined 13-wk studies
2,500 m g k g male rats, include: adrenal glands, brain,
one rat from the 313 mg/kg colon, esophagus, eyes (if
group, and one female rat grossly abnormal), gallbladder
from the 2,500 mg/kg group; (mice),gross lesions and tissue
histologic exams not masses, heart, kidneys, liver,
performed on mice lungs and mainstem bronchi,
mammary gland, mandibular
or mesenteric lymph nodes,
pancreas, parathyroids, pitui-
tary gland, prostateltestes or
ovaries/uterus, salivary glands,
small intestine, spleen, spinal
cord (ifneurologic signs
present), sternebrae or femur
or vertebrae including mar-
row, stomach, thymus, thy-
roid gland, trachea, and
urinary bladder

ANIMALS AND ANIMAL MAINTENANCE

Strain and Species

F344/N rata; B6C3F1 mice B6C3F1 mice F344/N rats; B6C3F1 mice F344/N rats; B6C3F1 mice

2-Mercaptobenzothiazole, NTP TR 332 28

TABLE 6. EXPERIMENTAL DESIGN AND MATERIALS AND METHODS IN THE GAVAGE STUDIES
O F 2-MERCAPTOBENZOTHIAZOLE (Continued)

First Sixteen-Day Second Sixteen-Day

Thirteen-Week
Two-year

Studies Studies
Studies
Studies

ANIMALS AND ANIMAL MAINTENANCE (Continued)

Animal Source

Charles River Breeding Harlan Industries

Charles River Breeding Charles River Breeding
Laboratories (Indianapolis, IN)
Laboratories (Portage, MI) Laboratories (Portage, MI)
(Kingston, NY)

Study Laboratory

Physiological Research Physiological Research

Physiological Research Physiological Research
Laboratories Laboratories
Laboratories Laboratories

Method of Animal Identification

Toe clip Toe clip

Toe clip Toe and ear clip

Time Held Before Study

14 d 19 d
Rats--19 d; mice-18 d 13 d
Age When Placed o n Study

Rats-6 wk; mice-6-8 wk 5-6 wk

7-8 wk Rats-6-7 wk; m i c e 4 wk

Age When Killed

Rats-8-9 wk; mice- 7-8 wk

20-21 wk Rats-111 wk;
9-11 wk
mice-112 wk

Necropsy Dates

Rats--2/27/80-2/28/80; 5/13/80
Rats--2/17/81; mice--11/17/80 Rats--7/11/83-7/13/83;
mice--2/28/80-2/29/80
mice--7/26/83-7/27/83

Method of Animal Distribution

Animals distributed to Same as first 16-d studies Same as first 16-dstudies Same as first 16-d studies
weight classes; assigned
to cages and then to groups
according to tables of
random numbers

Feed
NIH 07 Rat and Mouse Same as first 16-d studies Same as first 16-d studies Same as first 16-d studies
Ration (Zeigler Bros.,
Gardners, PA); available
ad libitum
Bedding
Aspen wood chips Same as first 16-d studies Same as first 16-d studies Same as first 16-d studies
(Minnesota Sawdust and
Shavings Co., Anoka, MN)

Water
Automatic watering Same as first 16-d studies Same as first 16-d studies Same as first 16-d studies;
system (Edstrom softened to < 1 graidgal
Industries, Waterford, hardness with sodium
WI); available ad libitum zeolite; then filtered
through spun polyethylene

Cages
Polycarbonate (Hazleton Same as first 16-d studies Same as first 16-d studies Same as first 16-d studies
Systems, Inc., Aberdeen, MD)

Cage Filters
Reemaye (Dupont, Style Same as first 16-d studies Same as first 16-d studies Same as first 16-d studies
2024) spun-bondedpolyester
(SnowFiltration Co.,
Cincinnati, OH)

29 2-Mercaptobenzothiaeole,NTP TR 332
TABLE 6. EXPERIMENTAL DESIGN AND MATERIALS AND METHODS IN THE GAVAGE STUDIES
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

First Sixteen-Day Second Sixteen-Day Thirteen-Week Two-year

Studies Studies Studies Studies

ANIMALS AND ANIMAL MAINTENANCE (Continued)

Animals per Cage
5 5 5 5

Other Chemicals on Study in the Same Room

None None None None

Animal Room Environment

Temp--22.2°-24.4"C; Temp--17.8"-25.5° C;
Ternp--22.2"-26.6"C; Temp--generally 21'-23" C;
hum--38%-50%; hum--35%-70%;
hurn--32%-50%; hum--generally 40%-60%;
fluorescent light 12 h/d fluorescent light 12 h/d
fluorescent light 12 h/d fluorescent light 12 h/d;
15 room air changes/h

A quality control skin grafting program has details of animal maintenance are given in Ta-
been in effect since early 1978 to monitor the ble 6.
genetic integrity of the inbred mice used to pro-
duce the hybrid B6C3F1 study animal. In mid- Clinical Examinations a n d Pathology
1981, data were obtained that showed incom-
patibility between the NIH C3H reference All animals were observed two times per day,
colony and the C3H colony from a Program sup- and clinical signs were recorded once per week.
plier. In August 1981, inbred parental lines of Body weights by cage were recorded once per
mice were further tested for genetic integrity via week for the first 12 weeks of the study and once
isozyme and protein electrophoresis profiles that per month thereafter. Mean body weights were
demonstrate phenotype expressions of known calculated for each group. Animals found mori-
genetic loci. bund and those surviving to the end of the stud-
ies were humanely killed. A necropsy was per-
The C57BW6N mice were homogeneous a t all formed on all animals including those found
loci tested. Eighty-five percent of the C3H mice dead, unless they were excessively autolyzed or
monitored were variant a t one to three loci, indi- cannibalized, missexed, or found missing. Thus,
cating some heterogeneity in the C3H line from the number of animals from which particular or-
this supplier. Nevertheless, the genome of this gans or tissues were examined microscopically
line is more homogeneous than that of randomly varies and is not necessarily equal to the
bred stocks. number of animals that were placed on study.

Male mice from the C3H colony and female mice During necropsy, all organs and tissues were ex-
from the C57BL/6N colony were used as parents amined for grossly visible lesions. Tissues were
for the hybrid B6C3F1 mice used in these stud- preserved in 10% neutral buffered formalin, em-
ies. The influence of the potential genetic non- bedded in paraffin, sectioned, and stained with
uniformity in the hybrid mice on these results is hematoxylin and eosin. Tissues examined mi-
not known, but results of the studies are not af- croscopically are listed in Table 6.
fected because concurrent controls were included
in each study. When the pathology evaluation was completed,
the slides, paraffin blocks, and residual wet tis-
Animal Maintenance sues were sent to the NTP Archives for inven-
tory, slide/block match, and wet tissue audit.
Animals were housed five per cage. Feed and The slides, individual animal data records, and
water were available ad libitum. Further pathology tables were sent to a n independent

2-Mercaptobenzothiazole, NTP TR 332 30

II. MATERIALS AND METHOD$

quality assessment laboratory. The individual Survival Analyses: The probability of survival
animal records and tables were compared for ac- was estimated by the product-limit procedure of
curacy, slides and tissue counts were verified, Kaplan and Meier (1958) and is presented in the
and histotechnique was evaluated. All tumor di- form of graphs. Animals were censored from the
agnoses, all target tissues, and all tissues from a survival analyses at the time they were found to
randomly selected 10% of the animals were eval- be missing or dead from other than natural
uated by a quality assessment pathologist. The causes; animals dying from natural causes were
quality assessment report and slides were sub- not censored. Statistical analyses for a possible
mitted to the Pathology Working Group (PWG) dose-related effect on survival used the method
Chairperson, who reviewed all target tissues of Cox (1972) for testing two groups for equality
and those about which there was a disagreement and Tarone's (1975) life table test for a dose-
between the laboratory and quality assessment related trend. When significant survival differ-
pathologists. ences were detected, additional analyses using
these procedures were carried out to determine
Representative slides selected by the Chairper- the time point at which significant differences in
son were reviewed by the PWG without knowl- the survival curves were first detected. All re-
edge of previously rendered diagnoses. When ported P values for the survival analysis are
the consennun diagnosis of the PWG differed two-sided.
from that of the laboratory pathologist, the labo-
ratory pathologist was asked to reconsider the Calculation of Incidence: The incidence of neo-
original diagnosis. This procedure has been de- plastic or nonneoplastic lesions is given as the
scribed, in part, by Maronpot and Boorman ratio of the number of animals bearing such le-
(1982) and Boorman et al. (1985). The final di- sions a t a specific anatomic site to the number of
agnoses represent a consensus of contractor pa- animals in which that site was examined. In
thologists and the NTP Pathology Working most instances, the denominators include only
Group. For subsequent analysis of pathology those animals for which the site was examined
data, the diagnosed lesions for each tissue type histologically. However, when macroscopic ex-
are combined according to the guidelines of amination was required to detect lesions (e.g.,
McConnell et al. (1986). skin or mammary tumors) prior to histologic
sampling, or when lesions could have appeared
Slidedtissues are generally not evaluated in a
at multiple sites (e.g., lymphomas), the denomi-
blind fashion (i.e., without knowledge of dose nators consist of the number of animals on which
group) unless the lesions in question are subtle a necropsy was performed.
or unless there is a n inconsistent diagnosis of le-
sions by the laboratory pathologist. Nonneo- Analysis of Tumor Incidence: Three statistical
plastic lesions are not examined routinely by the methods are used to analyze tumor incidence
quality assessment pathologist or PWG unless data. The two that adjust for intercurrent mor-
they are considered part of the toxic effect of the tality employ the classical method for combining
chemical. contingency tables developed by Mantel and
Haenszel (1959). Tests of significance included
pairwise comparisons of high dose and low dose
Statistical Methods groups with vehicle controls and tests for overall
dose-response trends.
Data Recording: Data on this experiment were
recorded in the Carcinogenesis Bioassay Data For studies in which compound administration
System (Linhart et al., 1974). The data elements has little effect on survival, the results of the
include descriptive information on the chemi- three alternative analyses will generally be
cals, animals, experimental design, survival, similar. When differing results are obtained by
body weight, and individual pathology results, the three methods, the final interpretation of the
a s recommended by the International Union data will depend on the extent to which the tu-
Against Cancer (Berenblum, 1989). mor under consideration is regarded as being the

31 2-Mercaptobenzothiazole,NTP TR 332
II. MATERIALS AND METHODS

cause of death. Continuity-corrected tests are animals in dosed and vehicle control groups
used in the analysis of tumor incidence, and re- were compared in each of five time intervals:
ported P values are one-sided. weeks 0-52, weeks 53-78, weeks 79-92, week 93
to the week before the terminal-kill period, and
Life Table Analysis-The first method of analy- the terminal-kill period. The denominators of
sis assumed that all tumors of a given type ob- these proportions were the number of animals
served in animals dying before the end of the actually examined for tumors during the time
study were “fatal”; i.e., they either directly or in- interval. The individual time interval com-
directly caused the death of the animal. Accord- parisons were then combined by the previously
ing to this approach, the proportions of tumor- described method to obtain a single overall re-
bearing animals in the dosed and vehicle control sult. (See Haseman, 1984, for the computational
groups were compared at each point in time a t details of both methods.)
which an animal died with a tumor of interest.
The denominators of these proportions were the
Unadjusted Analyses--Primarily, survival-ad-
total number of animals at risk in each group.
justed methods are used to evaluate tumor inci-
These results, including the data from animals
dence. In addition, the results of the Fisher
killed at the end of the study, were then com-
exact test for pairwise comparisons and the
bined by the Mantel-Haenszel method to obtain
Cochran-Armitage linear trend test (Armitage,
an overall P value. This method of adjusting for
1971; Gart e t al., 1979) are given in the appen-
intercurrent mortality is the life table method of
dixes containing the analyses of primary tumor
Cox (1972) and of Tarone (1975). The under-
incidence. Thesettwo tests are based on the over-
lying variable considered by this analysis is time
all proportion of &tumor-bearinganimals and do
to death due to tumor. If the tumor is rapidly
not adjust for suru?ival differences.
lethal, then time to death due to tumor closely
approximates time to tumor onset. In this case,
the life table test also provides a comparison of Historical Control Data: Although the concur-
the time-specific tumor incidences. rent control group is always the first and most
appropriate control group used for evaluation,
Incidental Tumor Analysis--The second method there are certain instances in which historical
of analysis assumed that all tumors of a given control data can be helpful in the overall as-
type observed in animals that died before the sessment of tumor incidence. Consequently,
end of the study were “incidental”; i.e., they control tumor incidences from the NTP his-
were merely observed at necropsy in animals torical control data base (Haseman et al., 1984,
dying of an unrelated cause. According to this 1985) are included for those tumors appearing to
approach, the proportions of tumor-bearing show compound-related effects.

2-Mercaptobenzothiazole,NTP TR 332 32
III. RESULTS

RATS
SIXTEEN-DAY STUDIES
THIRTEEN- WEEK STUDIES
TWO-YEAR STUDIES
Body Weights and Clinical Signs
Survival
Pathology and Statistical Anajyses of Results

MICE
SIXTEEN-DAY STUDIES
THIRTEEN-WEEK STUDIES
TWO-YEAR STUDIES
Body Weights and Clinical Signs
Survival
Pathology and Statistical Analyses of Results

33 2-Mercaptobenzothiazole,NTP TR 332
III. RESULTS: RATS

SIXTEEN-DAY STUDIES behavior that was more pronounced with in-

creasing dose and was characterized as resis-
Although there were no chemically related tance to gavage. Body weight gain was reduced
deaths in the 16-day studies (Table 71, mean with increasing dose, with a maximum change of
body weight gain in rats of each sex given the - 15% compared with vehicle controls. Liver
highest dose of 2-mercaptobenzothiazole (2,500 weight and liver weight to body weight ratios
mg/kg) was 6-7 g (8%-14%) less than that in ve- were increased in dosed rats with the greatest
hicle controls; for this reason, the highest dose change occurring at the two highest doses (750
chosen for the 13-week studies, 1,500 mg/kg, was and 1,500 mg/kg) (Table 9). No gross or mi-
between the two highest doses used in the 16- croscopic effects could be related to chemical
day studies (1,250 and 2,500 mg/kg). No administration.
compound-related gross pathologic effects were
observed.
Dose Selection Rationale: Because of lower
THIRTEEN-WEEK STUDIES weight gain at higher doses, doses selected for
rats for the 2-year studies were 375 and 750
No compound-related deaths occurred in rats mg/kg 2-mercaptobenzothiazole for males and
dosed with 2-mercaptobenzothiazole for 13 188 and 375 mg/kg for females, administered in
weeks (Table 8). The animals displayed irritable corn oil by gavage 5 days per week.

TABLE 7. SURVIVAL AND MEAN BODY WEIGHTS OF RATS IN THE SIXTEEN-DAY GAVAGE
STUDIES OF 2-MERCAPTOBENZOTHIAZOLE

Mean Body Weights (grams) Final Weight Relative

Dose Survival (a) Initial (b) Final Change (c) to Vehicle Controls
(mg/kg) (percent)

MALE
0 (d) 4/5 87 k 1 159 f 4 +72f 3 __
156 515 92 f 2 166 f 8 +74f 10 104
313 515 87 f 1 163 k 3 +76f 3 103
625 (e) 315 87 k 1 171 f 0 +85f 2 108
1,250 515 97 f 1 164 f 3 +67f 3 103

2,500 515 88 f 1 154 f 3 +66f 3 97

FEMALE
0 515 78 f 1 129 f 1 +51f 2 __
156 515 81 f 1 130 f 3 +49f 2 101
313 (e)415 75 f 1 126 f 1 +51f 1 98
625 515 71 f 1 121 f 1 +50f 1 94
1,250 515 83 f 1 134 f 1 +51f 1 104

2,500 515 84 f 2 128 f 3 +44f 3 99

(a)Number surviving/number initially in group

(b)Initial mean group body weight f standard error of the mean. Subsequent calculations are based on those animals
surviving to the end of the study.
(c)Mean body weight change of the survivors f standard error of the mean
(d)Day of death: 9
(e)Deaths due to gavage error

2-Mercaptobenzothiazole,NTP TR 332 34
TABLE 8. SURVIVAL AND MEAN BODY WEIGHTS O F RATS IN T H E THIRTEEN-WEEK GAVAGE
STUDIES O F 2-MERCAPTOBENZOTHIAZOLE

Mean Body Weights (grams) Final Wei ht Relative

Dose Survival (a) Initial (b) Final Change (c) to Vehicfe Controls
(mg/kg) (percent)

MALE
0 loll0 140 f 2 355 f 6 +215f 6 __

139 f 2 f 3
188
375
750
10110
9/10
10110
136 f 2
141 I3
357
336
342
f 8
F 10
+200
~ *
+218 f 2
- .
.
+201 f 10
8

101

95

96

1,500 8/10 142 It 2 325 f 9 +182 f 9 92

FEMALE
0 10/10 115 f 2 208 f 5 +93f 5 __

188 9/10 115 f 2 200 f 5 +84f 5 96

375 loll0 116 f 1 201 f 4 +85f 3 97

750 8110 115 f 2 191 f 3 +77f 2 92

1,500 10/10 115 f 2 195f 4 +80f 3 94

(a)Number surviving/number initially in group; all deaths due to avage error.

(b) Initial mean group body weight f standard error ofthe mean. subsequent calculations are based on those animals

survivin to the end of the stud ,

(c)Meantody weight change ofthe survivors f standard error of the mean

TABLE 9. ANALYSIS O F LIVER WEIGHTS FOR RATS IN THE THIRTEEN-WEEK GAVAGE

STUDIES O F 2-MERCAPTOBENZOTHIAZOLE (a)

Dose No. Final Mean Liver Weight Liver WeighVFinal

(mg/kg) Examined Body Weight (grams) (mg) Body Weight (mg/g)

MALE
0 10 355 f 19.1 13,593 f 2,121 38.4 f 6.07
188 10 357 f 8.5 15,661 f 793 (b)43.9 f 1.87
375 9 336 f 23.1 15,861 f 1,712 (d47.2 f 2.79
750 0 342 f 31.5 (c) 18.742 f 2.631 (c) 54.8 f 5.08
1,500 8 (b)325 f 26.8 (c) 161759 f 21660 (c) 51.3 f 5.42
FEMALE
0
188
375
0
9
0
208 f 15.9
200 f 15.6
201 f 11.3
6,606 f
(c)7.818 f
(c)8,027 f
795
814
688
*
31.8 f 3.28
(c)39.3 3.53
(d39.9 f 2.99
750 8 (b1191 f 8.8 (c)7,988 f 591 ( d 4 1 . 8 f 2.81
1,500 10 195 f 12.1 (~18,413f 652 (c)43.2 f 2.61

(a)Mean f standard deviation; P values are versus the vehicle controls by Dunnett’s test (Dunnett, 1955).

(b)P<0.05

(C)P<O.Ol

TWO-YEAR STUDIES

Body Weights and Clinical Signs controls (Table 10 and Figure 5). Mean body
weights of dosed female rats were generally
Mean body weights of dosed male rats were greater (up to 11%) than those of the vehicle con-
similar to or greater than those of the vehicle trols. Rats were lethargic after they were dosed.

35 2-Mercaptobenzothiazole, NTP TR 332

TABLE 10. MEAN BODY WEIGHTS AND SURVIVAL OF RATS IN THE TWO-YEAR GAVAGE STUDIES
OF 2-MERCAPTOBENZOTHIAZOLE

Weeks Vehicle Control Low Dose High Dose

on Av. Wt. No. of Av. Wt. Wt. (percent of No. of Av. Wt. Wt. (percent of No. of
Study (grams) Survivors (grams) veh. controls) Survivors (grams) veh. controls) Survivors

MALE 375 mg/kg 750 mg/kg

0 138 50 137 99 50 138 100 50
1 116 50 114 99 50 112 98 50
2 209 50 205 98 50 205 98 50
3 236 50 230 97 50 229 91 50
4 258 50 251 97 50 248 9% 50
5 219 50 213 98 50 210 97 50
6 288 50 288 100 50 285 99 50
I 312 50 303 91 50 301 96 50
8 325 50 315 97 50 314 97 50
9 338 50 321 91 50 325 96 50
10 341 50 335 97 50 332 96 50
11 356 50 345 97 50 342 96 50
12 389 50 350 96 50 346 96 50
16 391 50 381 91 50 316 96 50
20 419 50 405 97 50 391 95 50
25 443 50 426 96 50 411 94 50
31 455 50 438 96 50 431 95 50
35 473 50 456 96 50 444 94 50
39 484 50 412 98 50 489 96 50
44 492 50 488 99 50 418 97 50
46 501 50 497 98 50 486 96 50
53 509 50 502 99 50 493 91 50
51 510 50 507 99 50 494 91 49
62 515 50 514 100 50 498 97 48
66 511 50 511 100 50 504 91 41
IO 516 50 516 100 50 502 91 45
14 522 50 526 101 50 513 98 45
I9 523 50 528 101 48 518 99 44
a3 523 50 529 101 41 522 100 42
81 523 50 534 102 41 523 100 40
92 515 49 529 103 36 519 101 39
96 501 46 511 102 34 522 103 33
100 499 44 510 102 21 514 103 28
103 492 42 500 102 23 498 101 20

FEMALE 188 mg/kg 375 mg/kg

0 112 50 112 100 50 112 100 50
1 131 50 131 100 50 129 96 50
2 148 50 148 100 50 141 99 50
3 159 50 160 101 50 159 100 50
4 168 50 169 101 50 168 100 50
5 119 50 181 101 50 119 100 50
6 186 50 188 101 50 186 100 50
I 191 50 194 102 50 193 101 50
8 196 50 201 103 50 199 102 50
9 198 50 204 103 50 201 102 50
10 200 50 206 103 50 203 102 50
11 206 50 212 103 50 209 101 50
12 205 50 211 103 50 208 101 50
16 216 50 222 103 50 219 101 so
20 225 50 231 103 49 232 109 50
25 233 50 239 103 49 238 102 48
31 236 49 246 103 46 241 104 46
35 246 48 254 103 48 255 104 45
39 250 48 262 105 48 283 105 44
44 255 46 21 1 106 48 213 101 43
48 262 46 218 106 48 281 101 43
53 269 44 281 101 48 290 108 42
51 214 43 293 101 41 298 109 42
62 283 42 305 108 41 306 108 42
66 295 41 311 101 41 318 108 41
IO 300 41 325 108 46 326 109 41
74 309 40 331 109 46 338 109 41
I9 322 39 349 108 46 346 108 41
83 332 35 356 101 46 358 106 38
81 336 35 364 108 44 365 109 56
92 340 31 3ea 108 40 371 109 32
96 345 31 311 108 39 381 110 21
100 345 30 311 108 35 318 110 26
103 340 29 370 109 31 318 111 25

2-Mercaptobenzothiazole,NTP TR 332 36
550.0

500.0

450.0

400.0

sso.0

.................
....... 3 0 0 . 0

250.0

200.0

150.0

100.0
0 IS 30 4s 60 IS mo 10s
WEEKS ON STUDY

- 550.0
- 500.0

- 450.0

- 400.0

- S50.0

- 100.0

- 250.0

- 200.0

- 150.0

- 100.0
0 1s so 49 eo 7s 10 5
WEEKS ON STUDY

FIGURE 5. GROWTH CURVES FOR RATS ADMINISTERED

3-MERCAPTOBENZOTHIAZOLE IN CORN OIL BY GAVAGE FOR TWO YEARS

37 2-Mercaptobenzothiazole,NTP TR 332
III. RESULTS: RATS

Survival are summarized in Table A l . Table A2 gives the

survival and tumor status for individual male
Estimates of the probabilities of survival for rats. Table A3 contains the statistical analyses
male and female rats administered 2-mercapto- of those primary tumors that occurred with a n
benzothiazole a t the doses used in these studies incidence of at least 5 % in one of the three
and for vehicle controls are shown in Table 11 groups. The statistical analyses used are dis-
and in the Kaplan and Meier curves in Figure 6. cussed in Chapter II (Statistical Methods) and
Survival of the low dose group of male rats was Table A3 (footnotes). Historical incidences of tu-
significantly lower than that of the vehicle con- mors in corn oil vehicle control male rats are
trols after week 85. Survival of the high dose listed in Table A4. Findings on nonneoplastic le-
group of male rats was significantly lower than sions are summarized in Table A5.
that of the vehicle controls after week 83 (except
for weeks 94 and 95). Lesions in female rats are summarized in Ap-
pendix B. Histopathologic findings on neo-
Pathology and Statistical Analyses of
plasms are summarized in Table B1. Table B2
Results gives the survival and tumor status for individ-
This section describes the significant or note- ual female rats. Table B3 contains the statisti-
worthy changes in the incidences of rats with cal analyses of those primary tumors that oc-
neoplastic or nonneoplastic lesions of the hema- curred with an incidence of at least 5% in one of
topoietic system, pituitary gland, adrenal gland, the three groups. The statistical analyses used
pancreas, preputial gland, multiple organs, sub- are discussed in Chapter II (Statistical Methods)
cutaneous tissue, kidney, forestomach, and eye. and Table B3 (footnotes). Historical incidences
of tumors in corn oil vehicle control female rats
Lesions in male rats are summarized in Appen- are listed in Table B4. Findings on nonneo-
dix A. Histopathologic findings on neoplasms plastic lesions are summarized in Table B5.

TABLE 11. SURVIVAL OF RATS IN THE TWO-YEAR GAVAGE STUDIES OF

2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 188 mg/kg 375 mg/kg 750 mg/kg

MALE (a)
Animals initially in study 50 50 50
Nonaccidental deaths before termination (b) 8 28 29
Accidentally killed 0 0 1
Killed at termination 42 22 20
Survival P values (c) <0.001 <0.001 <0.001

FEMALE (a)
Animals initially in study 50 50 50
Nonaccidental deaths before termination (b) 21 18 25
Accidentally killed 1 1 0
Killed at termination 28 31 25
Survival P values (c) 0.535 0.415 0.639

(a)Terminal-kill period: week 104

(b) Includes animals killed in a moribund condition
(c)The result of the life table trend test is in the vehicle control column, and the resulb of the life table pairwise comparisons
with the vehicle controls are in the dosed columns.

2-Mercaptobenzothiazole, NTP TR 332 38

0 9

0 8

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0.5 .................
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375 MG/KG
...............
A = 7 5 0 MG/KG
I
r"""'................... .........-I."" ........
I_I_I. ................. 0.)

0.4
0
I I
I

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I

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WEEKS ON STUDY
i 75
I 10s
0.4

1.0 1.0
i
i
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1 '
0.9 .................& .......................... 0.9

4 I:1
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0.8
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0.7 0.7
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c- I

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m 0,s
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4s 00
1 i 90
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WEEKS ON STUDY

FIGURE 6. KAPLAN-MEIER SURVIVAL CURVES FOR RATS ADMINISTERED

2-MERCAPTOBENZOTHIAZOLE IN CORN OIL BY GAVAGE FOR TWO YEARS

39 2-Mercaptobenzothiazole,NTP TR 332
III. RESULTS: RATS

Hematopoietic System: The incidence of leuke- occurred with significant positive trends; the in-
mia in low dose male rats was significantly cidences of adenomas in low dose males and of
greater than that in the vehicle controls by the adenomas and adenomas or adenocarcinomas
life table test and exceeded the high value for (combined) in high dose females were signifi-
the historical corn oil vehicle control range cantly greater than those in the vehicle controls
(0/50-14/50) (Table 12). (Table 13). The incidence of hyperplasia of the
anterior pituitary was slightly increased in low
Pituitary Gland: Adenomas and adenomas or dose male rats.
adenocarcinomas (combined) in female r a t s

TABLE 12. ANALYSIS OF MONONUCLEAR CELL LEUKEMIA IN RATS IN THE TWO-YEAR

GAVAGE STUDIES OF 2-MERCAPTOBENZOTHIAZOLE (a)

Vehicle Control 188 mg/kg 375 mg/kg 750 mg/kg

MALE (b)

Overall Rates 7/50 (14%) 16/50 (32%) 3/50 (6%)

Adjusted Rates 15.1%
Terminal Rates 4/42 (10%) 6/22 (27%)
47.2% 2/20
12.3%( 10%)
Week of First Observation 91 78 91
Life Table Testa P = 0.475 P =0.002 P = 0.449N
Incidental Tumor Testa P=0.084N P=0.103 P = 0.157N

FEMALE (c)

Overall Rates 6/50 (12%) 14/50 (28%) 9/50 (18%)

Adjusted Rates 19.7% 35.4% 25.3%
Terminal Rates 4/28 (14%) 6/31 (19%) 2/25 (8%)
Week of First Observation 90 92 79
Life Table Testa P=0.221 P = 0.099 P = 0.279
Incidental Tumor Testa P = 0.399 P-0.215 P=0.415

(a)The statistical analyses used are discussed in Chapter II (Statistical Methods) and Appendix A, Table A3 (footnotes).
(b)Historical incidence ofleukemia in NTP studies (mean f SD): 202/1,450 (14% f 8%)
(c)Historical incidence of leukemia in NTP studies (mean f SD): 271/1,450 (19% f 9%)

2-Mercaptobenzothiazole,NTP TR 332 40
TABLE 13. ANALYSIS OF PITUITARY GLAND LESIONS I N RATS I N THE TWO-YEAR GAVAGE
STUDIES O F 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 188 mgkg 375 mg/kg 750 mgk g

MALE

Hyperplasia

Overall Rates 10150(20%) 17/50(34%) 12/48(25%)

Adenoma (a)

Overall Rates 14/50(28%) 21/50(42%) 12/48(25%)

Adjusted Rates 30.9% 59.9% 40.1%

Terminal Rates 11/42(26%) 10/22(45%) 5/20(25%)

Week of First Observation 94 82 82

Life Table Testa P = 0.106 P = 0.003 P=0.171

Incidental Tumor Testa P = 0.506N P=0.132 P = 0.482N

FEMALE

Hyperplasia

Overall Rates 8/49(16%) 10/50(20%) 6/50(12%)

Adenoma

Overall Rates 15/49(31%) 24/50(48%) 25/50(50%)

Adjusted Rates 44.6% 62.3% 73.2%

Terminal Rates 10128 (36%) 17/31(55%) 16/25(64%)

Week of First Observation 72 67 82

Life Table Testa P=0.014 P =0.146 P=O.O21

Incidental Tumor Testa P=0.015 P=0.139 P=0.027

Adenocarcinoma

Overall Rates 1/49(2%) 0150 (0%) 0150 (0%)

Adenoma or Adenocarcinoma (b)

Overall Rates 16/49(33%) 24/50(48%) 25/50(50%)

Adjusted Rates 46.2% 62.3% 73.2%

Terminal Rates 10128(36%) 17/31(55%) 16/25(64%)

Week of First Observation 72 67 82

Life Table Testa P = 0.024 P=0.206 P = 0.036

Incidental Tumor Testa P = 0.028 P=0.186 P=0.050

(a)Historical incidence of adenomas in NTP studies (mean f SD): 34411,411(24% f 8%)

(b) Historical incidence of adenomas, carcinomas, or adenocarcinomas (combined) in NTP studies (mean _+ SD): 56111,407

(40% f 8%)

Adrenal Gland: Pheochromocytomas in male The incidence of medullary hyperplasia was

and female rats occurred with significant posi- slightly increased in low dose male rats. The hy-
tive trends; the incidences in dosed male and perplasia was characterized by focal areas of
high dose female rats were significantly greater somewhat darker staining cells with relatively
than those in the vehicle controls by the life ta- larger nuclei; no invasion or compression of the
ble test (Table 14). The incidences for both low surrounding medulla or cortex was observed.
and high dose male rats exceeded the historical Benign pheochromocytomas were similar to the
corn oil vehicle control values (mean historical hyperplasia except that they were larger and
incidence, 33811,442, 23.4%;range, 2150-20149; compressed or displaced adjacent medulla and
Table A4c). cortex. Malignant pheochromocytomas invaded
the medulla and cortex and extended through
the adrenal capsule.

41 2-Mercaptobenzothiazole, NTP TR 332

TABLE 14. ANALYSIS OF ADRENAL GLAND LESIONS IN RATS I N T H E TWO-YEAR GAVAGE
STUDIES OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 188 mg/kg 373 mg/kg 730 mg/kg

MALE

Medullary Hyperplasia
Overall Rates 9/50(18%)
14/50(28%) 10/49(20%)
Pheochromocytoma
Overall Rates 18/50(36%) 25/50(50%) 22/49(45%)
Adjusted Rates 39.8% 70.3% 68.5%
Terminal Rates 15/42(36%) 12/22(55%) 11/20 (55%)
Week of First Observation 93 85 84
Life Table Tests P=0.002 PCO.001 P=0.002
Incidental Tumor Tests P =0.109 P=0.056 P=O.lll
Malignant Pheochromocytoma
Overall Rates 0/50(0%)
2/50(4%) 2/49(4%)
Pheochromocytoma or Malignant Pheochromocytoma (a)
Overall Rates 18/50(36%)
27/50(54%) 24/49(49%)
Adjusted Rates 39.8%
74.1% 75.5%
Terminal Rates 15/42(36%)
13/22(59%) 13/20 (65%)
Week of First Observation 93
85 84
Life Table Tests PCO.001
P co.00 1 PCO.001
Incidental Tumor Tests P=0.038
P=0.021 P=0.034
FEMALE

Medullary Hyperplasia
Overall Rates 5/50(10%)
8/50(16%) 2/50(4%)
Pheochromocytoma (b)
Overall Rates 1150 (2%) 5/50(10%) 6/50(12%)
Adjusted Rates 3.6% 14.6% 23.0%
Terminal Rates 1/28(4%) 3/31(10%) 5/25(20%)
Week of First Observation 104 96 97
Life Table Testa P = 0.030 P=0.137 P=0.041
Incidental Tumor Tests P=0.038 P=0.214 P=0.052

(a)Historical incidence in NTP studies (mean f SD): 347 11,442(24% f 9%)

(b)Historical incidence in NTP studies (mean f SD): 8211,443(6% f 4%)

Pancreas: The incidence of acinar cell adenomas primarily by their larger size and abnormal
in low dose male rats was significantly greater growth pattern.
than that in the vehicle controls by the inci-
dental tumor test (Table 15). The incidence of Preputial Gland: Adenomas in male rats oc-
pancreatic acinar cell hyperplasia was also in- curred with a significant positive trend by the
creased in the low dose group. Acinar cell hyper- incidental tumor test, and the incidences of ade-
plasia usually consisted of focal, circumscribed, nomas or carcinomas (combined) in dosed groups
round to oval lesions that slightly compressed were significantly greater than those in the ve-
the surrounding acini. The acinar pattern was hicle controls by the life table tests (Table 16).
prominent, and these a r e a s were clearly The number of tumors for any group did not ex-
demarcated from surrounding acinar tissue. ceed the historical corn oil vehicle control range
Adenomas generally were similar in appearance (0/50-9/50),
to the hyperplasia but were distinguished

2-Mercaptobenzothiazole, NTP TR 332 42

TABLE 15. ANALYSIS OF PANCREATIC ACINAR CELL LESIONS IN MALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 mg/kg 750 m g k g

Hyperplasia
Overall Rates 5/50(1 0%) 15/50(30%) 7/49(14%)
Adenoma (a)
Overall Rates 2/50(4%) 13/50(26%) 6/49(12%)

Adjusted Rates 4.5% 45.7% 23.0%

Terminal Rates 1/42(2%) 8/22 (36%) 3/20(15%)

Week of First Observation 94 88 98

Life Table Tests P = 0.017 P<O.OOl P = 0.030

Incidental Tumor Tests P=0.118 P <0.001 P = 0.160

(a)Historical incidence of acinar cell neoplasms in NTP studies (mean k SD): 8011,381(6% k 8%)

TABLE 16. ANALYSIS OF PREPUTIAL GLAND LESIONS IN MALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 mgkg 750 mgkg

Hyperplasia
Overall Rates 0150 (0%) 0150 (0%) 1/50(2%)
Adenoma
Overall Rates 0150(0%) 4/50(8%) 4/50(8%)

Adjusted Rates 0.0% 14.7% 14.4%

Terminal Rates 0142 (0%) 2/22(9%) 2/20(10%)

Week of First Observation 88 87

Life Table Tests P = 0.016 P=O.O19 P=O.O21

Incidental Tumor Tests P = 0.042 P = 0.076 P = 0.063

Carcinoma (a)
Overall Rates 1/50(2%) 2/50(4%) 1/50(2%)
Adenoma or Carcinoma (b)
Overall Rates 1/50(2%) 6/50(12%) 5/50(10%)
Adjusted Rates 2.2% 18.5% 19.2%
Terminal Rates 0142 (0%) 2/22(9%) 3/20(15%)
Week of First Observation 98 83 87
Life Table Tests P = 0.027 P=O.O21 P=0.030
Incidental Tumor Testa P = 0.094 P = 0.216 P=0.117

(a)Historical incidence in NTP studies (mean f SD): 3511,450(2% k 3%)

(b)Historical incidence in NTP studies (mean f SD): 6511,450(4% f 4%)

43 2-Mercaptobenzothiazole, NTP TR 332

III. RESULTS: RATS

Multiple Organs: Mesotheliomas in male rats Subcutaneous Tissue: Fibromas and fibromas,
occurred with a significant positive trend; the in- neurofibromas, sarcomas, or fibrosarcomas
cidences in the dosed groups were not signifi- (combined) in male rats occurred with signifi-
cantly greater than that in the vehicle controls cant positive trends by the life table test but not
(Table 17)and did not exceed the historical corn by the more appropriate incidental tumor test
oil vehicle control range for this neoplasm (0/50- (Table 18).
6/50).

TABLE 17. ANALYSIS O F MESOTHELIOMAS IN MALE RATS IN THE TWO-YEAR GAVAGE STUDY

OF 2-MERCAPTOBENZOTHIAZOLE (a)

Vehicle Control 375 mg/kg 750 m g k g

Overall Rates 0150 (0%) 2/50(4%) 3/50(6%)

Adjusted Rates 0.0% 6.6% 9.5%

Terminal Rates 0142(0%) 1/22(5%) 1/20(5%)

Week of First Observation 84 84

Life Table Tests P = 0.039 P=0.163 P = 0.066

Incidental Tumor Tests P=0.041 P=0.310 P = 0.158

(a)Historical incidence in NTP studies (mean k SD): 55/1,450(4% f 3%)

TABLE 18. ANALYSIS O F SUBCUTANEOUS TISSUE TUMORS IN MALE RATS IN THE TWO-YEAR

GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 mg/kg 750 mg/kg

Fibroma

Overall Rates 2/50(4%) 3/50(6%) 6/50(12%)

Adjusted Rates 4.8% 9.2% 19.6%

Terminal Rates 2/42(5%) 1/22(5%) 2/20(10%)

Week of First Observation 104 85 82

Life Table Tests P=0.024 P=0.299 P=0.033

Incidental Tumor Tests P= 0.064 P=0.612 P=0.153

Neurofibroma

Overall Rates 0150 (0%) 1/50(2%) 0150 (0%)

Sarcoma

Overall Rates 0150(0%) 1/50(2%) 1/50(2%)

Fibrosarcoma

Overall Rates 1/50(2%) 1/50(2%) 0150 (0%)

Fibroma, Neurofibroma, Sarcoma, or Fibrosarcoma (a)

Overall Rates 3/50(6%) 6/50(12%) 7/50( 14%)

Adjusted Rates 7.1% 17.7% 21.4%

Terminal Rates 3/42 (7%) 2/22(9%) 2/20(10%)

Week of First Observation 104 85 74

Life Table Tests P = 0.031 P=0.084 P=0.037

Incidental Tumor Tests P =0.129 P = 0.396 P = 0.237

(a)Historical incidence in NTP studies (mean k SD): 126/1,450(9% f 4%)

2-Mercaptobenzothiazole,NTP TR 332 44
III. RESULTS: RATS

Kidney: Nephropathy, characterized by tubular tubular cell neoplasms in male F344/N corn oil
degeneration and regeneration, was present in vehicle control rats is 811,448 (0.6%).
all male rats and in more than 75% of the female
Forestomach: Ulcers and inflammation were ob-
rats; a severity grade from minimal to severe (1-
served a t increased incidences in dosed rats, and
4) was recorded for each animal. The mean
epithelial hyperplasia and hyperkeratosis were
severity of nephropathy was increased in dosed
male rats (vehicle control: 2.3 [mild-moderate]; observed at increased incidences in dosed male
and low dose female rats (Table 20).
low dose and high dose: 3.4 [moderate-severe]).
Eye: Retinopathy and cataracts were observed
at increased incidences in low dose rats (retino-
Pelvic epithelial hyperplasia and transitional pathy--male: vehicle control, 0/50; low dose,
cell papillomas or carcinomas and tubular cell 10150; high dose, 0150; female: 1/50; 9/50; 0150;
hyperplasia and tubular cell adenomas were ob- cataracts--male: 1/50; 6/50; OJ50; female: 0150;
served in dosed male rats (Table 19). The his- 8/50; 0/50). Low dose groups were on the top two
torical incidence of transitional cell neoplasms rows of the racks near the fluorescent light
in male F344/N corn oil vehicle control rats is source. The cage racks were not rotated in these
111,448 ( < O . l % ) ; the historical incidence of studies.

