National Cancer Institute

CARCINOGENESIS

Technical Report Series

No. 93

1978

BIOASSAY OF

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE

FOR POSSIBLE CARCINOGEN ICITY

CAS No. 132-32-1

NCI-CG-TR-93

U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE

Public Health Service

National Institutes of Health
BIOASSAY OP
 

3-AMINO-9-ETHYLCARBAZOLE (HYDROCHLORIDE)
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
 

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-1337
 

BIOASSAY OF
 

3-AMINO-9-ETHYLCARBAZOLE (HYDROCHLORIDE)
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

FOREWORD; This report presents the results of the bioassay of
 

3-amino-9-ethylcarbazole (hydrochloride) conducted for the
 

Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute (NCI), National Institutes
 

of Health, Bethesda, Maryland. This is one of a series of
 

experiments designed to determine whether selected chemicals have
 

the capacity to produce cancer in animals. Negative results, in
 

which the test animals do not have a greater incidence of cancer
 

than control animals, do not necessarily mean that the test
 

chemical is not a carcinogen, inasmuch as the experiments are
 

conducted under a limited set of circumstances. Positive results
 

demonstrate that the test chemical is carcinogenic for animals
 

under the conditions of the test and indicate that exposure to
 

the chemical is a potential risk to man. The actual determination of the risk to man from animal carcinogens requires a wider
 

analysis.
 

CONTRIBUTORS; The bioassay of 3-amino-9-ethylcarbazole
 

(hydrochloride) was conducted by EG&G Mason Research Institute,
 

Worcester, Massachusetts, initially under direct contract to the
 

NCI and currently under a subcontract to Tracor Jitco, Inc.,
 

prime contractor for the NCI Carcinogenesis Testing Program.
 

The bioassay was conducted under the supervision of Drs. A.
 

Handler^ and E. Smith^, principal investigators, and Mr. G.
 

Wade-^. NCI project officers were Drs. E. K. Weisburger^, T.
 

Cameron^, and N. P. Page^»->. The program manager was Mr. J.
 

Baker^. Ms. A. Good^ supervised the technicians in charge of
 

animal care, and Ms. E. Zepp^ supervised the preparation of the
 

feed mixtures and collected samples of the diets for analysis.
 

Ms. D. Bouthot^ kept all daily records of the test, and Ms. R.
 

Monson^ prepared a report based on these records. Histopathologic examinations were performed by Drs. A. S. K. Murthy^
 

iii
 
and D. Hayden , and the diagnoses included in this report
 

represent their interpretations.
 

Animal pathology tables and survival tables were compiled at EG&G
 

Mason Research Institute, Rockville, Maryland". The statistical
 

analyses were performed by Dr. J. R. Joiner? and Ms. P. L. Yong',
 

using methods selected for the bioassay program by Dr. J. J.
 

Gart°.
 

The melting point of the test chemical and the analysis of the
 

dosed feed mixtures used in this bioassay were performed by Dr.
 

M. Hagopian-^. Chemicals were analyzed under the direction of Dr.
 

E. Murrill^. The results of the analyses were reviewed by Dr. C.
 

W. Jameson' and Dr. S. S. Olin?.
 

This report was prepared at Tracer Jitco? under the direction of
 

NCI. Those responsible for the report at Tracer Jitco were Dr.
 

L. A. Campbell, Director of the Bioassay Program; Dr. S. S. Olin,
 

Deputy Director for Science; Dr. J. F. Robens, toxicologist; Dr.
 

R. L. Schueler, pathologist; Dr. G. L. Miller, Ms. L. A. Waitz,
 

and Mr. W. D. Reichardt, bioscience writers; and Dr. E. W.
 

Gunberg, technical editor, assisted by Ms. Y. E. Presley and Ms.
 

P. J. Graboske.
 

The following scientists at NCI were responsible for evaluating
 

the bioassay, interpreting the results, and reporting the
 

findings: Dr. Kenneth C. Chu, Dr. Cipriano Cueto, Jr., Dr. J.
 

Fielding Douglas, Dr. Dawn G. Goodman^^ or. Richard A.
 

Griesemer, Dr. Morton H. Levitt, Dr. Harry A. Milman, Dr. Thomas
 

W. Orme, Dr. Robert A. Squire^, Dr. Sherman F. Stinson, Dr.
 

Jerrold M. Ward, and Dr. Carrie E. Whitmire.
 

'•Now with Abcor, Inc., 850 Main Street, Wilmington,
 

Massachusetts.
 

^Now with the University of Massachusetts Medical Center,
 

55 Lake Avenue, Worcester, Massachusetts.
 

iv
 
-^EG&G Mason Research Institute, 57 Union Street, Worcester,
 

Massachusetts.
 

^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

^Now with the Environmental Protection Agency, 401 M Street,
 

S.W., Washington, B.C.
 

^EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

7

Tracor Jitco, Inc., 1776 East Jefferson Street, Rockville,
 

Maryland.
 

^Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute,
 

National Institutes of Health, Bethesda, Maryland.
 

^Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

l^Now with Clement Associates, Inc., 1010 Wisconsin Avenue, N.W. ,
 

Suite 660, Washington, D.C.
 

1-^Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

v
 

SUMMARY
 

A bioassay of 3-amino-9-ethylcarbazole (hydrochloride) for
 

possible carcinogenicity was conducted by administering the test
 

chemical in feed to Fischer 344 rats and B6C3F1 mice. Both the
 

free amine form and the hydrochloride salt were used.
 

Groups of 50 rats of each sex and 50 mice of each sex were administered the test chemical at one of two doses, either 800 or
 

2,000 ppm for rats and either 800 or 1,200 ppm for mice, for 78
 

weeks. The r?ts were then observed for an additional 26-29
 

weeks, and the mice for an additional 16-17 weeks. Controls
 

consisted of groups of 50 untreated rats of each sex and 50
 

untreated mice of each sex; separate controls were used for the
 

groups of animals administered the different doses. All
 

surviving rats were killed at 104-110 weeks; all surviving mice
 

were killed at 94-97 weeks.
 

Since the suppliers of the low-dose rats and mice differed from
 

those of the corresponding low-dose controls, while the suppliers
 

for the high-dose rats and mice were the same as those of the
 

corresponding high-dose controls, comparisons of high-dose groups
 

with their corresponding controls were the more appropriate.
 

Furthermore, since the low-dose animals did not receive the same
 

regimen of administration of the test compound as that received
 

by the high-dose animals, and since tests using the low-dose
 

groups were not performed concurrently with those using the
 

high-dose groups, analyses of dose—related trends were not
 

possible. Although the interpretation of results of the study
 

was based primarily on comparisons of high-dose groups with their
 

respective controls, the results obtained with the low—dose 

groups, regardless of the indicated complicating factors, 

supported the interpretation. 

Neoplasms of the liver were observed in significant incidences in
 

rats and mice of both sexes. In male rats, hepatocellular
 

carcinomas alone were significantly higher (P _< 0.020) in both
 

the low- and high-dose groups. When neoplastic nodules of the
 

liver were combined with hepatocellular carcinomas, the
 

combination occurred at significant incidences (P £ 0.012) in the
 

low- and high-dose male rats and in the high-dose female rats
 

vii
 
(males: low-dose controls 0/36, low-dose 12/42; high-dose
 

controls 1/48, high-dose 22/48; females: high-dose controls 0/50,
 

high-dose 6/48). Hepatocellular carcinomas alone similarly
 

occurred at significant incidences (P < 0.001) in the low- and
 

high-dose male and female mice (males: low-dose controls 7/48,
 

low-dose 32/44; high-dose controls 6/44, high-dose 41/49;
 

females: low-dose controls 1/47, low-dose 37/43; high-dose
 

controls 1/45, high-dose 43/49).
 

Papillomas or carcinomas of the integumentary system occurred at
 

significant incidences (P _<_ 0.013) in both the low- and high-dose
 

male rats (low-dose controls 0/36, low-dose 8/44; high-dose
 

controls 0/48, high-dose 6/48); these tumors also occurred in
 

low- and high-dose female rats, but not at incidences high enough
 

to be statistically significant (low-dose controls 0/39, low-dose
 

4/44; high-dose controls 0/50, high-dose 4/49).
 

Carcinomas of the Zymbal's glands of the ear occurred at significant incidences (P < 0.045) in the low- and high-dose male and
 

female rats (males: low-dose controls 0/36, low-dose 5/44; high-


dose controls 0/48, high-dose 7/48; females: low-dose controls
 

0/39, low-dose 10/44; high-dose controls 0/50, high-dose 12/49).
 

These tumors were first observed as early as week 47 on study.
 

Adenocarcinomas of the uterus or endometrium occurred at a
 

significant incidence (P = 0.002) only in the high-dose female
 

rats (high-dose controls 1/50, high-dose 11/49); these tumors
 

also occurred in the low-dose females, but not at an incidence
 

high enough to be statistically significant (low-dose controls
 

4/38, low-dose 11/43).
 

It is concluded that under the conditions of this bioassay,
 

3-amino-9-ethylcarbazole (hydrochloride) was carcinogenic for the
 

liver, inducing hepatocellular carcinomas in Fischer 344 rats and
 

B6C3F1 mice of both sexes. Other tumors induced in the rats were
 

carcinomas or papillomas of the integumentary system in males,
 

carcinomas of the Zymbal's gland of the ear in males and females,
 

and adenocarcinomas of the uterus.
 

viii
 
TABLE OF CONTENTS
 

Page 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical.... 3
 

B. Dietary Preparation 4
 

C. Animals 5
 

D. Animal Maintenance 6
 

E. Subchronic Studies 11
 

F. Chronic Studies 12
 

G. Clinical and Pathologic Examinations 17
 

H. Data Recording and Statistical Analyses 18
 

III. Results - Rats 23
 

A. Body Weights and Clinical Signs (Rats) 23
 

B. Survival (Rats) 23
 

C. Pathology (Rats) 27
 

D. Statistical Analyses of Results (Rats) 34
 

IV. Results - Mice 37
 

A. Body Weights and Clinical Signs (Mice) 37
 

B. Survival (Mice) 37
 

C. Pathology (Mice) 41
 

D. Statistical Analyses of Results (Mice) 43
 

V. Discussion 45
 

VI. Bibliography 51
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in
 

Rats Fed 3-Amino-9-Ethylcarbazole Hydrochloride
 

in the Diet 53
 

Table Al Summary of the Incidence of Neoplasms in
 

Male Rats Fed 3-Amino-9-Ethylcarbazole
 

Hydrochloride in the Diet (Low Dose and Control)... 55
 

ix
 
Page 

Table A2 Summary of the Incidence of Neoplasms in 

Male Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 59 

Table A3 Summary of the Incidence of Neoplasms in 

Female Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (Low Dose and Control)... 63 

Table A4 Summary of the Incidence of Neoplasms in 

Female Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 68 

Appendix B Summary of the Incidence of Neoplasms in 

Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 73 

Table Bl Summary of the Incidence of Neoplasms in 

Male Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in .the Diet (Low Dode and Control)... 75 

Table B2 Summary of the Incidence of Neoplasms in 

Male Mice Fed 3-Amino-9-Ethycarbazole 

Hydrochloride in the Diet (High Dose and Control).. 78 

Table B3 Summary of the Incidence of Neoplasms in 

Female Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (Low Dose and Control)... 82 

Table B4 Summary of the Incidence of Neoplasms in 

Female Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 86 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 89 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (Low Dose and Control)... 91 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 97 

x
 
Page 

Table C3 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (Low Dose and Control)... 102 

Table C4 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 108 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 113 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (Low Dose and Control)... 115 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 121 

Table D3 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (Low Dose and Control)... 125 

Table D4 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet (High Dose and Control).. 130 

Appendix E Analyses of the Incidence of Primary Tumors 

in Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 135 

Table El Analyses of the Incidence of Primary Tumors 

in Male Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 137 

Table E2 Analyses of the Incidence of Primary Tumors 

in Female Rats Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 148 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 161 

xi
 
Page 

Table Fl Analyses of the Incidence of Primary Tumors 

in Male Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 163 

Table F2 Analyses of the Incidence of Primary Tumors 

in" Female Mice Fed 3-Amino-9-Ethylcarbazole 

Hydrochloride in the Diet 168 

Appendix G Analyses of Formulated Diets for 3-Amino-9­

Hydrochloride and Its Hydrochloride Salt 175 

TABLES 

Table 1 3-Amino-9-Ethylcarbazole Hydrochloride 

Chronic Feeding Studies in Rats 13 

Table 2 3-Amino-9-Ethylcarbazole Hydrochloride 

Chronic Feeding Studies in Mice 15 

FIGURES 

Figure 1 Growth Curves for Rats Fed 3-Amino-9­

Ethylcarbazole Hydrochloride in the Diet 24 

Figure 2 Survival Curves for Rats Fed 3-Amino-9­

Ethylcarbazole Hydrochloride in the Diet 25 

Figure 3 Growth Curves for Mice Fed 3-Amino-9­

Ethylcarbazole Hydrochloride in the Diet 38 

Figure 4 Survival Curves for Mice Fed 3-Amino~9­

Ethylcarbazole Hydrochloride in the Diet 39 

xii
 
I. INTRODUCTION
 

3-amino-9-ethylcarbazole (hydrochloride)
 

3-Amino-9-ethylcarbazole (CAS 132-32-1; NCI C01898), is an
 

aromatic amine dye intermediate that has been used industrially
 

in the manufacture of C. I. Pigment Violet 23 and C. I. Direct
 

Blue 108 (Society of Dyers and Colourists, 1971). It is also
 

used in histochemical laboratories in a colorimetric assay for
 

the enzyme peroxidase (Feinstein and Lindahl, 1973) and may be
 

supplanting benzidine as a biological stain (Kaplow, 1975).
 

Approximately 10,604 pounds were imported to the United States
 

for these uses in 1974 (United States International Trade
 

Commission, 1976), and over 1,000 pounds per year are now
 

manufactured domestically according to recent estimates (Stanford
 

Research Institute, 1977).
 

1 
3-Amino-9-ethylcarbazole in its free amine form or as the
 

hydrochloride salt (C03043) is one in a series of aromatic amine
 

dye intermediates that were selected for study in the
 

Carcinogenesis Testing Program because of the apparent increase
 

in the incidence of cancer among dye workers (Wynder et al.,
 

1963; Anthony and Thomas, 1970).
 

2 
II. MATERIALS AND METHODS
 

A. Chemical
 

3-Amino-9-ethylcarbazole and its hydrochloride salt were obtained
 

from Carroll Products, Wood River Junction, Rhode Island. The
 

melting point range of the batch of 3-amino-9-ethylcarbazole used
 

in the chronic studies was 80-85°C, significantly below that of
 

127° reported in the literature (Society of Dyers and Colourists,
 

1971). The hydrochloride salt used in the chronic studies
 

decomposed above 210°C. Perchloric acid titration of one amine
 

function also indicated a lower purity of 92.1 i 0.2% for the
 

free amine and 98.8 4- 0.4% purity for the hydrochloride salt.
 

The salt was shown to be the monohydrochlor ide using the
 

Buchler-Cotlove chloridometer. Gas-liquid chromatography
 

indicated three major impurities in the free amine, accounting
 

for 4.5% of the total peak area, and three major impurities in
 

the hydrochloride salt, accounting for 1.8% of the peak area.
 

Thin-layer chromatography showed two minor and four trace
 

impurities in the free amine and two minor and three trace
 

impurities in the hydrochloride salt. Infrared and nuclear
 

magnetic resonance spectra of both the free amine and the
 

hydrochloride salt were consistent with the structures.
 

Bulk chemicals were stored at 4°C in the original containers.
 

3 
In the chronic feeding studies, the free amine form of the test
 

chemical was administered to the low-dose rats through week 18
 

and to the low-dose mice through week 1 (see tables 1 and 2).
 

Thereafter, these animals received the hydrochloride salt of the
 

amine. The high-dose rats and mice received the hydrochloride
 

salt for the entire period of the study. Since it is considered
 

that the amine is converted systemically to the salt form, the
 

term 3-amino-9-ethylcarbazole hydrochloride is used to represent
 

the test chemical used in the chronic feeding studies.
 

B. Dietary Preparation
 

3-Amino-9-ethylcarbazole or its hydrochloride salt was
 

incorporated into powdered Wayne Lab Blox animal feed (Allied
 

Mills, Inc., Chicago, 111.) for administration during the
 

bioassay. The chemical was removed from its container, sifted,
 

and weighed out under an exhaust hood, then mixed by hand with an
 

aliquot of feed in a mortar until visual uniformity was reached.
 

This premix was then mixed with the remaining feed in a
 

Patterson-Kelly twin-shell blender for 20 minutes. Test diets
 

were prepared once per week and used within 1 week of
 

preparation. The diets were sealed in double plastic bags and
 

stored at 4°C.
 

4 
During the chronic studies, the concentration of 3-amino-9-ethylcarbazole or its hydrochloride salt was measured in selected
 

batches of formulated diets. The results are summarized in
 

Appendix G. At each dietary concentration, the mean of the
 

analytical concentrations for the samples tested was within 20%
 

of the theoretical concentration, and the coefficient of
 

variation was < 18%.
 

C. Animals
 

For the subchronic studies, Fischer 344 rats were obtained from
 

Charles River Breeding Laboratories, Inc, Wilmington,
 

Massachusetts, and C57BL/6 mice were obtained from Southern
 

Animal Farms, Prattville, Alabama.
 

For the chronic studies, Fischer 344 rats and B6C3F1 mice were
 

used. Charles River Laboratories supplied the male rats that
 

were administered the high dose of the test chemical, the female
 

rats that were administered both high and low doses, and the male
 

and female rats which were used as controls for the high-dose
 

groups. A. R. Schmidt, Madison, Wisconsin, supplied the group of
 

male rats that were administered the low dose of the test
 

chemical, and Laboratory Supply Company, Indianapolis, Indiana,
 

supplied the male and female rats which were used as the controls
 

for the low-dose groups.
 

5 
Charles River Laboratories supplied the male and female B6C3F1
 

mice that were administered the high dose of the test chemical
 

and the male and female mice that were used as controls. A. R.
 

Schmidt supplied the male and female mice that were administered
 

the low dose.
 