TABLE 19. NUMBER OF RATS WITH KIDNEY LESIONS IN THE TWO-YEAR GAVAGE STUDIES OF
2-MERC APTOBENZOTHIAZOLE

Male Female
Sitenesion 0 375 mg/kg 750 mg/kg 0 188 mg/kg 375 mg/kg

No. examined 50 50 49 50 50 50

Kidneylpelvis
Epithelial hyperplasia 0 4 1 1 0 0
Transitional cell papilloma 0 1 1 0 0 0
Transitional cell carcinoma 0 1 0 0 0 0
Kidneyhubule
Focal hyperplasia 0 3 3 1 0 0
Kidney
Tubular cell adenoma 0 1 1 0 0 0

TABLE 20. NUMBER OF RATS WITH FORESTOMACH LESIONS IN THE TWO-YEAR GAVAGE
STUDIES OF 2-MERCAPTOBENZOTHIAZOLE

Male Female
Lesion 0 375 mg/kg 750 mg/kg 0 188 mg/kg 375 mg/kg

No. examined 50 50 49 49 50 50

Ulcer 0 5 5 0 3 5
Inflammation 0 11 14 2 4 7
Epithelial hyperplasia 1 12 17 1 4 1
Hyperkeratosis 0 12 17 1 4 1

45 2-Mercaptobenzothiazole,NTP TR 332
III. RESULTS: MICE

SIXTEEN-DAY STUDIES 3,000 mg/kg were lethargic after day 1. Final

mean body weights were not adversely affected
An initial 16-day study was repeated because an by 2-mercaptobenzothiazole. No compound-rela-
excessive number of gavage accidents occurred. ted lesions were observed grossly. Since all but
In the second study, 4 5 males and 515 females one of the male and female mice dosed with
that received 3,000 mg/kg and 4/5 females that 3,000 mg/kg died, the highest dose used in the
received 1,500 mg/kg died before the end of the 13-week studies was 1,500 mg/kg for mice of
studies (Table 21). Mice that received 1,500 or each sex.

TABLE 21. SURVIVAL AND MEAN BODY WEIGHTS O F MICE I N T H E SECOND SIXTEEN-DAY

GAVAGE STUDIES O F 2-MERCAPTOBENZOTHIAZOLE

Mean Body Weights (grams) Final Weight Relative

Dose Survival (a) Initial (b) Final Change (c) to Vehicle Controls
(mg/kg) (percent)

MALE
0 515 23.2 f 0.8 24.1 f 0.8 +0.9 f 0.8 __

188 515 21.4 f 0.6 24.2 f 0.6 +2.8 f 0.4 100.4

375 515 21.4 f 0.7 25.0 f 0.6 +3.6 f 0.4 103.7

750 515 21.7 f 0.8 24.5 k 0.8 +2.8 f 0.3 101.7

1,500 515 22.9 2 0.3 25.0 f 0.3 t 2 . 1 f 0.2 103.7

3,000 (d) 115 22.1 f 0.4 27.0 + 3.4 112.0

FEMALE

0 515 19.0 f 0.2 20.0 f 0.1 + 1.0 f 0.1 .-

188 515 20.3 f 0.5 21.2 f 0.5 +0.9 f 0.2 106.0

375 515 20.3 f 0.7 21.8 f 0.7 +1.5 f 0.4 109.0

750 515 20.3 f 0.3 21.5 f 0.3 +1.2 f 0.3 107.5

1,500 (e) 115 19.8 f 0.4 22.5 +1.3 112.5

3,000 (e)015 18.6 f 0.3 (D (0 (D

(a)Number surviving/number initially in group

(b)Initial mean group body weight f standard error of the mean. Subsequent calculations are based on those animals

surviving to the end of the study.

(e)Mean body weight change of the survivors f standard error of the mean
(d)Day of d e a t h 2,2,2,3
(e)Day of death: all 2

(0No data are reported because of the 100% mortality in this group.

2-Mercaptobenzothiazole, NTP TR 332 46

III. RESULTS: MICE

THIRTEEN-WEEK STUDIES Lethargy and rough coats were observed in the

375 and-750 mg/kg groups. No compound-rela-
Five of 10 males and 7/10 females that received
ted gross or microscopic pathologic effects were
1,500mgkg died before the end of the studies
observed.
(Table 22). Two of the deaths were related to ga-
vage technique. Chemical administration did
not affect body weight gain. Liver weight to Dose Selection Rationale: Because of the deaths
body weight ratios of dosed groups were higher observed at 1,500 mg/kg, doses selected for mice
than those of the vehicle controls (Table 23). for the 2-year studies were 375 and 750 mg/kg 2-
Clonic seizures, lacrimation, and salivation were mercaptobenzothiazole, administered in corn oil
observed in the 750 and 1,500 mg/kg groups. by gavage, 5 days per week.

TABLE 22. SURVIVAL AND MEAN BODY WEIGHTS O F MICE IN THE THIRTEEN-WEEK GAVAGE

STUDIES O F 2-MERCAPTOBENZOTHIAZOLE

Mean Body Weights (grams) Final Weight Relative

Dose Survival (a) Initial (b) Final Change (c) to Vehicle Controls

(mgfltg) (percent)

MALE

0 10110 27.1 f 0.3 36.7 f 0.9 C9.6 f 0.7 __

94 loll0 25.8 f 0.4 37.0 f 0.8 + 11.2 f 0.7 100.8

188 10110 26.9 f 0.3 37.7 f 1.0 +10.8 -I 0.8 102.7

375 10/10 25.9 f 0.5 35.1 f 1.1 +9.2 k 0.9 95.6

750 10110 26.1 f 0.5 34.4 f 0.6 +8.3 f 0.3 93.7

1,500 (d) 5/10 26.7 f 0.4 35.2 f 1.3 +8.5 f 0.5 95.9
FEMALE

0 loll0 20.6 f 0.3 26.2 f 0.4 +5.6 f 0.4 __

94 10/10 20.4f 0.4 25.5 f 0.4 +5.1 f 0.3 97.3

188 loll0 20.3 f 0.4 25.9 k 0.6 +5.6 f 0.2 98.9

375 10110 20.0 f 0.3 25.8 f 0.4 +5.8 f 0.2 98.5

750 (e) 8/10 20.5 f 0.2 26.1 ?: 0.4 +5.5 f 0.4 99.6

1,500 (D 3/10 20.1 f 0.4 25.3 f 0.2 +4.6 f 0.4 96.6

(a)Number surviving/number initially in group

(b)Initial mean group body weight f standard error of the mean. Subsequent calculations are based on those animals

surviving to the end of the study.

(c) Mean body weight change of the survivors f standard error of the mean
(d)Week ofdeath: 1,2,3,4,6
(e)Week of death: 7,8

(0Week ofdeath: 1,1,1,1,6,8,10

47 2-Mercaptobenzothiazole, NTP TR 332

TABLE 23. ANALYSIS OF LIVER WEIGHTS FOR MICE IN THE THIRTEEN-WEEK GAVAGE

STUDIES OF 2-MERCAPTOBENZOTHIAZOLE (a)

Dose No. Final Mean Liver Weight Liver WeighUFinal

(mg/kg) Examined Body Weight (grams) (mg) Body Weight (mg/g)

MALE

0 f 2.8 1,821 f f 4.34

94
188
10
10
10
36.7
37.0
37.7
f 2.6
f 3.1
1,942*
2,034 f
213
208
184
49.6
52.5
(b)54.0
f 4.02
f 3.23
375 10 35.1 f 3.4 i,a55* 231 52.8 f 3.51
750 10 34.4 f 2.0 1,809 k ii5 52.6 f 3.15
1,500 5 35.2 f 2.8 2,090 184 (c) 59.5 f 3.94
FEMALE
0 10 26.2 f 1.3 1,129 f 242 42.9 f 7.71
94 10 25.5 f 1.3 1,237 f 123 48.6 5.03
1a8 10 25.9 f 1.7 1,238 f 113 47.9 f 3.61
375 10 25.8 f 1.3 1,232f 124 47.8 k 3.74
750 a 26.1 f 1.3 1,281 f 126 49.2 f 4.70
1,500 3 25.3 f 0.3 1,383 f 96 (d54.7 f 3.45

(a)Mean f standard deviation; Pvalues are versus the controls by Dunnett’s test (Dunnett, 1955).
(b)P<0.05
(c) P<O.Ol

TWO-YEAR STUDIES 4%-8% lower than those of the vehicle controls

from week 6 to week 64. Mean body weights of
Body Weights and Clinical Signs high dose female mice were within 6% of those of
the vehicle controls throughout the studies.
Mean body weights of high dose male mice were Mean body weights of low dose female mice were
6%-14% lower than those of the vehicle controls generally greater than those of the vehicle con-
from week 3 to week 64 (Table 24 and Figure 7). trols throughout the studies. Mice were lethar-
Mean body weights of low dose male mice were gic after they were dosed.

2-Mercaptobenzothiazole,NTP TR 332 48
TABLE 24. MEAN BODY WEIGHTS AND SURVIVAL OF MICE IN THE TWO-YEAR GAVAGE STUDIES
OF 2-MERCAPTOBENZOTHIAZOLE

Weeks Vehicle Control 375 mg/kg 750 m a g

on Av. Wt. No. of Av. Wt. Wt. (percent of No. of Av. Wt. Wt. (percent of No. of
Study (grams) Survivors (grams) veh. controls) Survivors (grams) veh. controls) Survivors

MALE
0 24.7 50 24.2 98 50 24.7 100 50
1 26.0 50 24.4 94 50 25.5 98 50
2 27.3 50 26.4 97 49 28.4 97 50
3 28.8 50 27.0 97 49 27.0 94 50
4 29.4 50 28.7 98 49 27.8 94 50
5 30.8 50 29.1 97 49 20.2 92 50
0 31.5 50 29.9 95 49 29.1 92 50
7 32.3 50 30.4 94 49 29.0 92 50
8 33.0 50 30.3 92 49 29.5 89 48
9 33.3 50 31.3 94 49 30.1 90 40
10 33.4 so 32.1 98 49 30.8 92 47
11 34.4 50 32.0 95 49 30.9 90 47
12 35.3 50 33.7 95 49 31.7 90 47
10 37.3 50 35.2 94 49 32.9 88 40
21 38.9 50 37.1 95 49 34.3 88 38
25 40.9 so 38.5 94 49 35.8 08 38
29 41.9 50 39.0 93 49 38.0 88 35
33 43.5 50 39.9 92 49 37.7 87 35
37 43.8 50 40.1 93 49 38.8 88 35
42 45.2 so 42.5 94 49 39.3 87 35
48 48.1 50 43.1 93 49 40.9 89 35
51 47.4 50 44.3 94 49 42.6 90 34
55 48.2 49 45.9 95 49 43.9 91 34
59 47.9 49 45.9 98 49 44.3 92 34
64 48.9 49 40.1 96 49 45.8 94 33
68 49.1 48 48.1 98 48 48.8 95 33
72 49.4 48 48.5 98 47 47.1 95 32
77 48.5 47 47.1 97 45 48.4 98 32
81 49.0 47 48.4 99 43 41.5 97 32
85 47.9 45 48.0 100 42 40.8 97 32
90 48.3 42 49.0 101 38 48.4 100 30
94 48.6 40 48.9 101 37 48.9 101 30
98 48.0 39 47.0 99 37 48.8 102 30
102 47.1 38 48.2 102 33 48.7 103 30

FEMALE
0 20.8 50 20.5 99 50 20.9 100 50
1 20.8 50 20.4 90 50 21.1 101 50
2 21.8 50 21.8 101 50 21.8 101 50
3 22.2 50 22.4 101 50 22.4 101 49
4 23.3 50 23.2 100 50 23.0 99 49
5 23.7 50 23.5 99 J0 22.9 97 49
8 24.0 50 23.7 99 50 23.9 100 49
7 23.9 50 24.1 101 50 24.1 101 49
8 24.0 50 24.5 102 50 24.2 101 49
9 24.4 50 24.4 100 50 24.8 101 49
10 25.2 50 24.9 99 50 24.9 99 49
11 28.1 50 25.8 99 50 25.7 98 45
12 28.8 50 28.0 98 50 20.0 98 49
10 27.7 50 27.6 100 50 27.0 97 44
21 29.1 50 28.4 98 50 28.1 91 40
25 30.8 50 30.3 99 50 29.5 90 40
29 30.3 50 31.0 102 50 30.2 100 39
33 31.1 50 31.8 102 50 30.9 99 39
37 32.0 50 33.0 101 50 32.2 99 39
42 34.8 50 33.3 98 50 32.8 94 39
46 35.6 50 34.8 97 50 33.4 94 37
51 37.4 50 38.7 98 50 35.0 94 35
55 39.4 50 38.0 98 50 37.2 94 34
59 39.9 49 39.3 98 50 38.9 91 33
e4 41.3 49 41.1 100 50 40.3 98 31
88 42.5 49 42.3 100 50 40.5 95 31
72 43.3 48 43.2 100 50 41.5 98 31
77 42.8 48 43.5 102 49 41.9 98 29
81 43.3 48 44.9 104 48 42.7 99 27
85 44.0 45 45.2 103 46 42.4 98 27
W 45.4 43 48.7 103 45 43.5 90 27
94 45.3 41 47.3 104 45 44.1 91 27
98 44.5 38 48.2 104 43 43.4 98 25
lo2 44.8 37 46.1 104 40 40.0 104 22

49 2-Mercaptobenzothiaaole,NTP TR 332
- '

j .

I
45.0 .............................. & ...........................
.......... a.... ...........................

I 45.0

40.0

35.0
35.0

30.0 30.0

MALE MICE
25.0 ............................
.....................................................................................
o x 575 MG/KG .................-

VEHICLE
25.0
A = 7 5 0 MG/KG

20.0 0 4-
4 - 15 20.0

50.0 50.0
i
Ij
4 5 0 ..................... I" .......&i ........................... 45.0
j
j
i e
!
40.0 ............................... I ..............

I
.......................................................

:
...............................
i ............

"
40.0

I .. 6~ i
...............
*
n

0
m
Z
35 0

30.0
.1 I
....................................................... "

B 8 BA
I
............................................

!
.......... ........."........-......................................................
1
..............

........................................................

35.0

30.0
4
W i

-
I

I
.......... ....................................................................... =VEHICLE ..............

1 /
25.0 ".I
25.0

20.0 20.0
0 lk 43 60 1 io 1
WEEKS ON STUOY

FIGURE 7, GROWTH CURVES FOR MICE ADMINISTERED

2-MERCAPTOBENZOTHIAZOLE IN CORN OIL BY GAVAGE FOR TWO YEARS

2-Mercaptobenzothiamle,NTP TR 332 50
III. RESULTS: MICE

Survival Lesions in male mice are summarized in Appen-

dix C. Histopathologic findings on neoplasms
are summarized in Table C l . Table C2 gives the
Estimates of the probabilities of survival for
survival and tumor status for individual male
male and female mice administered 2-mercapto-
mice. Table C3 contains the statistical analyses
benzothiazole a t the doses used in these studies
of those primary tumors that occurred with an
and for vehicle controls are shown in Table 25
incidence of a t least 5% in one of the three
and in the Kaplan and Meier curves in Figure 8.
groups. The statistical analyses used are dis-
Survival of the high dose group of female mice
was significantly lower than that of the vehicle cussed in Chapter II (Statistical Methods) and
Table C3 (footnotes). Findings on nonneoplastic
controls after week 27. Six high dose male and
lesions are summarized in Table C4.
four high dose female mice died on the same day
during week 13. Since they were mistakenly Lesions in female mice are summarized in Ap-
dosed twice within a 16-hour period, these mice pendix D. Histopathologic findings on neo-
were censored from the statistical incidence of plasms are summarized in Table D1. Table D2
survival after week 12. gives the survival and tumor status for individ-
ual female mice. Table D3 contains the statisti-
cal analyses of those primary tumors that oc-
Pathology and Statistical Analyses of
curred with an incidence of at least 5% in one of
Results
the three groups. The statistical analyses used
are discussed in Chapter II (Statistical Methods)
This section describes the significant or note- and Table D3 (footnotes). Historical incidences
worthy changes in the incidences of mice with of tumors in corn oil vehicle control female mice
neoplastic or nonneoplast,ic lesions of the liver, are listed in Table D4. Findings on nonneo-
pituitary gland, hematopoietic system, and lung. plastic lesions are summarized in Table D5.

TABLE 25. SURVIVAL OF MICE IN THE TWO-YEAR GAVAGE STUDIES OF

2-MERC APTOBENZOTHIAZOLE

Vehicle Control 375 mg/kg 750 mg/kg

MALE (a)
Animals initially in study 50 50 50
Nonaccidental deaths before termination (b) 11 17 14
Animals missing 1 0 0
Accidentally killed 0 0 6
Killed at termination 38 33 30
Survival P values (c) 0.204 0.262 0.254
FEMALE (a)
Animals initially in study 50 50 50
Nonaccidental deaths before termination (b) 13 10 24
Animals missing 0 1 0
Accidentally killed 0 0 4
Killed at termination 35 39 22
Died during termination period 2 0 0
Survival P values (c) 0.002 0.560 0.005

(a)Terminal-kill period: male, week 103; female, weeks 103-104

(b)Includes animals killed in a moribund condition
(c)The result of the life table trend test is in the vehicle control column, and the results of the life table pairwise comparisons
with the vehicle controls are in the dosed columns.

51 2-Mercaptobenzothiazole, NTP TR 332

1.0 1.0

0.9 0.9

J
4
L 0.8 0.8
>
a
3
v)
LL
0 0.7
0.7
cr
2
m
4
m 0.6 0.6
0
a

0.5 ............. p
A=

0.5

1 1 i I
0.4 I I I 1 0.4
30 4s 60 75 90
WEEKS ON STUDY

1.0 1.0

--t
0.9 ..........................
L.I
................................................................................ .........................

............
4
0.9

-J
4
L
>
K

3
v)
LL

0
*
k
J
0.8

0.7
..........................

..........................
..............................................................................

~ -
........................................................

. -
I
~ ~ . ~ t

..... .........a

0. 8

0.7

4
Lk.--.............
2-
m 0.6 ..................................................................................... .........................
0.6
0 I
K

0.5 .............. ............ .................. 0.5

0.4 -M- so 45

WEEKS ON S1 DY
! 1
0.4

FIGURE 8. KAPLAN-MEIER SURVIVAL CURVES FOR MICE ADMINISTERED

2-MERCAPTOBENZOTHIAZOLE IN CORN OIL BY GAVAGE FOR TWO YEARS

2-Mercaptobenzothiazole,NTP TR 332 52
III. RESULTS: MICE

Liver: The incidence of hepatocellular adenomas adenomas or carcinomas (combined) were seen
or carcinomas (combined) in low dose female in 16/49 vehicle control, 21/50 low dose, and
mice was significantly greater than that in the 14/50high dose male mice.
vehicle controls (Table 26). Hepatocellular

TABLE 26. ANALYSIS O F HEPATOCELLULAR TUMORS IN FEMALE MICE IN THE TWO-YEAR

GAVAGE STUDY O F 2-MERCAPTOBENZOTHIAZOLE (a)

Vehicle Control 375 mg/kg 750 mg/kg

Adenoma
Overall Rates 3/50 (6%) 7/49 (14%) 4/50 (8%)

Adjusted Rates 8.1% 17.9% 18.2%

Terminal Rates 3/37 (8%) 7/39(18%) 4/22 (18%)

Week of First Observation 103 103 103

Life Table Tests P = 0.159 P=0.178 P =0.231

Incidental Tumor Tests P = 0.159 P = 0.178 P = 0.23 1

Carcinoma

Overall Rates 1/50 (2%) 5/49 (10%) 0150 (0%)

Adjusted Rates 2.7% 12.2% 0.0%

Terminal Rates 1/37 (3%) 4/39 (10%) 0122 (0%)

Week of First Observation 103 89

Life Table Tests P = 0.590N P=0.116 P = 0.604N

Incidental Tumor Tests P = 0.552 P = 0.088 P = 0.604N

Adenoma or Carcinoma (b)

Overall Rates 4/50 (8%) 12/49 (24%) 4/50 (8%)

Adjusted Rates 10.8% 29.8% 18.2%

Terminal Rates 4/37 (11%) 11/39 (28%) 4/22 (18%)

Week of First Observation 103 89 103

Life Table Tests P=0.204 P = 0.035 P=O.343

Incidental Tumor Tests P=0.171 P = 0.028 P=0.343

(a)The statistical analyses used are discussed in Chapter II (Statistical Methods) and Appendix D, Table D3 (footnotes).
(b)Historical incidence in NTP studies (mean f SD): 116/1,489(8% k 6%)

53 2-Mercaptobenzothiazole, NTP TR 332

III. RESULTS: MICE

Pituitary Gland: Adenomas and adenomas or significantly lower than that in the vehicle con-
carcinomas (combined) in female mice occurred trols (Table 28).
with significant negative trends, and the inci-
Lung: The incidence of bronchopneumonia in all
dences in the dosed groups were significantly
groups of mice varied from 24% to 49% (male:
lower than those in the vehicle controls (Ta-
vehicle control, 12/49; low dose, 16/50; high dose,
ble 27).
16/50; female: 13/50; 24/49; 18/50). These le-
sions were of minimal to mild severity and con-
Hematopoietic System: Lymphomas in female sistent with those changes seen with viral infec-
mice occurred with a significant negative trend, tions. Serologic titers from sentinel animals
and the incidence in the low dose group was were positive for Sendai virus antibody.

TABLE 27. ANALYSIS OF PITUITARY GLAND LESIONS I N FEMALE MICE I N T H E TWO-YEAR

GAVAGE STUDY O F 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 mg/kg 750 mg/kg

Hyperplasia

Overall Rates 16/49 (33%) 14/49 (29%) 12/49 (24%)

Adenoma

Overall Rates 20149 (41%) 11/49 (22%) 3/49 (6%)

Adjusted Rates 51.1% 26.4% 12.5%

Terminal Rates 18/37 (49%) 9/39 (23%) 2/22 (9%)

Week of First Observation 92 87 94

Life Table Testa P=O.O02N P= 0.028N P = 0.004N

Incidental Tumor Testa P = 0.001N P = 0.035N P=0.003N

Carcinoma

Overall Rates 1/49 (2%) 0149 (0%) 0149 (0%)

Adenoma o r Carcinoma (a)

Overall Rates 21/49 (43%) 11/49 (22%) 3/49 (6%)

Adjusted Rates 52.1% 26.4% 12.5%

Terminal Rates 18/37 (49%) 9/39 (23%) 2/22 (9%)

Week of First Observation 71 87 94

Life Table Tests P <0.001N P = 0.019N P=0.003N

Incidental Tumor Testa P <0.00 1N P=0.024N P=0.001N

(a)Historical incidence of adenomas or carcinomas (combined) in NTP studies (mean k SD): 257/1,324 (19% f 9%)

TABLE 28. ANALYSIS OF HEMATOPOIETIC SYSTEM TUMORS I N FEMALE MICE IN T H E TWO-YEAR

GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 mg/kg 750 mg/kg

Malignant Lymphoma (a)

Overall Rates 19/50 (38%) 10149 (20%) 6/50 (12%)

Adjusted Rates 48.5% 23.2% 25.3%

Terminal Rates 17/37 (46%) 7/39 (18%) 5/22 (23%)

Week of First Observation 89 72 75

Life Table Testa P = 0.032N P=0.028N P=0.076N

Incidental Tumor Testa P=0.016N P=0.035N P = 0.057N

(a)Historical incidence of lymphomas or leukemia in NTP studies (mean f SD): 39311,494 (26% k 9%)

2-Mercaptobenzothiazole, NTP TR 332 54

IV. DISCUSSION AND CONCLUSIONS

55 2-Mercaptobenzothiazole,NTP TR 332

IV. DISCUSSION AND CONCLUSIONS

2-Mercaptobenzothiazole is used in automobile toxic doses of 2-mercaptobenzothiazole caused

tire production as a n accelerant for the rubber neurotoxicity (Johnson et al., 1970) and hepatox-
vulcanization process and as a preservative for icity (Guess and O’Leary, 1969; Litvinchuk,
textile or cordage materials (Santadonato et al., 1963; Vorob’eva and Mezentsera, 1968), there
1976). The chemical is also contained in rubber was no evidence from the present studies that
medical devices and in baby bottle nipples, and long-term exposure to 2-mercaptobenzothiazole
it can leach into aqueous media (Petersen et al., caused similar nonneoplastic lesions. Dosed
1981; Blosczyk and Doemling, 1982). Toxicity mice had some clinical signs of neurotoxicity
and carcinogenicity studies of 2-mercaptobenzo- characterized as postgavage lethargy (at 375
thiazole were conducted by the NTP because of and 750 mg/kg) and seizures (at 750 and 1,500
the high production volume (USITC, 19851, po- mg/kg) in the 13-week studies and as postgavage
tential human exposure, and use of the salts of 2- lethargy in rats and mice in the 2-year studies.
mercaptobenzothiazole a s fungicides and bac- Examination of tissues from the nervous system
tericides (Foltinova and Bloeckinger, 1970). did not reveal lesions that were attributable to
chemical administration.
There was no indication from the short-term
studies that the doses used in the 2-year studies Distribution studies after dermal application in-
would adversely affect survival of the rats. The dicated that the thyroid gland, liver, and kidney
dose selections were based on minimal toxic re- were the principal organs that accumulated 2-
sponses in the 13-week studies: minor decreases mercaptobenzothiazole ( N a g a m a t s u e t a l . ,
in body weight gain, small increases in liver 1979). In the present gavage studies, there was
weight to body weight ratios, and limited clin- no evidence of lesions in the thyroid gland,
ical observations. Despite this conservative ap- where neoplastic and nonneoplastic responses to
proach, the 2-mercaptobenzothiazole doses se- chemicals containing sulfur have most often oc-
lected proved to be toxic for both dose groups of curred (NCI, 1978a, 1979).
male rats and for high dose female mice, al- Although a variety of neoplasms occurred in rats
though survival a t 90 weeks ranged between dosed with 2-mercaptobenzothiazole, their inci-
70% and 100% for all dosed groups of rats. A re- dences were not always dose related. For ex-
view of the individual animal records indicated ample, the incidences of mononuclear cell leuke-
that tumors were observed in most of the rats mia and pancreatic acinar cell adenomas in male
that died before study termination. Lung hem- rats were increased only in the low dose groups.
orrhage and congestion were associated with Comparable numbers of male rats were a t risk
most of the mice that died early, and there was a at the end of the study (22 low dose and 20 high
consistent lack of tumors in these animals. dose), so it is doubtful that survival rates af-
However, final survival rates in these groups fected the dose-response relationship for neo-
were 40%-50%, so a sufficient number of animals plasms. Examples of neoplasms with dose-re-
remained at risk to permit determination of the lated trends included pituitary gland adenomas
presence or absence of carcinogenicity. in female rats and adrenal gland pheochromocy-
tomas in each sex of rats. These responses sug-
There was a documented incident of unusual gested that 2-mercaptobenzothiazole expressed
mortality in mice mistakenly dosed twice within some carcinogenic activity in rats a t doses suffi-
a 16-hour period during week 13 of the 2-year cient to accelerate mortality.
studies. These animals were censored from the
statistical analysis of survival after week 12; There was equivocal evidence for the carcino-
death may have been associated with the nar- genicity of 2-mercaptobenzothiazole in female
cotic effect of the bolus doses given at short mice as shown by an increased incidence of hepa-
intervals. tocellular adenomas or carcinomas (combined)
in the low dose group. It is possible that low sur-
The principal nonneoplastic lesions seen in these vival in the high dose group of female mice pre-
studies were nephropathy and inflammation and vented the expression of hepatocellular tumori-
ulceration of the forestomach in rats. Although genicity, since this is a late-appearing neoplasm
in earlier studies acute or short-term exposure to in mice.

2-Mercaptobenzothiazole, NTP TR 332 56

IV. DISCUSSION AND CONCLUSIONS

Some of the tumor responses to 2-mercapto- studies, even higher rates of mortality occurred
benzothiazole were comparable to those induced in each of the earlier studies, such that there
by other sulfur-containing chemicals in studies was either early termination of the studies or
evaluated by the NC1 (Griesemer and Cueto, early withdrawal of chemical exposure.
1980) and the NTP (Huff, 1982; Haseman et al.,
Metabolism studies in F344 rats indicated that
1984). Of these chemicals, 2-mercaptobenzothi-
the half-life for 2-mercaptobenzothiazole after
azole has the closest structural resemblance to
administration by gavage was less than 8 hours
4,4'-thiodianiline, When administered in feed,
and possibly as short as 4-6 hours (CMA, 1986a).
4,4'-thiodianiline caused hepatocellular carcino-
Absorption was rapid and unaffected by doses up
mas in male rats and in male and female mice
to 55 mg/kg. The major products of metabolism
(NCI, 1978a), whereas 2-mercaptobenzothiazole
were polar metabolites, a finding in agreement
induced hepatocellular adenomas i n female
with those from earlier dermal absorption stud-
mice. Other responses to thio chemicals com-
ies (Colucci and Buyske, 1965; Nagamatsu et al.,
parable to those induced by 2-mercaptobenzo-
1979) in which glucuronide and sulfate conju-
thiazole in the present studies included leuke-
gates of various proposed metabolites were dem-
mia in male rats induced by intraperitoneal
onstrated. In the CMA gavage study (1986a), 2-
injection of thio-TEPA (NCI, 1978b) and fore-
mercaptobenzothiazole-derived radioactivity in
stomach neoplasms and nephropathy in male
blood decreased very little between 24 and 96
rats after the administration of sulfallate in feed
hours, suggesting that residual 2-mercaptoben-
(NCI, 1978~).
zothiazole-derived material accumulated i n
blood; no data were available for other tissues, so
Neoplasms of the thyroid gland occurred in ani-
the potential accumulation of 2-mercaptobenzo-
mals dosed with the other thio compounds but
thiazole after long-term exposure is unknown.
not after 2-mercaptobenzothiazole exposure.
In a companion study, radiolabeled 2-mercapto-
Thioacetamide, thiourea, and thiouracil a r e
benzothiazole was administered intravenously
structurally similar to 2-mercaptobenzothiazole
to F344 r a t s (CMA, 198613). Whole blood,
and cause neoplasms of the thyroid gland and
plasma, urine, and feces were analyzed for radio-
sometimes the liver (Weisburger and Williams,
activity a t 5 and 15 minutes and a t 1, 2, 4, 24,
1980). For example, NJ'-diethylthiourea,
and 72 hours. Most of the radioactivity (91%-
which is structurally similar to the carcinogen
101%) was excreted in the urine and 4%-8% was
ethylene thiourea (IARC, 1974), caused thyroid
excreted in the feces by 72 hours. A small
gland tumors in rats of each sex when adminis-
amount (1.5%-2%) of the radioactivity remained
tered in feed (NCI, 1979). The mechanism of ac-
in the erythrocytes. The metabolites found in
tion was hypothesized to be interference with
the urine samples were the same as those found
thyroxine synthesis and subsequent stimulation
in the gavage study (CMA, 1986a).
of the pituitary gland-thyroid gland axis, caus-
ing enhanced secretion of thyrotropic hormone 2-Mercaptobenzothiazole was clearly clastogenic
and possible neoplasia of the thyroid gland. Pos- to cultured Chinese hamster ovary (CHO) cells
sible explanations for the lack of thyroid gland in the presence of S9 enzymes, inducing aberra-
tumor expression by 2-mercaptobenzothiazole tions at frequencies comparable to and even ex-
are the different route of administration or the ceeding those of the positive control chemical cy-
comparatively lower doses used in the present clophosphamide (Appendix E, Table E4). It also
studies. In the earlier studies, the thio chem- induced sister chromatid exchanges in CHO
icals were all given ad libitum in feed except for cells (Table E3) and thymidine kinase mutants
thio-TEPA, which was injected intraperitoneally in mouse L5178Y lymphoma cells in the pres-
three times per week. 4,4'-Thiodianiline, sulfal- ence of S9 (Table E2). In mouse lymphoma as-
late, and thio-TEPA were administered a t con- says, the frequency of thymidine kinase mutants
centrations high enough to affect the thyroid also was increased in the absence of $39 but only
gland, whereas this organ apparently was not af- a t toxic doses (Litton, 1985). Under these condi-
fected by 2-mercaptobenzothiazole administered tions, some of the mutant colonies produced were
by gavage a t lower concentrations. Although of small size, suggesting that 2-mercaptobenzo-
there was significant mortality in the present thiazole is capable of inducing chromosomal

57 2-Mercaptobenzothiazole, NTP TR 332

IV. DISCUSSION AND CONCLUSIONS

aberrations in this cell line as well. Although 2- Conclusions: Under the conditions of these 2-
mercaptobenzothiazole is clastogenic in vitro, year gavage studies, there was some evidence of
the only reported study for in vivo mutagenicity, carcinogenic activity* of 2-mercaptobenzothia-
a mouse bone marrow micronucleus test, did not zole for male F344/N rats, indicated by increased
show a n increase in the frequency of micronucle- incidences of mononuclear cell leukemia, pan-
ated polychromatic erythrocytes in these cells creatic acinar cell adenomas, adrenal gland
(Pharmakon, 1984). pheochromocytomas, and preputial gland adeno-
mas or carcinomas (combined). There was some
The experimental and tabulated data for the evidence of carcinogenic activity for female
NTP Technical Report on 2-mercaptobenzothia- F344/N rats, indicated by increased incidences of
zole were examined for accuracy, consistency, adrenal gland pheochromocytomas and pituitary
completeness, and compliance with Good Lab- gland adenomas. There was no evidence of car-
oratory Practice regulations. As summarized in cinogenic activity of 2-mercaptobenzothiazole for
Appendix H, the audit revealed no major prob- male B6C3F1 mice dosed with 375 or 750 mg/kg.
lems with the conduct of the studies or with col- There was equivocal evidence of carcinogenic ac-
lection and documentation of the experimental tivity for female B6C3F1 mice, indicated by in-
data. No discrepancies were found that influ- creased incidences of hepatocellular adenomas
enced the final interpretation of the results of or carcinomas (combined).
these studies.

*Explanation of Levels of Evidence of Carcinogenic Activity is on page 8.

A summary of the Peer Review commenta and the public discuseion on this Technical Report appears on pages 11-12,

2-Mercaptobenzothiazole,NTP TR 332 58
V. REFERENCES

59 2-Mercaptobenzothiazole, NTP TR 332

V. REFERENCES

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Eksp. Biol. Med. 93:87-88. and characterization of the L5178Y/TK+’-
mouse lymphoma mutagen assay system. Mu-
2. Armitage, P. (1971) Statistical Methods in tat. Res. 59:61-108.
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transformation of a sulfonamide to a mercaptan
3. Bauer, M. (1972) The environment and the and to mercapturic acid and glucuronide conju-
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sulfide-ring-UL-14 C in Fischer 344 male and fe- 17. Fisher, A.A. (1977) Allergen replacements in
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8. Chemical Manufacturers Association (CMA) 18. Foltinova, P.; Bloeckinger, G. (1970) Anti-
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ring-UL-14 C and 2-mercaptobenzothiazole di- its derivatives. Biologia (Bratislava) 25: 175-
sulfide-ring-UL-14 C in Fischer 344 male and fe- 180.
male rats dosed intravenously. Washington,
DC: Chemical Manufacturers Association. 19. Galloway, S.M.; Bloom, A.D.; Resnick, M.;
Margolin, B.H.; Nakamura, F.; Archer, P.;
9. Chiba, T. (1969) Effect of sulfur-containing Zeiger, E. (1985) Development of a standard pro-
compounds on experimental diabetes. VI. tocol for in vitro cytogenetic testing with Chi-
Screening for hypoglycemic action of sulfur-con- nese hamster ovary cells: Comparison of results
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20. Gart, J.J.; Chu, K.C.; Tarone, R.E. (1979) 29. Haseman, J.K.; Huff, J.; Rao, G.N.; Arnold,
Statistical issues in interpretation of chronic J.; Boorman, G.A.; McConnell, E.E. (1985) Neo-
bioassay tests for carcinogenicity. J. Natl. plasms observed in untreated and corn oil ga-
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63 2-Mercaptobenzothiazole,NTP TR 332
2-Mercaptobenaothiazole,NTP TR 332 64
APPENDIX A

SUMMARY OF LESIONS IN MALE RATS IN THE

TWO-YEAR GAVAGE STUDY OF

2-MERCAPTOBENZOTHIAZOLE

PAGE

TABLE A l SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE

TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 67

TABLE A2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR

GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 70

TABLE A3 ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR GAVAGE

STUDY OF 2-MERCAPTOBENZOTHIAZOLE 76

TABLE A4a HISTORICAL INCIDENCE OF LEUKEMIA IN MALE F344/N RATS ADMINISTERED

CORN OIL BY GAVAGE 80

TABLE A4b HISTORICAL INCIDENCE OF PITUITARY GLAND TUMORS IN MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE 80

TABLE A4c HISTORICAL INCIDENCE OF ADRENAL GLAND TUMORS IN MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE 81

TABLE A4d HISTORICAL INCIDENCE OF PANCREATIC ACINAR CELL TUMORS IN MALE

F344/N RATS ADMINISTERED CORN OIL BY GAVAGE 81

TABLE A4e HISTORICAL INCIDENCE OF PREPUTIAL GLAND TUMORS IN MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE 82

TABLE A4f HISTORICAL INCIDENCE OF SUBCUTANEOUS TISSUE TUMORS IN MALE

F344/N RATS ADMINISTERED CORN OIL BY GAVAGE 82

TABLE A4g HISTORICAL INCIDENCE OF MESOTHELIAL TUMORS IN MALE F344/N RATS

ADMINISTERED CORN OIL BY GAVAGE 83

TABLE A4h HISTORICAL INCIDENCE OF KIDNEY TUMORS IN MALE F344/N RATS

ADMINISTERED CORN OIL BY GAVAGE 83

TABLE A5 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS

IN THE TWO-YEAR GAVAGE STUDY OF SMERCAPTOBENZOTHIAZOLE 84

65 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole,NTP TR 332 66
TABLE A l . SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS NECROPSIED 50 50 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 50 50 50

INTEGUMENTARY SYSTEM
+Skin
(50) (50)
Squamous cell papilloma
2 (4%) 2 (4%)
Squamous cell carcinoma
1 (2%)
Basal cell tumor

Keratoacanthoma
2 (4%) 1 (2%)
*Subcutaneoustissue (50) (50)
Sarcoma, NOS 1 (2%) 1 (2%)
Fibroma 3 (6%) 6 (12%)
Fibrosarcoma 1 (2%)
Fibrous histiocytoma, malignant 1 (2%)
Lipoma
Neurofibroma 1 (2%)

RESPIRATORY SYSTEM
#Lung (50) (50) (50)
Alveolar/bronchiolar adenoma 2 (4%) 1 (2%)
Alveolar/bronchiolar carcinoma 1 (2%) 1 (2%)
C-cell carcinoma, metastatic 1 (2%)
Mucinous adenocarcinoma 1 (2%)
Pheochromocytoma, metastatic 1 (2%)

HEMATOPOIETIC SYSTEM
*Multiple organs (50) (50) (50)
Leukemia, mononuclear cell 7 (14%) 16 (32%) 3 (6%)
#Spleen (50) (50) (49)
Sarcoma, NOS 1 (2%)
#Thymus (50) (49) (48)
Thymoma, benign 1 (2%)

CIRCULATORY SYSTEM
#Spleen (50) (50) (49)
Hemangiosarcoma 1 (2%)
#Heart (50) (50) (50)
Pheochromocytoma, metastatic 1 (2%)
Neurilemoma, malignant 1 (2%)

DIGESTIVE SYSTEM
#Liver (50) (50) (50)
Neoplastic nodule 3 (6%) 2 (4%) 1 (2%)
Mixed hepato/cholangio carcinoma 1 (2%)
#Pancreas
(50) (50) (49)
Acinar cell adenoma
2 (4%) 13 (26%) 6 (12%)
#Duodenum
(50) (50) (49)
Leiomyosarcoma
1 (2%)

67 2-Mercaptobenzothiazole,NTP TR 332
TABLE A l . SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

URINARY SYSTEM
#Kidney (50) (50) (49)
Transitional cell carcinoma 1 (2%)
Tubular cell adenoma 1 (2%) 1 (2%)
#Kidney/pelvis (50) (50) (49)
Transitional cell papilloma 1 (2%) 1 (2%)
Leiomyosarcoma 1 (2%)

ENDOCRINE SYSTEM
#Pituitary intermedia (50) (50) (48)
Adenoma, NOS 1 (2%)
#Anterior pituitary (50) (50) (48)
Adenoma, NOS 14 (28%) 21 (42%) 12 (25%)
#Adrenal (50) (50) (49)
Cortical adenoma 1 (2%) 1 (2%)
#Adrenal medulla (50) (50) (49)
Pheochromocytoma 18 (36%) 25 (50%) 22 (45%)
Pheochromocytoma, malignant 2 (4%) 2 (4%)
#Thyroid (50) (50) (50)
Follicular cell adenoma 1 (2%)
Follicular cell carcinoma 1 (2%) 1 (2%)
C-cell adenoma 2 (4%) 3 (6%) 1 (2%)
C-cell carcinoma 5 (10%) 2 (4%)
#Pancreatic islets (50) (50) (49)
Islet cell adenoma 4 (8%) 2 (4%) 1 (2%)
Islet cell carcinoma 1 (2%) 1 (2%)

REPRODUCTIVE SYSTEM
*Mammary gland (50) (50) (50)
Fibroadenoma 2 (4%) 1 (2%) 1 (2%)
*Preputial gland (50) (50) (50)
Carcinoma, NOS 1 (2%) 2 (4%) 1 (2%)
Adenoma, NOS 4 (8%) 4 (8%)
#Testis (50) (50) (50)
Interstitial cell tumor 48 (96%) 48 (96%) 48 (96%)
Pheochromocytoma, metastatic 1 (2%)
#Tunica albuginea (50) (50) (50)
Mesothelioma, NOS 1 (2%) 1 (2%)

NERVOUS SYSTEM
#Brain (50) (50)
Astrocytoma 1 (2%)
Oligodendroglioma 1 (2%)

SPECIAL SENSE ORGANS

*Zymbal gland (50) (50) (50)
Carcinoma, NOS 1 (2%) 1 (2%)
Squamous cell carcinoma 1 (2%)

MUSCULOSKELETAL SYSTEM
None

BODY CAVITIES
*Mesentery (50) (50) (50)
Pheochromocytoma, invasive 1 (2%)

2-Mercaptobenzothiazole, NTP TR 332 68

TABLE Al. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ALL OTHER SYSTEMS

*Multiple organs (50) (50) (50)
Sarcoma, NOS 1 (2%)
Mesothelioma, malignant 1 (2%) 2 (4%)

ANIMAL DISPOSITION SUMMARY

Animals initially in study 50 50 50
Natural death 2 2 5
Moribund sacrifice 6 26 24
Terminal sacrifice 42 22 20
Dosing accident 1

TUMOR SUMMARY
Total animals with primary tumors** 49 50 48
Total primary tumors 123 169 125
Total animals with benign tumors 49 50 48
Total benign tumors 100 131 107
Total animals with malignant tumors 19 27 15
Total malignant tumors 20 35 16
Total animals with secondary tumors## 1 1
Total secondary tumors 1 4
Total animals with tumors uncertain--
benign or malignant 3 3 2
Total uncertain tumors 3 3 2

* Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
** Primary tumors: all tumors except secondary tumors
# Number of animals examined microscopically a t this site
# # Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

69 2-Mercaptobenzothiazole, NTP TR 332

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR GAVAGE
STUDY OF 2-MERCAPTOBENZOTHIAZOLE: VEHICLE CONTROL
ANIMAL

NUMBER

WEEKS ON
STUDY

Skin
Squamous cell papilloma
Sauamous mll carcinoma
~ k acell
l tumor
Keratoacanthoma
Subcutaneous tissue
Fibroma
Fibrosarcoma
Lipoma

Lungs and bronchi

Alveolarhmnchiolar adenoma
Alveolar/bmnchiolar carcinoma
Trachea

Bone marrow
Soleen

ymoma, benign

Heart
M
Salivary gland
Liver
Neoplastic nodule
Bile duct
Pancreas
Acinar cell adenorna
&eY

Small intestine
Large intestine

Kidney
Kidney/pelvis
Leiomyosarcoma
Urinary bladder

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Menoma. NOS X X X X X X
k + + + + + + + + + + + + + + + + + + + + + + + +
Pheochmmocytoma X X X X X X X X X X
Thyroid C + + + + + + + + + + + + + + + + + + + + + + + +
Follicular cell carcinoma
C-cell adenoma
Ccell carcinoma
Parathyroid
Pancreatic islets
Islet cell adenoma
Islet cell carcinoma
M
Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . .
Fibroadenoma
Testls
- --- - . . . . . . . . . . . . . . . . . . . . . . . . .

Interstitial cell tumor : X X X X X X X X X X X X X X X X X X X X X X X X

Prostate . . . . . . . . . . . . . . . . . . . . . . . . .

PreputiaUclitoral gland I N N N N N N N N N N N N N N N N N N N N N N N N
Carcinoma. NOS X

Brain
ALL P
Multiple organs, NOS I N N N N N N N N N N N N N N N N N N N N N N N N
Sarcoma. NOS X
Leukemia. mononuclear cell X X X X

+ . Tissue examlned microscop~cally : No tissue information submitted

-: Requimd tissue not axamlned m~croscop~cally C: Necropsy, no histology due to protocol
X: Tumor lnndence A: Autolysis
Necropsy, no autolysis. no mlcmscoplc examloatton M: Animal missing
Anlmal rn~ssexed B: No necropsy performed
Multlple m r m n m of morphology

2-Mercaptobenzothiazole, NTP TR 332

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS: VEHICLE CONTROL
(Continued)
ANIMAL 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
NUMBER 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 5
2 5 6 7 8 9 0 2 3 4 5 6 1 8 9 0 1 2 4 5 6 ~ 8 9 0
WEEKS ON 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Skin
Squamous cell papilloma
. . . . . . . . . . . . . . . . . . . . . . . . . * 50
1
Squamous cell carcinoma X 1
Basal cell tumor X 2
Keratoacanthoma 1
Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . . '50
Fibroma 2
Fibrosarcoma 1
Lipoma 1

Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . . 50

Alveolar/bronchiolar adenoma 2
Alveolar/bronchiolar carcinoma X 1
Trachea . . . . . . . . . . . . . . . . . . . . . . . . . 50

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . . 50
Spleen . . . . . . . . . . . . . . . . . . . . . . . . . 50
L mphnodes . . . . . . . . . . . . . . . . . . . . . . . . . 50
T$ys . . . . . . . . . . . . . . . . . . . . . . . . . 50
ymoma, benign X 1

Heart . . . . . . . . . . . . . . . . . . . . . . . . . 50
~

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . . 50
Liver . . . . . . . . . . . . . . . . . . . . . . . . . 50
Neoplastic nodule X 3
Bile duct . . . . . . . . . . . . . . . . . . . . . . . . . 50
Pancreas . . . . . . . . . . . . . . . . . . . . . . . . . 50
Acinar cell adenoma 2
. . . . . . . . . . . . . . . . . . . . . . . . .
E%,,
Small intestine
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. .
.
.
. .
.
.
.
.
.
.
.
.
.
.
.
.
.
50
50
50
Large intestine . . . . . . . . . . . . . . . . . . . . . . . . . 50

Kidney . . . . . . . . . . . . . . . . . . . . . . . . . 50
Kidneylpelvis . . . . . . . . . . . . . . . . . . . . . . . . . 50
Leiomyosarcoma X 1
Urinary bladder . . . . . . . . . . . . . . . . . . . . . . . . . 50
~~ ~~

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . . 50
Adenoma, NOS X x x x x x x x 14
Adrenal . . . . . . . . . . . . . . . . . . . . . . . . . 50
Pheochromocytoma x x x X x x x X 18
Thyroid . . . . . . . . . . . . . . . . . . . . . . . . . 50
Follicular cell carcinoma 1
C-cell adenoma 2
C m l l carcinoma
Parathyroid . . . . . . . .X . . . . . . . . . . . . . . X . . . X 5
41
Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . . 50
Islet cell adenoma X X 4
Islet cell carcinoma 1

Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . . '50

Fibroadenoma X X 2
Testis . . . . . . . . . . . . . . . . . . . . . . . . . SO
Interstitial cell tumor x x x x x x x x x x x x x x x x x x x x x x x 48
Prostate . . . . . . . . . . . . . . . . . . . . . . . . . 50
Preputial/clitoral gland N N N N N N N N N N N N N N N N N N N N N N N N N *50
Carcinoma, NOS 1

Brain . . . . . . . . . . . . . . . . . . . . . . . . . 50
ALL UTs-
Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N *50
Sarcoma, NOS 1
Leukemia, mononuclear cell X X 7

* Animals necropsied

71 2-Mercaptobenzothiazole, NTP TR 332

ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
NUMBER 2 4 1 2 4 1 4 4 2 2 4 4 3 0 4 4 1 3 0 1 3 0 1 5 2

5 1 3 3 7 6 8 2 9 6 3 0 0 8 9 5 1 8 5 9 2 1 0 0 7

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1

7 7 8 8 8 8 8 8 8 8 8 8 9 9 9 9 9 9 9 9 9 0 0 0 0

8 8 2 3 4 5 5 6 7 8 8 9 1 2 2 3 8 8 9 9 9 0 0 0 1

Skin . . . . . . . . . . . . . . . . . . . . . . . . .
Squamous cell papilloma
Keratoacanthoma x x
Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . .

Sarcoma, NOS X

Fibroma X X

Fibrosarcoma X

Neurofibroma

Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . .

Alveolar/bronchiolar adenoma X

Alveolar/bronchiolar Carcinoma

C cell carcinoma, metastatic X

Trachea . . . . . . . . . . . . . . . . . . . . . . . . .

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . .

Spleen . . . . . . . . . . . . . . . . . . . . . . . . .

Sarcoma, NOS X

Hemangiosarcoma x

L mphnodes . . . . . . . . . . . . . . . . . . . . . . . . .

Tgymua . . . . . . . . . . . . . . . . . . . . . . . . .

Heart . . . . . . . . . . . . . . . . . . . . . . . . .

Neunlemoma, malignant
X I

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Liver . . . . . . . . . . . . . . . . . . . . . . . . .

Neoplastic nodule X

Bile duct . . . . . . . . . . . . . . . . . . . . . . . . .

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . .

Acinar cell adenoma X x x x

. . . . . . . . . . . . . . . . . . . . . . . . .

:;?ax=
Small intestine
.
t
.
+
.
+
.
+
.
+
.
+
.
+
.
t
. . . . . . . . . . . . . . . . .
+ + + + + + + + + + + + + + t + +

Leiomyosarcoma
Large intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Kidney . . . . . . . . . . . . . . .X . . . . . . . . . . .

Transitional cell carcinoma

Tubular cell adenoma

Kidneylpelvis . . . . . . . . . . . . . . . . . . . . . . . . .

Transitional cell papilloma X

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .

M
Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma, NOS X x x x x x x x x x X

Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .

Cortical adenoma
Pheochromocytoma X x x x x x x x x X

Pheochromocytoma, malignant X

Thyroid . . . . . . . . . . . . . . . . . . . . . . . . .

Follicular cell adenoma X

Follicular cell carcinoma X

C cell adenoma

C-cell carcinoma X

Parathyroid + + + + + + + t - - - + t - + - + - + + + + + + +
Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . .

Islet cell adenoma X

Islet cell carcinoma

Mammary gland N N + + + N N + + N + + + N + + + + + + + + + + +

Fibroadenoma X

Testis . . . . . . . . . . . . . . . . . . . . . . . . .

Interstitial cell tumor x x x x x x x x x x x x x x x x x x x x x x x

Mesothelioma, NOS

Prostate . . . . . . . . . . . . . . . . . . . . . . . . .

Preputiaklitoral gland N N N N N N N N N N N N N N N N N N N N N N N N N

Carcinoma, NOS X X

Adenoma. NOS X

Brain . . . . . . . . . . . . . . . . . . . . . . . . . . .

E ORGANS

Zymbal gland N N N N N N N N N + N N N N N N N N N N N + N N N

Carcinoma, NOS X

Squamous cell carcinoma X

S
Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N

Mesothelioma, mall nant X

Leukemia, mononuc7ear cell X X X x x x x x

2-Mercaptobenzothiazole, NTP TR 332 72

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS: LOW DOSE
(Continued)
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

NUMBER 3 3 2 0 0 0 0 0 0 1 1 1 1 1 2 2 2 2 3 3 3 3 3 4 4

9 1 4 2 3 4 6 7 9 2 4 5 7 8 0 1 2 8 3 4 5 6 7 4 6

TOTAL:
1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 TISSUES

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 TUMORS

1 2 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

-I
Skin . . . . . . . . . . . . . . . . . . . . . . . . . $50

Squamous cell papilloma X X 2

Keratoacanthoma 2

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . . '50

Sarcoma. NOS 1

Fibroma X 3

Fibrosarcoma 1

Neurofibroma X 1

Lungs and bronchi 50

Alveolar/bronchiolar adenoma 1

Alveolar/bronchiolar carcinoma X 1

C-cell carcinoma. metastatic 1

Trachea . . . . . . . . . . . . . . . . . . . . . . . . . 49

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . . 50

Spleen . . . . . . . . . . . . . . . . . . . . . . . . . 50

Sarcoma, NOS 1

Hemang-iosarcoma 1

Lymph nodes . . . . . . . . . . . . . . . . . . . . . . . . . 50

Thymus . . . . . . . . . . . . . . . . . . . . . . . . . 49

-I
Heart . . . . . . . . . . . . . . . . . . . . . . . . .

Neunlemoma, malignant

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Liver . . . . . . . . . . . . . . . . . . . . . . . . .

Neoplastic nodule X

Bile duct . . . . . . . . . . . . . . . . . . . . . . . . . 50

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . . 50

Acinar cell adenoma X x x x x x X x x 13

. . . . . . . . . . . . . . . . . . . . . . . . . 50

%
::;Small
:Y intestine .
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. .
. . 50

50

Leiomyosarcoma X 1

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . . 50

Kidney . . . . . . . . . . . . . . . . . . . . . . . . . 50

Transitional cell carcinoma 1

Tubular cell adenoma X 1

Kidne yipelvis . . . . . . . . . . . . . . . . . . . . . . . . . 50

Transitional cell papilloma 1

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Pituitary
Adenoma, NOS
Adrenal
Cortical adenoma
I
.
.
.
.
. . . .
x
. . . .
.
x
.
.
.
x
. . . . . . . . .
x X x x
. . . . . . . . .
. . . . . . . .
x @X x
. . . . . . . .
x

-I
50

21

50

Pheochromocytoma x x x x x x x x x x x x x x x

Pheochromocytoma, malignant X

Thyroid . . . . . . . . . . . . . . . . . . . . . . . . .

Follicular cell adenoma

Follicular cell carcinoma
C cell adenoma x x X 3

C cell carcinoma X 2

Parathyroid . . . . . . . . . . . . . . . . . . . . . . . . . 43

Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . . 50

Islet cell adenoma X 2

Islet cell carcinoma X 1

Mammary gland
Fibroadenoma
I+ + + + + + + + + + + + + + + + + + + + + + + + + 1 '50
1

Testis . . . . . . . . . . . . . . . . . . . . . . . . . 50

Interstitial cell tumor X X X X X X X X X X X X X X X X X X X X X X X X X 48

Mesothelioma, NOS X I 1

Prostate

Preputialiclitoral gland

Carcinoma. NOS
Adenoma, NOS X X X 4

I
-I
Brain
m I A L SENSE ORGANS
Zymbal gland
Carcinoma, NOS

. . . . . . . . . . . . . . . . . . . . . . . . .
N N N N N N N N N N N N N N N N N N N N N N N N N

-I 50

'50

Squamous cell carcinoma

I i
ALL OTAFKSPSTEMS
Multiple organs, NOS

Mesothelioma, mali nant

Leukemia, mononucyear cell x x x x

* Animals necropsied

73 2-Mercaptobenzothiazole, NTP TR 332

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR GAVAGE
STUDY OF 2-MERCAPTOBENZOTHIAZOLE:HIGH DOSE
ANIMAL 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
NUMBER 0 7 5 8 7 9 6 8 7 6 9 9 5 7 6 6 9 6 7 7 5 5 7 9 5
0 4 1 8 5 6 0 2 0 5 9 4 7 2 1 0 1 8 9 1 4 5 3 7 9
WEEKS ON 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1
STUDY 5 5 6 6 6 7 8 8 8 8 9 9 9 9 9 9 9 9 9 9 0 0 0 0 0
7 9 2 6 8 4 2 2 4 7 1 4 5 5 6 6 6 8 8 9 0 0 0 0 1
M
Skin . . . . . . . . . . . . . . . . . . . . . . . . .

Squamous cell papilloma

Squamous cell carcinoma
Keratoacanthoma X
Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . .

Sarcoma, NOS X

Fibroma X X X X

Fibrous histiocytoma, malignant

Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . .

Mucinous adenocarcinoma

Pheochromocytoma, metastatic

Trachea . . . . . . . . . . . . . . . . . . . . . . . . .

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . .

Spleen . . . . . . . . . . . . . . . . . . . . . . . . .

L mphnodes . . . . . . . . . . . . . . . . . . . . . . . . .

Txymus . . . . . . . . . . . . . . . . . . . . . . . . .

Heart . . . . . . . . . . . . . . . . . . . . . . . . .

Pheochromocytoma, metastatic

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Liver . . . . . . . . . . . . . . . . . . . . . . . . .

Neoplastic nodule

Mixed hepatolcholanno carcinoma

Bile duct . . . . . . . . . . . . . . . . . . . . . . . . .

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . .

Acinar cell adenoma x x x

. . . . . . . . . . . . . . . . . . . . . . . . .

?:a
;Y
: . . . . . . . . . . . . . . . . . . . . . . . . .

Small intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Kidney . . . . . . . . . . . . . . . . . . . . . . . . .

Tubular cell adenoma

Kidneylpelvis . . . . . . . . . . . . . . . . . . . . . . . . .

Transitional cell papilloma X

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma, NOS x x X x x x
Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .

Cortical adenoma
Pheochromocytoma X x x x x x x x x x
Pheochromocytoma, malignant
Thyroid . . . . . . . . . . . . . . . . . . . . . . . . .

C cell adenoma
Parathyroid . . . . . . . . . . . . . . . . . . . . . . . . .

Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . .

Islet cell adenoma

Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Fibroadenoma

Testis . . . . . . . . . . . . . . . . . . . . . . . . .

Interstitial cell tumor i x x x x x x x x x x x x x x x x x x x x x x

Pheochromocytoma, metastatic
Mesothelioma, NOS
Prostate . . . . . . . . . . . . . . . . . . . . . . . . .
Preputiallclitoral gland i N N N N N N N N N N N N N N N N N N N N N N N N
Carcinoma, NOS
Adenoma, NOS x x
Brain . . . . . . . . . . . . . . . . . . . . . . . . .
Astrocytoma X
Oligodendroglioma c
ORGANS

Zymbal gland i N N N N N N N N N N N N N N N N N N N N N N N N
Carcinoma, NOS

Mesentery T N N N N N N N N N N N N N N N N N N N N N N N N
Pheochromocytoma, invasive

Multiple organs, NOS T N N N N N N N N N N N N N N N N N N N N N N N N

Mesothelioma. mall nant x x
Leukemia, mononucTear cell X

2-Mercaptobenzothiazole, NTP TR 332 74

ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
NUMBER 9 6 7 9 9 5 5 5 5 6 6 6 6 6 7 7 8 8 8 8 8 8 8 9 9

8 2 6 2 5 2 3 6 8 3 4 6 7 9 7 8 1 3 4 5 6 7 9 0 3

WEEKS ON 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1 2 2 2 2 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Skin . . . . . . . . . . . . . . . . . . . . . . . . . '50

Squamous cell papilloma X X 2

Squamous cell carcinoma X 1

Keratoacanthoma 1

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . . '50

S a m m a . NOS 1

Fibmma X X 6

Fibrous histiocytoma, malignant X 1

LungJand bronchi . . . . . . . . . . . . . . . . . . . . . . . . . 50

Munnous adenocarcinoma X 1

Pheochromocytoma. metastatic X 1

Trachea . . . . . . . . . . . . . . . . . . . . . . . . . 49

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . . 48

Spleen . . . . . . . . . . . . . . . . . . . . . . . . . 49

L mphnodes . . . . . . . . . . . . . . . . . . . . . . . . . 50

Tgymus + + - + + - + + + + + t + + t + + + + + + t +48
+ +

Heart . . . . . . . . . . . . . . . . . . . . . . . . . 50

Pheochromocytoma, metastatic X 1

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . . 50

Liver . . . . . . . . . . . . . . . . . . . . . . . . . 50

Neoplastic nodule X 1

M u d hepatolcholangio carcinoma X 1

Bile duct . . . . . . . . . . . . . . . . . . . . . . . . . 50

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . . 49

Acinar cell adenoma x x X 6

. . . . . . . . . . . . . . . . . . . . . . . . . 50

Eo:%Y
Small intestine
. . . . . .
. . . . . .
.
.
. . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . .
49

49

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . . 49

EM
Kidney . . . . . . . . . . . . . . . . . . . . . . . . . 49

Tubular cell adenoma X 1

Kidneylpelvis . . . . . . . . . . . . . . . . . . . . . . . . . 49

Transitional cell papilloma 1

Urinary bladder . . . . . . . . . . . . . . . . . . . . . . . . . 49

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . . 48

Adenoma, NOS X x x x X X 12

Adrenal . . . . . . . . . . . . . . . . . . . . . . . . . 49

Cortical adenoma X 1

Pheochromocytoma x x x x x x x x x x x X 22

Pheochromocytoma, malignant X X 2

Th mid . . . . . . . . . . . . . . . . . . . . . . . . . so

dr,11adenoma X

Parathyroid . . . . . . . . . . . . . . . . . . . . . . . . . 42

Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . . 49

Islet eel1 adenoma X 1

Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . . '50

Fibroadenoma X

Testis . . . . . . . . . . . . . . . . . . . . . . . . .

x x x x x x x x x x x x x x x x x x x x x x x x x

Interstitial cell tumor

X

+
N
t
N
+
N N
+
N
+
N
+
N
+
N
X
+
N
+
N N
+
N
+
N
+
N
t
N
+
N
t
N
+

X
N
+
N
+
N
+
N
X
+
N N
+
N
+
N
+
*i
+ +
1

. . . . . . . . . . . . . . . . . . . . . . . . . 50

N N + N N N N N N N N N N N N N N N N N N N N N N *50

X 1

N N N N N N N N N N N N N N N N N N N N N N N N N '50
X 1

Multiple organs NOS N N N N N N N N N N N N N N N N N N N N N N N N N '50

Mesothelioma,' mall nant 2

Leuhmra. mononuc7ear cell X X 3

* Animals necropsied

75 2-Mercaptobenzothiazole, NTP TR 332

TABLE A3. ANALYSIS O F PRIMARY TUMORS IN MALE RATS I N THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 m g k g 750 m g k g

Skin: Squamous Cell Papilloma or Carcinoma

Overall Rates (a) 2/50 (4%) 2/50 (4%) 3/50 (6%)
Adjusted Rates (b) 4.8% 9.1% 15.0%
Terminal Rates (c) 2/42 (5%) 2/22 (9%) 3/20 (15%)
Week of First Observation 104 104 104
Life Table Tests (d) P=0.132 P = 0.446 P=O.190
Incidental Tumor Tests (d) P=0.132 P = 0.446 P = 0.190
Cochran-ArmitageTrend Test (d) P = 0.406
Fisher Exact Test (d) P = 0.691 P = 0.500
Subcutaneous Tissue: Fibroma
Overall Rates (a) 2/50 (4%) 3/50 (6%) 6/50 (12%)

Adjusted Rates (b) 4.8% 9.2% 19.6%

Terminal Rates (c) 2/42 (5%) 1/22 (5%) 2/20(10%1

Week of First Observation 104 85 82

Life Table Tests (d) P = 0.024 P=O.299 P=0.033

Incidental Tumor Tests (d) P = 0.064 P =0.612 P = 0.153

Cochran-ArmitageTrend Test (d) P =0.090

Fisher Exact Test (d) P = 0.500 P=0.134

Subcutaneous Tissue: Fibroma or Neurofibroma

Overall Rates (a) 2/50 (4%) 4/50 (8%) 6/50 (12%)

Adjusted Rates (b) 4.8% 13.5% 19.6%

Terminal Rates (c) 2/42 (5%) 2/22 (9%) 2/20 (10%)

Week of First Observation 104 85 82

Life Table Tests (d) P=0.023 P=0.147 P = 0.033

Incidental Tumor Tests (d) P=0.060 P=0.353 P=0.153

Cochran-Armitage Trend Test (d) P = 0.099

Fisher Exact Test (d) P = 0.339 P=0.134

Subcutaneous Tissue: Fibroma or Fibrosarcoma

Overall Rates (a) 3/50 (6%) 4/50 18%) 6/50 (12%)

Adjusted Rates (b) 7.1% 11.6% 19.6%

Terminal Rates (c) 3/42 (7%) 1/22 (5%) 2/20 (10%)

Week of First Observation 104 85 82

Life Table Tests (d) P = 0.055 P=0.266 P = 0.064

Incidental Tumor Tests (d) P=0.143 P=0.671 P=0.237

Cochran-ArmitageTrend Test (d) P=0.187

Fisher Exact Test (d) P = 0.500 P=O.243

Subcutaneous Tissue: Fibroma, Neurofibroma, Sarcoma, o r Fibrosarcoma

Overall Rates (a) 3/50 (6%) 6/50 (12%) 7/50 (14%)

Adjusted Rates (b) 7.1% 17.7% 21.4%

Terminal Rates (c) 3/42 (7%) 2/22 (9%) 2/20 (10%)

Week of First Observation 104 85 74

Life Table Tests (d) P=0.031 P=0.084 P = 0.037

Incidental Tumor Tests (d) P=O.129 P = 0.396 P = 0.237

Cochran-ArmitageTrend Test (d) P=0.128

Fisher Exact Test (d) P=0.243 P=0.159

Lung: Alveolar/Bronchiolar Adenoma or Carcinoma

Overall Rates (a) 3/50 (6%) 2/50 (4%) 0150 (0%)
Adjusted Rates (b) 6.7% 6.5% 0.0%
Terminal Rates (c) 2/42 (5%) 1/22 (5%) 0120 (0%)
Week of First Observation 93 78
Life Table Tests (d) P = 0.190N P = 0.664 P=0.235N
Incidental Tumor Tests (d) P = 0.043N P=0.409N P = 0.124N
Cochran-ArmitageTrend Test (d) P = 0.082N
Fisher Exact Test (d) P=0.500N P = 0.121N

2-Mercaptobenzothiazole, NTP TR 332 76

TABLE A3. ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN T H E TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 mg/kg

Hematopoietic System: Mononuclear Cell Leukemia

Overall Rates (a)
7/50(14%)
16/50(32%) 3/50(6%)
Adjusted Rates (b)
15.1%
47.2% 12.3%
Terminal Rates (c)
4/42(10%)
6/22(27%) 2/20(10%)
Week of First Observation
91
78 91
Life Table Tests (d)
P = 0.475 P = 0.002 P = 0.449N
Incidental Tumor Tests (d)
P = 0.084N P = 0.103 P=0.157N
Cochran-Armitage Trend Test (d)
P=0.178N

Fisher Exact Test (d)

P = 0.028 P=0.159N

Liver: Neoplastic Nodule

Overall Rates (a) 3/50(6%)
2/50(4%) 1/50(2%)
Adjusted Rates (b) 6.8%
6.5% 5.0%
Terminal Rates (c) 2/42(5%)
1/22(5%) 1/20(5%)
Week of First Observation 94
78 104
Life Table Tests (d) P=0.431N
P = 0.663 P = 0.533N
Incidental Tumor Tests (d) P = 0.198N P=0.409N P = 0.401N
Cochran-Armitage Trend Test (d) P = 0.222N
Fisher Exact Test (d) P = 0.500N P=0.309N

Pancreas: Acinar Cell Adenoma

Overall Rates (a) 2/50(4%) 13/50(26%) 6/49(12%)

Adjusted Rates (b) 4.5% 45.7% 23.0%

Terminal Rates (c) 1/42(2%)

8/22(36%) 3/20(15%)

Week of First Observation 94

88 98

Life Table Tests (d) P = 0.017

P <0.001 P = 0.030

Incidental Tumor Tests (d) P=0.118

P <0.001 P = 0.160

Cochran-Armitage Trend Test (d) P = 0.146

Fisher Exact Test (d) P = 0.002 P = 0.128

Pituitary Gland: Adenoma

Overall Rates (a) 14/50(28%)
21/50(42%) 12/48(25%)

Adjusted Rates (b) 30.9%

59.9% 40.1%

Terminal Rates (c) 11/42(26%)

10122 (45%) 5/20(25%)

Week of First Observation 94

82 82

Life Table Tests (d) P=0.106

P = 0.003 P=0.171

Incidental Tumor Tests (d) P=0.506N

P=0.132 P = 0.482N

Cochran-Armitage Trend Test (d) P=0.426N

Fisher Exact Test (d) P = 0.104 P = 0.458N

Adrenal Gland: Pheochromocytoma

Overall Rates (a) 18/50(36%) 25/50(50%)
22/49(45%)

Adjusted Rates (b) 39.8% 70.3%

68.5%

Terminal Rates (c) 15/42(36%) 12/22(55%)

11/20(55%)

Week of First Observation 93 85

84

Life Table Tests (d) P = 0.002 P <0.001

P = 0.002

Incidental Tumor Tests (d) P = 0.109 P = 0.056

P=O.111

Cochran-Armitage Trend Test (d) P =0.213

Fisher Exact Test (d) P=0.113
P=0.243

Adrenal Gland: Pheochromocytoma or Malignant Pheochromocytoma

Overall Rates ( a )
18/50(36%) 27/50(54%)
24/49(49%)

Adjusted Rates (b)

39.8% 74.1%
75.5%

Terminal Rates (c)

15/42(36%) 13/22(59%)
13/20(65%)

Week of First Observation

93 85
84

Life Table Tests (d)

P<O.OOl P <0.001
P<O.OOl

Incidental Tumor Tests (d)

P = 0.038 P=0.021
P=0.034

Cochran-Armitage Trend Test (d)

P=0.115
Fisher Exact Test (d)
P = 0.054 P=0.135

77 2-Mercaptobenzothiazole, NTP TR 332

TABLE A3. ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)
~~ ~~ ~~~~ ~

Vehicle Control 375 mg/kg 750 mg/kg

Thyroid Gland: C-cell Adenoma

Overall Rates (a) 2/50(4%) 3/50(6%) 1/50(2%)

Adjusted Rates (b) 4.5% 12.5% 5.0%

Terminal Rates (c) 1/42(2%) 1/22(5%) 1/20(5%)
Week of First Observation 98 102 104
Life Table Tests (d) P= 0.526 P= 0.264 P=0.703N
Incidental Tumor Tests (d) P= 0.346N P= 0.594 P= 0.548N
Cochran-ArmitageTrend Test (d) P= 0.399N
Fisher Exact Test (d) P= 0.500 P=O.SOON
Thyroid Gland: C-cell Adenoma or Carcinoma
Overall Rates (a) 6/50(12%) 5/50(10%) 1/50(2%)

Adjusted Rates (b) 13.545 18.9% 5.0%

Terminal Rates (c) 4/42(10%) 2/22(9%) 1/20(5%)
Week of First Observation 98 92 104
Life Table Tests (d) P= 0.249N P= 0.388 P=0.219N
Incidental Tumor Tests (d) P=0.052N P=0.454N P= 0.082N
Cochran-ArmitageTrend Test (d) P = 0.049N
Fisher Exact Test (d) P=0.500N P= 0.056N
Pancreatic Islets: Islet Cell Adenoma
Overall Rates (a) 4/50(8%) 2/50(4%) 1/49(2%)

Adjusted Rates (b) 9.3% 6.5% 3.8%

Terminal Rates (c) 3/42(7%) 1/22(5%) 0120(0%)

Week of First Observation 102 78 101

Life Table Tests (d) P=0.318N P=0.591N P=0.425N
Incidental Tumor Tests (d) P=0.072N P= 0.295N P= 0.166N
Cochran-ArmitageTrend Test (d) P =0.122N
Fisher Exact Test (d) P= 0.339N P=0.187N

Pancreatic Islets: Islet Cell Adenoma or Carcinoma

Overall Rates (a) 5/50(10%) 3/50(6%) 1/49(2%)
Adjusted Rates (b) 11.6% 10.9% 3.8%
Terminal Rates (c) 4/42(10%) 2/22(9%) 0/20(0%)
Week of First Observation 102 78 101
Life Table Tests (d) P= 0.268N P=0.626 P= 0.324N
Incidental Tumor Tests (d) P= 0.069N P= 0.419N P= 0.118N
Cochran-ArmitageTrend Test (d) P=0.073N
Fisher Exact Test (d) P = 0.357N P=0.107N

Preputial Gland: Adenoma

Overall Rates (a) 0150 (0%) 4/50 ( 8 % ) 4/50 (8%)
Adjusted Rates (b) 0.0% 14.7% 14.4%
Terminal Rates (c) 0/42(0%) 2/22(9%) 2/20(10%)

Week of First Observation 88 87

Life Table Tests (d) P= 0.016 P= 0.019 P=0.021

Incidental Tumor Tests (d) P=0.042 P = 0.076 P= 0.063

Cochran-ArmitageTrend Test (d) P= 0.060
Fisher Exact Test (d) P=0.059 P=0.059

Preputial Gland: Adenoma or Carcinoma

Overall Rates (a) 1/50(2%) 6/50 (12%) 5/50(10%)

Adjusted Rates (b) 2.2% 18.5% 19.2%

Terminal Rates (c) 0/42(0%) 2/22(9%) 3/20(15%)

Week of First Observation 98 83 87

Life Table Tests (d) P=0.027 P = 0.021 P= 0.030

Incidental Tumor Tests (d) P= 0.094 P=0.216 P=0.117

Cochran-ArmitageTrend Test (d) P= 0.099

Fisher Exact Test (d) P = 0.056 P-0.102

2-Mercaptobenzothiazole, NTP TR 332 78

TABLE A3. ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN T H E TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 mg/kg

Testis: Interstitial Cell Tumor

Overall Rates (a) 48/50 (96%) 48/50 (96%) 48/50 (96%)
Adjusted Rates (b) 96.0% 100.0% 100.0%
Terminal Rates (c) 40142 (95%) 22/22 (100%) 20/20 (100%)
Week of First Observation 91 78 57
Life Table Tests (d) P<O.OOl P < 0.001 P <0.001
Incidental Tumor Tests (d) P=0.271 P=0.617 P = 0.412
Cochran-ArmitageTrend Test (d) P=0.601
Fisher Exact Test (d) P=0.691N P=0.691N

All Sites: Mesothelioma

Overall Rates (a) 0150 (0%) 2/50 (4%) 3/50 (6%)
Adjusted Rates (b) 0.0% 6.6% 9.5%
Terminal Rates (c) 0142 (0%) 1/22 (5%) 1/20 (5%)
Week of First Observation 84 84
Life Table Tests (d) P=0.039 P = 0.163 P = 0.066
Incidental Tumor Tests (d) P = 0.041 P = 0.3 10 P=0.158
Cochran-Armitage Trend Test (d) P = 0.082
Fisher Exact Test (d) P = 0.247 P=O.121

All Sites: Benign Tumors

Overall Rates (a) 49/50 (98%) 50150 (100%) 48/50 (96%)
Adjusted Rates (b) 98.0% 100.0% 100.0%
Terminal Rates (c) 41/42 (98%) 22/22 (100%) 20/20 (100%)
Week of First Observation 91 78 57
Life Table Tests (d) P <0.001 P < 0.001 P <0.00 1
Incidental Tumor Tests (d) P = 0.604 P = 0.629 P=0.648
Cochran-ArmitageTrend Test (d) P = 0.360N
Fisher Exact Test (d) P=0.500 P = 0.500N
All Sites: Malignant Tumors
Overall Rates ( a ) 19/50 (38%) 27/50 (54%) 15/50 (30%)
Adjusted Rates (b) 40.2% 63.9% 48.1%
Terminal Rates (c) 14/42 (33%) 8/22 (36%) 7/20 (35%)
Week of First Observation 91 78 57
Life Table Tests (d) P=0.137 P = 0.002 P = 0.199
Incidental Tumor Tests (d) P = 0.090N P = 0.43 1 P=0.209N
Cochran-ArmitageTrend Test (d) P=0.238N
Fisher Exact Test (d) P = 0.080 P = 0.264N

All Sites: All Tumors

Overall Rates (a) 49/50 (98%) 50150 (100%) 48/50 (96%)
Adjusted Rates (b) 98.0% 100.0% 100.0%
Terminal Rates (c) 41/42 (98%) 22/22 (100%) 20/20 (100%)
Week of First Observation 91 78 57
Life Table Tests (d) P <0.00 1 P <0.00 1 P<O.OOl
Incidental Tumor Tests (d) P = 0.604 P = 0.629 P = 0.648
Cochran-ArmitageTrend Test (d) P=0.360N
Fisher Exact Test (d) P = 0.500 P =0.500N

(a)Number oftumor-bearing animals/number ofanimals examined a t the site

(b) Kaplan-Meier estimated tumor incidences a t the end of the study after adjusting for intercurrent mortality
(c) Observed tumor incidence a t terminal kill
(d)Beneath the vehicle control incidence are the P values associated with the trend test. Beneath the dosed group incidence are
the P values corresponding to pairwise comparisons between that dosed group and the vehicle controls. The life table analysis
regards tumors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The incidental tumor
test regards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence
rates. A negative trend or lower incidence in a dosed group is indicated by (N).

79 2-Mercaptobenzothiazole, NTP TR 332

TABLE A4a. HISTORICAL INCIDENCE OF LEUKEMIA IN MALE F344M RATS ADMINISTERED CORN
OIL BY GAVAGE (a)

Incidence in Vehicle Controls

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence

TOTAL 202/1.450 (13.9%)

SD (b) 7.55%

Range (c)
High 14/50
LOW 1/50

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Standard deviation
(c)Range and SD are presented for groups of 35 or more animals.

TABLE A4b. HISTORICAL INCIDENCE OF PITUITARY GLAND TUMORS I N MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Adenoma Carcinoma or Adenoma, Carcinoma, or
Adenocarcinoma Adenocarcinoma

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence

TOTAL (b)344/1,411(24.4%) (c)26/1,411(1.8%) (b,c)37011,411 (26.2%)

SD (d) 7.92% 2.42% 8.34%

Range (e)
High 19/50 4/47 22/50
LOW 5/50 0150 6/50

(a)Data as of August 30,1985, for studies of at least 104 weeks

(b)Includes 34 chromophobe adenomas and 1 acidophil adenoma
(c)Includes four chromophobe carcinomas and two adenocarcinomas, NOS
(d)Standard deviation
(e)Range and SD are presented for groups of 35 or more animals.

2-Mercaptobenzothiazole,NTP TR 332 80
TABLE A4c. HISTORICAL INCIDENCE O F ADRENAL GLAND TUMORS IN MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Pheochromocytoma Malignant Pheochromocytoma
Pheochromocytoma o r Malignant
Pheochromocytoma

No 2-year studies by Physiological Research Laboratories are included in the historical data base.

Overall Historical Incidence

TOTAL 33811,442 (23.4%) 1311,442 (0.9%) 34711,442 (24.1%)

SD (b) 8.72% 1.27% 8.66%

Range (c)
High 20149 2150 20149
LOW 2150 0150 2/50

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Standard deviation
(c) Range and SD are presented for groups of 35 or more animals.

TABLE A4d. HISTORICAL INCIDENCE OF PANCREATIC ACINAR CELL TUMORS IN MALE F344/N
RATS ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence of Adenomas in Vehicle Controls

No 2-year studies by Physiological Research Laboratories are included in the historical data base.

Overall Historical Incidence

TOTAL (b)8011,381(5.8%)
SD (c) 8.00%

Range (d)
High 14/50
LOW 0150

(a)Data as of August 30,1985, for studies of a t least 104 weeks, An incidence of 22/50 for the benzyl acetate study for which
multiple sections were examined has been deleted.
(b)Includes two carcinomas
(c)Standard deviation
(d)Range and SD are presented for groups of 35 or more animals.

81 2-Mercaptobenzothiazole,NTP TR 332
TABLE A4e. HISTORICAL INCIDENCE O F PREPUTIAL GLAND TUMORS IN MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Adenoma Carcinoma Adenoma or Carcinoma
~~

No 2-year studies by Physiological Research Laboratories are included in the historical data base.

Overall Historical Incidence

TOTAL 30/1.450(2.1%) (b)35/1,450(2.4%) (b)65/1,450

(4.5%)
SD (c) 3.27% 2.53% 4.33%

Range (d)
High 7/50 5/50 9/50
Low 0/50 0/50 0150

(a)Data as of August 30,1985,for studies of a t least 104weeks

(b)Includes 26 carcinomas, NOS, 3 squamous cell carcinomas, and 6 adenocarcinomas, NOS
(c)Standard deviation
(d)Range and SD are presented for groups of 35 or more animals.

TABLE A4f. HISTORICAL INCIDENCE O F SUBCUTANEOUS TISSUE TUMORS I N MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Fibroma Fibrosarcoma Fibroma o r
Fibrosarcoma

No 2-year studies by Physiological Research Laboratories are included in the historical data base.

Overall Historical Incidence

TOTAL (b) 9311,460(6.4%) (c)33/1,450(2.3%) (b,c) 126/1,450(8.7%)

SD (d) 2.90% 2.86% 3.68%

Range (e)
High 6/50 6/50 8/50
Low 0/50 0150 1/50

(a)Data as of August 30,1985,for studies of a t least 104 weeks

(b) Includes five neurofibromas
(c)Includes 10 sarcomas, NOS,and 3 neurofibrosarcomas
(d)Standard deviation
(e)Range and SD are presented for groups of 35 or more animals.

2-Mercaptobenzothiazole,NTP TR 332 82
TABLE A4g. HISTORICAL INCIDENCE O F MESOTHELIAL TUMORS IN MALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Mesothelioma, Malignant All
NOS Mesothelioma Mesothelioma
~ -~ ~~

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence

TOTAL 4811,450 (3.3%) 811,450 (0.6%) 5511,450 (3.8%)

SD (b) 3.04% 1.30% 2.74%

Range (c)
High 6150 2150 6150
Low 0150 0150 0150

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Standard deviation
(c)Range and SD are presented for groups of 35 or more animals.