All animals were received at 28 days of age and were quarantined
 

for approximately 2 weeks prior to the start of the bioassay. At
 

the end of the quarantine period, animals were assigned to
 

control or dosed groups in such a way that the mean weights of
 

animals in each cage were approximately the same.
 

D. Animal Maintenance
 

All animals were housed by species in temperature-controlled
 

rooms. The temperature range was 23-34°C. Incoming air was
 

filtered through Tri-Dek® 15/40 denier Dacron filters, providing
 

six changes of room air per hour. Fluorescent lighting provided
 

illumination on a 12-hour daily cycle.
 

Rats were maintained separately according to sex, with five
 

animals per cage. During quarantine and for the first 13 months
 

of the bioassay, the rats were housed in galvanized or stainless
 

steel wire mesh cages (Fenco Cage Products, Boston, Mass.),
 

suspended above newspapers. The newspapers were replaced every
 

day, and the cages and racks were washed every week. For the
 

6 
remainder of the bioassay, the rats we>re housed in suspended
 

polycarbonate cages (Lab Products, Inc., Garfield, N. J.),
 

equipped with disposable nonwoven fiber filter sheets. Clean
 

bedding and cages were provided twice per week. Sanicel corncob
 

bedding (Paxton Processing Co., S. Lancaster, Mass.) was provided
 

for the first 8 months in the polycarbonate housing for the high-


dose rats, for the first 7 months for the high-dose control rats,
 

and for the entire period for the low-dose rats and their
 

controls. The high-dose rats and their controls were provided
 

with Aspen-bed® hardwood chip bedding (American Excelsior,
 

Summerville, Mass.) for the remainder of the bioassay. The racks
 

for the stainless steel cages were cleaned once every 2 weeks,
 

and the disposable filters were replaced at that time.
 

Mice were maintained separately according to sex, at first with
 

10 animals per cage, using shoe-box-type polycarbonate cages (Lab
 

Products, Inc.). During quarantine and the period of
 

administration of the test chemical, the cages were fitted with
 

perforated stainless steel lids. During the observation periods
 

that followed, stainless steel wire bar lids were used. Both
 

types of lids were obtained from Lab Products, Inc. Filtek®
 

nonwoven fiber filter bonnets (Lab Products, Inc.) were used over
 

the cage lids. After 12 months of the study, the high-dose mice
 

were housed five per cage, and after 11 months, their controls
 

7 
were housed five per cage; after 18 months of the study, the low-


dose mice and their controls were similarly housed five per cage.
 

The cages, lids, and bedding were changed three times per week
 

when the cage populations were 10 per cage and twice per week
 

when the cage populations were reduced to five per cage. Absorbdri hardwood chip bedding (Lab Products, Inc.) was used for 1
 

month for the high-dose mice and their controls and for 8 months
 

for the low-dose mice and their controls. Sanicel® corncob
 

bedding was used for 12 months. Bed-o-cob bedding (Anderson Cob
 

Mills, Inc., Maumee, Ohio) was then used for 8 months for the
 

high-dose mice and their controls and to the end of the bioassay
 

for the low-dose mice and their controls. The high-dose mice and
 

their controls were provided with Aspen-bed bedding for the
 

remainder of the study. Reusable filter bonnets and pipe racks
 

were sanitized every 2 weeks throughout the study.
 

All equipment that was sanitized was washed with Dubois Serve
 

Detergent and rinsed at 82°C.
 

Tap water (0.75-1.0 ppm chlorine) was provided ad libitum from
 

250-ml water bottles equipped with rubber stoppers and stainless
 

steel sipper tubes. The bottles were replaced twice per week,
 

and, for the rats only, filled as needed between changes.
 
Wayne® Lab diet was used throughout the bioassay. Pelleted
 

Wayne® Lab Blox was fed during the quarantine period to the
 

low-dose mice and their controls and to the low-dose rats and
 

their controls. Other groups were fed Wayne® Lab Blox meal
 

during quarantine. All animals received the meal during
 

administration of the test chemical. The meal was distributed in
 

Alpine aluminum feed cups (Curtin-Matheson Scientific, Inc.,
 

Woburn, Mass.) for the high-dose rats and their controls for the
 

first 11 months of the study and for all other groups for the
 

entire study. After 11 months, the high-dose rats and their
 

controls were fed from stainless steel gang-style hoppers
 

(Scientific Cages, Inc., Bryan, Tex.). During the observation
 

periods that followed administration of the test chemical, all
 

animals were fed Wayne® Lab Blox pellets on the floors of the
 

cages.
 

Rats and mice were housed in separate rooms. All rats except the
 

high-dose male and female controls were housed in a single room
 

with rats administered the following compounds in feed:
 

(CAS 99-55-8) 5-nitro-o-toluidine
 

(CAS 122-66-7) hydrazobenzene
 

(CAS 117-79-3) 2-aminoanthraquinone
 

(CAS 94-52-0) 5-nitro-lH-benzimidazole
 

(CAS 86-57-7) 1-nitronaphthalene
 

(CAS 615-05-4) 4-methoxy-m-phenylenediamine
 

(CAS 8003-03-0) aspirin: phenacetin: caffeine (APC)
 
The high-dose male and female control rats were housed in another
 

room with rats administered the following compounds in feed:
 

(CAS 61-82-5) 3-amino-s-triazole (positive control)
 

(CAS 129-15-7) 2-methyl-l-nitroanthraquinone
 

(C01978) 3'-nitro-p-acetophenetide
 

All control and low-dose male and female mice were housed in a
 

room with mice administered one of the following compounds in
 

feed:
 

(CAS 61-82-5) 3-amino-s-triazole (positive control)
 

(CAS 138-89-6) N,N-dimethyl-p-nitrosoaniline
 

(CAS 6369-59-1) toluene-2,5-diamine sulfate
 

(CAS 121-14-2) 2,4-dinitrotoluene
 

(CAS 117-79-3) 2-aminoanthraquinone
 

(CO1887) 3-amino-4-ethoxyacetanilide
 

(CAS 82-28-0) l-amino-2-methylanthraquinone
 

(CAS 99-59-2) 5-nitro-o-anisidine
 

(CAS 619-17-0) 4-nitroanthranilic acid
 

(CAS 86-57-7) 1-nitronaphthalene
 

(CAS 602-87-9) 5-nitroacenaphthene
 

(C01978) 3'-nitro-p-acetophenetide
 

(CAS 615-05-4) 4-methoxy-m-phenylenediamine
 

(CAS 8003-03-0) aspirin: phenacetin: caffeine (APC)
 

Male and female high—dose mice were housed in another room, with
 

mice administered one of the following compounds in feed:
 

(CAS 6369-59-1) toluene-2,5-diamine sulfate
 

(CAS 99-55-8) 5-nitro-o-toluidine
 

(CAS 122-66-7) hydrazobenzene
 

(CAS 94-52-0) 5-nitro-lH-benzimidazole
 

(CAS 99-59-2) 5-nitro-o-anisidine
 

(CAS 86-57-7) 1-nitronaphthalene
 

(CAS 615-05-4) 4-methoxy-m-phenylenediamine
 

10
 
E. Subchronic Studies
 

Six-week subchronic studies were conducted using Fischer 344 rats
 

and C57BL/6 mice to estimate the maximum tolerated doses of
 

3-amino-9-ethylcarbazole, on the basis of which two
 

concentrations (hereinafter referred to as "low doses" and "high
 

doses") were determined for the chronic studies. The free amine
 

form of the test chemical was administered in the diet for 4
 

weeks at concentrations of 40, 100, 250, 600, or 1,440 ppm. Five
 

males and five females of each species received the chemical at
 

one of each of the concentrations, and five males and five
 

females of each species were given basal diets. All animals were
 

killed and necropsied at the end of 6 weeks.
 

At the end of the studies, no differences in mean body weight
 

gains in various dosed male rats could be attributed to
 

administration of the test compound. Depressions in mean body
 

weight gains in dosed female rats compared to controls ranged
 

from 5-9% at doses of 40-600 ppm, and was 13% at 1,440 ppm. In
 

the mice, mean body weight gains were depressed from 7-12% in all
 

the male groups, but no consistent pattern of weight gain
 

depression was seen in the female groups. There were no deaths
 

in any of the animals.
 

11
 
On the basis of these data, initial doses of 300 and 600 ppm for
 

the free amine (equivalent to 400 and 800 ppm when calculated and
 

administered as the hydrochloride salt) were selected as the low
 

and high doses, respectively, to be administered to rats and mice
 

in the chronic studies.
 

F. Chronic Studies
 

The test groups, doses administered, and times on study of the
 

chronic studies are shown in tables 1 and 2.
 

Rats and mice that had been designated to be administered a low
 

dose of 300 ppm (free amine) were killed at weeks 49 and 32,
 

respectively, because these doses had no effect on the mean body
 

weights. The original high-dose groups then became the low-dose
 

groups, and new high-dose groups of rats and mice were started
 

concurrently at doses of 2,000 and 1,200 ppm (hydrochloride salt)
 

for the rats and the mice, respectively.
 