TABLE A4h. HISTORICAL INCIDENCE O F KIDNEY TUMORS I N MALE F344/N RATS ADMINISTERED
CORN OIL BY GAVAGE (a)
~~~ ~ ~~

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence
No. Examined No. of T u m o r s Diagnosis

1,448 1 Transitional cell papilloma

3 Tubular cell adenoma

2 Adenocarcinoma, NOS
3 Tubular cell adenocarcinoma

TOTAL 8 (0.6%) Tubular cell

l(O.18) Transitional cell

(a)Data asofAugust30,1985,for studiesofatleast 104weeks

(b)Standard deviation
(c)Range and SD are presented for groups of35 or more animals.

83 2-Mercaptobenzothiazole, NTP TR 332

TABLE A5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS NECROPSIED 50 50 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 50 50 50

INTEGUMENTARY SYSTEM
"Skin (50)
Mineralization
*Subcutaneous tissue (50)

Ulcer, NOS

Inflammation, active chronic

RESPIRATORY SYSTEM
*Nasal cavity (50) (50)
Hemorrhage (4%) 2 (4%)
Inflammation, acute (2%) 1 (2%)
Inflammation, chronic (2%) 6 (12%) 1 (2%)
Foreign material, NOS 1 (2%)
*Nasal turbinate (50) (50)
Inflammation, active chronic 2 (4%)
Inflammation, chronic 2 (4%) 4 (8%)
#Lung (50) (50) (50)
Mineralization 1 (2%)
Congestion, NOS 6 6 (12%) 3 (6%)
Edema, NOS 1 (2%)
Hemorrhage 6 (12%) 9 (18%)
Pneumonia, interstitial chronic 1 (2%) 5 (10%)
Bronchopneumonia, chronic 1 (2%) 2 (4%)
Granuloma, NOS 1 (2%)
Hyperplasia, alveolar epithelium 1 (2%) 1 (2%)
Histiocytosis 2 (4%) 4 (8%)

HEMATOPOIETIC SYSTEM
#Bone marrow (50) (50) (48)
Hemorrhage 1 (2%)
Fibrosis 3 (6%) 2 (4%)
Necrosis, NOS 2 (4%)
Hyperplasia, megakaryocytic 2 (4%)
#Spleen (50) (50) (49)
Fibrosis 3 (6%) 7 (14%) 7 (14%)
Pigmentation, NOS 44 (88%) 39 (78%) 46 (94%)
Atrophy, NOS 3 (6%)
Hyperplasia, lymphoid 2 (4%) 3 (6%)
Hematopoiesis 44 (88%) 41 (82%) 43 (88%)
#Splenic capsule (50) (50) (49)
Fibrosis 1 (2%)
#Lymph node (50) (50) (50)
Cyst, NOS 1 (2%)
#Mandibular lymph node (50) (50) (50)
Cyst, NOS 1 (2%)
Plasmacytosis 1 (2%)
Hyperplasia, lymphoid 8 (16%) 12 (24%) 3 (6%)
#Mesenteric lymph node (50) (50) (50)
Congestion, NOS 1 (2%) 2 (4%) 1 (2%)
Hyperplasia, reticulum cell 1 (2%)
#Liver (50) (50) (50)
Hematopoiesis 2 (4%) 1 (2%) 2 (4%)

2-Mercaptobenzothiazole, NTP TR 332 84

TABLE A5. SUMMARY OF T H E INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS I N T H E
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

HEMATOPOIETIC SYSTEM (Continued)

#Thymus (50) (49) (48)
Multiple cysts 1 (2%)
Congestion, NOS 1 (2%)
Hemosiderosis 1 (2%)

CIRCULATORY SYSTEM
*Multiple organs (50)
Periarteritis
#Heart
(50)
Mineralization
1 (2%)
Inflammation, chronic
46 (92%)
#Heart/atrium
(50)

Dilatation, NOS
1 (2%)
Thrombus, organized

*Pulmonary artery
(50)
(50)
Mineralization
3 (6%)
8 (16%)
*Pulmonary vein
(50)
(50)

Mineralization
1 (2%)
1 (2%)
#Pancreas
(50)
(49)
Periarteritis
1 (2%) 5 (10%)
*Mesentery
(50) (50)
Periarteritis
1 (2%)
#Testis
(50) (50)
Periarteritis
1 (2%)

DIGESTIVE SYSTEM
*Lip (50) (50) (50)
Inflammation, chronic 1 (2%)
*Tongue (50) (50)
(50)
Epidermal inclusion cyst 1 (2%)
#Salivary gland
(50)
(50)
(50)

Mineralization
1 (2%)
Inflammation, chronic
1 (2%)
Atrophy, NOS
1 (2%) 5 (10%)
#Liver
(50) (50)

Accessory structure
1 (2%)

Inflammation, chronic
1 (2%)

Granuloma, NOS
2 (4%)
2 (4%)

Necrosis, NOS
1 (2%)
2 (4%)

Metamorphosis, fatty
6 (12%)
4 (8%)
2 (4%)

Cytoplasmic vacuolization
3 (6%)
4 (8%)
2 (4%)

Focal cellular change

45 (90%)
24 (48%)
18 (36%)

Hepatocytomegaly
1 (2%)
2 (4%)

Hyperplasia, NOS
2 (4%)
2 (4%)

Angiectasis
1 (2%)

#Hepatic capsule
(50) (50)
Mineralization

#Liver/centrilobular
(50) (50)

Metamorphosis, fatty
2 (4%)
Cytoplasmic vacuolization
1 (2%) 1 (2%)
#Liver/periportal
(50) (50)
(50)

Inflammation, chronic
45 (90%) 46 (92%) 36 (72%)

Metamorphosis, fatty
5 (10%) 3 (6%)

Cytoplasmic vacuolization
1 (2%) 1 (2%)

#Bile duct
(50) (50) (50)
Multiple cysts
1 (2%)
Hyperplasia, NOS
46 (92%) 49 (98%) 47 (94%)

85 2-Mercaptobenzothiazole, NTP TR 332

TABLE A5. SUMMARY OF THE INCIDENCE OF fiONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)
~~ ~~ ~~

Vehicle Control Low Dose High Dose

DIGESTIVE SYSTEM (Continued)

#Pancreas (50) (50) (49)
Cystic ducts 1 (2%)
Inflammation, chronic 1 (2%) 1 (2%)
#Pancreatic acinus (50) (50) (49)
Focal cellular change 2 (4%) 3 (6%)
Atrophy, NOS 19 (38%) 27 (54%) 20 (41%)
Hyperplasia, NOS 5 (10%) 15 (30%) 7 (14%)
#Stomach (50) (50) (49)
Inflammation, active chronic 1 (2%)
#Gastric fundal gland (50) (50) (49)
Dilatation, NOS 38 (76%) 40 (80%) 34 (69%)
#Forestomach (50) (50) (49)
Edema, NOS 1 (2%)
Ulcer, NOS 5 (10%) 5 (10%)
Inflammation, acute 1 (2%)
Inflammation, active chronic 2 (4%) 7 (14%)
Inflammation, chronic 8 (16%) 7 (14%)
Hyperplasia, epithelial 1 (2%) 12 (24%) 17 (35%)
Hyperkeratosis 12 (24%) 17 (35%)
#Duodenum (50) (50) (49)
Ulcer, NOS 1 (2%)
Inflammation, acute 1 (2%)
Erosion 1 (2%)

URINARY SYSTEM
#Kidney (50) (50) (49)
Hemorrhage 1 (2%)
Nephropathy 50 (100%) 50 (100%) 49 (100%)
#Kidney/cortex (50) (50) (49)
Cyst, NOS 1 (2%) 2 (4%)
# Kidneykubule (50) (50) (49)
Mineralization 25 (50%) 24 (48%) 33 (67%)
Multiple cysts 1 (2%)
Inflammation, acute 2 (4%)
Pigmentation, NOS 46 (92%) 49 (98%) 42 (86%)
Hyperplasia, focal 3 (6%) 3 (6%)
#Kidney/pelvis (50) (50) (49)
Calculus, microscopic examination 2 (4%) 2 (4%)
Hemorrhage 1 (2%) 1 (2%) 1 (2%)
Inflammation, acute 1 (2%)
Hyperplasia, epithelial 4 (8%) 1 (2%)
#Urinary bladder (50) (49) (49)
Calculus, gross observation only 1 (2%)
Inflammation, hemorrhagic 1 (2%)
Inflammation, active chronic 1 (2%)
Inflammation, chronic 1 (2%)
*Urethra (50) (50) (50)
Calculus, microscopic examination 1 (2%) 3 (6%) 2 (4%)

ENDOCRINE SYSTEM
#Pituitary intermedia (50) (50) (48)
Cyst, NOS 3 (6%) 2 (4%)
Multiple cysts 1 (2%)
#Anterior pituitary (50) (50) (48)
Cyst, NOS 6 (12%) 5 (10%) 3 (6%)
Multiple cysts 1 (2%) 1 (2%)
Hemorrhage 1 (2%)
Focal cellular change 1 (2%)
Hyperplasia, NOS 10 (20%) 17 (34%) 12 (25%)

2-Mercaptobenzothiazole, NTP TR 332 86

TABLE A5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ENDOCRINE SYSTEM Continued)

#Adrenal cortex
(50)
(50) (49)
Accessory structure
2 (4%)
3 (6%)
Mineralization
1 (2%)
Degeneration, lipoid
7 (14%) 10 (20%)
9 (18%)
Metamorphosis, fatty
3 (6%) 4 (8%)
6 (12%)
Pigmentation, NOS
1 (2%)

Cytoplasmic vacuolization
1 (2%)

Hyperplasia, NOS
7 (14%) 10 (20%)
5 (10%)
Angiectasis
1 (2%)

#Adrenal medulla
(50) (50) (49)
Hyperplasia, NOS
9 (18%) 14 (28%) 10 (20%)
#Thyroid
(50)
(50) (50)

Embryonal duct cyst

1 (2%)
1 (2%) 1 (2%)

Mineralization
2 (4%)
Cystic follicles
6 (12%) 8 (16%) 12 (24%)
Pigmentation, NOS
1 (2%) 2 (4%) 2 (4%)
Hyperplasia, C-cell
28 (56%) 38 (76%) 34 (68%)
Hyperplasia, follicular cell
1 (2%)
#Thyroid follicle
(50) (50) (50)
Atrophy, NOS
1 (2%)
#Thyroid colloid
(50) (50)

Mineralization
1 (2%)
#Pancreatic islets
(50)
(49)

Hyperplasia, NOS
3 (6%)
1 (2%)

REPRODUCTIVE SYSTEM
*Mammary gland
(60)
(50) (50)

Cyst, NOS
3 (6%)
Multiple cysts
14 (28%) 11 (22%) 6 (12%)
Hyperplasia, cystic
2 (4%) 1 (2%)
*Preputial gland (50) 150) (50)
Cystic ducts 2 (4%) 1 (2%)
Lymphocytic inflammatory infiltration 1 (2%)
Inflammation, suppurative 1 (2%) 1 (2%)
1 (2%)
Abscess, NOS 1 (2%)

Inflammation, active chronic 11 (22%) 7 (14%)

5 (10%)
Inflammation, chronic 34 (68%) 34 (68%)
33 (66%)
Hyperplasia, NOS 1 (2%)
#Prostate
(50) (50) (50)
Mineralization
1 (2%)
Inflammation, active chronic
16 (32%) 20 (40%) 20 (40%)
Inflammation, chronic
10 (20%) 7 (14%) 7 (14%)
Hyperplasia, epithelial
1 (2%)
"Seminal vesicle
(50) (50) (50)
Dilatation, NOS
1 (2%)
Hemorrhage
1 (2%)
Inflammation, active chronic
1 (2%)
Inflammation, chronic
1 (2%)
Atrophy, NOS
3 (6%) 8 (16%) 4 (8%)
#Testis
(50) (50) (50)
Atrophy, NOS
48 (96%) 46 (92%) 44 (88%)
Hyperplasia, interstitial cell
46 (92%) 45 (90%) 45 (90%)
#Testidtubule
(50)
(50)
(50)
Mineralization
35 (70%) 30 (60%) 37 (74%)
Oligospermia
2 (4%)
*Epididymis
(50) (50)
Mineralization
1 (2%)
Inflammation, chronic

*Scrotum
(50) (50)
Steatitis
4 (8%) 2 (4%)

a7 2-Mercaptobenzothiazole, NTP TR 332

TABLE A5. SUMMARY OF T H E INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN T H E
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

NERVOUS SYSTEM
#Brain
(50) (50) (50)
Compression, NOS
1 (2%) 1 (2%)
Hemorrhage
1 (2%)
Malacia
1 (2%)

SPECIAL SENSE ORGANS

*Eye
(50) (50) (50)
Hemorrhage
1 (2%)
Inflammation, suppurative
1 (2%)
Retinopathy
10 (20%)
Phthisis bulbi
2 (4%)
*Eye/sclera (50) (50) 150)
Mineralization 1 (2%) 1 (2%)
*Eye/cornea (50) (50)
Inflammation, active chronic 2 (4%)
*Eye/crystalline lens (50) (50)
Cataract 1 (2%) 6 (12%)
*Nasolacrimal duct
(50) (50)'
Hemorrhage
9 (18%) 2 (4%)
Inflammation, acute

*Harderian gland (50)

Inflammation, chronic
*Ear canal (50)
Inflammation, active chronic

MUSCULOSKELETAL SYSTEM
None

BODY CAVITIES
*Epicardium (50) (50) (50)
Inflammation, active chronic 1 (2%)
*Mesentery
(50) (50) (50)
Ulcer, NOS
1 (2%)
Steatitis
1 (2%)

ALL OTHER SYSTEMS

None

SPECIAL MORPHOLOGY SUMMARY

None
~~ ~~

* Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
# Number of animals examined microscopically a t this site

2-Mercaptobenzothiazole,NTP TR 332 88
APPENDIX B

SUMMARY OF LESIONS IN FEMALE RATS IN THE

TWO-YEAR GAVAGE STUDY OF

2-MERCAPTOBENZOTHIAZOLE

PAGE
TABLE B1 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE
TWO-YEAR GAVAGE STUDY O F 2-MERCAPTOBENZOTHIAZOLE 91
TABLE B2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF' 2-MERCAPTOBENZOTHIAZOLE 94

TABLE B3 ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR GAVAGE

STUDY OF 2-MERCAF'TOBENZOTHIAZOLE 100

TABLE B4a HISTORICAL INCIDENCE OF LEUKEMIA IN FEMALE F344/N RATS ADMINISTERED

CORN OIL BY GAVAGE 104

TABLE B4b HISTORICAL INCIDENCE OF PITUITARY GLAND TUMORS IN FEMALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE 104

TABLE B4c HISTORICAL INCIDENCE O F ADRENAL GLAND TUMORS IN FEMALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE 104

TABLE B6 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE

RATS IN THE TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 106

89 2-Mercaptobenzothiazole,NTP TR 332
2-Mercaptobenzothiazole, NTP TR 332 90
TABLE B1. SUMMARY OF T H E INCIDENCE OF NEOPLASMS I N FEMALE RATS I N T H E TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS NECROPSIED 50 50 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 50 50 50

INTEGUMENTARY SYSTEM
*Skin (50) (50) (50)
Squamous cell carcinoma 1 (2%)
Basal cell tumor 1 (2%)
Keratoacanthoma 1 (2%)
"Subcutaneous tissue (50) (50) (50)
Sarcoma, NOS 1 (2%)
Fibroma 2 (4%) 3 (6%) 1 (2%)
Fibrosarcoma 1 (2%)
Fibrous histiocytoma, malignant 1 (2%)
Fibrous histiocytoma, metastatic 1 (2%)

RESPIRATORY SYSTEM
#Lung (50) (50) (50)
Alveolar/bronchiolar adenoma 2 (4%)
Alveolar/bronchiolar carcinoma 1 (2%)
Fibrous histiocytoma, metastatic 1 (2%)

HEMATOPOIETIC SYSTEM
*Multiple organs (50) (50) (50)
Leukemia, mononuclear cell 6 (12%) 13 (26%) 9 (18%)
#Spleen (50) (50) (50)
Leukemia, mononuclear cell 1 (2%)

CIRCULATORY SYSTEM
None

DIGESTIVE SYSTEM
'Oral cavity
Squamous cell papilloma
*Tongue
Squamous cell papilloma
#Salivary gland
Fibrous histiocytoma, metastatic
#Liver
Neoplastic nodule
Fibrous histiocytoma, metastatic
#Esophagus
Fibrous histiocytoma, metastatic

URINARY SYSTEM
None

ENDOCRINE SYSTEM
#Pituitary intermedia (49) (50) (50)
Adenoma, NOS 1 (2%)
#Anterior pituitary (49) (50) (50)
Adenoma, NOS 15 (31%) 24 (48%) 25 (50%)
Adenocarcinoma, NOS 1 (2%)
#Adrenal (50) (50) (50)
Cortical adenoma 2 (4%) 2 (4%)

91 2-Mercaptobenzothiazole, NTP TR 332

TABLE B1. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ENDOCRINE SYSTEM (Continued)

#Adrenal medulla (50) (50) (50)
Pheochromocytoma 1 (2%) 5 (10%) 6 (12%)
Ganglioneuroma 1 (2%)
#Thyroid (50) (50) (50)
Follicular cell adenoma 1 (2%)
C-cell adenoma 5 (10%) 2 (4%) 3 (6%)
C-cell carcinoma 1 (2%) 1 (2%)

REPRODUCTIVE SYSTEM
*Mammary gland (50) (50) (50)
Adenoma, NOS 1 (2%)
Adenocarcinoma, NOS 1 (2%)
Fibroadenoma 12 (24%) 17 (34%) 17 (34%)
*Clitoral gland (50) (50) (50)
Carcinoma, NOS 4 (8%) 8 (16%) 1 (2%)
Adenoma, NOS 5 (10%) 2 (4%) 3 (6%)
#Uterus (50) (50) (50)
Leiomyosarcoma 1 (2%)
Endometrial stromal polyp 13 (26%) 14 (28%) 8 (16%)
Endometrial stromal sarcoma 2 (4%) 2 (4%)
#Ovary (50) (50) (50)
Fibrous histiocytoma, metastetic 1 (2%)

NERVOUS SYSTEM
None

SPECIAL SENSE ORGANS

*Zymbal gland (50) (50) (50)
Carcinoma,NOS 1 (2%) 1 (2%)

MUSCULOSKELETAL SYSTEM
None

BODY CAVITIES
None

ALL OTHER SYSTEMS

None

ANIMAL DISPOSITION SUMMARY

Animals initially in study 50 50 50
Natural death 6 2 7
Moribund sacrifice 15 16 18
Terminal sacrifice 28 31 25
Dosing accident 1 1

2-Mercaptobenzothiazole, NTP TR 332 92

TABLE B1. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

TUMOR SUMMARY
Total animals with primary tumors++ 37 46 40
Total primary tumors 73 104 79
Total animals with benign tumors 31 41 36
Total benign tumors 55 78 64
Total animals with malignant tumors 14 21 13
Total malignant tumors 17 26 15
Total animals with secondary tumors## 1
Total secondary tumors 6
Total animals with tumors uncertain--
benign or malignant 1
Total uncertain tumors 1

Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
+*Primary tumors: all tumors except secondary tumors
# Number of animals examined microscopically a t thissite
# # Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

93 2-Mercaptobenzothiazole, NTP TR 332

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2.MERCAPTOBENZOTHIAZOLE: VEHICLE CONTROL
ANIMAL

I I
1 1 1 1 1 1 1 1 1 1 l l l l l l l l l l l l l l
NUMBER 4 0 2 2 3 1 1 2 0 3 2 3 2 3 1 1 2 4 1 3 1 0 0 0
8 4 1 7 9 5 5 1 8 8 4 4 3 6 0 1 9 7 2 3 5
WEEKS ON 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 l 1 1
STUDY 3 3 4 4 5 5 5 5 8 7 7 8 8 8 8 8 8 9 9 9 0 0 0 0 0
0 1 0 0 0 1 3 8 3 2 7 0 1 3 8 9 0 0 8 2 3 4 4 4
M

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . .

Sarcoma, NOS X

Fibroma

Fibrous histiocytoma malignant X

Fibrous histiocytoma: metastatic X

M
Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . .

Fibmus histiocytoma, metastatic X

Trachea . . . . . . . . . . . . . . . . . . . . . . . . .

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . .

Spleen . . . . . . . . . . . . . . . . . . . . . . . . .

L mphnodes . . . . . . . . . . . . . . . . . . . . . . . . .

Tiymus . . . . . . . . . . . . . . . . . . . . . . . . .

Heart + + t + + t + + + + + + + + + + + + + + + + t + +
M
Salivary land + t + + + + + + + + + + + + + + + + + + t + + t t
Fibrous$istiocytoma, metastatic X
Livar + + + t + + + + t + + + t + + + + + + + + + + + +
Neoplastic nodule X
Fibrous histiocytoma, metastatic X
Bile duct + + + + + + + + + + + + + + + + e + + + + + + + +
Pancreas . . . . . . . . . . . . . . . . . . . . . . . . .

Em hagus + e + + + + + + + + + + + + + + + + + + + + + + +
Fi!rous histiocytoma, metastatic X
Stomach . . . . . . . . . . . . . . . . . . . . . . . . .

Small intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Large intestine + + t + + + + + t + + + + + + + + + + + + + + + +

Kidney . . . . . . . . . . . . . . . . . . . . . . . . .

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma NOS X x x x x x
Adenocarhnoma, NOS X
Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .
Cortical adenoma
Pheochromocytoma X
Th mid . . . . . . . . . . . . . . . . . . . . . . . . .

&cell adenoma X
Parathyroid + - + - + + - + + + - - - + - - + + + - + + + + +
M

M a m m a y gland + + + + + + + + + + + + + + + + + N + + + + + + +
Adenocarcinoma, NOS X
Fibroadenoma X x x X
Preputial/chtoral gland N N N N N N N N N N N N N N N N N N N N N N N N N
Carcinoma. NOS X X
Adanoma, NOS X X X
UhNS + + + + + + + + + + + + Y + + + + + + + t + + + +
Endometnal stromal polyp x x x X x x
Endometnal stromal sarcoma X X
+ + + + + + + + + + + + + + e + + + + + + + + + +
%L%umua histiocytoma. rnataatatic X

Brain + + + t + + + + + + + + + + + t + t t + + + + + +
ANS
Zymbal gland
Carcinoma, NOS I N N N
~~
N N N
~
N N N N N N N N N Xt N N N N N N N N N I

Multi leorgans NOS N N N N N N N N N N N N N N N N N N N N N N N N N

LauEemia, mobonuciear cell X X
I I
+: Tissue examined micmscupically : No tissue information submitted
-. Required tissue not examined microscopically C: Necropdy, no histology due to protocol

Xi Tiimor incidence A: Autolysin

N: Necropsy, no autolysis, no microscopic examination M: Animal missing

S: Animal missexed 8: No nacropiy performed

2-Mercaptobenzothiazole, NTP TR 332 94

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: VEHICLE CONTROL
(Continued)
ANIMAL 1] 11 1 11 1 1 1 1 1 1 1 1| 1 1 1 1 1 1 1 1 1 1 1 1 1
NUMBER
6| 7j 8 9| 0 4 6 3 7 8 9 0 ) 1 2 3 5 1 2 3 4 5 7 8 9 0
TOTAL
WEEKS ON 11 1 1 1 1 1 1 1 1 1 1 11 11 1 1 11 1 1 1 11 1 1 1 1 1 1 TISSUES
STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 TUMORS
4 ] 4 4 4 4 4 4 4 4 ] 4 4 | 4 | 4 4 4 4 4 4 4 ) 4 4 4 4 4 4
INTEGUMENTARY SYSTEM
Subcutaneous tissue + + + + + ± + + + + + + + + + + + + + + + + + + + •50
Sarcoma, NOS I
Fibroma X X 2
Fibrous histiocytoma, malignant 1
Fibrous histiocytoma, metastatic 1
RESPIRATORY SYSTEM
Lungs and bronchi 50
Fibrous histiocytoma, metastatic 1
Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 50
HEMATOPOIETIC SYSTEM
Bone marrow 50
Spleen 50
Lymph nodes 50
Thymus + + + + + + + + + + + + + + + + + + + + + + + + + 50
CIRCULATORY SYSTEM
Heart + + + + + + + + + + + + + + + + + + + + + + + + + 50
DIGESTIVE SYSTEM
Salivary gland 50
Fibrous histiocytoma, metastatic 1
Liver 50
Neoplastic nodule 1
Fibrous histiocytoma, metastatic 1
Bile duct 50
Pancreas + + + + + + + + + + + + + + + + + + + + + + + 4- 50
Esophagus + + + + + + + + + + + + + + + + + + + + + + + -t- 50
Fibrous histiocytoma, metastatic 1
Stomach 49
Small intestine + + + + + + + + + + + + + + + + + + + + -I- + + + 50
Large intestine 50

URINARY SYSTEM
Kidney 50
Urinary bladder 50
ENDOCRINE SYSTEM
Pituitary 49
Adenoma, NOS X X X X X X X X X 15
Adenocarcinoma, NOS 1
Adrenal 50
Cortical adenoma X X 2
Pheochromocytoma 1
Thyroid 50
C cell adenoma X X X X 5
Parathyroid + + + + + + + + + - + + + + + - + + + + - + + + + 38

REPRODUCTIVE SYSTEM
Mammary gland + + + + + + + + + + + + + + + + + + + .(- + + + + + *50
Adenocarcinoma, NOS 1
Fibroadenoma X X X X X X X X 12
Preputial/chtoral gland N N N N N N N N N N N N N N N N N N N N N N N N N *50
Carcinoma, NOS X X 4
Adenoma, NOS X X 5
Uterus 50
Endometnal stromal polyp X X X X X X X 13
Endometnal stromal sarcoma 2
Ovary + + -t-4- + + + -t- + -t- + + + + + + + -l- + + + -t- + + + 50
Fibrous histiocytoma, metastatic 1
NERVOUS SYSTEM
Brain 50

SPECIAL SENSE ORGANS

Zymbal gland N N N N N N N N N N N N N N N N N N N N N N N N N *50
Carcinoma, NOS 1
ALL OTHER SYSTEMS
Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N »50
Leukemia, mononuclear cell X X X X 6

* Animals necropsied

95 2-Mercaptobenzothiazole, NTP TR 332

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR

GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE: LOW DOSE

ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

NUMBER 2 0 4 4 0 2 0 1 4 3 3 3 0 2 5 0 4 0 4 0 0 0 1 1 1

8 8 4 1 5 8 6 6 8 6 5 7 9 0 0 ' 2 6 1 3 3 4 7 0 1 2

b 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 ~ 1 ~

1 3 5 6 8 8 8 9 9 9 9 9 0 0 0 0 0 0 0 0 0 0 0 0 0

8 0 7 7 5 6 9 1 1 2 6 9 0 0 0 1 1 2 2 4 4 4 4 4 4

Skin + + + + + + + + + + + + + + + + + + + + N + + + +

Baaal cell tumor X

Keratoacanthoma

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . .

Fibroma X

Fibrosarcoma X

Lungs and bronchi + + + + t + + + + + + + + t t + + + + + + + + + +

Alveolar/bmnchiolar adenoma
Alveolar/bronchiolar carcinoma
Trachea . . . . . . . . . . . . . . . . . . . . . . . . .

~~ ~~~~~~

Bone marrow + + + + t + + t + t + + + + + t + + t + + + + + +

Spleen . . . . . . . . . . . . . . . . . . . . . . . . .

Leukemia, mononuclear cell X

L mphncdes . . . . . . . . . . . . . . . . . . . . . . . . .

Tgymus + + t + t + + t + + + + + + + + + + + + + + + + +
M

Heart + + + + t t + + + + + + + + + t + + t + + + + + +

Oral cavity N N N N N N N N N N N N N N N N N N N N N N N N N

Squamous cell papilloma X

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Liver . . . . . . . . . . . . . . . . . . . . . . . . .

Bile duct . . . . . . . . . . . . . . . . . . . . . . . . .

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . .

&%R""
Small intestine
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

L a r p intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Kidney . . . . . . . . . . . . . . . . . . . . . . . . .

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .
ENDOCRINESYSTEM
Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma, NOS X X x x x x x x x X

Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .

Cortical adenoma X

Pheochmmocytoma X X X

Ganglioneuroma

Thyroid + + + + t + + + + + + t + + + + + + t + + + + + +

Follicular cell adenoma X

C d l adenoma X

C d l carcinoma X

Parathy mid . . . . . . . . . . . . . . . . . . . . . . . . .

Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Fibroadenoma X X x x x x x x X

PmputiaUclitoral gland N N N N N N N N N N N N N N N N N N N N N N N N N

Carcinoma, NOS X X X

Adenoma, NOS Y

UteNS . . . . . . . . . . . . . . . . . . . . . . . . .

Endometrial stromal polyp x x X X

Ovary . . . . . . . . . . . . . . . . . . . . . . . . .

Brain + + t + t t + + + + + t + t + + + t + + + + + + +
~

ANS
Zymbal gland N N N N N t N N N N N N N N N N N N N N N N N N N

Carcinoma, NOS X

Multi le organs NOS N N N N N N N N N N N N N N N N N N N N N N N N N

LeuIemia, mohonuclear cell x x x x x x x x

2-Mercaptobenzothiazole,NTP TR 332 96

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: LOW DOSE
(Continued)
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

NUMBER 1 1 1 1 1 1 2 2 2 2 2 2 2 3 3 3 3 3 3 3 4 4 4 4 4

3 4 5 7 8 9 1 2 9 4 5 7 9 0 1 2 3 4 8 9 0 2 5 7 9

POTAL
WEEKS ON 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 'ISSUES
STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 'UMORS

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Skin + t t + + t + t + + t t t + + t + t + t + + + t t '50
Basal call tumor 1

Keratoacanthoma X 1

Subcutaneous tissue + t t t t t + t + + + + t + + t t t + t + + + + + '50

Fibroma X X 3

Fibrosarcoma 1

Lungs and bronchi t + + + + + + t t + t t t + + t t t t t t + t t t 50

Alveolar/bronchiolar adenoma X X 2

Alveolar/bronchiolar carcinoma X 1

Trachea t t + + + t + + t + + + + t t t + + + + + t + + + 50

Bone marrow
t + t t + t + t t + + + t + t + t + t + + t t t t 50

S leen + + + t t + + t + + + + t t t + + + + + + + + t t 50

Leukemia. mononuclear cell 1

L mphnodes + + t t t + + t + t + t t + + t t + t t t + + t t 50

T{ymus + + + + t + + t t t t t + + + t t + t + t + + t t 50

-
Heart t t t t + t t + + t + t t + + + t + t + t + + + + 50

Oral cavity N N N N N N N N N N N N N N N N N N N N N N N N N '50

Squamous cell papilloma X 2

Salivary gland + + t + t + + t + t + + + + + t + t + t + + + + + 50

Liver t + + t t t t + + + + + t + + t t t + t + + t + t 50

Bile duct + t + + + t t t + + + + + t + + + + + t t + + + + 50

Pancreas + + + t + t + + t t t + + t t t + t t + + + + + + 50

t t + + + + t + t + t + t + t t + + t + t + + + + 50

:Small
s%Yintestine +
t
+
t
+
+
+
+
+
+
+
+
+
t
+
+
+ + t t t +
+ + + t t +
+
t
t
t
+
t
t
t
+
+
+
t
+
+
+
t
t
t
+
t
t
+
50

50

Large intestine t + t t t + + + + t t t t t + + + + t + t + t t + 50

Kidney t + + t + t + + + + t t t t t t t + + + t t + + + 50

Unnary bladder t t t + t t + + + + t + t + + + t t t t + + t + + 50

Pituitary t + + t t + t t + t + + + t + + t + t + + + + t + 50

Adenoma, NOS x x x x x x x x x x x x x x x 25

Adrenal + t + + + + t + + + + t t t + t + t t t + + t + t 50

Cortical adenoma X 2

Pheochromocytoma x x 5

Ganglioneuroma X 1

Thyroid t t + + t + + + t + + + + + + t t + + + t t + t t 50

Follicular call adenoma 1

C.cell adenoma X 2

C-cell carcinoma 1

Parathyroid t + - + + + + + + + + + + - t - + + + + + + t + + 43

+ + + + + + t + + + t t + + + t t + + + + t + t + '50

x x x X X X x x 17

N N N N N N N N N N N N N N N N N N N N N N N N N '50
X x x x X 8

1 2

Uterus t + t t + t + t + + t + + t + + + t + + + + + + t SO

Endometnal stromal polyp X x x x x x x x x x 14

Ovary + + + + + + + + + + + t t + + + t + t t + + + + + 50

Brain + + + + + + t + t + + + + + + t + t + + + t t + + 50

ANS

Zymbal gland N N N N N N N N N N N N N N N N N N N N N N N N N '50

Carcinoma, NOS 1

Multi leorgans NOS N N N N N N N N N N N N N N N N N N N N N N N N N '50

Leuemia. mobonuclaar cell X X X x x 13

* Animals necropsied

97 2-Mercaptobenzothiazole,NTP TR 332

TABLE 82. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE:HIGH DOSE
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0
NUMBER 8 5 5 9 7 5 6 8 6 8 5 1 6 0 6 9 6 7 8 9 1 5 1 1 5
0 1 2 1 2 9 0 5 9 8 8 1 5 0 6 3 1 4 7 6 3 1 6 0 3
WEEKSON 3-0 o o 0 0 0 o o o o D o 0 0 o o 0 0 0 0 0 0 0 r
STUDY 2 2 2 2 3 3 4 5 6 7 8 8 8 8 9 9 9 9 9 9 9 9 9 9 0
2 5 5 5 1 9 4 2 4 9 2 2 3 6 0 0 2 2 2 2 4 5 6 1 1

Skin t t t t + t t t t t t t t t + t t t t t t t t t t
Squamous call carcinoma X
Subcutaneous tiisur t t t t t t t t t t + + t t + t + + t t t t t t t
Fibroma
M

t u n s a n d bronchi t t t t t t t t t t t t t t + + + + t t t t t t t
Tracaea t t t t t t t t t t t t t t t + + t t t t t t t t

M
Bone marrow t t t t t t t t t t t + t + t t t t + t t t t t +
Spleen + t + t t t t t t t + + t t t + t t t t t t t t t
L mphnodes t t t + t t t + t t t t t t t t t + + t t t t t t
Jymus t t t t t t t t t + t + + + t + t + t t t t t t t

Heart t + t t t t t t t + t t t t t t t t + t t t t t t

Salivary gland t t t t t t t t + t t + + t + t t t t t + t t t t

Liver + + t t + t t t t t t t t t + t t t t t t t t t t
Bile duct t t t t t t t + t + + t + + t + t t + t + t t t t
Pancreas t t t t + t + + + t + + t + + + t t t + t t t t t
t t t t t + t t t t t + t t t t + + + t t + + t t

:Small
tsoo% Y
intestine
t
t
t t
t t
t
t
t
t
t
t
t t t t t t t t
+ + + t + t t t
t
t
t t t t
t t t +
t t + +
+ t t t t
t
t
t

Large intestine t + + t t t + t t + + t + t t + + t + + t t t t +

Kidney t + t + t t + + t t t t + + + t + + t + t t t + +
Unnary bladder t t t t + t t + t + t t t t t t - t t t t + + t t

P
Pituitary t t t t t t t t t + t t t + t t + t t t + t t t t
Adenoma, NOS X X X x x x x x x
Adrenal t t t t t t t t t t + t t + t + + t t t t + + t t
Pheochromocytoma X
Thyroid t t t t t t + t + + t t t t t + t t t t t t t t t
C cell adenoma X
C cell carcinuma
Parathyroid - - - t - - - t t t t t - t t t - t t t t - - t t

Mammary gland + t t + t t t t + t t t + + t t t t + t t t + t t
Adenoma, NOS
Fibroadenoma X x x x x x x x
Preputial/clitoral land N N N N N N N N N N N N N N N N N N N N N N N N N

Carcinoma NO$
Adenoma, kOS X
Uterus t t t t t t t t t t t t + + + + t t t t t t t + t

Leiomyosarcoma X
Endometnal stromal polyp X X X
Endometnal stromal sarcoma X
Ovary t + t t t t t t t t t t t t t + + t t t t t t t t

Brain + t t t t t t t t t t t t t t t t t t t t t t t t
ALL -EMS
Multi leorgans NOS N N N N N N N N N N N N N N N N N N N N N N N N N

Leu&mia, mobonuciear call x x x x x x X

2-Mercaptobenzothiazole, NTP TR 332 98

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE R A T S HIGH DOSE
(Continued)
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

NUMBER 5 5 5 6 6 6 6 6 7 7 7 7 8 8 8 8 6 8 9 9 9 9 9 9 9

4 5 6 2 3 4 7 8 5 7 8 9 1 2 3 4 8 9 0 2 4 5 7 8 9

fOTAL:

WEEKS ON 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 ISSUES

STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 'UMORS

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Skin . . . . . . . . . . . . . . . . . . . . . . . . . '50

Squamous cell carcinoma 1

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . . * 60

Fibroma X 1

EM
Lun and bronchi . . . . . . . . . . . . . . . . . . . . . . . . . 50

Tracyea . . . . . . . . . . . . . . . . . . . . . . . . . 50

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . . 50

Spleen . . . . . . . . . . . . . . . . . . . . . . . . . 50

L mphnodea . . . . . . . . . . . . . . . . . . . . . . . . . 50

dymus . . . . . . . . . . . . . . . . . . . . . . . . . 50

Heart + + + + + + t + + + + + + + + + + t + + + + + + t 50

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . . 50

Liver . . . . . . . . . . . . . . . . . . . . . . . . . 50

Bile duct . . . . . . . . . . . . . . . . . . . . . . . . . 50

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . . 50

2:kr
Small intestine
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. . . . . . . . . . . . . . . . . . . . . . . . .
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. . . . . . .
. . . . . . .
50

50

50

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . . 50

Kidney . . . . . . . . . . . . . . . . . . . . . . . . . 50

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . . 49

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . . 50

Adenoma. NOS x x x x x x x x x x x x x x x X 25

Adrenal . . . . . . . . . . . . . . . . . . . . . . . . . 50

Pheochromocytoma X X X x x 6

Thyroid . . . . . . . . . . . . . . . . . . . . . . . . . 50

C.cell adenoma X X 3

C.cell carcinoma X - 1

Parathyroid . . . . . . . . . . . . . . . . . . . . . . . . . 35

Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . . *50

Adenoma, NOS X 1

Fibroadenoma x x x x x x X X X 17

Preputial/clitoral gland N N N N N N N N N N N N N N N N N N N N N N N N N '50

Carcinoma NOS X 1

Adsnoma. NOS X X 3

Uterus . . . . . . . . . . . . . . . . . . . . . . . . . 50

Leiomyosarcoma 1

Endometnal stromal polyp x x X X X 8

Endometnal stromal sarcoma X 2

Ovary
. . . . . . . . . . . . . . . . . . . . . . . . . 50

Brain + + t + + + + + t + + + + + t + + + + + + + + t + 50

Multi le organs NOS N N N N N N N N N N N N N N N N N N N N N N N N N *50

~euRRmia,mohonuclear cell X X 9

*Animals necropsied

99 2-Mercaptobenzothiazole, NTP TR 332

TABLE B3. ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 188 mg/kg 375 mg/kg

Subcutaneous Tissue: Fibroma

Overall Rates (a) 2/50 (4%)
3/50 (6%) 1/50 (2%)
Adjusted Rates (b) 7.1%
9.1% 4.0%
Terminal Rates (c) 2/28 (7%)
2/31 (6%) 1/25(4%)
Week of First Observation 104
101 104
Life Table Tests (d) P = 0.443N
P = 0.551 P = 0.540N
Incidental Tumor Tests (d) P = 0.407N
P=0.629 P=0.540N
Cochran-ArmitageTrend Test (d) P = 0.400N

Fisher Exact Test (d)

P = 0.500 P = 0.500N
Subcutaneous Tissue: Fibroma or Fibrosarcoma
Overall Rates (a) 2/50 (4%)
4/50 (8%) 1/50 (2%)
Adjusted Rates (b) 7.1%
11.2% 4.0%
Terminal Rates (c) 2/28 (7%)
2/31 (6%) 1/25 (4%)
Week of First Observation 104
91 104
Life Table Tests (d) P=0.442N
P = 0.408 P=0.540N
Incidental Tumor Tests (d) P=0.393N
P=0.426 P=0.540N
Cochran-ArmitageTrend Test (d) P = 0.407N

Fisher Exact Test (d)

P = 0.339 P = 0.500N
Subcutaneous Tissue: Fibroma, Sarcoma, or Fibrosarcoma
Overall Rates (a) 3/50 (6%)
4/50 (8%) 1/50 (2%)
Adjusted Rates (b) 9.2%
11.2% 4.0%
Terminal Rates (c) 2/28 (7%)
2/31 (6%) 1/25 (4%)
Week of First Observation 51
91 104
Life Table Tests (d) P=0.280N
P=0.572 P=0.341N
Incidental Tumor Tests (d) P=0.228N
P = 0.510 P =0.3 ION
Cochran-ArmitageTrend Test (d) P=0.253N

Fisher Exact Test (d)

P = 0.500 P = 0.309N

Lung: AlveolarlBronchiolar Adenoma or Carcinoma

Overall Rates (a) 0150 (0%) 3/50 (6%)
0/50 (0%)
Adjusted Rates (b) 0.0% 9.7%
0.0%
Terminal Rates (c) 0128 (0%) 3/31 (10%)
0125 (0%)
Week of First Observation 104

Life Table Tests (d) P = 0.613 P=0.139

(e)
Incidental Tumor Tests (d) P = 0.613 P = 0.139
(e)
Cochran-ArmitageTrend Test (d) P = 0.638
Fisher Exact Test (d) P=0.121
(e)
Hematopoietic System: Mononuclear Cell Leukemia
Overall Rates (a)
6/50 (12%) 14/50 (28%)
9/50 (18%)

Adjusted Rates (b)

19.7% 35.4%
25.3%

Terminal Rates (c)

4/28 (14%) 6/31 (19%)
2/25 (8%)

Week of First Observation

90 92
79

Life Table Tests (d)

P=0.221 P = 0.099
P = 0.279

Incidental Tumor Tests (d)

P = 0.399 P = 0.215
P=0.415

Cochran-ArmitageTrend Test (d)

P = 0.263
Fisher Exact Test (d)
P = 0.039 P=0.288

Pituitary Gland: Adenoma

Overall Rates (a)
15/49 (31%) 24/50 (48%)
25/50 (50%)

Adjusted Rates (b)

44.6% 62.3%
73.2%

Terminal Rates (c)

10128 (36%) 17/31 (55%)
16/25 (64%)

Week of First Observation

72 67
82

Life Table Tests (d)

P = 0.014 P = 0.146
P=0.021

Incidental Tumor Tests (d)

P = 0.015 P=0.139
P = 0.027

Cochran-Armitage Trend Test (d)

P=0.033
Fisher Exact Test (d)
P = 0.059
P=0.039

2-Mercaptobenzothiazole, NTP TR 332 100

TABLE B3. ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN T H E TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 188 mg/kg 375 mg/kg

Pituitary Gland: Adenoma or Adenocarcinoma

Overall Rates (a) 16/49(33%) 24/50(48%) 25/50(50%)
Adjusted Rates (b) 46.2% 62.3% 73.2%
Terminal Rates (c) 10/28(36%) 17/31(55%) 16/25(64%)
Week of First Observation 72 67 82 '
Life Table Tests (d) P=0.024 P=0.206 P = 0.036
Incidental Tumor Tests (d) P=0.028 P=0.186 P = 0.050
Cochran-ArmitageTrend Test (d) P=0.051
Fisher Exact Test (d) P = 0.088 P = 0.061
Adrenal Gland: Pheochromocytoma
Overall Rates (a) 1/50(2%) 5/50(10%) 6/50(12%)
Adjusted Rates (b) 3.6% 14.6% 23.0%
Terminal Rates (c) 1/28(4%) 3/31(10%) 5/25(20%)
Week of First Observation 104 96 97
Life Table Tests (d) P=0.030 P=0.137 P=0.041
Incidental Tumor Tests (d) P=0.038 P=0.214 P=0.052
Cochran-ArmitageTrend Test (d) P = 0.049
Fisher Exact Test (d) P=0.102 P = 0.056
Thyroid Gland: C-cell Adenoma
Overall Rates (a) 5/50(10%) 2/50(4%) 3/50(6%)
Adjusted Rates (b) 16.4% 6.5% 10.3%
Terminal Rates (c) 4/28(14%) 2/31(6%) 2/25(8%)
Week of First Observation 77 104 82
Life Table Tests (d) P=0.302N P=0.175N P = 0.395N
Incidental Tumor Tests (d) P = 0.365N P = 0.227N P=0.477N
Cochran-Armitage Trend Test (d) P=0.274N
Fisher Exact Test (d) P=0.218N P = 0.357N
Thyroid Gland: C-cell Adenoma o r Carcinoma
Overall Rates (a) 5/50(10%) 3/50(6%) 4/50(8%)
Adjusted Rates (b) 16.4% 9.1% 14.2%
Terminal Rates (c) 4/28(14%) 2/31 (6%) 3/25(12%)
Week of First Observation 77 101 82
Life Table Tests (d) P = 0.473N P=0.297N P = 0.546N
Incidental Tumor Tests (d) P=0.512N P=0.302N P = 0.628N
Cochran-ArmitageTrend Test (d) P = 0.427N
Fisher Exact Test (d) P=0.357N P=0.500N
Mammary Gland: Fibroadenoma
Overall Rates (a) 12/50(24%) 17/50(34%) 17/50(34%)
Adjusted Rates (b) 37.5% 43.5% 50.4%
Terminal Rates (c) 9/28(32%) 10131 (32%) 9/25(36%)
Week of First Observation 63 67 64
Life Table Tests (d) P=0.121 P = 0.336 P = 0.150
Incidental Tumor Tests (d) P=0.117 P30.289 P = 0.144
Cochran-ArmitageTrend Test (d) P=0.164
Fisher Exact Test (d) P = 0.189 P = 0.189
Clitoral Gland: Adenoma
Overall Rates (a) 5/50(10%) 2/50(4%) 3/50(6%)
Adjusted Rates (b) 15.6% 5.8% 10.7%
Terminal Rates (c) 3/28(11%) 1/31 (3%) 2/25(8%)
Week ofFirst Observation 77 100 92
Life Table Tests (d) P = 0.297N P = 0.166N P=0.383N
Incidental Tumor Tests (d) P = 0.319N P=0.198N P = 0.445N
Cochran-Armitage Trend Test (d) P = 0.274N
Fisher Exact Test (d) P=0.218N P=0.357N

101 2-Mercaptobenzothiazole, NTP TR 332

TABLE B3. ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 188 mg/kg 375 mg/kg

Clitoral Gland: Carcinoma

Overall Rates la)
4/50(8%)
8/50(16%) 1150(2%)
Adjusted Rates (b)
12.3%
22.3% 4.0%
Terminal Rates (c)
2/28 (7%) 5/31(16%) 1125 (4%)
Week of First Observation
53
57
Life Table Tests (d)
P=0.231N
P = 0.247 P =0.218N
Incidental Tumor Tests (d)
P =0.273N
P=0.258 P = 0.249N
Cochran-ArmitageTrend Test (d)
P = 0.188N
Fisher Exact Test (d)
P=0.178
P=0.181N

Clitoral Gland: Adenoma or Carcinoma

Overall Rates (a)
9/50(18%)
10150 (20%) 3/50(6%)
Adjusted Rates (b)
26.6%
27.2% 10.7%
Terminal Rates (c)
5/28(18%)
6/31(19%) 2/25 (8%)
Week of First Observation
53
57 92
Life Table Tests (d)
P=0.084N
P = 0.574N P = 0.085N
Incidental Tumor Tests (d)
P = 0.100N P=0,592N P=O.lllN
Cochran-ArmitageTrend Test (d)
P = 0.060N
Fisher Exact Test (d)
P = 0.500 P=0.061N

Uterus: Endometrial Stromal Polyp

Overall Rates (a)
13/50(26%) 14/50(28%) 8/50(16%)

Adjusted Rates (b)

38.0% 40.0% 26.8%
Terminal Rates (c)
8/28 (29%) 11/31 (35%) 5/25(20%)
Week of First Observation
63 89 82
Life Table Tests (d)
P = 0.184N P = 0.509N P = 0.206N
Incidental Tumor Tests (d)
P=0.181N P=0.490 P = 0.226N
Cochran-ArmitageTrend Test (d)
P =0.144N
Fisher Exact Test (d)
P = 0.500 P = 0.163N
All Sites: Benign Tumors
Overall Rates (a)
31/50(62%) 41/50(82%)
36/50(72%)
Adjusted Rates (b)
81.3% 95.3%
94.7%
Terminal Rates (c)
21/28(75%) 29/31(94%)
23/25(92%)
Week of First Observation
63 67
64
Life Table Tests (d)
P=0.081 P=O.219
P = 0.123
Incidental Tumor Tests (d)
P = 0.045 P =0.127
P = 0.070
Cochran-ArmitageTrend Test (d)
P=0.157
Fisher Exact Test (d)
P = 0.022 P = 0.198
All Sites: Malignant Tumors
Overall Rates (a) 14/50(28%) 21/50(42%) 13/50(26%)

Adjusted Rates (b) 37.0% 49.3% 37.2%

Terminal Rates (c) 5/28 (18%) 10131 (32%) 5/25 (20%)

Week of First Observation 51 57 79

Life Table Tests (d) P=0.538N P=0.274 P=0.547N

Incidental Tumor Tests (d) P=0.329N P = 0.273 P=0.335N

Cochran-ArmitageTrend Test (d) P = 0.459N

Fisher Exact Test (d) P = 0.104 P = 0.500N
All Sites: All Tumors
Overall Rates (a)
37/50(74%) 46/50(92%) 40150 (80%)
Adjusted Rates (b)
86.0% 97.9% 95.2%
Terminal Rates (c)
22/28 (79%) 30131 (97%) 23/25(92%)
Week of First Observation
51 57 64
Life Table Testa (d)
P= 0.180 P = 0.347 P =0.239
Incidental Tumor Testa (d)
P=0.088 P=O.O91 P=0.173
Cochran-ArmitageTrend Test (d)
P = 0.257
Fisher Exact Test (d)
P = 0.015 P = 0.317

2-Mercaptobenzothiazole,NTP TR 332 102

TABLE B3. ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YFAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

(a)Number of tumor-bearing animals/number of animals examined a t the site

(b) Kaplan-Meier estimated tumor incidences at the end of the study after adjusting for intercurrent mortality
(c)Observed tumor incidence a t terminal kill
(d)Beneath the vehicle control incidence are the P values associated with the trend tgst. Beneath the dosed group incidepce are
the P values corresponding to pairwise comparisons between that dosed group and the vehicle controls. The life table analysis
regards tumors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The incidental tumor
test regards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence
rates. A negative trend or lower incidence in a dosed group is indicated by (N).
(e) No P value is reported because no tumors were observed in the dosed and vehicle control groups.

103 2-Mercaptobenzothiazole, NTP TR 332

TABLE B4a. HISTORICAL INCIDENCE O F LEUKEMIA IN FEMALE F344/N RATS ADMINISTERED CORN
OIL BY GAVAGE (a)

Incidence in Vehicle Controls

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence
TOTAL 27 111,450 ( 18.7%)

SD (b) 8.52%

Range (c)
High 21/50
LOW 2150

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Standard deviation
(c)Range and SD are presented for groups of 35 or more animals.
TABLE B4b. HISTORICAL INCIDENCE O F PITUITARY GLAND TUMORS IN FEMALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Adenoma Carcinoma o r Adenoma, Carcinoma, or
Adenocarcinoma Adenocarcinoma

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence
TOTAL (b) 52011,407 (37.0%) (c)4311,407 (3.1%) (b,c)56111,407 (39.9%)

SD (d) 8.35% 2.90% 8.47%

Ran e ( e )
Ifigh 27/49 5/47 30149
LAW 9150 0150 11/50

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Includes 449 adenomas, NOS,and 72 chromophobe adenomas
(c)Includes 33 carcinomas, NOS,6 adenocarcinomas, NOS, and 4 chromophobe carcinomas
(d)Standard deviation
(e)Range and SD are presented for groups of 35 or more animals.
TABLE B4c. HISTORICAL INCIDENCE O F ADRENAL GLAND TUMORS I N FEMALE F344/N RATS
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Pheochromocytoma Malignant Pheochromocytoma
Pheochromocytoma o r Malignant
Pheochromocytoma
~~~~ ~~~~ ~

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence
TOTAL 8211,443 (5.7%) 511,443 (0.3%) 8611,443 (6.0%)

SD (b) 3.59% 0.77% 3.56%

Ran e(c)
I-figh 7150 1/50 8/50

LOW 0150 0/50 1/50

(a) Data as of August 30,1985, for studies of a t least 104 weeks

(b)Standard deviation
(c) Range and SD are presented for groups of 36 or more animals.

2-Mercaptobenzothiazole, NTP TR 332 104

TABLE B5. SUMMARY OF T H E INCIDENCE OF NONNEOPLASTIC LESIONS I N FEMALE RATS I N T H E
TWO-YEAR GAVAGE STUDY OF 8-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS NECROPSIED 50 50 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 50 50 50

INTEGUMENTARY SYSTEM
*Subcutaneous tissue (50) (50) (50)
Cyst, NOS 1 (2%)
Steatitis 1 (2%)
Inflammation, chronic 1 (2%) 1 (2%)

RESPIRATORY SYSTEM
*Nasal cavity (50) (50) (50)
Hemorrhage 2 (4%) 4 (8%)
Inflammation, acute 1 (2%) 2 (4%)
Inflammation, chronic 4 (8%) 1 (2%) 2 (4%)
Foreign material, NOS 1 (2%) 1 (2%)
*Nasal turbinate (50) (50) (50)
Inflammation, active chronic 2 (4%)
Inflammation, chronic 2 (4%) 3 (6%)
#Lung/bronchiole (50) (50) (50)
Inflammation, acute 1 (2%)
#Lung (50) (50) (50)
Mineralization 1 (2%)
Congestion, NOS 3 (6%) 16 (32%) 7 (14%)
Edema, NOS 2 14%) 3 (6%)
Hemorrhage 10 (20%) 10 (20%) 9 (18%)
Pneumonia, interstitial chronic 3 (6%) 2 (4%) 1 (2%)
Bronchopneumonia, chronic 2 (4%)
Granuloma, pyogenic 1 (2%)
Foreign material, NOS 1 (2%) 1 (2%)
Hyperplasia, alveolar epithelium 1 (2%)
Histiocytosis 2 (4%) 5 (10%) 3 (6%)

HEMATOPOIETIC SYSTEM
#Spleen (50) (50)
Hematoma, NOS
Pigmentation, NOS 50 1100%) 49 (98%)
Hyperplasia, lymphoid 2 (4%) 1 (2%)
Hematopoiesis . 38 (76%) 38 (76%) 41 (82%)
#Splenic capsule (50) (50) (50) ,
Fibrosis 1 (2%)
#Mandibular lymph node (50) (50) (50)
Congestion, NOS 1 (2%)
Plasmacytosis 1 (2%)
Hyperplasia, lymphoid 4 (8%) 4 (8%) 2 (4%)
#Mesenteric lymph node (50) (50) 150)
Congestion, NOS 1 (2%) 2 (4%) 1 (2%)
Hyperplasia, lymphoid 2 (4%)
#Liver (50) (50) (50)
Hematopoiesis 3 (6%) 1 (2%)
#Thymus (50) (50) (50)
Embryonal duct cyst 1 (2%)
Multiple cysts 1 (2%)
Congestion, NOS 3 (6%) 1 (2%)
Hemorrhage 1 (2%)

105 &Mercaptobenzothiazole, NTP TR 332

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

CIRCULATORY SYSTEM
#Heart (50) (50) (50)
Inflammation, chronic 46 (92%) 47 (94%) 47 (94%)
*Aorta (50) (50) (50)
Periarteritis 1 (2%)
*Pulmonary artery (50)
(50) (50)
Mineralization 4 (8%)
2 (4%) 7 (14%)
*Pulmonary vein (50)
(50) (50)
Mineralization 2 (4%)

DIGESTIVE SYSTEM
*Intestinal tract (50) (50) (50)
Bezoar 1 (2%)
*Tongue (50)
(50) (50)
Hyperplasia, epithelial 1 (2%)

#Salivary gland
(50) (50) (50)
Inflammation, chronic
1 (2%)
Atrophy, NOS
5 (10%) 2 (4%) 5 (10%)
#Liver
(50) (50) (50)

Accessory structure
6 (12%) 8 (16%) 1 (2%)

Bile stasis
1 (2%)

Hemorrhage
1 (2%)

Inflammation, acute
1 (2%)

Inflammation, chronic
1 (2%)
Granuloma, NOS
5 (10%) 12 (24%) 2 (4%)
Necrosis, NOS
1 (2%) 1 (2%) 3 (6%)
Pigmentation, NOS
1 (2%)
Cytoplasmic vacuolization
5 (10%) 4 (8%)
4 (8%)
Focal cellular change
43 (86%) 42 (84%)
39 (78%)
Hepatocytomegaly
1 (2%) 1 (2%)

Hyperplasia, NOS
3 (6%)

Angiectasis
1 (2%) 2 (4%)
2 (4%)
#Liver/periportal
(50) (50) (50)
Inflammation, chronic
42 (84%) 45 (90%) 45 (90%)
#Bile duct
(50) (50) (50)
Hyperplasia, NOS
34 (68%) 42 (84%) 45 (90%)
#Pancreas (50)
(50) (50)
Cystic ducts 1 (2%)

Lymphocytic inflammatory infiltration 1 (2%)

Inflammation, chronic 1 (2%) 1 (2%)
#Pancreatic acinus
(50)
(50) (50)
Focal cellular change
1 (2%)
2 (4%)
Atrophy, NOS
15 (30%) 27 (54%) 16 (32%)
Hyperplasia, NOS
6 (12%) 4 (8%) 4 (8%)
#Esophagus
(50) (50) (50)
Ulcer, NOS
1 (2%)
Inflammation, acute
1 (2%) 1 (2%)
Necrosis, NOS
1 (2%)
#Stomach
(49)
(50) (50)
Bezoar
1 (2%)

#Gastric fundal gland

(49)
(50) (50)
Dilatation, NOS
31 (63%)
41 (82%) 31 (62%)
#Glandular stomach
(49) (50) (50)
Inflammation, active chronic
1 (2%)
Dysplasia, epithelial
1 (2%)
#Forestomach
(49) (50) (50)
Ulcer, NOS
3 (6%) 5 (10%)
Inflammation, acute
1 (2%) 2 (4%) 6 (12%)
Inflammation, active chronic
1 (2%) 2 (4%) 1 (2%)
Necrosis, NOS
1 (2%)
Hyperplasia, epithelial
1 (2%) 4 (8%) 1 (2%)
Hyperkeratosis
1 (2%) 4 (8%) 1 (2%)

2-Mercaptobenzothiazole, NTP TR 332 106

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

DIGESTIVE SYSTEM (Continued)

#Small intestine
Inflammation, acute
#Duodenum
Bezoar
#Ileum
Inflammation, chronic
#Cecum
Necrosis, NOS

URINARY SYSTEM
#Kidney (50)
(50)
(50)
Nephropathy 38 (76%) 42 (84%) 41 (82%)
# Kidneyltubule (50)
(50) (50)
Mineralization 46 (92%) 44 (88%) 46 (92%)
Pigmentation, NOS 46 (92%) 48 (96%) 46 (92%)
Hyperplasia, focal 1 (2%)
# Kidneylpelvis (50)
(50)
(50)
Calculus, microscopic examination 1 (2%)
Mineralization 1 (2%)
Hemorrhage 1 (2%)
Hyperplasia, epithelial 1 (2%)

ENDOCRINE SYSTEM
#Pituitary (49) (50)
(50)

Multiple cysts 1 (2%)

Hematoma, organized 1 (2%)
#Pituitary intermedia (49) (50)
(50)
Cyst, NOS 2 (4%) 1 (2%) 1 (2%)
Multiple cysts 1 (2%)
Hemorrhagic cyst 1 (2%)
#Anterior pituitary (49) (50) (60)
Cyst, NOS 5 (10%)
6 (12%) 10 (20%)
Multiple cysts 20 (41%) 22 (44%) 11 (22%)
Congestion, NOS 1 (2%)
Hemorrhage 1 (2%)
Hemorrhagic cyst 5 (10%) 1 (2%)
Pigmentation, NOS 1 (2%)
Hyperplasia, NOS 8 (16%) 10 (20%) 6 (12%)
Angiectasis 2 (4%) 1 (2%) 1 (2%)
#Adrenal (50)
(50)
(50)
Accessory structure 1 (2%)
#Adrenal cortex (50)
(50)
(50)
Accessory structure 2 (4%)

Congestion, NOS 1 (2%)

1 (2%)

Hemorrhagic cyst 1 (2%)

Inflammation, chronic 1 (2%)

Degeneration, lipoid 8 (16%)

19 (38%)
15 (30%)

Necrosis, NOS 1 (2%)

1 (2%)

Metamorphosis, fatty 1 (2%)

Pigmentation, NOS 3 (6%)

1 (2%) 1 (2%)

Hypertrophy, focal 1 (2%)

2 (4%)

Hyperplasia, NOS 11 (22%)

8 (16%) 9 (18%)

#Adrenal medulla (50)

(50)
(50)
Necrosis, NOS 1 (2%)
Hyperplasia, NOS 5 (10%) 8 (16%) 2 (4%)

107 2-Mercaptobenzothiazole,NTP TR 332

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ENDOCRINE SYSTEM (Continued)

#Thyroid
(50) (50) (50)
Embryonal duct cyst
2 (4%) 1 (2%)
Mineralization
1 (2%)
Cystic follicles
4 (8%) 3 (6%) 6 (12%)
Inflammation, chronic
1 (2%)
Hyperplasia, C-cell
30 (60%) 42 (84%) 34 (68%)
#Thyroid follicle
(50) (60) (60)
Atrophy, focal
1 (2%)
#Pancreatic islets
(50) (50) (50)
Focal cellular change
1 (2%)

REPRODUCTIVE SYSTEM
*Mammary gland
(50) (50) (50)
Multiple cysts
26 (52%) 40 (80%) 33 (66%)
Hyperplasia, NOS
1 (2%)
*Preputial gland
(50) (50) (50)
Inflammation, chronic
1 (2%)
*Clitoral gland (50) (50 1 (50)
Cystic ducts 1 (2%)
Inflammation, suppurative 3 (6%) 1 (2%)

Inflammation, active chronic 7 (14%) 7 (14%) 4 (8%)

Inflammation, chronic 18 (36%) 25 (50%) 18 (36%)

#Uterus
(50)
(50) (50)

Dilatation, NOS
2 (4%)
3 (6%)

Hydrometra
1 (2%) 1 (2%)

Hematoma, NOS
1 (2%)
Hematoma, organized
1 (2%) 1 (2%)
Inflammation, chronic
1 (2%)
Hyperplasia, epithelial
1 (2%)
#Cervix uteri
(50) (50) (50)
Polyp, NOS
1 (2%)
#Uterus/endometrium
(50) (50) (50)
Inflammation, chronic
1 (2%)
Hyperplasia, cystic
9 (18%) 14 (28%) 6 (12%)
#Ovary
(50)
(50) (50)
Parovarian cyst
2 (4%)
5 (10%) 2 (4%)
Inflammation, chronic
1 (2%)
#Mesovarium
(50) (50) (50)
Steatitis
1 (2%)

NERVOUS SYSTEM
#Brain
(50)
(50) (50)
Compression, NOS
6 (12%)
2 (4%) 5 (10%)
Mineralization
1 (2%)
Malacia
1 (2%)

SPECIAL SENSE ORGANS

*Eye (50)
(50) (50)
Retinopathy 1 (2%)
9 (18%)
*Eye/sc le r a (50) (50) (50)
Mineralization 1 (2%)
*Eye/crystalline lens (50) (50) (50)
Cataract 8 (16%)
*Nasolacrimal duct (50)
(50) (50)
Hemorrhage 3 (6%)
1 (2%) 1 (2%)
Inflammation, active chronic 1 (2%) 1 (2%)
*Harderian gland
(50) (50) (50)
Inflammation, chronic
1 (2%)

2-Mercaptobenzothiazole, NTP TR 332 108

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

MUSCULOSKELETAL SYSTEM
None

BODY CAVITIES
*Mesentery (50) (50) (50)
Steatitis 6 (12%) 7 (14%) 7 (14%)

ALLOTHERSYSTEMS
*Multiple organs (50) (50) (50)
Inflammation, chronic 1 (2%)
Adipose tissue
Steatitis 1

SPECIAL MORPHOLOGY SUMMARY

None

* Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
# Number of animals examined microscopically a t thissite

109 2-Mercaptobenzothiazole, NTP T R 332

2-Mercaptobenzothiazole, NTP TR 332 110
APPENDIX C

SUMMARY OF LESIONS IN MALE MICE IN THE

TWO-YEAR GAVAGE STUDY OF

2-MERCAPTOBENZOTHIAZOLE

PAGE

TABLE C l SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE

TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 113
TABLE C2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 116

TABLE C3 ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR GAVAGE

STUDY OF 2-MERCAPTOBENZOTHIAZOLE 122

TABLE C4 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE

IN THE TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 126

111 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole, NTP TR 332 112
TABLE C l . SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS MISSING 1
ANIMALS NECROPSIED 49 50 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 49 50 50

INTEGUMENTARY SYSTEM
*Skin (49) (50) (50)
Keratoacanthoma 1 (2%)
*Subcutaneous tissue
(49) (50) (50)
Sarcoma, NOS
1 (2%) 2 (4%)
Fibroma
1 (2%) 3 (6%) 1 (2%)
Fibrosarcoma
2 (4%) 2 (4%) 1 (2%)

RESPIRATORY SYSTEM
*Nasal cavity (49) (50) (50)
Fibroma 1 (2%)
#Lung (49) (50) (50)
Adenocarcinoma, NOS, metastatic 1 (2%)
Hepatocellular carcinoma, metastatic 1 (2%)
Alveolar/bronchiolar adenoma 3 (6%) 4 (8%) 4 (8%)
Alveolar/bronchiolar carcinoma 5 (10%) 5 (10%) 1 (2%)
Sarcoma, NOS, metastatic 1 (2%)
Neurilemoma, metastatic 1 (2%)

HEMATOPOIETIC SYSTEM
*Multiple organs (49) (50) (50 )
Malignant lymphoma, lymphocytic type 1 (2%)
Malignant lymphoma, histiocytic type 3 (6%) 1 (2%)
Malignant lymphoma, mixed type 2 (4%) 1 (2%) 2 (4%)
#Mesenteric lymph node (49) (50) (48)
Malignant lymphoma, mixed type 1 (2%)
#Liver (49) (50) (50)
Malignant lymphoma, NOS 1 (2%)
XPeyer’s patch (49) (50) (50)
Malignant lymphoma, mixed type 1 (2%)

CIRCULATORY SYSTEM
#Spleen (49)
Hemangiosarcoma
#Liver
(49)
Hemangioma

Hemangiosarcoma

#Testis

Hemangioma

DIGESTIVE SYSTEM
#Salivary gland (49) (50) (50)
Adenocarcinoma, NOS, metastatic 1 (2%)
#Liver
(49) (50) (50)
Hepatocellular adenoma
11 (22%) 14 (28%) 9 (18%)
Hepatocellular carcinoma
5 (10%) 9 (18%) 6 (12%)
Sarcoma, NOS
2 (4%)

113 2-Mercaptobenzothiazole,NTP TR 332

TABLE C l . SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

URINARY SYSTEM
#Kidney (49) (50) (50)
Tubular cell adenoma 1 (2%)
#Kidney/cortex (49) (50) (50)
Adenocarcinoma, NOS, metastatic 1 (2%)

ENDOCRINE SYSTEM
#Anterior pituitary (49) (48)
Adenoma, NOS 1 (2%)
#Adrenal (49) (50)
Cortical adenoma 1 (2%)
#Adrenal medulla (49) (50)
Pheochromocytoma 3 (6%)
#Thyroid (49) (50)
Follicular cell adenoma 2 (4%)
#Pancreatic islets (49) (50)
Islet cell adenoma

REPRODUCTIVE SYSTEM
*Mammary gland (49) (50) (50)
Adenocarcinoma, NOS 1 (2%)
#Testis (49) (50) (50)
Interstitial cell tumor 1 (2%)

NERVOUS SYSTEM
None

SPECIAL SENSE ORGANS

*Harderian gland (49) (50) (50)
Adenoma, NOS 3 (6%) 2 (4%) 2 (4%)

MUSCULOSKELETAL SYSTEM
None

BODY CAVITIES
*Mediastinum (49) (50 ) (50)
Alveolar/bronchiolar carcinoma, invasive 1 (2%)
Neurilemoma, metastatic 1 (2%)
*Pleura (49) (50) (50)

Alveolar/bronchiolar carcinoma, invasive 2 (4%)

ALL OTHER SYSTEMS

Orbital region
Neurilemoma, malignant 1

ANIMAL DISPOSITION SUMMARY

Animals initially in study
50 50
50

Natural death
4 6
12

Moribund sacrifice
7 11
2

Terminal sacrifice
38
33
30

Accidentally killed, NOS

6

Animal missing
1

2-Mercaptobenzothiazole, NTP TR 332 114

TABLE C l . SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

TUMOR SUMMARY

Total animals with primary tumors** 31 39 25

Total primary tumors 42 55 34

Total animals with benign tumors 20 24 16

Total benign tumors 22 32 19

Total animals with malignant tumors 20 21 14

Total malignant tumors 20 23 15

Total animals with secondary tumors## 2 3 1

Total secondary tumors 4 5 1

* Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
** Primary tumors: all tumors except secondary tumors
# Number of animals examined microscopically at this site
# # Secondary tumors: metastatic tumors or tumors invasive into a n adjacent organ

115 2-Mercaptobenzothiazole,NTP TR 332

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE : VEHICLE CONTROL
ANIMAL 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

NUMBER 2 0 0 0 2 1 1 4 1 1 3 2 0 0 0 0 0 0 1 1 1 1 1 1 2

0 1 9 8 7 7 8 5 1 2 1 1 2 3 4 5 6 7 0 3 4 5 6 9 2

WEEKS ON
STUDY

Skm
+ + + + + + + + + + + M + + + + + + + + + + + + +

Keratoacanthoma

Subcutaneous tissue
+ + + + + + + + + + + M + + + + + + + + + + + + +

Sarcoma. NOS
X

F~hroma

F~brosarcoma
X X

Lungs and bronchi

+ + + + + + + + + + + M + + + + + + + + + + + + +

Adenocarcinoma. NOS, metastatic

X

Alveolarhronchiolar adenoma
X

Alveolarhronch~olarcarcinoma
X

Sarcoma. NOS, metastat~c

X

Trachea + + + + + + + + + + + M + + + + + + + + + + + + +

Bone marrow
+ + + + + + + + + + + M + + + + + + + + + + + + +

Spleen
+ + + + + + + + + + + M + + + + + + + + + + , + c

L mph nodes
+ + + + + + + + + + + M + + + + + + + + + + + + +

TKY~US + + + + + + + + + + - M - + + + + + + - + + + - +

Heart + + + + + + + + + + + M + + + + + + + + + + + + +

Salivary gland
+ t t + + + + + + + + M + + + + + + + + + + + + +

Adenocarcmoma, NOS, metastatic

X
Llver
+ + + + + + + + + + + M + + + + + + + + + + + + +

Hepatocellular adenoma
X X
X X X

Hepatocellular carcinoma
X X X X X
Blle duct
+ + + + + + + + + + + M + + + + + + + + + + + + +

Gallbladder & common bile duct

+ + + + + + + + + + + M + + + + + + + + + + + + +

Pancreas
+ + + + + + + + + + + M + + + + + + + + + + + + +

:22r
Small intestlne

+ + + + + + + + + + + M + + + + + + + + + + + + +
+ + + + + + + + + + + M + + + + + + + + + + + + +
+ + + + + + + + + + + M + + + + + + + + + + + + +

Malignant lymphoma. mixed type

Large Intestme
. . . . . . . . . . . . . . . . . . . . . . . . .

K~dney . . . . . . . . . . . . . . . . . . . . . . . . .

Adenocarcinoma, NOS, metastatic X

Unnary bladder + + + + + + + + + + + M + + + + + + + + + + + + +

Pituitary + + + + + + + + + + + M + + + + + + + + + + + + +
Adrenal + t + + + + + + + + + M + + + + + + + + + + + + +

Thyroid + + + + + + + + + + + M + + + + + + + + + + + + +

Follicular cell adenoma

Parathyroid - + + + - - + - + - X M - + + + + + + - + + + - +

Mammary gland N N N N N N N N + N N M N N N N N N N N + N N N +
Adenocarcmoma, NOS X

Testis + + + + + + + + + + + M + + + + + + + + + + + + +

Interstitial cell tumor

Prostate + + + + + + + + + + + M + + + + + + + + + + + + +

Bram + + + + + + + + + + + M + + + + + + + + + + + + +

Harderian gland N N N N N N N N N N N M N N N N N N N N N N N N N

Adenoma. NOS X

Mult~pleo rgans. NOS N N N N N N N N N N N M N N N N N N N N N N N N N

Malignant lymphoma, histiocyt~ctype X

Malignant lymphoma, mixed type X

+: Tissue examined m~croscopically No tissue information submitted

-: Required t ~ s s u enot examined m~croscoplcally C ; Necropsy. no histology due to protocol
X: Tumor ~ n c ~ d e n c e A: Autolysis
N: Necropsy, no autolys~s,no microscopic exammation M: Animal missing
S: Animal missexed B: No necropsy performed

2-Mercaptobenzothiazole, NTP TR 332

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: VEHICLE CONTROL
(Contin ued)
ANIMAL 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

NUMBER
2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 5

3 4 5 6 8 9 0 2 3 4 5 6 7 8 9 0 1 2 3 4 6 7 8 9 0

WEEKS ON Y 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 l l l l l

STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3

Skin t t t t t t t t t t t t t t t t t + t t t + t t + *49

Keratoacanthoma X 1

Subcutaneous tissue t t t + t + t t + t t t t t t t t t t + + + + t t 149

Sarcoma, NOS 1

Fibroma X 1

Fibrosarcoma 2

Lunpr and bronchi t t t t t t t t t t t t t t t t t + t t t t t + t 49

Adenocarcinoma NOS metastatic 1

Alveolarlbmnchlblar ahenoma x x 3

Alveolarlbmnchiolar carcinoma x x X X 5

Sarcoma, NOS,metastatic 1

Trachea t t t t t + t t t + t + t t t + + + t t + + + + + 49

Bone marrow + t t t t t t t t t t t t t t t + t + t + + t + + 49

Spleen t t t t t t + t t t + t t t t t + t t + t + t + + 49

L mphncdes + t t t t t t t t + + t t + t + t + + t + + + + + 49

Tiymus t t t t t t t t + t t + t t + t + t + t + + t + t 45

Heart t t t + t t t t t t t + t t + t t + t + t + t + t 49

Salivary gland t t t + t + + t t t t + t t + t + t t + + + + + + 49

Adenocarcinoma, NOS,metastatic 1

Liver t + t t t t t t t + t t t + t + t t + t + + t t + 49

Hepatocellular adenoma x x x x x X 11

Hepatocellular carcinoma 5

Bile duct t t t t t t + t t + t + t + t t t t + t + + t + + 49

Gallbladder & common bile duct t t t t t t t t t t + t + t t t t + t t + t + t + *49

Pancreas t t t t t t t t t t t t t t t t + + + t t + + t + 49

+ t t t t t t t t t t t t t t t t + t t + + t + t 49

%kY
Small intestine
t t t t t t t t t t t t t t t t + + + t + + + + + 49

t t t t t t t t t t t t t + t + + t t + + + t + + 49

Malignant lymphoma, mixed type X 1

Large intestine t t t t t + t t t t t t t t + t + t t t t + + t + 49

Kidney t t t t t t t t t t t + + t t t + t t t + + t t + 49

Adenocarcinoma, NOS,metestatic 1

Unnary bladder t t t t + t t t t t t + + t t t + t t + t + + t + 49

Pituitary t t t t t t t t + t t + t t + t t t + t + + t + + 49

Adrenal t t t + t t t t t t t t t t t + t t + + + + t + + 49

Thyroid t t t t t t t t t + t t + + t t t + t t + + t + t 49

Follicular cell adenoma X 2

Parathy
Parathymid
mid - t t - t t t - - t - + - - + t t t - + - - t + - 29

Mammary gland N N N N N N N N N N N N N N N N N N N N N N N N N *49

Adenocarcinoma, NOS 1

Testis t t t t + t t t + t + t + t + + + + + t + + t + t 49

Interstitial cell tumor X 1

Prostate t t t t t t t t t t t t t + t t t t t t + t + + + 49

Brain + t + + + + + t + + t t t t + t + t t t + + + + + 49

ANS

Harderian gland N N N N N N N N N N N N N N N N N N N N N N N N N *49

Adenoma, NOS x x 3

~ ~ ~~ ~~ ~ _ _ _ _ _ ~ ~

Multiple organs NOS N N N N N N N N N N N N N N N N N N N N N N N N N *49

Malignant lymphoma, histiocytic type X X 3

Malignant lymphoma, mixed type - - X 2

Animals necropsied

117 2-Mercaptobenzothiazole, NTP TR 332

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE: LOW DOSE
ANIMAL

NUMBER

WEEKS ON 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1
STUDY 0 8 6 7 7 7 8 8 8 8 8 8 9 9 9 0 0 0 0 0 0 0 0 0 0
1 8 9 6 6 7 0 3 6 7 9 9 2 8 8 0 1 3 3 3 3 3 5 9 3

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . .

Sarcoma, NOS X X

Fibroma

Fibrosarcoma X X

M
Lungs and bmnchi . . . . . . . . . . . . . . . . . . . . . . . . .

Alveolarlbmnchiolar adenoma x x
Alveolarlbmnchiolar carcinoma X X " X
Neurilemoma, metastatic n
Trachea + + + + + + + t + + + + t + + + + + t + + + t + +
Nasal cavity . . . . . . . . . . . . . . . . . . . . . . . . .
Fibroma X

Bone marmw . . . . . . . . . . . . . . . . . . . . . . . . .

Spleen . . . . . . . . . . . . . . . . . . . . . . . . .

Hemangiosarcoma
L mphncdes . . . . . . . . . . . . . . . . . . . . . . . . .
Tgymus . . . . . . . . . . . . . . . . . . . . . . . . .

~~

Heart . . . . . . . . . . . . . . . . . . . . . . . . .

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Liver . . . . . . . . . . . . . . . . . . . . . . . . .
Hepatocellular adenoma x x x x x x x
Heoatocellular carcinoma x x x x x x
Hemanpoma
Malignant lymphoma, NOS X
Bile duct . . . . . . . . . . . . . . . . . . . . . . . . .
Gallbladder & common bile duct N + + + + t + + + + + + t t + + + N N + + + + + +
Pancreas . . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . .

:t%kY
Small intsstine
. . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . .

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Kidney . . . . . . . . . . . . . . . . . . . . . . . . .
Tubular cell adenoma
Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma. NOS
Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .

Cortical adenoma

Pheochromocytoma X X

Thyroid . . . . . . . . . . . . . . . . . . . . . . . . .

Parathyroid + + + + - - - - - - - + - + + + + - - - - + + + -
M
Mammary gland N N N N N N N N N N N N N N N N N N N N N N N N N
Testis . . . . . . . . . . . . . . . . . . . . . . . . .

Hemangioma
Prostate . . . . . . . . . . . . . . . . . . . . . . . . .

Brain . . . . . . . . . . . . . . . . . . . . . . . . .

mCIAL ORGANS
Hardenan gland Y N N N N N N N N N N N N N N N N N N N N N N N N
Adenoma, NOS X X

Pleura Y N N N N N N N N N N N N N N N N N N N N N N N N
Alveolar/bronchiolar carcinoma, invasive X
Mediastinum Y N N N N N N N N N N N N N N N N N N N N N N N N
Alveolar/bronchiolar carcinoma, invasive
Neunlemoma, metastatic X
~~

S
Multiple organs NOS Y N N N N N N N N N N N N N N N N N N N N N N N N
Malignant lydphoma, lymphocytlc type X
Mali nant lymphoma, mixed type
Orbitay repon
Neunlemoma. malignant X

2-Mercaptobenzothiazole, NTP TR 332 118

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: LOW DOSE
(Continued)
ANIMAL
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

NUMBER 1 1 1 1 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 4 4 4 4 5

2 3 4 6 0 1 3 4 5 6 7 9 0 2 3 4 6 1 8 9 4 5 6 7 0

TOTAL:
WEEKSON 1 1 1 1 1 1 1 1 1 1 1 ~ ~ 1 1 1 1 1 1 1PISSUES
l ~
~ ~

STUDY 0 0 0 0 0 0 ~ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0I'UMORE

3 3 3 3 3 4 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3

-
Subcutaneous tissue t t t t t t t t t t t t t t t t t t + + + + + t t '50

Sarcoma, NOS 2

Fibroma x x X 3

Fibrosarcoma 2

Lungs and bronchi + t t t t t t t t t t t + + t t t t + t + + + t t 50

Alveolarfbronchiolar adenoma X X 4

Alveolarfbronchiolar carcinoma X X 5

Neunlemoma, metastatic 1

Trachea + t t t t t t t t t + t + t t t + t t + + + + t t 50

Nasal cavity t + t + t t + + t + t + t t t t t t + N + + + t t *50

Fibroma 1

Bone marrow + t t t t t t t t t t t t + t t t + t t t + + t + 48

Spleen t t t t + t t t t t t t t t t t + t t + t t + t + 50

Hemanposarcoma X 1

L mphnodea + t t t t t + t t + t t t t t + + t t t t t t t + 50

Tgymus t t + t t - + t t t t t - + + t t t + t - + t t t 44

Heart t t t t t t t t t t t t + t t t t + + t + + + t + 50

Salivary gland t t t t t + t t t t t t + t + + + t t + + + t t t 50

Liver + + t t t t t t + t t t + t t t + t + t + t t t t 50

Hepatocellular adenoma X X x x x x X 14

Hepatocellular carcinoma X X X 9

HemanPoma X 1

Malignant lymphoma, NOS 1

Bile duct t t t t + t t t t t t t + t + t t t t t t + t t t 50

Gallbladder & common bile duct t t t t + t + t + + + t + + t + t t + t N + + t + * 50

Pancreas t t t t t t t t t t t + t t t t t t + t t + + t + 50

t t t t t t t t + t t + + t t + t t t + t t + t t 50

t t t t t t t t t t t t + + t t + + + t t t t t t SO

t t t t t t t t t t + + + t t t t + + t t t + t t 50

+ t t t t t t t + t t t + t t t t t t + + + + t t 50

Kidney + t + t t t t t t t t t t + t t + + t + + + + t t 50

Tubular cell adenoma X 1

Urinary bladder t t t t t t + + t t t + t t + t + + + + + + + + + 50

-
Pituitary + t t t t t t + + t t t t + + + t + + - + + + + + 48

Adenoma, NOS X 1

Adrenal t t t t t t t t t t t t t t t + + t + + t t t + t 50

Cortical adenoma X 1

Pheochromocytoma X 3

Thyroid + t t t t t t t + t t t t t t t + t t t + + t t t 50

Parathyroid t t t + - - t + t t t t t t - t t - + + - t + t - 31

Mammary gland N N N N N N N N N N N N N N N N N N N N N N N N N '50

Testis t + t t t t t + t t t t t t t t t t + + + + t t t 50

Hemannoma X 1

Prostate t t t t + + + t t t t t t t + + + t + t + + + t + 50

-
Brain t t t t t t + t t + + t + t + t + + + + + + + + + 50

~~ ~ -
ANS
Hardanan gland V N N N N N N N N N N N N N N N N N N N N N N N N '50

Adenoma, NOS 2

-
Pleura i N N N N N N N N N N N N N N N N N N N N N N N N $50

Alveolar/bronchiolar carcinoma. inv c 2

Mediastinum P N N N N N N N N N N N N N N N N N N N N N N N N *50

Alveolar/bronchiolar carcinoma, inv c 1

Neunlemoma, metastatic 1

-
S
Multiple organs NOS i N N N N N N N N N N N N N N N N N N N N N N N N *50
Malig lym h o k a lymphocytic type 1

Mali nant yymphbma. mixed type X 1

Orbitay m o o n
Neunlemoma, malignant 1

* Animals necropsied

119 2-Mercaptobenzothiazole, NTP TR 332

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE: HIGH DOSE
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
NUMBER 5 8 5 5 6 6 8 8 9 9 9 7 9 8 8 9 8 9 5 8 5 5 5 5 5
5 3 4 8 2 5 8 9 7 9 8 6 0 6 7 3 2 1 3 4 1 2 6 7 9
WEEKS ON ~ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1
STUDY 0 0 0 1 1 1 1 1 1 1 1 1 2 2 2 4 6 7 8 8 0 0 0 0 0
7 7 9 3 3 3 3 3 3 4 6 7 6 7 7 7 1 1 9 9 3 3 3 3 3

Subcutaneous tissue
. . . . . . . . . . . . . . . . . . . . . . . . .

Fibroma

Fibrosarcoma

Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . .

Hepatocellular carcinoma metastatic X

Alveolar/bronchiolar admoma X
Alveolar/bronchiolar carcinoma
Trachea . . . . . . . . . . . . . . . . . . . . . . . . .

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . .

Soleen . . . . . . . . . . . . . . . . . . . . . . . . .

L*m hnodes . . . . . . . . . . . . . . . . . . . . . . . . .

%a!ignant lymphoma, mixed type

Thymus . . . . . . . . . . . . . . . . . . . . . . . . .