The free amine, which was administered to the low-dose rats
 

through week 18 and to the low-dose mice during week 1, was
 

replaced by equimolar concentrations of the hydrochloride salt
 

because of concern that the free amine might be unstable in the
 

feed. The high-dose rats and mice received the hydrochloride
 

salt for the entire period of administration of the chemical.
 

12
 
Table 1. 3-Amino-9-Ethylcarbazole Hydrochloride
 

Chronic Feeding Studies in Rats
 

Sex and
 

Test
 

Group
 

Male
 

Low-Dosec»d
 

Low-Dose Control6
 

High-Dosef »8
 

High-Dose Controls
 

Female
 

Low-Dosec»n
 

Low-Dose Control6
 

High-Dosef »8
 

High-Dose Controls
 

Initial
 

No. of
 

Animals3.
 

50
 

50
 

50
 

50
 

50
 

50
 

50
 

50
 

3-Amino-9­

Ethylcarbazole
 

Hydrochloride
 

in Dietb
 

(ppm)
 

800
 

0
 

2,000
 

0
 

800
 

0
 

2,000
 

0
 

Time
 

Dosed
 

(weeks )
 

78
 

78
 

78
 

78
 

on Study
 

Observed
 

(weeks)
 

26-27
 

107
 

28-29
 

109
 

27-28
 

107-108
 

29
 

109-110
 

a

Rats were 41-44 days of age when placed on study.
 

"Diets were available jid_ libitum 7 days per week.
 

c

The test chemical was administered to the low-dose groups as the
 

free amine (600 ppm) for the first 18 weeks, then as the
 

hydrochloride salt for the remaining 60 weeks. The dose in the
 

above table is calculated on the basis of the weight of the
 

hydrochloride salt.
 

"The rats used in the low-dose male group were obtained from
 

A. R. Schmidt, Madison, Wisconsin.
 

13
 
Table 1. 3-Amino-9-Ethylcarbazole Hydrochloride
 

Chronic Feeding Studies in Rats
 

(continued)
 

e

The rats used in the low-dose control groups were obtained
 

from Laboratory Supply Company, Indianapolis, Indiana.
 

^The test chemical was administered to the high-dose groups
 

as the hydrochloride salt for the entire study.
 

§The rats used in the high-dose and corresponding control
 

groups were obtained from Charles River Breeding Laboratories,
 

Inc., Wilmington, Massachusetts. The high-dose and
 

corresponding control groups were started 50-51 weeks after
 

the low-dose and corresponding control groups.
 

^The female rats used in the low-dose group were obtained from
 

Charles River Breeding Laboratories, Inc., Wilmington,
 

Massachusetts.
 

14
 
Table 2. 3-Amino-9-Ethylcarbazole Hydrochloride
 

Chronic Feeding Studies in Mice
 

3-Amino-9­

Ethylcarbazole 

Sex and Initial Hydrochloride Time on Study 

Test No. of in Dietb Dosed Observed 

Group Animals3 (ppm) (weeks) (weeks) 

Male
 

Low-Do sec 50 800 78 16-17
 

Low-Dose Control^ 50 0 96
 

e
 High-Dosed

» 50 1,200 78 17
 

High-Dose Controld 50 0 96
 

Female
 

Low-Dosec 50 800 78 17
 

Low-Dose Control^ 50 0 97
 

e
 High-Dosed

» 50 1,200 78 17
 

High-Dose Controld 50 0 96
 

a

Mice were 41-44 days of age when placed on study.
 

t>Diets were available ad libitum 7 days per week.
 

c

The test chemical was administered to the low-dose groups as the
 

free amine (600 ppm) for the first week, then as the
 

hydrochloride salt for the remaining 77 weeks. The dose in the
 

above table is calculated on the basis of the weight of the
 

hydrochloride salt. The mice used in the low-dose groups were
 

obtained from A. R. Schmidt, Madison, Wisconsin.
 

15
 
Table 2. 3-Amino-9-Ethylcarbazole Hydrochloride
 

Chronic Feeding Studies in Mice
 

(continued)
 

^The mice used in the high-dose groups and in all control groups
 

were obtained from Charles River Breeding Laboratories, Inc.,
 

Wilmington, Massachusetts. The high-dose and corresponding
 

control groups were started about 33 weeks after the low-dose
 

and corresponding control groups were placed on study.
 

e

The test chemical was administered as the hydrochloride salt
 

for the entire study.
 

16
 
G. Clinical and Pathologic Examinations
 

Inspection for mortality and morbidity was carried out twice
 

daily. Body weights were recorded every 2 weeks for the first 12
 

weeks and every month thereafter. Clinical observations were
 

recorded every month.
 

Moribund animals and animals that survived to the end of the
 

bioassay were killed using CC>2 anesthesia and necropsied.
 

Necropsies were also performed on all animals found dead, unless
 

precluded by autolysis or severe cannibalization. The following
 

tissues were examined microscopically: tissue masses, abnormal
 

regional lymph nodes, skin, mandibular lymph nodes, mammary
 

gland, salivary gland, thigh muscle, sciatic nerve, bone marrow,
 

costochondral junction (rib), thymus, larynx, trachea, lungs and
 

bronchi, heart, thyroid, parathyroid, esophagus, stomach,
 

duodenum, jejunum, ileum, colon, mesenteric lymph nodes, liver,
 

gall bladder (mice), pancreas, spleen, kidney, adrenal, bladder,
 

seminal vesicles/prostate/testis (males), ovary/uterus (females),
 

nasal cavity, brain, pituitary, eyes, external and middle ear,
 

and spinal cord. Peripheral blood smears were prepared from each
 

animal whenever possible. The different tissues were preserved
 

in 10% buffered formalin, embedded in paraffin, sectioned, and
 

stained with hematoxylin and eosin. Special staining techniques
 

were utilized when indicated for more definitive diagnoses.
 

17
 
A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals may have
 

been missing, cannibalized, or judged to be in such an advanced
 

state of autolysis as to preclude histopathologic evaluation.
 

Thus, the number of animals from which particular organs or
 

tissues were examined microscopically varies, and does not
 

necessarily represent the number of animals that were placed on
 

study in each group.
 

H. Data Recording and Statistical Analyses
 

Pertinent data on this experiment have been recorded in an automatic data processing system, the Carcinogenesis Bioassay Data
 

System (Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design,
 

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

Against Cancer (Berenblum, 1969). Data tables were generated for
 

verification of data transcription and for statistical review.
 

These data were analyzed using the appropriate statistical
 

techniques described in this section. Those analyses of the
 

experimental results that bear on the possibility of
 

carcinogenicity are discussed in the statistical narrative
 

sections.
 

18
 
Probabilities of survival were estimated by the product-limit
 

procedure of Kaplan and Meier (1958) and are presented in this
 

report in the form of graphs. Animals were statistically
 

censored as of the time that they died of other than natural
 

causes or were found to be missing; animals dying from natural
 

causes were not statistically censored. Statistical analyses for
 

a possible dose-related effect on survival used the method of Cox
 

(1972) for testing two groups for equality and Tarone's (1975)
 

extensions of Cox's methods for testing for a dose-related trend.
 

One-tailed P values have been reported for all tests except the
 

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

given as the ratio of the number of animals bearing such lesions
 

at a specific anatomic site (numerator) to the number of animals
 

in which that site is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic examination was required to detect lesions prior to histologic sampling
 

(e.g., skin or mammary tumors), or when lesions could have
 

appeared at multiple sites (e.g., lymphomas), the denominators
 

consist of the numbers of animals necropsied.
 

19
 
The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

significantly higher proportion of tumors than did the control
 

animals. As a part of these analyses, the one-tailed Fisher
 

exact test (Cox, 1970) was used to compare the tumor incidence of
 

a control group with that of a group of dosed animals at each
 

dose level. When results for a number of dosed groups (k) are
 

compared simultaneously with those for a control group, a
 

correction to ensure an overall significance level of 0.05 may be
 

made. The Bonferroni inequality (Miller, 1966) requires that the
 

P value for any comparison be less than or equal to 0.05/k. In
 

cases where this correction was used, it is discussed in the
 

narrative section. It is not, however, presented in the tables,
 

where the Fisher exact P values are shown.
 

The Cochran-Armitage test for linear trend in proportions
 

(Armitage, 1971), was not used.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

of tumors. In this analysis, deaths that occurred before the
 

first tumor was observed were excluded by basing the statistical
 

tests on animals that survived at least 52 weeks, unless a tumor
 

was found at the anatomic site of interest before week 52. When
 

such an early tumor was found, comparisons were based exclusively
 

20
 
on animals that survived at least as long as the animal in which
 

the first tumor was found. Once this reduced set of data was
 

obtained, the standard procedures for analyses of the incidence
 

of tumors were followed.
 

When appropriate, life-table methods were used to analyze the
 

incidence of tumors. Curves of the proportions surviving without
 

an observed tumor were computed as in Saffiotti et al. (1972).
 