~~

Heart . . . . . . . . . . . . . . . . . . . . . . . . .

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .

Liver .........................

Hepatocellular adenoma A

Hepatocellular carcinoma x x x
Sarcoma, NOS
HemanGosarcoma X
Bile duct . . . . . . . . . . . . . . . . . . . . . . . . .

Gallbladder & common bile duct + + N + + + + + + + + + + + N N + + + + + + + N +

Pancreas . . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . .

. . . . . . . . . . . . . . . . . . . . . . . . .

Small intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . .

EM
Kidney . . . . . . . . . . . . . . . . . . . . . . . . .

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma, NOS X
Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .

Thyroid - - . . . . . . . . . . . . . . . . . . . . . . .
Follicular cell adenoma
Parathyroid - - _ t + + + + - - - + - - - - + + + + - + + + +
Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . .

Islet call adenoma X

Mammary gland N N N N N N N N N N N N N N N N N N N N N N N N N
Testis . . . . . . . . . . . . . . . . . . . . . . . . .

Prostate . . . . . . . . . . . . . . . . . . . . . . . . .

Brain . . . . . . . . . . . . . . . . . . . . . . . . .

ORGANS

~~ ~

Harderian gland N N N N N N N N N N N N N N N N N N N N N N N N N
Adenoma, NOS
S

Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N

Malignant lymphoma, histiocytic type
Malignant lymphoma, mixed type

2-Mercaptobenzothiazole, NTP TR 332 120

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: HIGH DOSE
(Continued)
ANIMAL Ol 0 Ol Ol til Ol Ol Ol Ol 0 01 01 01 0 01 0 Ol Ol Ol Ol Ol Ol Ol Ol 1
NUMBER 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 8 8 8 9 9 9 9 0
0| l| 3\ 4| 6| 7] 8| 9| 0| 1 2| 3| 41 5 ?| 8 9| 0| 1| 5\ 2| 4\ 5| e| 0
TOTAL
WEEKS ON 11 11 11 11 11 1 1 11 iril 11 11 1 11 11 11 1 11 11 11 11 1 11 11 I TISSUES
STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 TUMORS
3| 3| 3| 3\ 3| 3 3 3| 3| 3\ 3\ 3\ 3 3| 3\ 3| 3 3| 3| 3| 3| 3 3| s| 3
INTEGUMENTARY SYSTEM
Subcutaneous tissue •50
Fibroma X 1
Fibrosarcoma X 1
HESKlHA'l'OKy SYSTEM
Lungs and bronchi
Hepatocellutar carcinoma, metastatic
+ + + + «. + + + + + + + + + + + + + + + + + + + + SO
1
Alveolar/bronchiolar adenoma X X X 4
Alveolar/bronchiolar carcinoma X 1
Trachea 49
HEMATOPOIETTC SYSTEM
Bone marrow SO
Spleen SO
Lymph nodes 48
Malignant lymphoma, mixed type X 1
Thymus + + + + + + + + + + + + ­ + + + + + + + + + + + + 46
CIRCULATORY SYSTEM
Heart 50
DIGESTIVE SYSTEM
Salivary gland 50
Liver 50
Hepatocellular adenoma X X X X X X X X 9
Hepatocellular carcinoma X X X 6
Sarcoma, NOS X X 2
Hemangiosarroma 1
Bile duct 50
Gallbladder & common bile duct •50
Pancreas 50
Esophagus 50
Stomach 50
Small intestine 50
Large intestine 50
URINARY SYSTEM
Kidney 50
Unnary bladder 48
ENDOCRINE SYSTEM
Pituitary 50
Adenoma, NOS 1
Adrenal 48
Thyroid 47
Follicular cell adenoma X 1
Parathyroid — + + + + + — — — + + — + — + + — — + + + -*- — — + 29
Pancreatic islets 50
Islet cell adenoma 1
REPRODUCTIVE SYSTEM
Mammary gland N N N N N N N N N N N N N N N N N N N N N N N N N •50
Testis + + -t- + -h + + -f- + + + + + -t- + + + + + + + -»- + + + SO
Prostate t + t H - + -)- + + + + 4-1- + 4- + + + -h + + + 1- + t + 50
NERVOUS SYSTEM
Brain + + + . + + + + + + + + + + + + + + + + + , + + + 50
SPECIAL SENSE ORGANS
Hardenan gland N N N N N N N N N N N N N N N N N N N N N N N N N •50
Adenoma, NOS X X 2
ALL OTHER SYSTEMS
Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N •50
Malig lymphoma, histiocytic type X 1
Malignant lymphoma, mixed type X X 2

* Animals necropsied

121 2-Mercaptobenzothiazole, NTP TR 332

TABLE C3. ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 mg/kg 750 mg/kg

Subcutaneous Tissue: Fibroma

Overall Rates (a)
1/49 (2%) 3/50 (6%)
1/50 (2%)
Adjusted Rates (b)
2.6% 9.1%
3.3%
Terminal Rates (c)
1/38 (3%) 3/33 (9%)
1/30 (3%)
Week of First Observation
103 103
103
Life Table Tests (d)
P = 0.523 P = 0.256
P = 0.708
Incidental Tumor Tests td)
P=0.523 P=0.256
P = 0.708
Cochran-ArmitageTrend Test (d)
P=0.603N
Fisher Exact Test (d)
P =0.316
P = 0.747N
Subcutaneous Tissue: Fibroma or Fibrosarcoma
Overall Rates (a)
3/49 (6%) 5/50(10%)
2/50 (4%)
Adjusted Rates (b)
6.8% 13.4%
6.7%
Terminal Rates (c)
1/38 (3%) 3/33 (9%)
2/30 (7%)
Week of First Observation
81 76
103
Life Table Tests (d)
P=0.568N P = 0.305
P = 0.627N
Incidental Tumor Tests (d)
P = 0.498 P=0.351
P = 0.605
Cochran-Armitage Trend Test (d)
P = 0.410N
Fisher Exact Test (d)
P = 0.369
P = 0.490N

Subcutaneous Tissue: Sarcoma or Fibrosarcoma

Overall Rates (a)
3/49 (6%) 4/50 (8%)
1/50 (2%)
Adjusted Rates (b)
6.2% 8.8%
3.3%
Terminal Rates (c)
0138 (0%) 0133 (0%)
1/30 (3%)
Week of First Observation
54 69
103
Life Table Tests (d)
P=0.393N P=0.472
P=0.433N
Incidental Tumor Tests (d)
P=0.528N P = 0.630
P = 0.595N
Cochran-Armitage Trend Test (d)
P = 0.244N
Fisher Exact Test (d)
P=0.511
P=0.301N

Subcutaneous Tissue: Fibroma, Sarcoma, or Fibrosarcoma

Overall Rates (a)
4/49 (8%) 7/50 (14%)
2/50 (4%)
Adjusted Rates (b)
8.7% 17.1%
6.7%
Terminal Rates (c)
1/38 (3%) 3/33 (9%)
2/30 (7%)
Week of First Observation
54 69
103
Life Table Tests (d)
P = 0.470N P = 0.227
P=0.484N
Incidental Tumor Tests (d)
P = 0.574 P = 0.304
P=0.616N
Cochran-Armitage Trend Test (d)
P=0.286N
Fisher Exact Test (d)
P = 0.274 P=0.329N
Lung: Alveolar/Bronchiolar Adenoma
Overall Rates (a)
3/49 (6%) 4/50 (8%)
4/50 (8%)
Adjusted Rates (b)
7.9% 12.1%
13.3%
Terminal Rates (c)
3/38 (8%) 4/33 (12%)
4/30 (13%l
Week of First Observation
103 103
103
Life Table Tests (d)
P=0.297 P = 0.423
P = 0.371
Incidental Tumor Tests (d)
P=0.297 P = 0.423
P=0.371
Cochran-Armitage Trend Test (d)
P = 0.435
Fisher Exact Test (d)
P = 0.51 1
P = 0.51 1

Lung: Alveolar/Bronchiolar Carcinoma

Overall Rates ( a )
5/49(10%) 5/50 (10%)
1/50 (2%)
Adjusted Rates (b)
13.2% 13.4%
3.3%
Terminal Rates (c)
5/38(13%)
3/33 (9%)
1/30 (3qo)
Week of First Observation
103 68
103
Life Table Tests (d)
P = 0.160N P = 0.555
P=0.163N
Incidental Tumor Tests (d)
P=0.158N P=0.595N
P =0.163N
Cochran-ArmitageTrend Test (d)
P = 0.085N
Fisher Exact Test (d)
P=0.617N
P=0.098N

2-Mercaptobenzothiazole, NTP TR 332 122

TABLE C3. ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN T H E TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 m g k g

Lung: Alveolar/Bronchiolar Adenoma o r Carcinoma

Overall Rates (a) 7/49(14%) 9/50(18%)
5/50(10%)

Adjusted Rates (b) 18.4% 24.9%

16.7%

Terminal Rates (c) 7/38(18%) 7/33(21%)

5/30(17%)

Week of First Observation 103 68

103

Life Table Tests (d) P=0.524N P=0.292

P=0.552N

Incidental Tumor Tests (d) P = 0.537N P=0.376

P=0.552N

Cochran-ArmitageTrend Test (d) P=0.318N

Fisher Exact Test (d) P=0.410
P=0.365N

Hematopoietic System: Malignant Lymphoma, Histiocytic Type

Overall Rates (a)
3/49(6%)
0/50(0%) 1/50(2%)
Adjusted Rates (b)
7.9%
0.0%
3.3%
Terminal Rates (c)
3/38(8%)
0133 (0%) 1/30(3%)
Week of First Observation
103
103
Life Table Tests (d)
P=0.231N
P = 0.147N P =0.393N
Incidental Tumor Tests (d)
P=0.231N
P = 0.147N P = 0.393N
Cochran-Armitage Trend Test (d)
P=0.171N

Fisher Exact Test (d)

P=0.117N
P=0.301N

Hematopoietic System: Malignant Lymphoma, Mixed Type

Overall Rates (a) 3/49(6%) 1/50(2%)
3/50(6%)

Adjusted Rates (b) 7.4% 3.0%

10.0%

Terminal Ratesk) 2/38(5%) 1/33(3%)

3/30(10%)

Week of First Observation 89 103 103

Life Table Tests (d) P = 0.484 P = 0.356N P=0.541

Incidental Tumor Tests (d) P = 0.445 P = 0.345N P=0.475

Cochran-ArmitageTrend Test (d) P = 0.585N

Fisher Exact Test (d) P=0.301N
P =0.651N
Hematopoietic System: Lymphoma, All Malignant
Overall Rates (a) 6/49( 12%) 3/50(6%)
4/50(8%)
Adjusted Rates (b) 15.1% 8.3%
13.3%
Terminal Rates (c) 5/38(13%) 1/33(3%)
4/30(13%)
Week of First Observation 89 98
103
Life Table Tests (d) P=0.440N P=0.311N
P=0.531N
Incidental Tumor Tests (d) P = 0.501N P=0.155N
P = 0.583N
Cochran-ArmitageTrend Test (d) P=0.286N
Fisher Exact Test (d) P=0.233N
P = 0.357N
Circulatory System: Hemangioma or Hemangiosarcoma
Overall Rates (a) 0149 (0%) 3/50(6%) 1/50(2%)
Adjusted Rates (b)
0.0% 9.1% 3.3%
Terminal Rates (c)
0138 (0%) 3/33(9%) 1/30(3%)
Week of First Observation
103 103
Life Table Tests (d)
P = 0.306 P = 0.097 P = 0.453
Incidental Tumor Tests (d)
P =0.306 P=0.097 P = 0.453
Cochran-ArmitageTrend Test (d)
P = 0.384
Fisher Exact Test (d)
P =0.125
P = 0.505
Liver: Hepatocellular Adenoma
Overall Rates (a) 11/49(22%)
14/50(28%)
9/50(18%)
Adjusted Rates (b) 27.0%
38.4%
30.0%
Terminal Rates (c) 9/38(24%)
11/33(33%)
9/30(30%)
Week of First Observation 84
89
103
Life Table Tests (d) P=0.468
P=0.203
P = 0.555
Incidental Tumor Tests (d) P=0.376
P =0.255
P = 0.476
Cochran-ArmitageTrend Test (d) P=0.339N

Fisher Exact Test (d) P = 0.343 P = 0.382N

123 2-Mercaptobenzothiazole, NTP TR 332

TABLE C3. ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 m g k g

Liver: Hepatocellular Carcinoma

Overall Rates (a)
5/49 (10%) 9/50 (18%)
6/50 (12%)
Adjusted Rates (b)
11.6% 21.1%
18.6%
Terminal Rates (c)
2/38 (5%) 3/33 (9%)
4/30 (13%)
Week of First Observation
75 76
71
Life Table Tests (d)
P = 0.243 P = 0.164
P=0.312
Incidental Tumor Tests (d)
P=0.120 P = 0.295
P=0.130
Cochran-ArmitageTrend Test (d)
P = 0.457
Fisher Exact Test (d)
P = 0.205
P =0.514
Liver: Hepatocellular Adenoma or Carcinoma
Overall Rates (a)
16/49 (33%) 21/50 (42%)
14/50 (28%)

Adjusted Rates (b)

36.6% 50.1%
43.6%

Terminal Rates (c)

11/38(29%) 13/33 (39%)
12/30 ( 40%)

Week of First Observation

75 76
71

Life Table Tests (d)

P=0.343 P = 0.126
P = 0.422

Incidental Tumor Tests (d)

P = 0.196 P=0.233
P=O.219

Cochran-ArmitageTrend Test (d)

P=0.348N
Fisher Exact Test (d)
P=0.226
P=0.388N

Adrenal Gland: Pheochromocytoma

Overall Rates (a)
0149 (0%) 3/50 (6%)
0148 (0%)

Adjusted Rates (b)

0.0% 9.1%
0.0%
Terminal Rates (c)
0138 (0%) 3/33 (9%)
0130 (0%)
Week of First Observation
103

Life Table Tests (d)

P=0.574 P = 0.097
(e)
Incidental Tumor Tests (d)
P = 0.574 P = 0.097
(e)
Cochran-Armitage Trend Test (d)
P = 0.635
Fisher Exact Test (d)
P=0.125
(e)
Harderian Gland: Adenoma
Overall Rates (a)
3/49 (6%) 2/50 (4%)
2/50 (4%)

Adjusted Rates (b)

7.4% 5.5%
6.7%

Terminal Rates (c)

2/38 (5%) 1/33 (3%)
2/30 (7%)

Week of First Observation

89 89
103

Life Table Tests (d)

P=0.522N P = 0.559N
P=0.616N

Incidental Tumor Tests (d)

P = 0.594 P=0.539N
P = 0.678

Cochran-ArmitageTrend Test (d)

P = 0.398N
Fisher Exact Test (d)
P = 0.490N
P = 0.490N

All Sites: Benign Tumors

Overall Rates (a)
20149 (41%) 24/50 (48%)
16/50 (32%)

Adjusted Rates(b)
47.2% 64.6%
5 1.6%

Terminal Rates (c)

16/38 (42%) 20133 (61%)
15/30 (50%)

Week of First Observation

84 89
89

Life Table Tests (d)

P = 0.460 P=0.121
P = 0.545

Incidental Tumor Tests (d)

P=0.322 P =0.215
P=0.332

Cochran-ArmitageTrend Test (d)

P=0.212N
Fisher Exact Test (d)
P = 0.303 P=0.241N

All Sites: Malignant Tumors

Overall Rates (a)
20149 (41%) 21/50 (42%)
14/50 (28%)

Adjusted Rates (b)

42.9% 45.0%
43.6%

Terminal Rates (c)

12/38 (32%) 8/33 (24%)
12/30 (40%)

Week of First Observation

54 68
71

Life Table Tests (d)

P = 0.467N P = 0.363
P = 0.466N

Incidental Tumor Tests (d)

P=0.409 P = 0.379N
P = 0.430

Cochran-ArmitageTrend Test (d)

P=O.lllN
Fisher Exact Test (d)
P=0.534
P=0.129N

2-Mercaptobenzothiazole, NTP TR 332 124

TABLE C3. ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 mg/kg

All Sites: All Tumors

Overall Rates (a) 31/49 (63%) 39/50 (78%) 25/50 (50%)
Adjusted Rates (b) 65.6% 82.9% 75.8%
Terminal Rates (c) 22/38 (58%) 25/33 (76%) 22/30 (73%)
Week of First Observation 54 68 71
Life Table Tests (d) P=0.372 P = 0.037 P=0.479
Incidental Tumor Tests (d) P=O.119 P=0.104 P=0.158
Cochran-ArmitageTrend Test (d) P =0.100N
Fisher Exact Test (d) P = 0.082 P = 0.130N

(a)Number of tumor-bearing animals/number of animals examined a t the site

(bJ Kaplan-Meier estimated tumor incidences at the end of the study after adjusting for intercurrent mortality
(c)Observed tumor incidence a t terminal kill
(d)Beneath the vehicle control incidence are the Pvalues associated with the trend test. Beneath the dosed group incidence are
the P values corresponding to pairwise comparisons between that dosed group and the vehicle controls. The life table analysis
regards tumors in animals dying prior to terminal kill as being (directly or indirectly) the cause ofdeath. The incidental tumor
test regards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence
rates. A negative trend or lower incidence in a dosed group is indicated by (N).
(e)No P value is reported because no tumors were observed in the 750 mgkgand vehicle control groups.

125 2-Mercaptobenzothiazoie, NTP TR 332

TABLE C4. SUMMARY OF T H E INCIDENCE OF NONNEOPLASTIC LESIONS I N MALE MICE IN T H E
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dbse High Dose

ANIMALS INITIALLY IN STUDY so 50 50

ANIMALS MISSING 1
ANIMALS NECROPSIED 49 50 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 49 50 50

INTEGUMENTARY SYSTEM
*Skin
(49) (50) (SO)
Mineralization
1 (2%)
Ulcer, NOS
3 (6%) 1 (2%)
Inflammation, chronic
3 (6%) 3 (6%)
Exfoliative dermatitis
1 (2%)
Hyperkeratosis
1 (2%)
*Subcutaneous tissue
(49) (50) (50)
Steatitis
1(2%)
Abscess, NOS
1(2%)
Inflammation, chronic
2 (4%)
Granuloma, NOS
1(2%)
Granuloma, foreign body
2 (4%)
Granulation tissue
1 (2%)

RESPIRAT0 RY SYSTEM
*Nasal cavity (49) (50) (50)
Hemorrhage 10 (20%) 7 (14%) 7 (14%)
Lymphocytic inflammatory infiltration 2 (4%) 1 (2%) 1 (2%)
Inflammation, acute 3 (6%)
*Nasal turbinate
(49) (50) (50)
Inflammation, chronic
1 (2%)
#Lung (49) (50) (50)
Mineralization 1 (2%) 2 (4%) 3 (6%)
Emphysema, alveolar 1 (2%)
Congestion, NOS 1 (2%) 8 (16%)
Hemorrhage 11 (22%) 7 (14%) 16 (32%)
Bronchopneumonia, NOS 12 (24%) 16 (32%) 16 (32%)
Lymphocytic inflammatory infiltration 1 (2%) 1 (2%) 10 (20%)
Pneumonia, interstitial chronic 1 (2%)
Cholesterol deposit 2 (4%) 5 (10%) 8 (16%)
Hemosiderosis 1 (2%)
Hyperplasia, alveolar epithelium 12 (24%) 19 (38%) 13 (26%)
Histiocytosis 13 (27%) 20 (40%) 13 (26%)

HEMATOPOIETIC SYSTEM
#Bone marrow
(49) (48) (50)
Hemorrhage
1 (2%)
Fibrosis
2 (4%) 1 (2%)
Hyperplasia, granulocytic
40 (82%) 40 683%) 28 (56%)
#Spleen
(49) (50) (50)
Pigmentation, NOS
4 (8%) 5 (10%) 4 (8%)
Hyperplasia, lymphoid
12 (24%) 6 (12%) 10 (20%)
Hematopoiesis
12 (24%) 10 (20%) 4 (8%)
#Lymph node
(49) (50) (48)
Hyperplasia, lymphoid
3 (6%) 1 (2%) 1 (2%)
#Mandibular lymph node
(49) (50) (48)
Pigmentation, NOS
1 (2%) 2 (4%)
Hyperplasia, lymphoid
3 (6%) 2 (4%) 4 (8%)
#Mesenteric lymph node
(49) (50) (48)
Congestion, NOS
13 (27%) 2 (4%) 1 (2%)
Hemorrhage
1 (2%)
Inflammation, acute
1 (2%)
Inflammation, active chronic
1 (2%)
Hyperplasia, lymphoid
3 (6%) 7 (14%) 2 (4%)

2-Mercaptobenzothiazole,NTP TR 332 126

TABLE C4. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)
~~ ~~

Vehicle Control Low Dose High Dose

HEMATOPOIETIC SYSTEM (Continued)

#Liver (49) (50) (50)
Hematopoiesis 2 (4%) 1 (2%) 2 (4%)
#Peyer’s patch (49) (50) (50)
Hyperplasia, lymphoid 1 (2%)
#Thymus (45) (44) (46)
Embryonal duct cyst 1 (2%)
Cyst, NOS 1 (2%) 1 (2%)
Necrosis, NOS 1 (2%)
Hyperplasia, reticulum cell 1 (2%)
Hyperplasia, lymphoid 2 (4%) 1 (2%)

CIRCULATORY SYSTEM
#Heart (49) (50) (50)
Lymphocytic inflammatory infiltration 1 (2%)
Inflammation, chronic 4 (8%) 2 (4%) 1 (2%)
*Artery (49) (50) (50)
Mineralization 1 (2%)
Periarteritis 1 (2%)
*Aorta (49) (50) (50)
Mineralization 1 (2%)

DIGESTIVE SYSTEM
*Root oftooth (49) (50) (50)
Inflammation, active chronic 1 (2%)
*Pulp of tooth (49) (50) (50)
Dysplasia, NOS 1 (2%) 1 (2%)
#Salivary gland (49) (50) (50)
Mineralization 1 (2%)
Lymphocytic inflammatory infiltration 1 (2%) 1 (2%) 2 (4%)
Inflammation, chronic 1 (2%) 5 (10%)
Atrophy, NOS 1 (2%) 3 (6%) 3 (6%)
#Liver (49 ) (50) (50)
Mineralization 1 (2%)
Cyst, NOS 1 (2%)
Congestion, NOS 1 (2%)
Lymphocytic inflammatory infiltration 2 (4%)
Inflammation, acute 1 (2%) 1 (2%) 1 (2%)
Inflammation, active chronic 2 (4%)
Necrosis, coagulative 1 (2%)
Cytoplasmic vacuolization 7 (14%) 7 (14%) 9 (18%)
Focal cellular change 2 (4%) 3 (6%) 2 (4%)
Hepa tocytomegaly 1 (2%) 3 (6%) 1 (2%)
#Liver/centrilobular (49 (50) (50)
Necrosis, coagulative 1 (2%)
#Liver/periportal (49) (50) (50)
Inflammation, chronic 1 (2%)
*Gallbladder (49) (50) (50)
Cyst, NOS 2 (4%) 1 (2%)
Inflammation, acute 1 (2%)
#Pancreas (49) (50) (50)
Cyst, NOS 1 (2%)
Necrosis, NOS 1 (2%)
#Pancreatic acinus (49) (50) (50)
Focal cellular change 1 (2%) 3 (6%) 3 (6%)
Atrophy, NOS 2 (4%) 1 (2%)
Hyperplasia, NOS 1 (2%) 1 (2%)
#Stomach (49) (50) (50)
Inflammation, chronic 1 (2%)
#Gastric fundal gland (49) (50) (50)
Dilatation, NOS 6 (12%) 1 (2%) 2 (4%)

127 2-Mercaptobenzothiazole, NTP TR 332

TABLE C4. SUMMARY OF T H E INCIDENCE OF NONNEOPLASTIC LESIONS I N MALE MICE I N T H E
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

DIGESTIVE SYSTEM (Continued)

#Glandular stomach (49) (50) (50)
Mineralization 1 (2%)
Lymphocytic inflammatory infiltration 1 (2%)
Inflammation, acute 2 (4%)
Inflammation, active chronic 1 (2%)
Inflammation, chronic 1 (2%)
Metaplasia, squamous 1 (2%)
Dysplasia, epithelial 1 (2%) 3 (6%)
#Forestomach
(49) (50) (50)
Mineralization
1 (2%) 1 (2%)
Inflammation, acute
1 (2%) 1 (2%)
Inflammation, active chronic
2 (4%) 1 (2%)
Hyperplasia, epithelial
1 (2%)
Hyperkeratosis
2 (4%) 1 (2%) 1 (2%)
Acanthosis
1 (2%)

URINARY SYSTEM
#Kidney (49) (50) (50)
Lymphocytic inflammatory infiltration 1 (2%) 8 (16%)
Inflammation, chronic 3 (6%) 8 (16%) 3 (6%)
# Kidneykortex
(49) (50)
(50)
Cyst, NOS
4 (8%) 3 (6%)
Multiple cysta
4 (8%)
Metaplasia, osseous
1 (2%) 2 (4%)
# Kidneykubule
(49) (50)
(50)

Mineralization
32 (65%) 29 (58%) 26 (52%)
Dilatation, NOS
5 (10%) 5 (10%) 6 (12%)
Cyst, NOS
1 (2%)
Necrosis, NOS
9 (18%) 6 (12%) 5 (10%)
Cytoplasmic vacuolization
3 (6%)
Regeneration, NOS
39 (80%) 42 (84%) 34 (68%)
#Kidney/pelvis
(49) (50) (50)
Hemorrhage
1 (2%)
#Urinary bladder (49) (50) (48 )

Calculus, gross observation only 2 (4%)

Calculus, microscopic examination 1 (2%)

Mineralization 1 (2%)

Cast, NOS 1 (2%)

Lymphocytic inflammatory infiltration 2 (4%)
Inflammation, acute 1 (2%)
Inflammation, chronic 2 (4%)
*Urethra
(49) (50) (50)
Cast, NOS
19 (39%) 13 (26%) 24 (48%)

ENDOCRINE SYSTEM
#Pituitary intermedia (49) (48) (50)

Cyst, NOS 1 (2%)

#Anterior pituitary
(49) (48) (50)
Cyst, NOS
1 (2%) 3 (6%)
Multiple cysts
1 (2%) 1 (2%) 1 (2%)
Hyperplasia, NOS
1 (2%) 3 (6%) 1 (2%)
Hyperplasia, focal
1 (2%)
# Adrenakapsule
(49) (50) (48)
Hyperplasia, NOS
48 (98%) 45 (90%) 39 (81%)
#Adrenal cortex
(49) (50) (48)
Accessory structure
1 (2%)
Cyst, NOS
1 (2%)
Focal cellular change

Hypertrophy, focal
1 (2%)
Hyperplasia, NOS 5 (10%) 4 (8%)

2-Mercaptobenzothiazole,NTP TR 332 128

TABLE C4. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ENDOCRINE SYSTEM (Continued)

#Adrenal medulla (49) (50) (48)
Hyperplasia, NOS 2 (4%) 3 (6%)
#Thyroid (49) (50) (47)
Cystic follicles 14 (29%) 1 1 (22%) 11 (23%)
Inflammation, chronic 1 (2%)
Hyperplasia, C-cell 4 (8%) 1 (2%)
Hyperplasia, follicular cell 2 (4%)
#Thyroid follicle (49) (50) (47)
Atrophy, NOS 1 (2%)
#Parathyroid (29) (31) (29)
Metaplasia, osseous 1 (3%)
#Pancreatic islets (49) (50) (50)
Hyperplasia, NOS 20 (41%) 20 (40%) 13 (26%)

REPRODUCTIVE SYSTEM
*Preputial gland (49) (50) (50)
Cystic ducts 8 (16%) 6 (12%) 4 (8%)
Inflammation, suppurative 1 (2%)
Abscess, NOS 1 (2%)
Inflammation, active chronic
Inflammation, chronic 10 (20%)
#Prostate (49)
Inflammation, chronic 2 (4%)
*Seminal vesicle (49)
Dilatation, NOS 2 (4%)
Cyst, NOS
Inflammation, chronic
#Testis (49)
Atrophy, NOS 1 (2%)
Hyperplasia, interstitial cell
#Testidtubule (49) (50)
Mineralization 3 (6%)
*Epididymis (49) (50)
Inflammation, chronic 1 (2%) 1 (2%)
Granuloma, spermatic 2 (4%)
*Scrotum (49) (50)
Steatitis 1 (2%) 2 (4%)

NERVOUS SYSTEM
*Choroid plexus (49) (50) (50)
Mineralization 1 (2%)
#Brain (49) (50) (50)
Compression, NOS 1 (2%)
Mineralization 37 (76%) 27 (54%) 22 (44%)
Congestion, NOS 1 (2%)

SPECIAL SENSE ORGANS

*Eye (49)
Cataract
+Eyeicornea (49)
Inflammation, active chronic
*Eyelid (49)
Inflammation, chronic 1 (2%)
*Nasolacrimal duct (49)
Hemorrhage 4 (8%)
Lymphocytic inflammatory infiltration 1 (2%)
Inflammation, acute 1 (2%)

129 2-Mercaptobenzothiazole, NTP TR 332

TABLE C4. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS I N MALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

MUSCULOSKELETAL SYSTEM
*Kneejoint (49) (50) (50)
Osteoarthritis 1 (2%)
*Tarsal joint (49) (50) (50)
Ankylosis 18 (37%) 15 (30%) 12 (24%)

BODY CAVITIES
*Peritoneum (49) (50) (50)
Steatitis 1 (2%)
*Pericardium (49) (50) (50)
Inflammation, chronic 1 (2%)
*Mesentery (49) (50) (50)
Cyst, NOS 1 (2%)
Steatitis 3 (6%) 4 (8%) 3 (6%)

ALL OTHER SYSTEMS

*Multiple organs (49) (50) (50)
Lymphocytic inflammatory infiltration 43 (88%) 40 (80%) 31 (62%)

SPECIAL MORPHOLOGY SUMMARY

Animal missingho necropsy 1

* Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
d Number of animals examined microscopically a t this site

2-Mercaptobenzothiazole,NTP TR 332 130

APPENDIX D

SUMMARY OF LESIONS IN FEMALE MICE IN THE

TWO-YEAR GAVAGE STUDY OF

2-MERCAPTOBENZOTHIAZOLE

PAGE

TABLE D1 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE

TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 133
TABLE D2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 136

TABLE D3 ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR GAVAGE

STUDY OF 2-MERCAPTOBENZOTHIAZOLE 142

TABLE D4a HISTORICAL INCIDENCE OF HEPATOCELLULAR TUMORS IN FEMALE B6C3F1

MICE ADMINISTERED CORN OIL BY GAVAGE 145

TABLE D4b HISTORICAL INCIDENCE OF PITUITARY GLAND TUMORS IN FEMALE B6C3F1

MICE ADMINISTERED CORN OIL BY GAVAGE 145

TABLE D4c HISTORICAL INCIDENCE OF HEMATOPOIETIC SYSTEM TUMORS IN FEMALE

B6C3F1 MICE ADMINISTERED CORN OIL BY GAVAGE 146

TABLE D5 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE

MICE IN THE TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE 147

131 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole,NTP TR 332 132
TABLE D1. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS MISSING 1
ANIMALS NECROPSIED 50 49 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 50 49 50

INTEGUMENTARY SYSTEM

*Subcutaneoustissue
(50) (49) (50)
Fibrosarcoma
t 2 (4%) 1 (2%)

RESPIRATORY SYSTEM

*Nasal cavity
(50) (49) (50)
Carcinoma, NOS, invasive
1 (2%)
#Lung (50) (49) (50)
Carcinoma, NOS, metastatic 1 (2%)
Adenocarcinoma, NOS, metastatic 1 (2%)
Alveolar/bronchiolar adenoma 2 (4%) 1 (2%) 1 (2%)
Alveolar/bronchiolar carcinoma 1 (2%) 1 (2%)
Endometrial stromal sarcoma, metastatic 1 (2%)

HEMATOPOIETIC SYSTEM
*Multiple organs (50) (49) (50)
Malignant lymphoma, histiocytic type 1 (2%)
Malignant lymphoma, mixed type 17 (34%) 6 (12%) 6 (12%)
#Spleen (50) (49) (50)
Malignant lymphoma, mixed type 1 (2%) 1 (2%)
#Lymph node (50) (47) (48)
Malignant lymphoma, mixed type 1 (2%)
#Liver (50) (49) (50)
Malignant lymphoma, mixed type 1 (2%)
#Jejunum (50) (49) (48)
Malignant lymphoma, mixed type 1 (2%)

CIRCULATORY SYSTEM
#Liver
Hemangiosarcoma
#Uterus
Hemangioma

DIGESTIVE SYSTEM
*Tongue (50) (49) (50)
Squamous cell carcinoma 1 (2%)
#Liver (50) (49) (50)
Carcinoma, NOS, metastatic 1 (2%)
Hepatocellular adenoma 3 (6%) 7 (14%) 4 (8%)
Hepatocellular carcinoma 1 (2%) 5 (10%)
Endometrial stromal sarcoma, metastatic 1 (2%)
#Stomach
(50) (48) (50)
Sarcoma, NOS
1 (2%)
#Jejunum
(50) (49) (48)
Carcinoma, NOS
1 (2%)

URINARY SYSTEM
None

133 2-Mercaptobenzothiazole, NTP TR 332

TABLE D1. SUMMARY O F T H E INCIDENCE OF NEOPLASMS IN FEMALE MICE IN T H E TWO-YEAR
GAVAGE STUDY O F 2-MERCAFTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ENDOCRINE SYSTEM
#Anterior pituitary (49) (49) (49)
Carcinoma, NOS 1 (2%)
Adenoma, NOS 20 (41%) 11 (22%) 3 (6%)
# Adrenallcapsule (50) (47) (50)
Carcinoma, NOS 1 (2%)
Adenoma, NOS 1 (2%)
#Adrenal medulla (50)
Pheochromocytoma 1 (2%)
#Thyroid (50)
Follicular cell adenoma
Follicular cell carcinoma
#Pancreatic islets (50)
Islet cell adenoma 1 (2%)

REPRODUCTIVE SYSTEM
*Mammary gland (50) (49) (50)
Adenocarcinoma, NOS 2 (4%)
*Vagina (50) (49) (50)
Squamous cell carcinoma 1 (2%)
#Uterus (50) (49) (50)
Endometrial stromal polyp 1 (2%)
Endometrial stromal sarcoma 1 (2%)
#Ovary (50) (48) (46)
Cystadenoma, NOS 1 (2%) 2 (4%)
Granulosa cell tumor 1 (2%)

NERVOUS SYSTEM
#Braidmeninges
Fibrosarcoma, invasive
*Spinal dura mater
Fibrosarcoma, invasive
#Brain
Carcinoma, NOS,invasive

SPECIAL SENSE ORGANS

*Harderian gland (50) (49) (50)
Carcinoma, NOS 1 (2%)
Adenoma, NOS 2 (4%) 5 (10%) 2 (4%)

MUSCULOSKELETAL SYSTEM
*Vertebra (50) (49) (50)
Fibrosarcoma, invasive 1 (2%)

BODY CAVITIES
*Mesentery (50) (49) (50)
Sarcoma, NOS, invasive 1 (2%)

ALL OTHER SYSTEMS

*Multiple organs (60) (49) (50)
Sarcoma, NOS 1 (2%)
Fibrosarcoma, metastatic 1 (2%)

2-Mercaptobenzothiazole,NTP TR 332 134

TABLE D1. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICF IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ANIMAL DISPOSITION SUMMARY

Animals initially in study 50 50 50
Natural death 4 2 22
Moribund sacrifice 11 8 2
Terminal sacrifice 35 39 22
Accidentally killed, NOS 4
Animal missing 1

TUMOR SUMMARY
Total animals with primary tumors** 38 33 15
Total primary tumors 63 46 22
Total animals with benign tumors 25 21 11
Total benign tumors 32 26 12
Total animals with malignant tumors 27 18 9
Total malignant tumors 31 19 10
Total animals with secondary tumors## 5 2 2
Total secondary tumors 8 2 2
Total animals with tumors uncertain--
benign or malignant 1
Total uncertain tumors 1

* Number of animals receiving complete necropsy examinations; all grsss lesions including massesexarnined microscopically.
t Multiple occurrence of morphology in the same organ; tissue is counted once only.
** Primary tumors: all tumors except secondary tumors
# Number of animals examined microscopically a t this site
# # Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

135 2-Mercaptobenzothiazole, NTP TR 332

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE: VEHICLE CONTROL

WEEKS ON

STUDY

Subcutaneous tissue + + t + t + + + + + + t t + + + + + + + + + + + t
Fibrosarcoma x @X
RESPIRATORY SYSTEM
Lungs and bronchi + + t + + t + + + + + + + + t + + + + t + + + + +
Carcinoma, NOS, metastatic X
Alveolar/bronchiolar adenoma X
Alveolar/bronchiolar carcinoma
Endometrial stromal sarcoma, metastatic
Trachea . . . . . . . . . . . . . . . . . . . . . . . . .
Nasal cavity N N + + + + t + + + + + t + t + + + + + + + + + +
Carcinoma, NOS,invasive X

Bone marrow + + + + t t + t t t + + t + + + + t + + t t + t t
Spleen + + + t + + + + t t + t t + + + + + t t + t + t +
Malignant lymphoma, mixed type
L mphnodes + + + t t + + + + t + + + + + + + + + + + + + + +
dymus . . . . . . . . . . . . . . . . . . . . . . . . .

Heart t + + + + + t + + t + + + + + + t + t t + t + + +

Oral cavity N N N N N N N N N N N N N N N N N N N N N N N N N
Squamous cell carcinoma X
Salivary gland + t + t + + + t t + + + + + + + t + + + t t + + +
Liver t + + + + + + + t + + t + t + + + t + + + t t + t
Hepatocellular adenoma X
He atocellular carcinoma X
Entlometrial stromal sarcoma metastatic
Malignant lymphoma, mixed iype
Bile duct + t + + + + + + + + + + + + + + + + t + + + + + t
Gallbladder & common bile duct + t + t + + + t + t + + + + + + t + + + + + t + t
Pancreas + + + t + + + t + + + + + + + + + + + + t + + + +

:;::2Y

Small intestine
+
+
+
+
+
t
+
t
t
+
t
t
t
t
+
+
+
+
+
+
t
+
+
+
t
t
t
t
t
+
+
t
t
+
t
+
+
+
+
+
+
+
+
t
+
+
t
+
+
+
+
t
+
+
+
+
+
+ + + + + t
+ + + + + t
t t + + + t
Carcinoma, NOS
Large intestine t + + t + t + + + t + + + t + t t t + + t + t t t
M
Kidney + + t t + t + + + + + t + + + + + + + + + t t + +
Urinary bladder t t t t + t + + + + + + t + t + + + + + + + + + t

Pituitary + + + + t t t + - + + + + + + + + + + + + + + + +
Carcinoma, NOS X
Adenoma, NOS X X x x
Ad re na I + + + + t t + + + + + t + + + + + + + + + + + + t
Adenoma, NOS
Pheochromocytoma
Thyroid t + + t + + + + + + + + + + + + + + + + + t + + t
Parathyroid + - + - + - - - - + + + + + + + + + + + + + - + -
Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . .
Islet-cell adenoma

Mammary gland N N + + + + t + + + + + + + + + + + + t + + + + +
Vamna N N N N N N N N N N N N N N N N N N N N N N N N N
+ + + + + + t + + t t + + + t + + + + + + + + + +

t + t + + t t + t + + + + + t + + + + + + t + + +

Brain
Carcinoma. NOS, invasive
Spinal cord
Fibrosarcoma, invasive

Hardenan gland N N N N N N N N N N N N N N N N N N N N N N N N N
Carcinoma. NOS X
Adenoma, NOS
- _.
ETAL SYSTEM
Bone N N N N N N N N N N N N N N N N N N N N N N N N N
Fibrosarcoma, invasive X

Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N

Sarcoma, NOS
Fibrosamoma metastatic X
Malignant IyAphoma, mixed type X x x x x x x x
+: Tissue examined micm6copically ' No tissue information submitted
-: Required tissue not examined micmrcopically CI Necropsy, no histology due to protocol
X: Tumor incidenci A: Autolysis
N: Nscropay, no autolysis, no microlcopic examination M: Animal missing
S Animal miassxed B: No necropry performed

@. Multiple occurrence of morphology

2-Mercaptobenzothiazole, NTP TR 332 136

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: VEHICLE CONTROL
(Continued)
ANIMAL
NUMBER
1
1
5
1
6
1
1
7
1
1
8
1
2
0
1 1 1
2 2 2 2
3 5 6 7
1
2
8
1 1 1
3 3 3 3
0 1 2 3
1
3
5
3
6
1
3
9
1
4
0
1
1
4
1 1 1
4 4 4 4
2 3 4 7
1
4
9
1

0

1- ~ ~ ~ ~ ~

WEEKS ON 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 ~ ~ ~ ~ ~ ~
STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Subcutaneous tissue . . . . . . . . . . . . . . . . . . . . . . . . .

Fibrosarcoma
KESPIRATORY SYSTEM
Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . . 50

Carcinoma, NOS, metastatic 1

Alveolar/bronchiolar adenoma X 2

Alveolar/bronchiolar carcinoma X 1

Endometrial stromal sarcoma, meta X 1

Trachea . . . . . . . . . . . . . . . . . . . . . . . . . 50

Nasal cavity . . . . . . . . . . . . . . . . . . . . . . . . . '50

Carcinoma, NOS, invasive 1

Bone marmw . . . . . . . . . . . . . . . . . . . . . . . . . 50

S lean . . . . . . . . . . . . . . . . . . . . . . . . . 50

&alignant lymphoma, mixed type X 1

LvmDh nodes . . . . . . . . . . . . . . . . . . . . . . . . . 50

Thymus . . . . . . . . . . . . . . . . . . . . . . . . . 44

Heart . . . . . . . . . . . . . . . . . . . . . . . . . 50

Oral cavity N N N N N N N N N N N N N N N N N N N N N N N N N *50

Squamous cell carcinoma 1

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . . 50

Liver . . . . . . . . . . . . . . . . . . . . . . . . . 50

Hepatocellular adenoma x x 3

He atocellular carcinoma
En2ometnal stromal sarcoma, meta
Malignant lymphoma, mixed type
Bile duct
I
. . .
X
. . . . . . . . . . . . . . . . . . . .
X
. .
1

50

Gallbladder 81 common bile duct . . . . . . . . . . . . . . . . . . . . . . . . . * 50

Pancreas + + + + + + + e + + + + + + + + + + + + + + + + 50+

. . . . . . . . . . . . . . . . . . . . . . . . .
2lk:BCfls
Small intestine
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
. .
. .
.
.
.
.
.
.
50

50

50

Carcinoma, NOS X 1

Large intestine . . . . . . . . . . . . . . . . . . . . . . . . . 50

7
Unnary bladder
. . . . . . . . . . . . . . . . . . . . . . . . .
. . . . . . . . . . . . . . . . . . . . . . . . .
50

50

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . . 49

Carcinoma, NOS 1

Adenoma, NOS x x x x x x x x x x x x x x x x 20

Adrenal . . . . . . . . . . . . . . . . . . . . . . . . . 50

Adenoma, NOS X 1

Pheochromocytoma X 1

Thyroid . . . . . . . . . . . . . . . . . . . . . . . . . 50

Parathyroid + + - + - + + + + - + + - + + + + + + + + - + + - 36

Pancreatic islets . . . . . . . . . . . . . . . . . . . . . . . . . 50

Islet cell adenoma X 1

EM
Mammary gland . . . . . . . . . . . . . . . . . . . . . . . . . * 50

Vagma N N N N N N N N N N N N N N N N N N N N N N N N N '50

Sauamous cell carcinoma X 1

uterus . . . . . . . . . . . . . . . . . . . . . . . . . 50

Endometrial stromal polyp 1

Endometrial stromal sarcoma X 1

Hemangioma X 1

Ovary . . . . . . . . . . . . . . . . . . . . . . . . . 50

Brain
M

I+++++ + + + + + + + + + + + + + + + + + + + + I-- 50

I:
Carcinoma, NOS,invasive
Spinal cord N N N N N N N N N N N N N N N N N N N N N N N N N *50
Fibrosarcoma, invasive

Hardenan gland N N N N N N N N N N N N N N N N N N N N N N N N N '50

Carcinoma. NOS 1

Adenoma, NOS X x 2

Bone N N N N N N N N N N N N N N N N N N N N N N N N N *50

Fibrosarcoma, invasive 1

Multiple or ans. NOS N N N N N N N N N N N N N N N N N N N N N N N N N '50

Sarcoma, ROS
Fibmsarcoma, metastatic
X 1

Malignant lymphoma, mixed type X X x x x X x x x 17

1-1
* Animals necropsied

137 2-Mercaptobenzothiazole,NTP TR 332

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE: LOW DOSE
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
NUMBER 2 0 3 4 4 4 3 4 1 1 2 0 0 0 0 0 0 0 0 1 1 1 1 1 1
4 7 5 0 7 3 1 8 2 3 3 1 2 3 4 5 6 8 9 0 1 4 5 6 7
WEEKS ON 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
STUDY 7 7 8 8 8 9 9 9 9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
2 7 7 7 9 7 8 8 9 2 2 3 3 3 3 3 3 3 3 3 3 3 3 3 3

Subcutaneous tissue + + + + + + + + + + M + + + + + + + + + + + + + +
Fibrosarcoma X
RESPIRATORY SYSTEM
Lungs and bronchi + + + + + + + + + + M + + + + + + + + + + + + + +
Adenocarcinoma, NOS, metastatic X
Alveolar/bronchiolar adenoma
Trachea + + + + + + + + + + M + + + + + + + + + + + + + +

Bone marrow + + + + + + + + + + M + + + + + + + + + + + + + +
Spleen + + + + + + + + + + M + + + + + + + + + + + + + +
Malignant lymphoma, mixed type
Lymph nodes . . . . . . . . . . . . . . . . . . . . . . . . .
Malignant lymphoma, mixed type
Thymus + + + - + + + + + + M + + + + + - + + + + + + + +

Heart . . . . . . . . . . . . . . . . . . . . . . . . .

TEM
Salivary gland t + + t + t + + t + M + + + + + + + + + + + + + +
Liver . . . . . . . . . . . . . . . . . . . . . . . . .
Hepatocellular adenoma X X X

Hepatocellular carcinoma X x x

Hemanaosarcoma X

Bile duct + + + + + + + + + + M + + + + + + + + + + + + + t

:,,,,
Gallbladder & common bile duct
Pancreas

Small intestine
Malignant lymphoma, mixed type
Large intestine
+ + + + + + + + + +
. . . . . . . . . . . . . . . . . . . . . . . . .
+
+
+
+
+
+
+
+
+ + + + + + + + M + + + + + + + + + + + + + +
- + + + + + + + M + + + + + + + + + + + + + +
+ + + + + + + + M + + + + + + + + + + + + + +
+ + + + + + + +
M

M
+

+
+ + + + + + + + + +

+ + + + +
X
+ + + + +
+

+
+ +

+ +

Kidney + + + + + + + + + + M + + + + + + + + + + + + + +
Unnary bladder + + + + + + + + + + M + + + + + + + + + + + + + +

Pituitary + + + + + + + + + + M + + + + + + + + + + + + + +
Adenoma, NOS X X x x X
Adrenal + + + + + + + + + + M + + + + + + + + + + + + + -
Thyroid + + + + + + + + + + M + + + + + + + + + + + + + +
Follicular cell adenoma X
Parathyroid + + - + - + + - + - M - + + + + + - + - - + + - +

Mammary gland + + + + + + + + + + M + + + + + + + + + + + + + +
Adenocarcinoma. NOS X
Uterus + + + + + + + + + + M + + + + + + + + + + + + + +
Ovary + + + + + + + + + + M + + + + + + + + + + + + + +
Cystadenoma, NOS X

Granulosa cell tumor

Brain + + + + + + + + + + M + + + + + + + + + + + + + +
Fibrosarcoma. invasive X
SPECIAL
Hardenan gland N N N N N N N N N N M N N N N N N N N N N N N N N
Adenoma, NOS X X x x
S

Multiple organs, NOS N N N N N N N N N N M N N N N N N N N N N N N N N

Malignant lymphoma, histiocytic type X
Malignant lymphoma, mixed type X x x X X

2-Mercaptobenzothiazole, NTP TR 332 138

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: LOW DOSE
(Continued)
ANIMAL 01 01 01 01 01 01 Of 01 01 01 01 0] 01 01 01 01 01 01 01 01 01 01 01 01 0
NUMBER 1 1 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 4 4 4 4 4 4 5
8| 9| 0| l| 2| 5| 6| 7\ 8| 9| 0| 2| 3| 4| 6| 7| 8\ 9\ l\ 2| 4| S| 6\ 9\ 0
TOTAL:
WEEKS ON TISSUES
STUDY 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 TUMORS
3| 3| 3 3| 3| 3| 3| 3 3 3 3 3| 3| 3| 3| 3 3| 3 3 3| 3 3\ 3 S\ 3
INTEGUMENTARY SYSTEM
Subcutaneous tissue •49
Fibrosarcoma 1
RESPIRATORY SYSTEM
Lungs and bronchi 49
Adenocarcinoma, NOS, metastatie 1
Alveolar/bronchiolar adenoma X 1
Trachea 49
HEMATOPOIETl'lC bl^FEM
Bone marrow 49
Spleen 49
Malignant lymphoma, mixed type X 1
Lymph nodes 47
Malignant lymphoma, mixed type X 1
Thymus 44
CIRCULATORY SYSTEM
Heart 49
DJUESTIVE stSTEM
Salivary gland 49
Liver 49
Hepatocellular adenoma X X X X 7
Hepatocellular carcinoma X X 5
Hemangiosarcoma 1
Bile duct 49
Gallbladder & common bile duct •49
Pancreas 49
Esophagus 49
Stomach 48
Small intestine 49
Malignant lymphoma, mixed type 1
Large intestine 49
URINARY SYSTEM
Kidney 49
Urinary bladder 49
ENDOCK1N& SYSTEM
Pituitary 49
Adenoma, NOS X X X X X X 11
Adrenal 47
Thyroid 49
Follicular cell adenoma 1
Parathyroid + -f + + + + + _ + + _ + _ - + < . _ + + _-- + + + 32
REPRODUCTIVE SYSTEM
Mammary gland *49
Adenocarcinoma, NOS X 2
Uterus 49
Ovary 48
Cystadenoma, NOS 1
Granulosa celt tumor X 1
NERVOUS SYSTEM
Brain + + + + + + ^ + 4. + + ^ + 1. + + + 4. + + + + + H . + 49
Fibrosarcoma, invasive 1
SPECIAL SENSE ORGANS
Harderian gland N N N N N N N N N N N N N N N N N N N N N N N N N . •49
Adenoma, NOS X 5
ALL OTHER SYSTEMS
Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N •49
Malignant lymphoma, histiocytic type I
Malignant lymphoma, mixed type X 6

* Animals necropsied

139 2-Mercaptobenzothiazole, NTP TR 332

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR
GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE:HIGH DOSE
: IMA
NUMBER 6 0 7 7 9 9 5 6 5 9 5 8 6 5 9 7 7 8 8 8 7 5 6 6 6
5 0 6 8 4 R 2 6 6 1 3 0 9 8 2 3 5 8 9 1 7 5 3 2 4

WEEKSON 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ~ ~ ~ ~ ~
STUDY 0 1 1 1 1 1 1 1 2 2 2 4 4 4 4 5 5 9 6 7 7 ? 8 9 9
2 2 3 3 3 3 0 8 0 0 8 9 5 8 9 2 8 9 4 3 5 8 1 4 7

Lungs and bronchi . . . . . . . . . . . . . . . . . . . . . . . . .

Alveolarbronchiolar adenoma X
Alveolar/bronchiolar carcinoma
Trachea . . . . . . . . . . . . . . . . . . . . . . . . .

Bone marrow . . . . . . . . . . . . . . . . . . . . . . . . .

Spleen . . . . . . . . . . . . . . . . . . . . . . . . .

L mphncdes . . . . . . . . . . . . . . . . . . . . . . . . .

Txymus . . . . . . . . . . . . . . . . . . . . . . . . .

Heart . . . . . . . . . . . . . . . . . . . . . . . . .

Salivary gland . . . . . . . . . . . . . . . . . . . . . . . . .
Liver + + + + + + + + + + + + + + + + + + + + + t t t t
Carcinoma, NOS,metastatic
Hepatocellular adenoma
Bile duct . . . . . . . . . . . . . . . . . . . . . . . . .

Gallbladder & common bile duct + + + + + + + + + + + + + + + + N + + + + t + + t

Pancreas + + + + + + + + t + + t + + + + + + + + + + t t +
%2rsNOS
Sarcoma,
. . . .
+ + + +
. . . .
+ + + +
. . . . .
+ + C + +
. . . . .
+ + + t
. . . .
+ + + t
. . . .
+ + + t
x
Small intestine + + + + + t + + + + + + + + - + + + + - + t t + t
Large intestine . . . . . . . . . . . . . . . . . . . . . . . . .

Kidney . . . . . . . . . . . . . . . . . . . . . . . . .

Unnary bladder . . . . . . . . . . . . . . . . . . . . . . . . .

Pituitary . . . . . . . . . . . . . . . . . . . . . . . . .

Adenoma, NOS X
Adrenal . . . . . . . . . . . . . . . . . . . . . . . . .
Carcinoma, NOS
Thyroid . . . . . . . . . . . . . . . . . . . . . . . . .

Follicular cell carcinoma

Parathyroid + + + + + + + - + + + - - - - - - + - + + - - + -
Mammary gland N N + + + + + N + + + + + + + + + + + + t + + + +
Uterus . . . . . . . . . . . . . . . . . . . . . . . . .
Ovary . . . . . . . . . . . . . . . . . . . . . . . . .

Cystadenoma, NOS

Brain

Hardenan gland N N N N N N N N N N N N N N N N N N N N N N N N N
Adenoma, NOS

Mesentery N N N N N N N N N N N N N N N N N N N N N N N N N
Sarcoma, NOS,invasive X
ALL
Multiple organs, NOS N N N N N N N N N N N N N N N N N N N N N N N N N
Malignant lymphoma, mixed type X
I I

2-Mercaptobenzothiazole,NTP TR 332 140

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: HIGH DOSE
(Continued)
ANIMAL 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

NUMBER 9 9 6 5 5 5 5 6 6 6 7 7 7 7 7 8 8 8 8 8 8 9 9 9 9

0 7 1 1 4 7 9 0 7 8 0 1 2 4 9 2 3 4 5 6 7 3 5 8 9

TOTAL:
WEEKS ON
0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 TISSUES
STUDY 9 9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 TUMORS
a 8 0 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Lungs and bronchi + + + + t t + + + + t t + t t t t t + + + t t + t 50

Alveolar/bronchiolar adenoma 1

Alveolar/bronchiolar carcinoma X 1

Trachea t + + t + + + + + + t + + + + + + + t + + + t + + 50

Bone marrow + + + + + + + t t t + t + + + + t + + t + + + + + 50

Spleen + + t + + + t t t t t t + + t t t + t t + + t + t 50

L mphncdes t + + + + + + + + + + t + + + + t + + t + + + + t 48

Txymus . . . . . . . . . . . . . . . . . . . . . . . . . 49

Heart t + + + + + + t + + + + + + t t + t + + t + + + + 50

TEM
Salivary gland + t t t + + t t + + + t + + + + t + t + t + + + + 50

Liver + + t + + + + t + + + + + + t t t t + t t t + + + 50

Carcinoma, NOS,metastatic X 1

Hepatocellular adenoma x x X X 4

Bile duct t + + + t + t + + + + t + + + + + + + + + t t + + 50

Gallbladder & common bile duct t + + + + + + + + + + t + + t + t t + + t t + t t 850

Pancreas + t + + + + + + + + t t + + + t t t t + t t t t + 50

:Ekrs
Sarcoma, NOS
t
+
+
t
t
+
t
t
+
+
+
+
+
+
+
+
t + + t + + t t t + t + t t t t +
+ t + t + t t + t + + + + t t + +
49

50

Small intestine t + + + + + + + t + + t + t t t + + + + + t t + t 48

Large intestine + + + + + + + + + t t + t t + + t t t t t t + + t 48

Kidney + + + + + + + + + + t t t + t + + + + + t + +
50
+ +
Unnary bladder + + + + + t + + + + t + + t t + t + + + + t t t + 49

Pituitary t + + t + + t + t + + t + t + t + t + + + + + + t 49

Adenoma, NOS X X 3

Adrenal + + t + + + + t + t t + + + t + + t + + + + + + + 50

Carcinoma, NOS X 1

Thyroid + t t + t + + + + + t t t + t + + + + t t + + t + 49

Follicular cell carcinoma X 1

Parathyroid t + + t - t + t - + - - t t - t t t t - + - + + + 32

Mammary gland + + + + + t t + t + + t + t + + t + + + + t + + + *50

Uterus + + t + t t + + + + + + t t + t t + + t t + t t t 50

Ovary + + t + + t t t t t - + t + t + + + + - + + t + + 46

Cystadenoma, NOS X X 2

-
Brain t + + t t + t + + + + + t t + + + t t t t + t + t 50

ORGANS
Hardenan gland N N N N N N N N N N N N N N N N N N N N N N N N N $50
Adenoma, NOS X X 2

M esentery N N N N N N N N N N N N N N N N N N N N N N N N N
Sarcoma, NOS,invasive
ALL O T m
Multiple organs. NOS N N N N N N N N N N N N N N N N N N N N N N N N N
Malignant lymphoma, mixed type X X x x X

I-

* Animals necropsied

141 2-Mercaptobenzothiazole, NTP TR 332

TABLE D3. ANALYSIS O F PRIMARY TUMORS IN FEMALE MICE IN T H E TWO-YEAR GAVAGE STUDY
O F 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control 375 m g k g 750 mgkg

Lung: AlveolarlBronchiolar Adenoma or Carcinoma

Overall Rates (a) 3/50 (6%) 1/49 (2%) 2/50 (4%)
Adjusted Rates (b) 7.7% 2.6% 8.0%
Terminal Rates (c) 2/37 (5%) 1/39 (3%) 1/22 (5%)
Week of First Observation 93 103 81
Life Table Tests (d) P=0.587N P = 0.287N P=0.638
Incidental Tumor Tests (d) P=0.584 P=0.293N P = 0.601
Cochran-Armitage Trend Test td) P = 0.400N
Fisher Exact Test td) P = 0.316N P = 0.500N

Hematopoietic System: Malignant Lymph.oma, Mixed Type

Overall Rates ( a )
19/50 (38%) 9/49 ( 18%) 6/50 (12%)
Adjusted Rates (b)
48.5% 20.8% 25.3%
Terminal Rates (c)
17/37 (46%) 6/39 (15%) 5/22 (23%)
Week of First Observation
89 72 75
Life Table Tests (d)
P = 0.028N P = 0.016N P =0.076N
Incidental Tumor Tests (d)
P =0.014N P=0.020N P = 0.057N
Cochran-Armitage Trend Test (d)
P=O.OOlN
Fisher Exact Test (d)
P = 0.025N P=0.003N
Hematopoietic System: Lymphoma, All Malignant
Overall Rates (a)
19/50 (38%) 10149 (20%) 6/50 (12%)
Adjusted Rates (b)
48.5% 23.2% 25.3%
Terminal Rates (c)
17/37 (46%) 7/39 (18%) 5/22 (23%)
Week of First Observation
89 72 75
Life Table Tests (d)
P = 0.032N P = 0.028N P = 0.076N
Incidental Tumor Tests (d)
P=0.016N P=0.035N P=0.057N
Cochran-Armitage Trend Test (d)
P = 0.002N
Fisher Exact Test (d)
P = 0.044N P=0.003N
Liver: Hepatocellular Adenoma
Overall Rates (a) 3/50 (6%) 7/49(14%) 4/50 (8%)
Adjusted Rates (b) 8.1% 17.9% 18.2%
Terminal Rates (c) 3/37 (8%) 7/39 (18%) 4/22(18%J
Week of First Observation 103 103 103
Life Table Tests (d) P=0.159 P = 0.1 78 P =0.231
Incidental Tumor Tests (d) P=0.159 P=0.178 P = 0.231
Cochran-Armitage Trend Test (d) P = 0.432
Fisher Exact Test (d) P = 0.15 1 P = 0.500

Liver: Hepatocellular Carcinoma

Overall Rates ( a ) 1/50 (2%) 5/49 (10%) 0/50 (0%)
Adjusted Rates (b) 2.7% 12.2% 0.0%
Terminal Rates (c) 1/37 (3%) 4/39 (10%) 0122 (0%)
Week of First Observation 103 89
Life Table Tests (d) P=0.590N P=0.116 P = 0.604N
Incidental Tumor Tests (d) P=0.552 P = 0.088 P=0.604N
Cochran-Armitage Trend Test (d) P=0.400N
Fisher Exact Test (d) P=0.098 P=0.500N
Liver: Hepatocellular Adenoma or Carcinoma
Overall Rates (a) 4/50 (8%) 12/49 (24%) 4/50 (8%)
Adjusted Rates (b) 10.8% 29.8% 18.2%
Terminal Rates (c) 4/37 (11%) 11/39 (28%) 4/22(18%)
Week of First Observation 103 89 103
Life Table Tests (d) P = 0.204 P=0.035 P=0.343
Incidental Tumor Tests (d) P=0.171 P = 0.028 P = 0.343
Cochran-Armitage Trend Test (d) P=0.558
Fisher Exact Test (d) P = 0.024 P = 0.643

2-Mercaptobenzothiazole, NTP TR 332 142

TABLE D3. ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 mg/kg

Pituitary Gland: Adenoma

Overall Rates (a)
20/49 (41%) 11/49 (22%) 3/49 (6%)
Adjusted Rates (b)
51.1% 26.4% 12.5%
Terminal Rates (c)
18/37 (49%) 9/39 (23%) 2/22 (9%)
Week of First Observation
92 87 94
Life Table Tests (d)
P=0.002N P=0.028N P=0.004N
Incidental Tumor Tests (d)
P=O.OOlN P=0.035N P=0.003N
Cochran-Armitage Trend Test (d)
P<0.001N
Fisher Exact Test (d)
P = 0.041N PC0.001N
Pituitary Gland: Adenoma or Carcinoma
Overall Rates (a)
21/49 (43%) 11/49 (22%) 3/49 (6%)
Adjusted Rates (b)
52.1% 26.4% 12.5%
Terminal Rates (c)
18/37 (49%) 9/39 (23%) 2/22 (9%)
Week of First Observation
71 87 94
Life Table Tests (d)
P <0.001 N P =0.019N P=0.003N
Incidental Tumor Tests (d)
P <0.001 N P =0.024N P=O.OOlN
Cochran-Armitage Trend Test (d)
P<O.OOl N
Fisher Exact Test (d)
P = 0.026N P C 0.00 1N

Harderian Gland: Adenoma

Overall Rates (a)
2/50 (4%) 5/49 (10%) 2/50 (4%)
Adjusted Rates (b)
5.4% 12.3% 9.1%
Terminal Rates (c)
2/37 (5%) 4/39 (10%) 2/22 (9%)
Week of First Observation
103 98 103
Life Table Tests (d)
P=0.351 P = 0.245 P = 0.496
Incidental Tumor Tests (d)
P=0.372 P=0.237 P = 0.496
Cochran-Armitage Trend Test (d)
P = 0.583
Fisher Exact Test (d)
P=0.210 P =0.69 1

Harderian Gland: Adenoma or Carcinoma

Overall Rates (a)
3/50 (6%) 5/49 (10%) 2/50 (4%)
Adjusted Rates (b)
7.8% 12.3% 9.1%
Terminal Rates (c)
2/37 (5%) 4/39 (10%) 2/22 (9%)
Week of First Observation
96 98 103
Life Table Tests (d)
P = 0.505 P = 0.398 P=0.642
Incidental Tumor Tests (d)
P = 0.547 P=0.378 P=0.678
Cochran-Armitage Trend Test (d)
P=0.421N
Fisher Exact Test (d)
P = 0.346 P = 0.500N

All Sites: Benign Tumors

Overall Rates (a)
25/50 (50%) 21/49 (43%) 11/50 (22%)
Adjusted Rates (b)
62.3% 49.7% 45.1 %
Terminal Rates ( c )
22/37 (59%) 18/39 (46%) 9/22 (41%)
Week of First Observation
92 87 81
Life Table Tests (d)
P =0.129N P=0.193N P =0.178N
Incidental Tumor Tests (d)
P = 0.143N P = 0.242N P =0.186N
Cochran-Armitage Trend Test td)
P=0.003N
Fisher Exact Test td)
P =0.305N P=0.003N

All Sites: Malignant Tumors

Overall Rates ( a )
27/50 (54%) 18/49 (37%) 9/50 (18%)
Adjusted Rates ( b )
61.1% 39.6% 36.5%
Terminal Rates (c)
20137 (54%) 12/39 (31%) 7/22 (32%)
Week of First Observation
71 72 75
Life Table Tests (d)
P=0.023N P=0.048N P = 0.044N
Incidental Tumor Tests (d)
P=0.005N P=0.054N P = 0.013N
Cochran-Armitage Trend Test (d)
P<O.OOl N
Fisher Exact Test (d)
P=0.064N P < 0.001N

143 2-Mercaptobenzothiazole, NTP TR 332

TABLE D3. ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR GAVAGE STUDY
OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control 375 mg/kg 750 mg/kg

All Sites: All Tumors

Overall Rates (a) 38/50 (76%) 33/49 (67%) 15/50 (30%)
Adjusted Rates (b) 82.6% 70.1% 59.2%
Terminal Rates (c) 29/37 (78%) 25/39 (64%) 12/22 (55%)
Week of First Observation 71 72 75
Life Table Tests (d) P = 0.025N P=0.142N P = 0.034N
Incidental Tumor Tests (d) P = 0.006N P = 0.197N P =0.009N
Cochran-Armitage Trend Test (d) P <0.001N
Fisher Exact Test (d) P=0.2'32N P <0.001 N

(a)Number of tumor-bearing animals/number of animals examined a t the site

(b)Kaplan-Meier estimated tumor incidences a t the end of the study after adjusting for intercurrent mortality
(c)Observed tumor incidence a t terminal kill
(d)Beneath the vehicle control incidence are the P values associated with the trend test. Beneath the dosed group incidence are
the P values corresponding to pairwise comparisons between that dosed group and the vehicle controls. The life table analysis
regards tumors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The incidental tumor
test regards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence
rates. A negative trend or lower incidence in a dosed group is indicated by (N).

2-Mercaptobenzothiazole, NTP TR 332 144

TABLE D4a. HISTORICAL INCIDENCE OF HEPATOCELLULAR TUMORS I N FEMALE B6C3F1 MICE
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Adenoma Carcinoma Adenoma o r Carcinoma

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence
TOTAL 7111,489 (4.8%) 4611,489 (3.1%) 116/1,489(7.8%)
SD (b) 4.29% 2.62% 5.56%
Range (c)
High 9/50 5/50 (d) 14/50
Low 0150 0150 0149

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Standard deviation
(c) Range and SD are presented for groups of 35 or more animals.
(d)Second highest: 9/50

TABLE D4b. HISTORICAL INCIDENCE OF PITUITARY GLAND TUMORS IN FEMALE B6C3F1 MICE
ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Adenoma Carcinoma Adenoma o r Carcinoma
~~ ~~ ~~~

No 2-year studies by Physiological Research Laboratories are included in the historical data base.
Overall Historical Incidence

TOTAL (b)237/1,324(17.9%) (c)20/1,324(1.5%) (b,c)257/1,324(19.4%)

SD (d) 8.44% 2.79% 8.95%

Range (e)
High 18/49 5/47 18/49
LOW 2/44 0149 2/44

(a)Data as of August 30,1985, for studies of a t least 104 weeks

(b)Includes 198 adenomas, NOS, 38 chromophobe adenomas, and 1 acidophil adenoma
(c)Includes 14 carcinomas, NOS, 5 adenocarcinomas, NOS, and 1 acidophil carcinoma
(d)Standard deviation
(e) Range and SD are presented for groups of 35 or more animals.

145 2-Mercaptobenzothiazole,NTP TR 332

TABLE D4c. HISTORICAL INCIDENCE OF HEMATOPOIETIC SYSTEM TUMORS IN FEMALE B6C3F1
MICE ADMINISTERED CORN OIL BY GAVAGE (a)

Incidence in Vehicle Controls

Lymphoma Lymphoma or Leukemia

No 2-year studies by Physiological Research Laboratories are included in the historical data base.

Overall Historical Incidence

TOTAL 37911,494 (25.4%) 39311,494 (26.3%)
SD (b) 9.16% 9.25%

Range (c)
High 21/50 21/50
Low 4/50 4/50

(a)Data as of August 30,1985, for studies of at least 104 weeks

(b) Standard deviation
(c)Range and SD are presented for groups of 35 or more animals.

2-Mercaptobenzothiazole,NTP TR 332 146

TABLE D5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE

Vehicle Control Low Dose High Dose

ANIMALS INITIALLY IN STUDY 50 50 50

ANIMALS MISSING 1
ANIMALS NECROPSIED 50 49 50
ANIMALS EXAMINED HISTOPATHOLOGICALLY 50 49 50

INTEGUMENTARY SYSTEM
*Skin
(50) (49) (50)
Ulcer, NOS
1 (2%)
Inflammation, chronic
1 (2%)

RESPIRATORY SYSTEM
*Nasal cavity (50) (49) (50)
Hemorrhage 6 (12%) 1 (2%) 1 (2%)
Lymphocytic inflammatory infiltration 3 (6%) 3 (6%) 8 (16%)
Inflammation, acute 1 (2%) 1 (2%)
Inflammation, chronic 2 (4%)
Metaplasia, squamous 1 (2%)
*Nasal turbinate
(50) (49) (50)
Hemorrhage
1 (2%)
Inflammation, chronic
3 (6%)
#Trachea
(50) (49) (50)
Mineralization
1 (2%)
#Lunghronchiole
(50) (49) (50)
Hyperplasia, NOS
1 (2%)
#Lung (50) (49) (50)
Mineralization 1 (2%) 1 (2%)
Congestion, NOS 1 (2%) 11 (22%)
Hemorrhage 12 (24%) 3 (6%) 12 (24%)
Bronchopneumonia, NOS 13 (26%) 24 (49%) 18 (36%)
Lymphocytic inflammatory infiltration 2 (4%) 1 (2%) 9 (18%)
Pneumonia, interstitial chronic 1 (2%)
Cholesterol deposit 4 (8%) 5 (10%) 6 (12%)
Hyperplasia, alveolar epithelium 11 (22%) 28 (57%) 16 (32%)
Histiocytosis 16 (32%) 30 (61%) 17 (34%)

HEMATOPOIETIC SYSTEM
#Brain (50) (49) (50)
Lymphocytosis 1 (2%)
*Multiple organs (50) (49) (50)
Hyperplasia, lymphoid 1 (2%)
#Bone marrow
(50) (49) (50)
Fibrosis
15 (30%) 16 (33%) 12 (24%)
Hyperplasia, granulocytic
32 (64%) 28 (57%) 10 (20%)
#Spleen
(50) (49) (50)
Pigmentation, NOS
5 (10%) 4 (8%) 5 (10%)
Hyperplasia, lymphoid
19 (38%) 15 (31%) 6 (12%)
Hematopoiesis
10 (20%1 9 (18%) 2 (4%)
#Lymph node
(50) (47) (48)
Hyperplasia, lymphoid
1 (2%)
#Mandibular lymph node
(50) (47) (48)
Hemosiderosis
1 (2%)
Hyperplasia, lymphoid
8 (16%) 4 (9%) 1 (2%)
#Mesenteric lymph node
(50) (47) (48)
Congestion, NOS
1 (2%)
Inflammation, acute
2 (4%)
Hyperplasia, reticulum cell
1 (2%)
Hyperplasia, lymphoid
8 (16%) 6 (13%)
Hematopoiesis
1 (2%)

147 2-Mercaptobenzothiazole,NTP TR 332

TABLE D5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

HEMATOPOIETIC SYSTEM (Continued)

#Renal lymph node

Congestion, NOS

Hyperplasia, lymphoid

#Liver

Hematopoiesis

#Ovary/parovarian

Hyperplasia, lymphoid

#Thymus

Cyst, NOS

Hyperplasia, lymphoid

CIRCULATORY SYSTEM
*Multiple organs (50)
Periarteritis 2 (4%)
#Mesenteric lymph node (50)
Thrombosis, NOS 1 (2%)
#Heart
(50)
(49) (50)
Mineralization
1 (2%) 1 (2%)
Thrombosis, NOS
1 (2%)
Embolus, septic
1 (2%)
Inflammation, acute
1 (2%)
Inflammation, chronic
7 (14%) 2 (4%) 1 (2%)
*Pulmonary vein
(50)
(49) (50)
Mineralization
1 (2%)
Thrombosis, NOS
1 (2%)
#Ovary
(50) (48) (46)
Thrombosis, NOS
1 (2%)

DIGESTIVE SYSTEM
*Pulp of tooth (50) (49) (50)

Inflammation, chronic 1 (2%)

#Salivary gland (50) (49) (50)
Lymphocytic inflammatory infiltration 1 (2%) 1 (2%)
Inflammation, chronic 2 (4%) 2 (4%)
#Liver (50)
(49) (50)
Accessory structure 1 (2%)
Bile stasis 1 (2%)
Congestion, NOS 1 (2%)
Lymphocytic inflammatory infiltration 2 (4%)
Inflammation, acute 1 (2%) 1 (2%)
Inflammation, chronic 1 (2%)
Necrosis, NOS 2 (4%) 2 (4%)
Hemosiderosis 1 (2%)
Cytoplasmic vacuolization 5 (10%) 8 (16%) 9 (18%)
Focal cellular change 1 (2%) 1 (2%) 1 (2%)
Hepatocytomegaly 1 (2%)
#Liver/periportal
(50) (49)
Inflammation, chronic
2 (4%)
*Gallbladder
(50)
(49)
Multiple cysts
1 (2%)
#Bile duct
(50)
(49)
Hyperplasia, NOS
1 (2%)
#Pancreas
(50) (49)
Cystic ducts
2 (4%) 1 (2%)
Inflammation, chronic
2 (4%) 2 (4%)
Atrophy, NOS
1 (2%)

2-Mercaptobenzothiazole, NTP TR 332 148

TABLE D5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

DIGESTIVE SYSTEM (Continued)

#Pancreatic acinus (50) (49) (50)
Cytoplasmic vacuolization 1 (2%)
Focal cellular change 1 (2%) 1 (2%)
Atrophy, NOS
Hyperplasia, NOS 3 (6%) 2 (4%)
#Esophagus (50) (49)
Hyper keratosis 1 (2%)
#Stomach (50) (50)
Inflammation, acute 1 (2%)
#Gastric fundal gland (50) (50)
Dilatation, NOS 7 (14%) 3 (6%)
#Glandular stomach (50) (50)
Mineralization 1 (2%) 1 (2%)
Cyst, NOS 1 (2%) 1 (2%)
Inflammation, acute 1 (2%)
Inflammation, chronic 2 (4%)
#Forestomach (50) (48) (50)
Ulcer, NOS 1 (2%)
Inflammation, acute 1 (2%)
Inflammation, active chronic 1 (2%) 1 (2%)
Hyperkeratosis 2 ' (4%)
Acanthosis 1 (2%)
#Cecum (50) (49) (48)
Edema, NOS 1 (2%)
*Rectum (50) (49) (50)
Infection, protozoan 1 (2%)

URINARY SYSTEM
#Kidney (50) (49) (50)
Lymphocytic inflammatory infiltration 1 (2%) 3 (6%) 9 (18%)
Pyelonephritis, acute 1 (2%)
Inflammation, chronic 3 (6%) 3 (6%) 2 (4%)
Glomerulonephritis, chronic 1 (2%)
Infarct, focal 1 (2%)
#Kidney/cortex (50) (49) (50)
Metaplasia, osseous 1 (2%) 1 (2%) 1 (2%)
#Kidney/glomerulus (50) (49) (50)
Amy Ioidosis 1 (2%)
# Kidneykubule (50) (49) (50)
Mineralization 1 (2%) 6 (12%) 4 (8%)
Dilatation, NOS 2 (4%) 5 (10%) 5 (10%)
Nephrosis, cholemic 1 (2%)
Necrosis, NOS 4 (8%) 7 (14%) 4 (8%)
Pigmentation, NOS 1 (2%)
Regeneration, NOS 18 (36%) 15 (31%) 14 (28%)
#Urinary bladder (50) (49) (49)
Calculus, microscopic examination 1 (2%)
Hemorrhage 1 (2%)
Lymphocytic inflammatory infiltration 1 (2%) 2 (4%)
Inflammation, chronic
Hyperplasia, epithelial

149 2-Mercaptobenzothiazole, NTP TR 332

TABLE D5. SUMMARY OF THE INCIDENCE OF'NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

ENDOCRINE SYSTEM
#Anterior pituitary (49) (49) (49)
Cyst, NOS 2 (4%) 1 (2%)
Multiple cysts 1 (2%)
Hemorrhagic cyst 2 (4%)
Focal cellular change 1 (2%)
Hyperplasia, NOS 16 (33%) 14 (29%) 12 (24%)
Angiectasis 1 (2%)
#Adrenal (50)
(47) (50)

Degeneration, lipoid 1 (2%)

#AdrenaYcapsule (47) (50)

Hyperplasia, NOS 46 (98%) 49 (98%)

#Adrenal cortex (47) (50)

Accessory structure 1 (2%)

Congestion, NOS 1 (2%) 1 (2%)
Degeneration, lipoid 1 (2%) 2 (4%)
Necrosis, NOS 1 (2%)
Amyloidosis 1 (2%)
Metamorphosis, fatty 2 (4%)
Pigmentation, NOS 1 (2%)
Focal cellular change 1 (2%)
Hyperplasia, NOS 3 (6%) 2 (4%)
#Adrenal medulla
(50)
(47)
Hyperplasia, NOS
1 (2%) 1 (2%)
Hyperplasia, focal
1 (2%)
#Thyroid (50)
(49)
Embryonal duct cyst 1 (2%)
Cystic follicles 20 (40%)
15 (31%) 6 (12%)
Hyperplasia, C-cell 1 (2%)
1 (2%)
Hyperplasia, follicular cell 2 (4%)
1 (2%)
#Parathyroid (36) (32) (32)
Embryonal duct cyst 1(3%)
Hyperplasia, NOS 1 (3%)
#Pancreatic islets
(50)
(49) ' (50)
Hyperplasia, NOS
4 (8%) 7 (14%) 3 (6%)

REPRODUCTIVE SYSTEM
*Mammary gland (50)
(49) (50)

Multiple cysts 11 (22%) 5 (10%) 1 (2%)

Inflammation, acute 1 (2%)
Inflammation, chronic 1 (2%)
#Uterus (50) (49) (50)
Hydrometra 3 (6%) 5 (10%)
Hematoma, organized 1 (2%)
Inflammation, acute 2 (4%)
#Uterudendometrium (50)
(49) (50)

Inflammation, acute 3 (6%)

Hyperplasia, cystic 47 (94%) 42 (86%) 25 (50%)
#Ovary (50)
(48) (46)
Follicular cyst, NOS 1 (2%) 2 (4%) 2 (4%)
Parovarian cyst 5 (10%) 5 (10%) 1 (2%)
Hemorrhagic cyst 1 (2%)
Abscess, NOS 1 (2%)
Amyloidosis 1 (2%)
Cytomegaly 1 (2%)
Hyperplasia, epithelial 1 (2%) 2 (4%)
Angiectasis 1 (2%)

2-Mercaptobenzothiazole, NTP TR 332 150

TABLE D5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR GAVAGE STUDY OF 2-MERCAPTOBENZOTHIAZOLE (Continued)

Vehicle Control Low Dose High Dose

NERVOUS SYSTEM
#Brain (50) (49) (50)
Compression, NOS 2 (4%) 1 (2%)
Mineralization 23 (46%) 26 (53%) 9 (18%)
Congestion, NOS 1 (2%)
Infarct, NOS 1 (2%)
#Braidthalamus (50) (49) (50)
Malacia 1 (2%)

SPECIAL SENSE ORGANS

*Nasolacrimal duct (50) (49) (50)
Hemorrhage 13 (26%) 4 (8%) 3 (6%)
Lymphocytic inflammatory infiltration 1 (2%) 6 (12%)
Inflammation, acute 1 (2%)
Inflammation, chronic 1 (2%)
-

MUSCULOSKELETAL SYSTEM
*Bone (50) (49) (50)
Fibrous osteodystrophy 1 (2%)

BODY CAVITIES
+Pleura (50) (49) (50)
Vegetable foreign body 1 (2%)
*Mesentery (60) (49) (50)
Steatitis 6 (12%) 7 (14%) 2 (4%)
Inflammation, chronic 1 (2%)

ALL OTHER SYSTEMS

*Multiple organs (50) (49) (50)
Lymphocytic inflammatory infiltration 43 (86%) 42 (86%) 29 (58%)

SPECIAL MORPHOLOGY SUMMARY

Animal missingho necropsy 1

* Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.
# Number of animals examined microscopically a t thissite

151 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole,NTP TR 332 152
APPENDIX E

GENETIC TOXICOLOGY OF

2-MERCAPTOBENZOTHIAZOLE

PAGE

TABLE E l MUTAGENICITY OF 2-MERCAPTOBENZOTHIAZOLE IN SALMONELLA

TYPHIMURIUM 154

TABLE E2 MUTAGENICITY OF 2-MERCAPTOBENZOTHIAZOLE IN MOUSE L5178Y

LYMPHOMA CELLS 156
TABLE E3 INDUCTION OF SISTER CHROMATID EXCHANGES IN CHINESE HAMSTER
OVARY CELLS BY 2-MERCAPTOBENZOTHIAZOLE 158

TABLE E4 INDUCTION OF CHROMOSOMAL ABERRATIONS IN CHINESE HAMSTER

OVARY CELLS BY 2-MERCAPTOBENZOTHIAZOLE 159

153 2-Mercaptobenzothiazole, NTP TR 332

TABLE E l . MUTAGENICITY OF 2-MERCAPTOBENZOTHIAZOLE IN SALMONELLA TYPHlMURlUM (a)

Dose Revertants/plate (b)

Strain (pglplate) -s 9 t 10% S9 (hamster) + 10% S 9 (rat)
~~ ~

Study Performed at E G t G Mason Research Institute

TA100 0 136 f 10.5 133 f 3.2 106 f 7.4

3.3 125 f 9.9 __ -_
10 133 f 10.4 138 f 3.8 103 f 13.3
33 118 f 9.8 115 f 9.8 131 f 7.2
100 131 f 5.6 109 f 3.8 123 f 10.1
333 (c)63 f 0.9 105 f 11.9 130 f 4.9
1,000 __ (Cl53 f 7.9 (c)67 f 9.0
Trial summary Negative Negative Negative
Positive
control (d) 1,109 f 29.4 927 f 16.4 878 f 4.5
TA1535 0 23 f 1.0 10 f 0.9 9f 1.2
3.3 25 k 1.2 __ __
10 29 f 5.6 9f 2.1 9f 0.9
33 23 f 0.6 9k 0.9 11 f 0.0
100 28 f 3.6 11 f 1.3 8f 0.6
333 (c)20 f 2.8 6f 2.0 6f 0.7
1,000 __ (c)9f 0.9 (c)4 f 0.7
Trial summary Negative Negative Negative
Positive
control (d) 866 f 2.8 89 f 5.9 78 f 7.5

TA1537 0 8f 0.3 6f 0.3 8f 2.0

3.3 6f 1.2 __ __
10 5f 0.9 5f 0.6 7f 1.0
33 6f 0.9 5k 1.3 10 f 1.5
100 6f 0.3 11 f 2.3 7f 2.6
333 (c)6 f 0.7 7f 0.7 7f 1.5
1,000 __ (c)O f 0.3 (c)O f 0