The week during which an animal died naturally or was sacrificed
 

was entered as the time point of tumor observation. Cox's
 

methods of comparing these curves were used for two groups;
 

Tarone's extension to testing for linear trend was used for three
 

groups. The statistical tests for the incidence of tumors which
 

used life-table methods were one-tailed and, unless otherwise
 

noted, in the direction of a positive dose relationship. Significant departures from linearity (P < 0.05, two-tailed test) were
 

also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared to its control was calculated
 

from the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as Pt/Pc where pt is the true binomial
 

probability of the incidence of a specific type of tumor in a
 

dosed group of animals and pc is the true probability of the
 

spontaneous incidence of the same type of tumor in a control
 

21
 
group. The hypothesis of equality between the true proportion of
 

a specific tumor in a dosed group and the proportion in a control
 

group corresponds to a relative risk of unity. Values in excess
 

of unity represent the condition of a larger proportion in the
 

dosed group than in the control.
 

The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical
 

analyses. The interpretation of the limits is that in
 

approximately 95% of a large number of identical experiments, the
 

true ratio of the risk in a dosed group of animals to that in a
 

control group would be within the interval calculated from the
 

experiment. When the lower limit of the confidence interval is
 

greater than one, it can be inferred that a statistically significant result (P < 0.025 one-tailed test when the control incidence
 

is not zero, P < 0.050 when the control incidence is zero) has
 

occurred. When the lower limit is less than unity, but the upper
 

limit is greater than unity, the lower limit indicates the
 

absence of a significant result while the upper limit indicates
 

that there is a theoretical possibility of the induction of
 

tumors by the test chemical, which could not be detected under
 

the conditions of this test.
 

22
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of high-dose male and female rats were lower
 

than those of corresponding controls throughout the bioassay
 

(figure 1). The mean body weight of the low-dose males was
 

slightly lower than that of the corresponding controls during the
 

latter part of the bioassay, and the mean body weight of the
 

low-dose females was essentially unaffected. Fluctuation in the
 

growth curve may be due to mortality; as the size of a group
 

diminishes, the mean body weight may be subject to variation. No
 

other clinical signs related to administration of the test
 

chemical were observed.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats fed 3-amino-9-ethylcarbazole
 

hydrochloride at the doses used in this bioassay, together with
 

those of the corresponding controls, are shown in figure 2. In
 

each sex, the result of the Cox test comparing the survival of
 

the low-dose group with its controls is significant (P <_ 0.002),
 

indicating increased mortality in the dosed groups. In females,
 

the result of the Cox test comparing the high-dose group with its
 

controls is also significant (P < 0.001), but in male rats the
 

23
 
Figure 1. Growth Curves For Rats Fed 3-Amino-9-Ethylcarbazole Hydrochloride In The Diet 

24 
Figure 2. Survival Curves For Rats Fed 3-Amino-9-Ethylcarbazole Hydrochloride In The Diet 

25 
survival of the high-dose group was not significantly different
 

from that of the controls.
 

In male rats, 37/50 (74%) of the low-dose group and 37/50 (74%)
 

of the low-dose control group were alive at week 78 on study. In
 

the low-dose control group, 10 animals were killed at week 29 and
 

5 at week 78; in the low-dose group, 3 animals were reported as
 

missing, 1 at week 1 and 2 at week 2; 5 low-dose animals were
 

killed at week 78. Forty-six out of 50 (92%) of the high-dose
 

group and 48/49 (98%) of the high-dose control group were alive
 

at week 78 on study; 5 animals of each group were killed at week
 

78.
 

In female rats, 39/50 (78%) of the low-dose animals and 38/50
 

(76%) of the low-dose controls were alive at week 78 on study.
 

In the low-dose control group, 10 animals were killed at week 29
 

and 5 at week 78; in the low-dose group, 5 animals were killed at
 

week 78. Forty-five out of 50 (90%) of the high-dose group and
 

49/50 (98%) of the high-dose control animals were alive at week
 

78 on study; 5 animals of each group were killed at week 78.
 

Sufficient numbers of rats of each sex were at risk for
 

development of late-appearing tumors.
 

26
 
C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

Appendix A, tables A1-A4; findings on nonneoplastic lesions are
 

summarized in Appendix C, tables C1-C4.
 

The occurrence of neoplasms of the skin, auditory sebaceous
 

gland, preputial/clitoral glands, liver, mammary gland, uterus,
 

and intestine in Fischer 344 rats appeared to be related to
 

administration of 3-amino-9-ethylcarbazole hydrochloride.
 

Epithelial neoplasms of the skin not localized to any particular
 

site were found in dosed rats of each sex, but in only one
 

control animal — a female. The types and incidences of the
 

tumors of the skin are summarized in the following tabulation:
 

Low-Dose High-Dose Low High
 

MALE RATS Control Control Dose Dose
 

Number of animals
 

necropsied (46) (48) (44) (48)
 

Skin and Subcutis
 

Squamous-Cell
 

Papilloma 3 2
 

Squamous-Cell
 

Carcinoma 3 3
 

Basal-Cell
 

Carcinoma 4 1
 

Sebaceous Adenoma 1
 

Adenoma, NOS*
 2
 

*Not otherwise specified
 

27
 
Low-Dose High-Dose Low High
 

FEMALE RATS Control Control Dose Dose
 

Number of animals
 

necropsied (49) (50) (44) (49)
 

Skin and Subcutis
 

Squamous-Cell
 

Papilloma 1
 1
 

Squamous-Cell
 

Carcinoma 3 3
 

Basal-Cell
 

Carcinoma 1
 

Sebaceous Adenocarcinoma 1
 

Squamous-cell papilloma was a papillary growth of the squamous
 

epithelium with hyperkeratosis. Squamous-cell carcinomas in a
 

few dosed rats had grown into the subcutaneous tissue. An
 

occasional cell nest was seen in the lumen of lymphatics or blood
 

vessels. The neoplastic cells formed keratin pearls which varied
 

in size and shape. Many of the cells were anaplastic, and their
 

cytoplasm was eosinophilic. Keratohyalin granules were present
 

only in some cells. Nuclei were large, and mitotic figures
 

numerous. This tumor metastasized to the lung in two rats (one
 

high-dose male, one control female). Basal-cell carcinomas
 

extended down to the panniculus adiposus and were ulcerated at
 

the surface. Tumor cells were lobulated or closely packed, and
 

the cytoplasm was basophilic. Nuclei were hyperchromatic, and
 

there were many mitotic figures. Keratin pearls and hair
 

follicles in some tumors suggested differentiation. Unlike the
 

28
 
squamous-cell carcinoma, cells in this tumor did not exhibit much
 

pleomorphism.
 

Neoplasms of the auditory sebaceous gland occurred in 5/44 low-


dose males and 7/48 high-dose males; 10/44 low-dose females and
 

12/49 high-dose females; and in none of the controls. Thirteen
 

tumors were considered to be carcinomas, NOS, and 21 to be
 

squamous-cell carcinomas. The morphologies of these tumors were
 

similar to those described by Pliss (1973). Sebaceous carcinoma
 

in one rat metastasized to the lung.
 

Adenocarcinomas of the uterus were found in 5/98 combined control
 

and 22 dosed female rats (11/43 low-dose, 11/49 high-dose).
 

Papillary ingrowths of the epithelium filled the lumen and had
 

invaded into the myometrium. Transformed glands were adjacent to
 

each other with little supporting stroma. Tumor cells were
 

arranged in acini or syncitia. The cytoplasm of these cells was
 

basophilic. Vacuolization and/or bright eosinophilic droplets
 

were seen in some cells. Nuclear pleomorphism was not marked.
 

Nuclei were vesicular with prominent nucleoli. Both normal and
 

abnormal mitotic figures were numerous. Large areas of necrosis
 

and inflammatory cells replaced part of the tumor parenchyma.
 

The uterine adenocarcinomas in three rats had metastasized to the
 

lung.
 

29
 
Carcinomas and adenocarcinomas of the mammary gland occurred in
 

10 dosed female rats (8/44 low-dose, 3/49 high-dose) and 2/89
 

control female rats. These tumors were solitary or multiple and
 

were comprised of glandular structures of varying sizes or of
 

closely packed cells. Stroma was scant between the transformed
 

acini. Secretory material was found in the lumen of some glands.
 

Tumor cells were cuboidal and the cytoplasm was eosinophilic or
 

vacuolated. Both normal and abnormal mitotic figures were
 

numerous. In a few tumors, central necrosis was found in nests
 

of densely packed cells. A mammary adenocarcinoma had
 

metastasized to the lung and liver in one dosed rat.
 

Tumors of the preputial gland were found in 2/44 low-dose, 5/48
 

high-dose, and in none of the control male rats. Tumors of the
 

clitoral gland were found in 6/44 low-dose, 7/49 high-dose, and
 

3/99 control female rats.
 

The incidences of neoplastic nodules of the liver and hepatocellular carcinomas in control and dosed rats are summarized in
 

the following tabulation:
 

30
 
Low-Dose High-Dose Low High
 

MALE RATS Control Control Dose Dose
 

Number of Animals
 

with Tissue
 

Examined
 

Microscopically (46) (48) (42) (48)
 

Liver
 

Hepatocellular
 

Carcinoma 1 6 8
 

Hepatocellular
 

Carcinoma and
 

Neoplastic Nodule i 1 12 22
 

FEMALE RATS
 

Number of Animals
 

with Tissue
 

Examined
 

Microscopically (49) (50) (43) (48)
 

Liver
 

Hepatocellular
 

Carcinoma 2 1 3
 

Hepatocellular
 

Carcinoma and
 

Neoplastic Nodult 2 1 6
 

Neoplastic nodules of the liver were small, and compressed the
 

adjacent parenchyma in areas. The cells were large with acidophilic cytoplasm and hyperchromatic nuclei. A few mitotic
 

figures were present. Hepatocellular carcinomas involved a part
 

or an entire lobe of the liver. The lobular architecture was
 

distorted. Liver plates were several cells thick. There was a
 

pleomorphism in the size of transformed hepatocytes. The cytoplasm of the cells was acidophilic or vacuolated. The nuclei
 

were large, and the nucleoli were prominent. Mitotic figures
 

31
 
were numerous. Cholangiolar tumors were associated with hepatocellular carcinomas in two male rats of the high-dose group. No
 

metastases were found.
 

The incidences of adenomatous polyps and adenocarcinomas of the
 

small intestine and colon are summarized in the following
 

tabulation:
 

Low-Dose High-Dose Low High
 

MALE RATS Control Control Dose Dose
 

Number of Animals
 

with Tissue
 

Examined
 

Microscopically (43) (46) (41) (46)
 

Small Intestine
 

Adenomatous Polyp 1
 

Adenocarcinoma 3 1
 

Colon
 

Adenomatous Polyp 1
 

Adenocarcinoma 1 1
 

Adenocarcinoma in
 

Adenomatous Polyp 1
 

FEMALE RATS
 

Number of Animals
 

with Tissue
 

Examined
 

Microscopically (47) (48) (40) (46)
 

Small Intestine 

Adenomatous Polyp 2 

Adenocarcinoma 2 

Adenomatous polyps had a pedicle and were supported by fibrovascular stroma. Glands varied in size in these polyps. Cells
 

lining these glands were piled up in areas, and the cytoplasm was
 

32
 
more basophilic than in cells of the adjacent normal mucosa.
 

Adenocarcinomas were either polypoid or sessile and infiltrated
 

into the submucosa. Glands were closely packed and exhibited
 

pleomorphism in shape and size; a few were cystic. Neoplastic
 

cells were either columnar or cuboidal in shape, and the cytoplasm was basophili :. Only a few of the neoplastic cells stained
 

positive with the PAS stain, and Paneth cell granules stained
 

bright red in tumors of the small intestine. Nuclei were hyper-


chromatic and mitotic figures were numerous. In some of the
 

tumors, there were areas of necrosis and clusters of inflammatory
 

cells.
 

A variety of nonneoplastic lesions occurred in both dosed and
 

control animals. The incidence, distribution, and severity of
 

these lesions were similar to those known to occur spontaneously
 

in aged Fischer 344 rats.
 

Based on the histopathologic examination, 3-amino-9-ethylcarbazole hydrochloride was considered to be carcinogenic to
 

Fischer 344 rats under the conditions of this study, since there
 

was an increased incidence of neoplasms r-.f the skin in males,
 

mammary gland and uterus in females, and auditory sebaceous
 

glands, preputial/clitoral glands, liver, and intestines in males
 

and females.
 

33
 
D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

least two animals in one group and with an incidence of at least
 

5% in one or more than one group. No Cochran-Armi tage tests were
 

made, because the low- and high-dose groups were not tested
 

concurrently. The 10 low-dose control animals of each sex that
 

were killed at week 29 on study were deducted from the
 

denominators of the incidences in the controls.
 

When the incidences of all animals with carcinomas or papillomas
 

in the integumentary system were combined for analysis, the
 

results of the Fisher exact test indicated a significantly higher
 

incidence of these tumors in both the male high-dose (6/48 [13%],
 

P = 0.013) and the male low-dose (8/44 [18%], P = 0.006) groups
 

than in the respective controls. These tumors did not appear in
 

statistically significant incidences in female dosed groups,
 

although they were present in both the female high-dose group
 

(4/49 [8%]) and the female low-dose group (4/44 [9%]), compared
 

with 0/50 and 0/39 in the two control groups.
 

The results of the Fisher exact test on the incidence in the male
 

rats of hepatocellular carcinomas show that the incidences in the
 

low- and high-dose groups are significantly higher than those in
 

34
 
the corresponding control groups (P = 0.020 and P = 0.015,
 

respectively). The incidences of the combination of neoplastic
 

nodules and hepatocellular carcinomas in the low- and high-dose
 

groups are also significantly higher than those in the
 

corresponding control groups (P < 0.001). In the female rats,
 

the combined incidence of neoplastic nodules of the liver and
 

hepatocellular carcinomas is significantly higher (P = 0.012) in
 

the high—dose group than in the corresponding control group.
 

The Fisher exact comparison of the incidence of adenocarcinomas
 

of the uterus or endometrium in the high-dose group of female
 

rats with that in the corresponding control group shows a P value
 

of 0.002.
 

In either the male or female rats, carcinomas or squamous-cell
 

carcinomas of the Zymbal's gland occurred exclusively in the
 

dosed groups. In the male rats, the results of the Fisher exact
 

test show that the incidences in the low- and high-dose groups
 

are significantly higher than those in their corresponding
 

controls (P = 0.045 and P = 0.006, respectively). In the
 

females, the results of the Fisher exact test comparing the
 

incidence in each dosed group with that in its control group are
 

significant (P < 0.001).
 

35
 
No other tumors occurred at significantly higher incidences in
 

dosed groups of rats than in controls. A significant result (P =
 

0.018) in the negative direction is observed in the comparison of
 

the incidence of interstitial-cell tumors in low-dose male rats
 

with that in its control group. Historical records show that
 

groups of this strain of rat usually show incidences of this
 

tumor from 75 to 100%.
 

In summary, the statistical analysis of the data for rats
 

indicates an association of the administration of this chemical
 

with the incidences of squamous-cell carcinomas or papillomas of
 

the integumentary system in both sexes, hepatocellular carcinomas
 

in male rats, the combination of neoplastic nodules of the liver
 

and hepatocellular carcinomas in both sexes of rats, and
 

carcinomas of the Zymbal's gland in both sexes. In addition, the
 

high-dose female rats had a significant increase in
 

adenocarcinomas of the uterus.
 

36
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of high-dose male and female mice were lower
 

than those of corresponding controls throughout the bioassay
 

(figure 3). The mean body weight of the low-dose females was
 

slightly lower than that of the controls during the latter part
 

of the bioassay, but the mean body weight of the low-dose males
 

was essentially unaffected. Fluctuation in the growth curve may
 

be due to mortality; as the size of a group diminishes, the mean
 

body weight may be subject to variation. No other clinical signs
 

related to administration of the test chemical were observed.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice fed 3-amino-9-ethylcarbazole
 

hydrochloride at the doses used in this bioassay, together with
 

those of the corresponding controls, are shown in figure 4. In
 

male mice, the results of the Cox tests comparing the survival of
 

the low- and high-dose groups with their respective controls were
 

both significant (low-dose P = 0.034; high-dose P = 0.006). In
 

female mice, the result of the Cox test comparing the low-dose
 

group with its control is not significant, but the result of the
 

37
 
Figure 3, Growth Curves For Mice Fed 3-Amino-9-Ethylcarbazole Hydrochloride In The Diet 

38 
TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves For Mice Fed 3-Amino-9-Ethylcarbazole Hydrochloride In The Diet 

39 
comparison of the high-dose group with its controls indicates
 

increased mortality in that dosed group (P = 0.004).
 

In male mice, 42/50 (84%) of the low-dose group and 44/50 (88%)
 

of the low-dose control group were alive at week 94. In the
 

low-dose control group, five animals were killed at week 78, and
 

in the low-dose group, one animal was reported missing at week
 

16. Thirty-eight out of 50 (76%) • of the high-dose group and
 

40/50 (80%) of the high-dose control group were alive at week 95.
 

In the high-dose control group, five animals were killed at week
 

49 and five at week 78; in the high-dose group, five animals were
 

killed at week 78.
 

In female mice, 35/50 (70%) of the low-dose group and 39/50 (78%)
 

of the low-dose control group were alive at week 95. In the
 

low-dose control group, five animals were killed at week 78, and
 

in the low-dose group, two animals were reported as missing, one
 

at week 20 and one at week 76. Thirty-two out of 50 (64%) of the
 

high-dose group and 38/50 (76%) of the high-dose control group
 

were alive at week 95. In the high-dose control group, five
 

animals were killed at week 49 and five at week 78; in the
 

high-dose group, five animals were killed at week 78 and one
 

animal was reported as missing at week 11.
 

Sufficient numbers of mice of each sex were at risk for
 

development of late-appearing tumors.
 

40
 
C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

Appendix B, tables B1-B4; findings on nonneoplastic lesions are
 

summarized in Appendix D, tables D1-D4.
 

A dose-related increase in the incidence of hepatocellular
 

carcinomas was found in mice administered 3-amino-9-ethylcarbazole hydrochloride. The following tabulation summarizes the
 

occurrence of these tumors in the various groups of animals and
 

the numbers of animals in which pulmonary metastases were found:
 

MALE MICE 

Low-Dose 

Control 

High-Dose 

Control 

Low 

Dose 

High 

Dose 

Number of Animals 

with Tissue 

Examined 

Microscopically (48) (48) (44) (49) 

Hepatocellular 

Carcinoma 

Pulmonary 

Metastases 

7 6 

1 

32 41 

3 

FEMALE MICE 

Number of Animals 

with Tissue 

Examined 

Microscopically (47) (50) (43) (49) 

Hepatocellular 

Carcinoma 

Pulmonary 

Metastases 

1 1 37 

2 

43 

5 

41
 
Hepatic tumors involved a part or an entire lobe of the liver.
 