Trial summary Negative Negative Negative

Positive
control (d) 563 f 42.3 74 f 13.6 75 f 2.2

Revertants/plate (b)
-s 9 t 10% S9 (hamster) t 10% S9(rat)
Trial 1 Trial 2 Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3

TA98 0 22 f 3.5 21 f 1.9 28 f 2.9 26 f 2.9 36 f 2.6 24 f 3.2 28 f 3.1 33 f 0.9

3.3 15 f 2.2 18 f 3.4 .- __ -_ __
10 21 f 2.3 25 f 4.1 29 f 6.1 29 +_ 1.2 31 f 0.7 24 f 1.5 28 f 0.6 31 k 3.4
33 19 k 2.6 18 f 4.4 26 f 1.3 __ __ 28 f 5.9 __ _-
100 14 f 3.1 17 f 3.8 35 f 3.2 33 f 2.3 23 f 3.2 36 f 2.6 32 f 2.6 36 f 4.4
200 __ -. __ 44 f 1.3 29 f 4.1 __ 31 f 2.5 33 f 2.2
333 12 k 1.9 15 f 1.2 42 f 1.2 49 f 2.3 26 f 3.2 49 f 2.9 40 f 2.3 33 k 2.5
400 -_ __ 43 f 3.6 32 f 5.4 __ 48 f 2.2 28 f 5.3
500 -_ _. __ 46 f 8.8 27 f 5.3 -_ 37 k 3.3 30 f 5.9
600 __ 43 f 4.3 18 f 2.6 __ 38 k 3.5 30 f 5.5
700 _. 40 f 0.9 28 f 4.1 __ 29 f 2.0 23 f 0.9
1,000 21 f 3.8 22 f 0.9 14 f 0.3 21 f 1.2 34 f 4.8 14 f 2.3
Trial summary Weakly Weakly
Negative Negative Equivocal Positive Negative Positive Equivocal Negative
Positive
control(d) 1,399 f 48.1 1,408 f 141.7 944 f 37.8 1,187 f 62.3 553 f 39.4 818 f 8.4 1,001 f 58.5 418 f 21.8

2-Mercaptobenzothiazole, NTP TR 332 154

TABLE E l . MUTAGENICITY OF 2-MERCAPTOBENZOTHIAZOLE I N SALMONELLA TYPHlMURlUM (Continued)

R e v e r t a n t s h l a t e (b)
Strain Dose - 59 t 10% S9 (hamster) t 10% S9 (rat)
(pdplate) Trial 1 Trial 2 Trial 1 Trial 2 Trial 1 Trial 2

Study performed at C a s e Western Reserve University

TAlOO 0 92 f 1.3 104 f 8.1 127 f 6.0 109 f 7.9 129 f 5.5 92 f 7.1
10 -_ 97 f 4.3 ___- 104 f 4.6 -_
--
89 f 4.1
33 97 f 7.6 100 f 6.2 89 f 4.0 98 -+ 6.5
100 92 f 6.7 109 f 4.6 85 f 7.8 93 f 7.3 91 f 15.1 81 f 11.7
333 67 f 3.8 56 f 12.9 95 f 8.8 61 f 15.0 82 -+ 14.8 72 f 1.9
1,000 (e)20 f 10.2 -- 74 f 2.8 (e)O k 0.0 22 f 11.7 (e)O f 0.0
3,333 (e)O f 0.0 _- (e)ll f 1.2 _- (e)O f 0.0 _-
10,000 __ (e)O f 0.0 (e)O f 0.0 (e10 f 0.0
Trial summary Negative Negative Negative Negative Negative Negative
Positive
control (d) 420 f 16.9 440 f 30.4 2,102 f164.8 2,286 f 43.9 1,660 f 54.6 2,162 f 59.6
TA1535 0 6 f 1.2 4 f 1.5 11 f 0.9 4 f 1.3 11 f 1.2 8 f 2.2
10 _- 4 f 2.3 __ 4 f 1.0 __ 7 f 1.0
33 8f 2.2 2 f 0.3 _- 2 f 0.7 -_ 5 f 1.3
100 5f 1.5 1 f 0.3 8 f 3.1 6 f 2.8 6 f 0.9 5 f 1.2
333 4f 0.9 3 f 1.5 6 f 1.8 6 f 3.0 10 f 3.0 2 f 0.9
1,000 7f 3.5 Toxic Toxic Toxic Toxic Toxic
3,333 (e)O f 0.0 __ (e)O f 0.0 _- (e)O f 0.0 __
10,000 __ __ (e)O f 0.0 __ (e)O f 0.0 __
Trial summary Negative Negative Negative Negative Negative Negative
Positive
control (d) 409 k192.4 100 f 14.9 67 f 6.6 86 f 7.5 148 f 11.8 95 k 13.1
TA1537 0 6 f 0.0 5f 2.7 9 f 1.5 7 f 1.5 8 f 3.5
3.3 __ 4f 0.0 __ __ _-
10 __ 3f 2.0 _- 11 f 2.6 7 f 1.9
33 6 f 1.9 3f 0.6 _- 8f 2.4 __ 5 f 0.6
f 0.3 5 k 1.8 3 f 0.9
100
333
1,000
3 f 1.0
Toxic
Toxic
4f
6f
--
0.0
1.5
9
6
6f
f 0.3
3.0
6f
6f
(e)O f
1.7
0.9
0.0
5 *
2 L 0:3
0.7 4 f 0.7
(e)O f 0.0
3,333 (e)O f 0.0 -- (e)O f 0.0 __ (e)O f 0.0 __
10,000 __ -- (e)O f 0.0 (e)O k 0.0
Trial summary Negative Negative Negative Negative Negative Negative
Positive
control (d) 182 f 55.1 271 f 32.8 306 f 94.4 164 f 21.8 431 f 13.6 365 f 17.4
TA98 0 16 f 0.6 14 f 4.4 21 k 2.2 20 f 1.9 19 f 2.4 13 f 0.7
10 -_ 8 f 1.2 -_ 17 f 1.5 __ 18 f 3.1
33 13 f 1.5 17 f 1.5 _- 14 k 4.2 _- 15 f 3.5
100 18 f 1.0 15 f 3.0 15 k 1.2 23 f 2.0 18 f 3.8 18 f 3.5
333 20 k 2.3 12 -c 2.0 15 f 1.9 12 f 2.1 12 f 1.9 17 f 0.9
1,000 7 f 6.5 (e)O f 0.0 Toxic Toxic
__ Toxic __-_
3,333 (e)O f 0.0 (e)O f 0.0 (e)O f 0.0
10,000 (eJO f 0.0 _- (e)O -+ 0.0 Toxic
Trial summary Negative Negative Negative Negative Negative Negative
Positive
control (d) 277 f 22.0 169 f 31.3 1,937 f 32.6 1,126 f 71.3 1,388 f 78.5 1,153 f 72.4

(a)The detailed protocol is presented in Haworth et al. (1983). Cells and study compound or solvent (dimethyl sulfoxide) were

incubated in the absence of exogenous metabolic activation t -S9) o r with Aroclor 1254-induced S9 from male Syrian hamster

liver or male Sprague Dawley rat liver. High dose was limited by toxicity or solubility but did not exceed 10 mg/plate;

0 pg/plate dose is the solvent control.

tb) Revertants are presented a s mean f standard error from three plates.

(c) Slight toxicity

(d) Positive control; 2-aminoanthracene was used on all strains in the presence of S9. In the absence of metabolic activation 4-
nitro-o-phenylenediamine was used with TA98, sodium azide was used with TAlOO and TA1535, and 9-aminoacridine was used
with TA1537.
(e)Precipitate on plate

155 2-Mercaptobenzothiazole, NTP TR 332

TABLE E2. MUTAGENICITY OF 2-MERCAPTOBENZOTHIAZOLE IN MOUSE L5178Y LYMPHOMA
CELLS (a,b)

Cloning Relative Mutant Mutant

Compound Concentration Efficiency Total Growth Count Fraction (c)
(pglml) (percent) (percent)

- s9
Trial 1
Ethanol (d) 91.5 f 3.6 100.0 f 3.4 61.5 f 3.7 22.5 f 1.7
2-Mercaptobenzothiazole 30 84.3 f 5.5 86.7 f 5.5 68.0 f 4.0 27.0 f 2.1
40 76.0 f 7.2 74.3 f 3.4 62.0 f 2.1 28.0 f 2.5
50 81.0 f 11.0 57.0 f 5.0 74.5 f 8.5 31.5 f 7.5
60 85.7 f 4.4 63.3 f 3.8 54.0 f 5.3 21.3 f 3.2
80 82.7 f 4.7 51.0 f 5.6 81.7 f 8.3 32.7 f 2.9
100 76.7 f 3.5 33.7 f 0.7 77.3 f 8.1 34.0 f 5.0
150 Lethal __
Methyl methanesulfonate 5 47.0 f 7.9 28.0 f 6.2 264.3 f 21.3 (e)197.0 k 34.6
Trial 2
Ethanol (d) 103.5 f 4.6 100.3 4 2.4 80.5 f 4.7 26.0 f 0.8
2-Mercaptobenzothiazole 40 84.0 f 12.8 26.3 4 10.3 63.3 f 3.8 26.7 f 4.1
60 91.3 f 11.5 18.0 f 3.2 51.7 f 12.7 19.0 f 4.0
80 89.7 4 12.8 14.3 ? 2.4 100.3 f 28.3 36.0 f 5.5
(f) 100 63.0 f 3.0 6.0 f 0.0 64.5 f 4.5 34.5 f 0.5
120 Lethal __ __ __
Methyl methanesulfonate 5 56.7 f 2.7 31.0 f 2.1 662.7 k 16.2 (e)393.0 f 19.3

+sg (g)
Trial 1
Ethanol (d) 84.3 f 6.8 100.0 f 4.4 196.5 f 8.9 79.0 f 7.4
2-Mercaptobenzothiazole 1.25 68.0 f 4.5 65.3 f 1.5 137.3 f 33.5 65.7 f 13.0
2.5 67.3 f 4.7 69.7 f 2.0 103.7 f 6.4 51.7 f 1.3
5 90.0 f 12.1 56.7 f 3.9 101.0 f 19.1 38.0 f 5.9
7.5 88.7 f 3.8 33.3 f 2.4 192.7 f -6.0 72.7 f 4.3
10 86.7 f 3.3 24.0 f 1.0 226.7 f 12.3 87.7 f 7.3
15 79.0 f 4.2 12.0 f 2.0 307.3 f 55.5 (e) 130.3 k 24.3
Methylcholanthrene 2.5 36.0 f 10.7 19.0 f 10.0 528.3 k 74.0 (e)537.0 k 74.1
Trial 2
Ethanol 81.0 f 3.0 100.3 ? 6.1 86.3 4 5.2 35.7 k 0.9
2-Mercaptobenzothiazole 5 79.0 f 6.1 85.0 f 10.5 155.7 f 5.0 (e)66.0 f 4.2
6 73.3 f 4.7 59.7 f 10.7 149.0
~ f 10.1
~
~ - - (e)69.7
,._ ... f
- 9.7
8 81.0 f 8.7 48.3 f 4.2 140.0 f 16.0 (e)58.0 f 316
10 83.0 f 9.9 35.7 f 2.2 250.3 f 9.0 (e) 104.0 f 14.4
12 75.0 f 9.0 29.0 f 4.5 218.3 f 15.2 ( e ) 100.0 k 15.6
16 68.3 f 4.8 21.3 f 2.3 144.0 f 57.4 (e169.3 f 28.0
Methylcholanthrene 2.5 73.0 f 13.1 46.0 f 9.0 589.0 f 55.5 (e)277.3 f 25.1
Trial 3
Ethanol (d) 97.5 k 3.7 100.3 4 4.8 113.8 f 1.0 39.0 f 1.8
2-Mercaptobenzothiazole 4 82.7 f 2.2 71.0 f 5.5 139.3 f 5.9 56.0 f 1.5
8 87.3 f 13.3 59.0 f 8.0 140.7 f 24.5 53.3 f 2.7
146.7 i 56.7 3 5.5
10
12
16
87.3 f
70.3 f
82.3 f
10.5
4.1
3.9
49.0
33.7
30.3
**
k 4.2
0.9
2.2
141.3 f
i8.2
18.7 (e)66.7 f 7.2
184.3 5 15.0 (e)74.7 k 2.8
20 87.7 f 8.3 21.0 f 1.7 189.0 f 16.7 (e172.0 f 4.0
Methylcholanthrene 2.5 72.7 f 4.2 42.3 f 3.4 706.7 f 81.4 (e)324.7 k 31.4

2-Mercaptobenzothiazole, NTP TR 332 156

TABLE E2. MUTAGENICITY OF 2-MERCAPTOBENZOTHIAZOLE IN MOUSE L5178Y LYMPHOMA CELLS
(Continued)

(a)Study performed at Litton Bionetics, Inc. The experimental protocol is presented in detail by Myhr et al. (1985)and follows
the basic format of Clive e t al. (1979).The highest dose ofstudy compound is determined by solubility or toxicity and may not
exceed 5 mg/ml. All doses are tested in triplicate; unless otherwise specified, the average for the three tests is presented in the
table. Cells (6 X 10Vml) were treated for 4 hours a t 37” C in medium, washed, resuspended in medium, and incubated for 48
hours at 37”C. After expression, 3 X 106cells were plated in medium and soft agar supplemented with trifluorothymidine for
selection of cells that were mutant a t the thymidine kinase (TK) locus, and 600 cells were plated in nonselective medium and
soft agar to determine the cloning efficiency.
(b)Mean k standard error of replicate trials for approximately 3 X 106 cells each. All data are evaluated statistically for both
trend and peak response (P<0.05 for a t least one of the three highest dose sets). Both responses must be significantly (P<0.05)
positive for a chemical to be considered mutagenic. If only one of these responses is significant, the call is “questionable”; the
absence of both trend and peak response results in a “negative” call.
(c)Mutant fraction (frequency)is a ratio of the mutant count to the cloning efficiency, divided by 3 (to arrive at MF per 1 x 108
cells treated); MF = mutant fraction.
(d) Data presented are the average of four tests.
(e) Significant positive response; occurs when the relative mutant fraction (average MF of treated culture/average MF of sol-

vent control) is greater than or equal to 1.6.

(D Data presented are the average oftwo tests; doses in one test were lethal.

(g) Tests conducted with metabolic activation were performed as described in (a) except that S9, prepared from the liver of
Aroclor 1254-inducedF344 rats, was added at the same time a s the study chemical and/or solvent (ethanol).

157 2-Mercaptobenzothiazole, NTP TR 332

TABLE E3. INDUCTION OF SISTER CHROMATID EXCHANGES IN CHINESE HAMSTER OVARY CELLS
BY 8-MERCAPTOBENZOTHIAZOLE (a)

No. OF SCEs/ Relative

Total Chromo. No. of Chromo- SCEs/ Hours SCEs/Cell
Compound Dose Cells some8 SCEs dome Cell in BrdU (percent)
(Wrnl) (b)

-89 (c)
Trial No. 1.-Summary: Negative
Dimethyl sulfoxide 50 1,035 500 0.48 10.0 25.3 ..
2-Mercaptobenzothiazole 12.5 50 1,036 47 1 0.46 9.4 (d132.6 94.0
14.9 50 1,027 515 0.50 10.3 (d)32.6 103.0
20.1
24.8
50
0 __
1,025 568
..
0.55
..
11.4
..
(d)32,6
-.
114.0
-.
Mitomycin C 0.001 50 1,027 741 0.72 14.8 26.3 148.0
0.010 5 104 205 1.97 41.0 26.3 410.0

t S 9 (e)
Trial No. 1--Summary: Positive
Dimethyl sulfoxide 50 1,038 477 0.46 9.5 25.3 __
2-Mercaptobenzothia2ole 99.2 50 1,028 53 1 0.52 10.6 25.3 111.6
247.5 50 1,026 536 0.52 10.7 25.3 112.6
501.6
750
50
0 __
1,045 640
.. __
0.61 12.8
.- __
(d)32.6 134.7
.-
Cyclophosphamide 0.4 50 1,020 634 0.62 12.7 25.3 133.7
2.0 5 104 142 1.37 28.4 26.3 298.9

Trial No. %-Summary: Positive

Dimethyl sulfoxide BO 1,025 454 0.44 9.1 26.6 .-
2-Mercaptobenzothiamle 351.6 50 1,032 558 0.64 11.2 (d136.6 123.1
401.6 50 1,036 624 0.60 12.5 (d)36.6 137.4
445.3
502.3
50
0
1,041
-. __
588 0.66
-.
11.8
..
(d)36.6
-.
129.7
..
Cyclophosphamide 0.4 50 1,035 702 0.68 14.0 26.6 153.8
2 .o 5 108 183 1.69 36.6 25.6 402.2

(a) Study performed a t Litton Bionetics, Inc. SCE = sister chromatid exchange; BrdU = bromodeoxyuridine4 A detailed de-
scription ofthe SCE protocol is presented by Galloway e t al. (1985). Briefly, Chinese hamster ovary cells were incubated with
study compound or solvent (dimethyl sulfoxide) a8 described in (c) or (d) below and cultured for sufncient time to reach second
metaphase division. Cells were then collected by mitotic shake-off, fixed, air-dried, and stained.
(b)SCEs/cell in treated culture expressed a s a percent of the SCEs/cell in the control culture
(c)In the absence ofS9, Chinese hamster ovary cella were incubated with s t d y compound or solvent for 2 hours a t 37' C. Then
BrdU was added, and incubation was continued for 24 hours. Cells were washed, fresh medium containing BrdU and colcemid
wan added, and incubation was continuedfor 2.3 hours.
(d) Because of significant chemically induced cell cycle delay, incubation time before addition of colcemid was lengthened to
provide sufficient metaphases a t harvest.
(e) In the presence of SS,cells were incubated with study compound or solvent for 2 hours at 37'C. Then cells were washed, and
medium containing BrdU was added. Cells were incubated for a further 26 hours, with colcemid preeent for the final 2-3 houre.
S9 was from the liver ofAroclor 1254-inducedmale Sprague Dawley rata.

2-Mercaptobenzothiazole,NTP TR 332 158

TABLE E4. INDUCTION OF CHROMOSOMAL ABERRATIONS IN CHINESE HAMSTER OVARY CELLS
BY 2-MERCAPTOBENZOTHIAZOLE (a)

-S9 (b) ts9 (Cl

Dose Total No. of Absl Percent Dose Total No. of Absl Percent
(pg/ml) Cells Abs Cell Cells (pg/ml) Cells Abs Cell Cells
with Abs with Abs

Trial 1--Harvesttime 20.5 h (d) Trial 1--Harvesttime 20.5 h (d)

Dimethyl sulfoxide Dimethyl sulfoxide
100 0 0.00 0 100 1 0.01 1

2-Mercaptobenzothiazole 2-Mercaptobenzothiazole

10 100 1 0.01 I 351.8 100 18 0.18 9

14.9 100 1 0.01 1 400.8 100 14 0.14 9
19.9 loo 2 0.02 2 451.0 50 24 0.48 16
30.1 0 500.5 0
Summary: Negative Summary: Positive

Mitomycin C Cyclophosphamide

0.025 100 12 0.12 10 2.5 100 4 0.04 4

0.062 25 14 0.56 10 12.5 25 12 0.48 36

Trial 2-Harvest time 20.5 h (d)

Dimethyl sulfoxide

50 1 0.02 2

2-Mercaptobenzothiazole
373.5 25 12 0.48 24
399 25 17 0.68 28
425 25 21 0.84 28
450 0

Summary: Positive

Cyclophosphamide
3.8 50 3 0.06 6
12.5 25 9 0.36 20

(a)Study performed a t Litton Bionetics, Inc. Abs = aberrations. A detailed presentation ofthe technique for detecting chromo-
somal aberrations is found in Galloway et al. (1985). Briefly, Chinese hamster ovary cells were incubated with study compound
or solvent (dimethyl sulfoxide) a s indicated in ( b )or (c). Cells were arrested in first metaphase by addition of colcemid and har-
vested by mitotic shake-off, fixed, and stained in 6%Giemsa.
(b)In the absence ofS9, Chinese hamster ovary cells were incubated with study compound or solvent (dimethyl sulfoxide) for 8-
10 hours a t 37" C. Cells were then washed, and fresh mediutn containing colcemid was added for a n additional 2-3 hours fol-
lowed by harvest.
(c)In the presence ofS9, cells were incubated with study compound or solvent (dimethyl sulfoxide) for 2 hours a t 37OC. Cells
were then washed, medium was added, and incubation was continued for 8-10 hours. Colcemid was added for the last 2-3 hours
of incubation before harvest, S9 was from the liver ofAroclor 1254-induced male Sprague Dawley rats.
td) Because of significant chemically induced cell cycle delay, incubation time before addition of colcemid was lengthened to
provide sufficient metaphases a t harvest.

159 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole,NTP TR 332 160
APPENDIX F

SENTINEL ANIMAL PROGRAM

PAGE

TABLE F1 MURINE ANTIBODY DETERMINATIONS FOR RATS AND MICE IN THE TWO-YEAR
GAVAGE STUDIES OF 2-MERCAPTOBENZOTHIAZOLE 163

161 2-Mercaptobenzothiazole, NTP TR 332

APPENDIX F. SENTINEL ANIMAL PROGRAM

I. Methods

Rodents used in the Carcinogenesis Program of the National Toxicology Program are produced in op-
timally clean facilities to eliminate potential pathogens that may affect study results. The Sentinel
Animal Program is part of the periodic monitoring of animal health that occurs during the toxicologic
evaluation of chemical compounds. Under this program, the disease state of the rodents is monitored
via serology on sera from extra (sentinel) animals in the study rooms. These animals are untreated,
and these animals and the study animals are both subject to identical environmental conditions. The
sentinel anirrials come from the same production source and weanling groups as the animals used for
the studies of chemical compounds.

Fifteen B6C3F1 mice and 15 F344/N rats of each sex are selected at the time of randomization and al-
location of the animals to the various study groups. Five animals of each designated sentinel group
are killed a t 6,12, and 18 months on study. Data from animals surviving 24 months are collected
from 5/50 randomly selected control animals of each sex and species. The blood from each animal is
collected and clotted, and the serum is separated. The serum is cooled on ice and shipped to Microbio-
logical Associates’ Comprehensive Animal Diagnostic Service for determination of the antibody ti-
ters. The following tests are performed:

Hemagglutination Complement
Inhibition Fixation ELISA

Mice PVM (pneumonia virus of mice) M. Ad. (mouse adenovirus) MHV (mouse
Reo 3 (reovirus type 3) LC M (1ymphocy tic hepatitis virus)
GDVII (Theiler’s choriomeningitis virus) M . pul. (Mycoplasma
encephalomyelitis virus) pulrnonis) (24 mo)
Poly (polyoma virus)
MVM (minute virus of mice)
Ectro (infectious ectromelia)
Sendai

Rats PVM RCV (rat coronavirus) M . pul. (24 mo)

KRV (Kilham rat virus)
H-1 (Toolan’s H-1 virus)
Sendai

II. Results
Results are presented in Table F1.

2-Mercaptobenzothiazole, NTP TR 332 162

TABLE F1. MURINE ANTIBODY DETERMINATIONS FOR RATS AND MICE IN THE TWO-YEAR
GAVAGE STUDIES OF 2-MERCAPTOBENZOTHIAZOLE (a)

No. of Positive Serologic

Interval (months) Animals Reaction for

RATS
6 8/10 Sendai

12 10110 Sendai

18 1/10 Sendai

24 10110 (b)M.pul.
4110 Sendai

MICE

6 9/10 Sendai

12 10/10 Sendai

18 5/10 Sendai

24 6/10 Sendai

(a) Blood samples were taken from sentinel animals a t 6,12, and 18 months aRer the start ofdosing and from the vehicle control
animals just before they were killed; samples were sent to Microbiological Associates (Bethesda, MD) for determination of anti-
body titers.
(b) Further evaluation of this assay indicated that it is not specific for M.pulrnonis, and these results are considered false
positive.

163 2-Mercaptobenzothiazole, NTP TR 332

2-Mercaptobenzothiazole, NTP TR 332 164
APPENDIX G

INGREDIENTS, NUTRIENT COMPOSITION, AND

CONTAMINANT LEVELS

IN NIH 07 RAT AND MOUSE RATION

Pelleted Diet: July 1981 to July 1983

(Manufactured by Zeigler Bros.* Inc., Gardners, PA)

PAGE
TABLE 6 1 INGREDIENTS OF NIH 07 RAT AND MOUSE RATION 166

TABLE C2 VITAMINS AND MINERALS IN NIH 07 RAT AND MOUSE RATION 166

TABLE G3 NUTRIENT COMPOSITION OF NIH 07 RAT AND MOUSE RATION 167

TABLE C4 CONTAMINANT LEVELS IN NIH 07 RAT AND MOUSE RATION 168

165 2-Mercaptobenzothiazole,NTP TR 332

TABLE G l . INGREDIENTS OF NIH 07 RAT AND MOUSE RATION (a)

Ingredients (b) Percent by Weight

Ground X2 yellow shelled corn 24.50

Ground hard winter wheat 23.00
Soybean meal (49% protein) 12.00
Fish meal (60% protein) 10.00
Wheat middlings 10.00
Dried skim milk 5.00
Alfalfa meal (dehydrated, 17% protein) 4.00
Corn gluten meal (60% protein) 3.00
Soy oil 2.50
Brewer's dried yeast 2.00
Dry molasses 1.50
Dicalcium phosphate 1.25
Ground limestone 0.50
Salt 0.50
Premixes (vitamin and mineral) 0.25

(a)NIH, 1978; NCI, 1976

(b)Ingredients ground to pass through a U.S. Standard Screen No. 16 before being mixed

TABLE G2. VITAMINS AND MINERALS IN NIH 07 RAT AND MOUSE RATION (a)

Amount Source

Vitamins
A 5,500,000 IU Stabilized vitamin A palmitate or acetate
4,600,000 IU D-activated animal sterol

K3 2.8 g Menadione activity

d-a-Tocopheryl acetate 20,000 IU

Choline 560.0 g Choline chloride

Folic acid 2.2 g

Niacin 30.0 g

d-Pantothenic acid 18.0 g d-Calcium pantothenate

Riboflavin 3.4 g

Thiamine 10.0 g Thiamine mononitrate

B12 4,000 pg

Pyridoxine 1.7 g Pyridoxine hydrochloride

Biotin 140.0 mg d-Biotin

Minerals
Iron 120.0 g Iron sulfate

Manganese 60.0 g Manganous oxide

Zinc 16.0 g Zinc oxide

Copper 4.0 g Copper sulfate

Iodine 1.4 g Calcium iodate

Cobalt 0.4 g Cobalt carbonate

~~~

(a) Per ton (2,000 lb) of finished product

2-Mercaptobenzothiazole,NTP TR 332 166

TABLE G3. NUTRIENT COMPOSITION OF NIH 07 RAT AND MOUSE RATION

Mean k Standard
Nutrients Deviation Range Number of Samples

Crude protein (percent by weight) 23.6 f 0.87 22.2-25.3

25

Crude fat (percent by weight) 4.92 f 0.54 3.3-5.7

25

Crude fiber (percent by weight) 3.30 f 0.26 2.9-3.8

25

Ash (percent by weight) 6.43 f 0.39 5.7-7.2

25

Amino Acids ( p e r c e n t of total diet)

Arginine 1.323 f 0.830 1.21-1.39

4

Cystine 0.310 f 0.099 0.218-0.400

4

Glycine 1.155 f 0.069 1.06-1.21

4

Histidine 0.572 f 0.030 0.530-0.603

4

Isoleucine 0.910 f 0.033 0.881-0.944

4

Leucine 1.949 f 0.065 1.85-1.99

4

Lysine 1.275 f 0.076 1.20-1.37

4

Methionine 0.422 f 0.187 0.306-0.699

4

Phenylalanine 0.909 f 0.167 0.665-1.04

4

Threonine 0.844 k 0.029 0.824-0.886

4

Tryptophan 0.187 0.171-0.21 1

3

Tyrosine 0.631 f 0.094 0.566-0.769

4

Valine 1.11 f 0.050 1.05-1.17

4

Essential Fatty Acids ( p e r c e n t of total diet)

Linoleic 2.44 2.37-2.52
3

Linolenic 0.274 0.256-0.308

3

Arachidonic 0.008 __ 1

Vitamins

Vitamin A (IUkg) 12,088 f 4,119 7,500-24,000 25

Vitamin D (IUkg) 4,650 3,000-6,300 2

a-Tocopherol (ppm) 41.53 f 7.52 31 .l-48.9 4

Thiamine (ppm)(a) 16.2 f 2.30 12.0-21.o 24

Riboflavin (ppm) 7.5 f 0.96 6.1-8.2 4

Niacin (ppm) 85.0 f 14.2 65.0-97.0 4

Pantothenic acid (ppm) 29.3 f 4.6 23.0-34.0 4

Pyridoxine (ppm) 7.6 f 1.5 5.6-8.8 4

Folic acid (ppm) 2.8 f 0.88 1.8-3.7 4

Biotin (ppm) 0.27 f 0.05 0.21-0.32 4

Vitamin BIz (ppb) 21.0 f 11.9 1 1 .O-38.0 4

Choline (ppm) 3,302.0 f 120.0 3,200.0-3,430.0 4

Minerals

Calcium (percent) 1.23 k 0.10 1.08-1.44

25

Phosphorus (percent) 0.98 f 0.05 0.88-1.11

25

Potassium (percent) 0.862 f 0.100 0.772-0.974

3

Chloride (percent) 0.546 f 0.100 0.442-0.635

4

Sodium (percent) 0.311 f 0.038 0.258-0.350

4

Magnesium (percent) 0.169 k 0.133 0.151-0.181

4

Sulfur (percent) 0.316 f 0.070 0.270-0.420

4

Iron (ppm) 447.0 f 57.3 409.0-523.0

4

Manganese (ppm) 90.6 f 8.20 81.7-95.5

4

Zinc (ppm) 53.6 f 5.27 46.1-58.6

4

Copper (ppm) 10.77 f 3.19 8.09-15.39

4

Iodine (ppm) 2.95 f 1.05 1.52-3.82

4

Chromium (ppm) 1.81 f 0.28 1.44-2.09

4

Cobalt (ppm) 0.68 f 0.14 0.49-0.80

4

~~

(a) One batch (7/22/81)

not analyzed for thiamine.

167 2-Mercaptobenzothiazole,NTP TR 332

TABLE G4. CONTAMINANT LEVELS I N NIH 07 RAT AND MOUSE RATION

Mean f Standard

Contaminants Deviation
Range N u m b e r of Samples

Arsenic (ppm) 0.50 f 0.13 0.29-0.77 26

Cadmium (ppm) <0.10 <0.10-0.10 25
Lead (ppm) (a) 0.74 f 0.42 0.33-1.97 23
Lead (ppm) (b) 0.92 f 0.75 0.33-3.37 25
Mercury (ppm) (c) <0.05 25
Selenium (ppm) 0.29 f 0.07 0.14-0.40 25

Aflatoxins (ppb) < 10 <5.0- < 10.0 25

Nitrate nitrogen (ppm) (d)

9.22 f 4.39 1.9-17.0 25
Nitrite nitrogen (ppm) (d)
2.19 f 1.55 <0.6-6.9 25
BHA (ppm) (e)
5.86 f 4.87 2.0-17.0 25
BHT (ppm) (e)
3.0 f 2.7 < 1.0-12.0 25

Aerobic plate count (CFU/g) (0

43,936 f 31,267 4,900-1 10,000 25
Coliform (MPN/g) (g)
14.96 f 22.36 <3-93 24
Coliform (MPN/g) (h)
32.76 f 91.66 <3-460 25
E. coli (MPN/g) (i) <3 25

Total nitrosamines (ppb) 3.42 f 2.72 0.8-9.3 26

N-Nitrosodimethylamine(ppb) 2.68 f 2.37 0.8-8.3 25
N-Nitrosopyrrolidine (ppb) 1.14 f 0.48 <0.5-2.9 25

Pesticides (ppm)

a-BHC (c,j) <0.01 25

B-BHC (c) <0.02 26
y-BHC-Lindane (c) <0.01 25
6-BHC (c) <0.01 25
Heptachlor (c) <0.01 25
Aldrin (c) <0.01 25
Heptachlor epoxide (c) <0.01 25
DDEW <0.01 25
DDD(c) <0.01 25
DDTW <0.01 25
HCBW eo.01 25
Mirex (c) <0.01 25
Methoxychlor (k) <0.05 25
Dieldrin (c) <0.01 25
Endrin (c) <0.01 25
Telodrin (c) <0.01 26
Chlordane (c) <0.05 25
Toxaphene (c) <0.1 25
Estimated PCBs (c) <0.2 25
Ronnel (c) <0.01 25
Ethion (c) <0.02 25
Trithion (c) <0.05 25
Diazinon (c) <0.1 25
Methyl parathion (c) <0.02 25
Ethyl parathion (c) <0.02 26
Malathion (1) 0.09 f 0.06 <0.05-0.27 25
Endosulfan I (m) <0.01 20
Endosulfan II (m) <0.01 20
Endosulfan sulfate (m) <0.03
20

2-Mercaptobenzothiazole, NTP TR 332 168

TABLE 6 4 . CONTAMINANT LEVELS I N NIH 07 RAT AND MOUSE RATION (Continued)

(a) Mean, standard deviation, and range exclude two high values of 2.66 ppm and 3.37 ppm obtained for batches produced on

8/26/81 andon7/21/82.

(b)Mean,standard deviation, and range include the high values given in (a).

(c)All values were less than the detection limit. The detection limit is given as the mean.
(d) Sources of contamination: alfalfa, grains, and fish meal
(e) Sources of contamination: soy oil and fish meal
(0CFU =colony forming unit
(g)MPN = most probable number; mean, standard deviation, and range exclude one high value of 460 MPN/g obtained for the
batch produced on 9/23/82.
(h) Mean, standard deviation, and range include the high value listed in (g).
(i)All values were less than 3 MPNlg.
(i)BHC = hexachlorocyclohexane or benzene hexachloride
(k)Two observations were above the detection limit. The values and the d a b 8 they were obtained are given under the range.
(1)Eleven batches contained more than 0.06 ppm.
(m)Four batches (7/22/81-11/25/81) were not analyzed for endosulfan I, endosulfan II, or endorulfan rulfate.

169 2-Mercaptobenzothiazole,NTP TR 332

2-Mercaptobenzothiazole,NTP TR 332 170

APPENDIX W

AUDIT SUMMARY

171 2-Mercaptobenzothiazole,NTP TR 332

APPENDIX H. AUDIT SUMMARY

The experimental data, pathology materials, and draft NTP Technical Report for the 2-year studies of
2-mercaptobenzothiazole in F344/N rats and B6C3F1 mice were examined for accuracy, consistency,
and completeness. The studies were conducted for the NTP by Physiological Research Laboratories
(Minneapolis, Minnesota) under a subcontract with Tracor Jitco, Inc. (Rockville, Maryland), until
February 28, 1983, and then under a contract with the National Institute of Environmental Health
Sciences (NIEHS). Animal exposures for the 2-year studies began in July 1981, about 3 months prior
to the date (October 1, 1981) when the NTP required studies to be conducted in full compliance with
the FDA Good Laboratory Practice regulations for nonclinical laboratory studies. The retrospective
audit was conducted for the NIEHS a t the NTP Archives in September and October 1986 by Dynamac
Corporation, J.C. Bhandari, D.V.M., Ph.D., Principal Investigator. Other individuals who conducted
the audit are listed in the full report, which is on file a t the NIEHS. The data audit included a review

All records concerning animal receipt, quarantine, randomization, and disposition prior to

study start.

All chemistry records.

Body weight and clinical observation data for a random 10%sample of the study animals.

A random 50% percent sample of the dose records.

All inlife records concerning environmental conditions, palpable masses, mortality, and ani-

mal identification.

All postmortem records for individual animals concerning identification, disposition codes,

condition codes, and correlation between necropsy observations and histopathologic findings.

Wet tissues from a random 10% sample of the study animals to verify animal identification

and to examine for untrimmed potential lesions.

Slides and blocks of tissues from all vehicle control and high dose animals to examine for

proper match and inventory.

The audit showed that inlife procedures were documented in the Materials and Methods Report sub-
mitted by the study laboratory and by archival records with the exception of periodic animal room
procedures for cage and rack changes, equipment sanitization, light cycle, twice daily morbidity and
moribundity checks, and animal dosing for the first several months. The analytical chemistry records
from the study laboratory were complete and accurate, but raw data for the initial characterization of
2-mercaptobenzothiazole by Midwest Research Institute were not present at the Archives for the au-
dit. Review of the pathology documents resulted in a change in disposition code for 10 mice from nat-
ural death or moribund kill to accidental death because of gavage trauma. Review of the pathology
specimens revealed only miscellaneous findings that were not significant to the interpretation of the
study results.

In summary, the findings of the data audit were adequately resolved or were considered not to affect
the interpretation of these studies. Thus, the retrospective audit, coupled with audit of the draft
Technical Report, shows that the records and specimens for the 2-year studies of 2-mercaptobenzo-
thiazole support the data and results presented in this NTP Technical Report.

2-Mercaptobenzothiazole, NTP TR 332 172

Q U.S. GOVERNMENT PRINTING OFFICE: 1 9 8 G 2 0 1 -7 7 8 8 0 3 0 5
NATIONAL TOXICOLOGY PROGRAM TECHNICAL REPORTS

PUBLISHED AS OF APRIL 1988

TR No. CHEMICAL TR No. CHEMICAL

200 2,6-Toluenediamine Dihydrochloride 263 1,2-Dichloropropane

20 1 2,3,7,8-Tetrachlorodibenzo-p-dioxin
(Dermal) 267 Propylene Oxide
202 1,2,3,6,7,8-Hexachlorodibenzo-p-dioxinand
269 Telone II@
(Dermal)
1,2,3,7,8,9-Hexachlorodibenzo-p-dioxin 27 1 HC Blue No. 1

203 Phenol 272 Propylene

204 Benzoin 273 Trichloroethylene (Four strains of rats)
205 4,4’-Oxydianiline 274 Tris(2-ethylhexy1)phosphate
206 Dibromochloropropane 275 2-Chloroethanol
207 Cytembena 276 8-Hydroxyquinoline
208 FD & C Yellow No. 6 281 H.C. Red No. 3
209 2,3,7,8-Tetrachlorodibenzo-p-dioxin(Gavage) 282 Chlorodibromomethane
210 1,2-Dibromoethane (Inhalation) 284 Diallylphthalate (Rats)
211 C.I. Acid Orange 10 285 C.I. Basic Red 9 Monohydrochloride
212 Di(2-ethy1hexyl)adipate 287 Dimethyl Hydrogen Phosphite
213 Butylbenzyl Phthalate 288 1,3-Butadiene
214 Caprolactam 289 Benzene
216 Bisphenol A 29 1 Isophorone
216 11-Aminoundecanoic Acid 293 HC Blue No. 2
217 Di(2-ethylhexy1)phthalate 294 Chlorinated Trisodium Phosphate
219 2,6-Dichloro-p-phenylenediamine 295 Chrysotile Asbestos (Rats)
220 C.I. Acid Red 14 296 Tetrakis(hydroxymethy)phosphonium Sulfate and
221 Locust Bean Gum Tetrakis(hydroxymethy)phosphonium Chloride
222 C.I. Disperse Yellow 3 298 Dimethyl Morpholinophosphoramidate
223 Eugenol 299 C.I. Disperse Blue 1
224 Tara Gum 300 3-Chloro-2-methylpropene
225 D & C RedNo. 9 301 o-Phenylphenol
226 C.I. Solvent Yellow 14 303 4-Vinylcyclohexene

227 Gum Arabic 304 Chlorendic Acid

228 Vinylidene Chloride 305 Chlorinated Paraffins ( C u , 43% chlorine)

229 Guar Gum 306 Dichloromethane

230 Agar 307 Ephedrine Sulfate

231 Stannous Chloride 308 Chlorinated Paraffins (C12,60% chlorine)

232 Pentachloroethane 309 Decabromodiphenyl Oxide

233 2-Biphenylamine Hydrochloride 310 Marine Diesel Fuel and JP-5 Navy Fuel

234 Allyl Isothiocyanate 311 Tetrachloroethylene (Inhalation)

235 Zearalenone 312 n-Butyl Chloride

236 D-Mannitol 314 Methyl Methacrylate

237 1,1,1,2-Tetrachloroethane 315 Oxytetracycline Hydrochloride

238 Ziram 316 1-Chloro-2-methylpropene

239 Bis(2-chloro-1methy1ethyl)ether 317 Chlorpheniramine Maleate

240 Propyl Gallate 318 Ampicillin Trihydrate

242 Diallyl Phthalate (Mice) 319 1,4-Dichlorobenzene

244 Polybrominated Biphenyl Mixture 320 Rotenone

245 Melamine 321 Bromodichloromethane

247 L-Ascorbic Acid 322 Phenylephrine Hydrochloride

248 4,4’-Methylenedianiline Dihydrochloride 323 Dimethyl Methylphosphonate

249 Amosite Asbestos 324 Boric Acid

250 Benzyl Acetate 325 Pentachloronitrobenzene

25 1 Toluene Diisocyanate 326 Ethylene Oxide

252 Geranyl Acetate 327 Xylenes (Mixed)

253 Allyl Isovalerate 328 Methyl Carbamate

255 1,2-Dichlorobenzene 329 1,2-Epoxybutane

257 Diglycidyl Resorcinol Ether 333 N-Phenyl-2-naphthylamine

259 Ethyl Acrylate 334 2-Amino-5-nitrophenol

26 1 Chlorobenzene

These NTP Technical Reporta are available for sale from the National Technical Information Service, U.S.Department of
Commerce, 5285 Port Royal Road, Springfield, VA 22161 (703-487-4650). Single copies of this Technical Report are avail-
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