The normal lobular architecture of the liver was not preserved.
 

Sinusoids in these areas were distended. A moderate pleomorphism
 

in the size of transformed hepatocytes was evident. The
 

cytoplasm was acidophilic, and in some animals it was vacuolated,
 

suggesting fatty infiltration. Nuclei were hyperchromatic and
 

were occasionally bizarre. Mitotic figures were numerous. Small
 

areas of necrosis, accompanied by inflammatory cells, were
 

scattered in the tumor parenchyma.
 

Adenomatous polyps of the gall bladder were found in three dosed
 

male mice, one in the low-dose group and two in the high-dose
 

group. None were found in the controls. These polyps were
 

characterized by papillary-like fronds of the columnar epithelium
 

supported by a thin fibrovascular stroma. Many of the epithelial
 

cells contained secretory material on the luminal side, and
 

piling of cells was noted in areas.
 

A variety of nonneoplastic lesions were observed, but the
 

incidence, distribution, and severity of these lesions were
 

similar to those known to occur spontaneously in aged B6C3F1
 

mice.
 

Based on the histopathologic examination, 3-amino-9-ethylcarbazole hydrochloride was considered to be carcinogenic in
 

42
 
B6C3F1 mice under the conditions of this study, since there was a
 

dose-related increase in the incidence of hepatocellular
 

carcinomas in both the males and the females.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

least two animals of one group and at an incidence of at least 5%
 

in one or more than one group. No Cochran-Armitage tests were
 

made, because the low- and high-dose groups were not tested
 

concurrently. The results of the five high-dose control animals
 

of each sex that were killed at week 49 are not included in the
 

analysis.
 

In both the male and female mice, the results of the Fisher exact
 

test show that the incidence of hepatocellular carcinomas in each
 

of the dosed groups is significantly higher (P < 0.001) than that
 

in the corresponding control group.
 

The historical records of the bioassay program at this laboratory
 

indicate that the mean incidence of hepatocellular carcinoma in
 

control male B6C3F1 mice is 49/350 (14%) with 24/54 (44%) being
 

the highest incidence of this tumor in any control group of male
 

mice, compared with 32/44 (73%) in the low-dose group and 41/49
 

(84%) in the high-dose group of this bioassay.
 

43
 
This historical record also indicates that the mean incidence of
 

hepatocellular carcinoma is 13/350 (4%) with 7/54 (13%) being the
 

highest incidence of this tumor in any control group of female
 

mice, compared with 37/43 (86%) in the low-dose group and 43/49
 

(88%) in the high-dose group of this bioassay. The incidences in
 

both the low- and high-dose groups are significantly high (P <
 

0.001) when they are compared with that in the historical-control
 

group.
 

The statistical conclusion is that the incidence of hepatocellular carcinomas in mice is associated with the administration of
 

3-amino-9-ethylcarbazole hydrochloride. There is no other
 

incidence of tumors in either sex that shows a statistical
 

difference in any dosed group compared with its control group.
 

44
 
V. DISCUSSION
 

The toxicity of 3-amino-9-ethylcarbazole hydrochloride for
 

Fischer 344 rats and B6C3F1 mice was shown by markedly lower mean
 

body weights in high-dose male and female rats and mice and by
 

slightly lower mean body weights in low-dose male mice and
 

low-dose female mice, when comparisons are made with
 

corresponding untreated controls. The survival rate of the dosed
 

groups of rats and mice was lower than that of the untreated
 

controls; however, a sufficient number of animals survived to be
 

at risk for development of late-appearing tumors. No other
 

clinical signs were observed that could be related to
 

administration of the test chemical.
 

Since the suppliers of the low-dose rats and mice differed from
 

those of the corresponding low-dose controls, while the suppliers
 

for the high-dose rats and mice were the same as those of the
 

corresponding high-dose controls, comparisons of high-dose groups
 

with their corresponding controls were the more appropriate.
 

Furthermore, since the low-dose animals did not receive the same
 

regimen of administration of the test compound as that received
 

by the high-dose animals, and since tests using the low-dose
 

groups were not performed concurrently with those using the
 

high-dose groups, analyses of dose-related trends were not
 

possible. Although the interpretation of results of the study
 

45
 
was based primarily on comparisons of high-dose groups with their
 

respective controls, the results obtained with the low-dose
 

groups, regardless of the indicated complicating factors,
 

supported the interpretation.
 

Neoplasms of the liver were observed in significant incidences in
 

rats and mice of both sexes. In male rats hepatocellular
 

carcinomas alone were significantly higher (P < 0.020) in both
 

the low- and high-dose groups. When neoplastic nodules of the
 

liver were combined with hepatocellular carcinomas, the
 

combination occurred at significant incidences (P <^ 0.012) in the
 

low- and high-dose male rats and in the high-dose female rats
 

(males: low-dose controls 0/36, low-dose 12/42; high-dose
 

controls 1/48, high-dose 22/48; females: high-dose controls 0/50,
 

high-dose 6/48). Hepatocellular carcinomas alone similarly
 

occurred at significant incidences (P < 0.001) in the low- and
 

high-dose male and female mice (males: low-dose controls 7/48,
 

low-dose 32/44; high-dose controls 6/44, high-dose 41/49;
 

females: low-dose controls 1/47, low-dose 37/43; high-dose
 

controls 1/45, high-dose 43/49).
 

Papillomas or carcinomas of the integumentary system occurred at
 

significant incidences (P _£ 0.013) in both low- and high-dose
 

male rats (low-dose controls 0/36, low-dose 8/44; high-dose
 

controls 0/48, high-dose 6/48); these tumors also occurred in
 

46
 
low- and high-dose female rats, but not at incidences high enough
 

to be statistically significant (low-dose controls 0/39, low-dose
 

4/44; high-dose controls 0/50, high-dose 4/49).
 

In both male and female rats, carcinomas of the Zymbal's glands
 

of the ear occurred only in the dosed groups. In male rats the
 

incidences in the low- and high-dose groups were higher (P =
 

0.045 and P = 0.006, respectively) than in their corresponding
 

controls (males: low-dose controls 0/36, low-dose 5/44; high-dose
 

controls 0/48, high-dose 7/48). In females the incidences in
 

each group were significant (P £ 0.001), compared with
 

corresponding controls (females: low-dose controls 0/39, low-dose
 

10/44; high-dose controls 0/50, high-dose 12/49). These tumors
 

were first observed as early as week 47 on study.
 

Adenocarcinomas of the uterus or endometrium occurred at a
 

significant incidence (P = 0.002) only in the high-dose female
 

rats (high-dose controls 1/50, high-dose 11/49); these tumors
 

also occurred in the low-dose females, but not at an incidence
 

high enough to be statistically significant (low-dose controls
 

4/38, low-dose 11/43).
 

Carcinomas and adenocarcinomas of the mammary gland were observed
 

in 7/44 low-dose female rats and 3/49 high-dose females, but in
 

only 2/39 low-dose female controls and 0/50 high-dose female
 

47
 
controls. The incidences of the tumors in the dosed groups were
 

not high enough, however, to be statistically significant when
 

compared with incidences in corresponding control groups.
 

Adenomatous polyps or adenocarcinomas of the small intestine were
 

observed in 4/41 low-dose male rats, 1/46 high-dose males, and
 

4/40 low-dose females. The same tumors were observed in the
 

colons of 2/41 low-dose and 2/46 high-dose males. None were
 

observed at either site in the control male or female rats.
 

Similarly, adenomas of the preputial gland were observed in 1/44
 

(2%) low-dose and 4/48 (8%) high-dose male rats, but in none of
 

the controls, and adenomas of the clitoris or clitoral gland
 

occurred in 3/44 (7%) low-dose and 3/49 (6%) high-dose females,
 

but in only 2/99 (2%) of the controls. Although the incidences
 

of these different tumors were not statisticaly significant,
 

their historical spontaneous incidences in similar bioassays
 

performed on other test chemicals at the same laboratory were
 

very low (1/334 [0.3%] for adenomatous polyps or adenocarcinomas
 

of the small intestine or colon in males and 0/336 in females,
 

only 4/334 [1%] for adenomas of the preputial gland in males, and
 

only 3/336 [0.9%] for adenomas of the clitoris or clitoral gland
 

in females). Thus, the occurrence of these tumors in the dosed
 

groups of rats in the present bioassay may have been associated
 

with administration of 3-amino-9-ethylcarbazole hydrochloride.
 

48
 
It is concluded that under the conditions of this bioassay,
 

3-amino-9-ethylcarbazole hydrochloride was carcinogenic for the
 

liver, inducing hepatocellular carcinomas in Fischer 344 rats and
 

B6C3F1 mice of both sexes. Other tumors induced in the rats were
 

carcinomas or papillomas of the integumentary system in males,
 

carcinomas of the Zymbal's gland of the ear in males and females,
 

and adenocarcinomas of the uterus.
 

49
 

VI. BIBLIOGRAPHY
 

Anthony, H. M. and Thomas, G. M., Tumors of the urinary bladder:
 

an analysis of the occupations of 1,030 patients in Leeds,
 

England. J. Natl. Cancer Inst. 45(5):879-895, 1970.
 

Armitage, P. , Statistical Methods in Medical Research, John Wiley
 

& Sons, INc., New York, 1971, pp. 362-365.
 

Berenblum, I., ed., Carcinogenicity Testing; _A Report of the
 

Panel on Carcinogenicity of the Cancer Research Commission
 

of UICC, Vol. 2_, International Union Against Cancer, Geneva,
 

1969.
 

Cox, D. R. , Regression models and life tables. J. R. Statist.
 

Soc. B 34 (2);187-220. 1972.
 

Cox, D. R. , Analysis of Binary Data, Methuen & co., Ltd., London,
 

1970, pp. 48-52.
 

Feinstein, R. N. and Lindahl, R. , Detection of oxidases on
 

polyacrylamide gels. Analyt. Biochem. 56:353-360, 1973.
 

Gart, J. J., The comparison of proportions: a review of
 

significance tests, confidience limits and adjustments for
 

stratification. Rev. Int. Stat. Inst. 39 (2):148-169, 1971.
 

Kaplan, E. L. and Meier, P., Nonparametrie estimation from
 

incomplete observations. J_,_ Am. Statist. Assoc. 53:457-481,
 

1958.
 

Kaplow, L. S. , Substitute for benzidine in myeloperoxidase
 

stains. Amer. J. Clin. Pathol. 63, 451, 1975.
 

Linhart, M. S. , Cooper, J. A., Martin, R. L. , Page, N. P., and
 

Peters, J. A., Carcinogenesis bioassay data system. Comp.
 

and Biomed. Res. 7:230-248, 1974.
 

Miller, R. G., Jr., Simultaneous Statistical Inference,
 

McGraw-Hill Book co., New York, 1966, pp. 6-10.
 

Pliss, G. B. , Tumours of the auditory sebaceous glands. Pathology
 

of Tumours in Laboratory Animals, Vol. 1- Tumours of the
 

Rat, Turusov, V. S., ed,, International Agency for Research
 

on Cancer, Lyon, 1973, pp. 23-27.
 

51
 
Saffiotti, U. , Montesano, R., Sellakumar, A. R., Cefis, F., and
 

Kaufman, D. G., Respiratory tract carcinogenesis in hamsters
 

induced by different numbers of administrations of benzo (a)
 

pyrene and ferric oxid. Cancer Res. 32;1073-1081, 1972.
 

Society of Dyers and Colourists, Colour Index, Vol. j4, 1971,
 

pp. 4754, 4794, 4467.
 

Stanford Research Institute, 1977 Directory of_ Chemical
 

Producers, U.S.A., Menlo Park, Calif., 1977, p. 407.
 

Tarone, R. E., Tests for trend in life table analysis.
 

Biometrika 62 (3):679-682, 1975.
 

United States International Trade Commission, Imports of
 

Benzenoid Chemicals and Products, 1974. USITC Publication
 

762, Washington, D.C., 1976, p. 9.
 

Wynder, E. L. , Onderdonk, J., and Mantel, N., An epidemiological
 

investigation of cancer of the bladder. Cancer
 

16:1388-1407, 1963.
 

52
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS FED 3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

IN THE DIET
 

53
 

TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

LOW DOSE
 

CONTROL
 LOW DOSE
 

ANIHALS IHITIAILY IN STUDY 50 50
 

ANIMALS MISSING 3
 

ANIHALS NECROPSIED 46 nn
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 46 43
 

INTEGUMENTARY SYSTEM
 

*SKIN (46) (44)
 

NEOPLASM, NOS 1 (2*)
 

SgUAMODS CELL PAPILLOMA 3 (7%)
 

SQOAMOUS CELL CARCINOMA 3 (7«)
 

BASAL-CELL CARCINOMA 4 (9%)
 

SEBACEOUS ADENOMA 1 (2%)
 

*SUBCUT TISSUE (46) (44)
 

FIBROMA 1 (2X)
 

FIBBOSARCOMA 1 (215)
 

RESPIRATORY SYSTEM 

fTRACHEA 

ADENOCARCINOMA, NOS, METASTATIC 

(U5) 

1 (2*) 

('40) 

fLUNG/BRONCHUS 

PAPILLOMA, NOS 

(46) (43) 

1 (2*) 

#LUNG 

ADENOCARCINOMA, NOS, METASTATIC 

ALVEOLAR/BRONCHIOLAR ADENOMA 

ALVEOLAR/BBONCHIDLAR CARCINOMA 

(t6) 

1 (2%) 

(43) 

2

1

 (5«) 

 (2%) 

HEHATOPOIETIC SYSTEM
 

*HULTIPLE ORGANS (<46) (44)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

ONDIFFERENTIATED LEUKEMIA 1 (2%)
 

MYELOMONOCYTIC LEUKEMIA 4 (9%)
 

HONOCYTIC LEUKEMIA 1 J2X) . 1 J2S1 .
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECSOPSIED
 

55
 
A1. MALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

LOW DOSE
 

CONTROL
 

ILYMPH NODE (38)
 

ADENOCAPCINOMA, NOS, BETASTATIC 1 (3%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

ILIVER (1*6)
 

NEOPLASTIC NODULE
 

HEPATOCE1LULAR CARCINOMA
 

*SMALL INTESTINE (t3)
 

ADENOCARCINOMA, NOS
 

ADENOHATOUS POLYP, NOS
 

MUCINOUS CYSTADENOCARCINOMA
 

LEIOMYOSARCOMA
 

f JEJUNUM (43)
 

CYSTADENOCARCINOMA, NOS
 

LEIOMYOSARCOMA
 

#COLON (43)
 

ADENOCARCINOMA, NOS
 

ADENOCA IN ADENOMATOUS POLYP
 

URINARY SYSTEM
 

NONE
 

ENDOCBINE SYSTEM
 

tPITUITAPY (41)
 

ADENOMA, NOS 2 (5%)
 

CHROMOPHOBE ADENOMA 10 (24?,)
 

((ADRENAL (43)
 

ADENOCARCINCMA, NOS, METASTATIC 1 (2%)
 

PHEOCHROMOCYTOMA 6 J1U%L
 

LOW DOSE
 

(3U)
 

(42)
 

6 (14%)
 

6 (14%)
 

(41)
 

1 (2?)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(41)
 

1 (1%)
 

1 (2%)
 

(38)
 

1 (3%)
 

1 (3*)
 

<17)
 

^ (14%)
 

7 (19%)
 

(41)
 

^ 1«L
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

56
 
A1. MALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

GANGLIONEUROM A 1 (2% ) 

fTHYROI D 

ADENOMA , NOS 

ADENOCARCINOMA , (JOS 

FOLLICULAR-CELL ADENOM A 

FOLLICDLAR-CELL CARCINOM A 

C-CELL ADENOH A 

(«5) 

1 (2% ) 

2 («X) 

1 (2%) 

(40) 

1 (3%) 

1 (3K) 

7 (5%) 

1 (3S) 

#PANCREATIC ISLSTS 

ISLET-CELL ADENOM A 

(142) 

2 (5%) 

(3-5) 

4 (10% ) 

REPRODUCTIV E SYSTEM 

*MAMMAR Y GLAN E (46) (44) 

FIBROADENOM A 1 (2%) 

*PREPUTIA L GLAN D (1*6) (44 ) 

ADENOMA , NCS 1 (2X) 

SEBACEOUS ADENOH A 1 (2%) 

IPROSTATE (H5 ) (41) 

PARAGANGLIOMA , NO S 1 (2%) 

tTESTIS (1*5) (43 ) 

INTERSTITIAL-CELL TOMO R 33 (73%) 32 (TUX ) 

NERVOUS SYSTEM 

#BRAI N ((*!*) (42 ) 

ASTROCYTOMA 1 (2% ) 

SPECIAL SENSE ORGAN S 

*ZYMBAL' S GLAN D (46) (44 ) 

CARCINOMA , NOS 1 (2% ) 

SQOAMOUS CELL CARCINOH A 4 (9%) 

HUSCULOSKELETAL SYSTEH 

NONE. _ _ 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NOBBER OF ANIHALS NECROPSIED
 

57
 
A1. MALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

BODY CAVITIES
 

*BODY CAVITIES
 

NESOTHELIOHA, NOS
 

ALL OTHER SYSTEMS
 

TAIL
 

BASAL-CELL CARCINOMA
 

ANIHAL DISPOSITION SUBBAEY
 

ANIHALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERHISAL SACRIFICE
 

ANIHAL HISSING
 

9 INCLUDES AUTOLYZED ANIHALS
 

TUHOR SUHMARY
 

TOTAL ANIHALS WITH PRIMARY TOHORS*
 

TOTAL P8IHARY TUMORS
 

TOTAL ANIHALS WITH BENIGN TUHORS
 

TOTAL BENIGN TUHORS
 

TOTAL ANIHALS WITH HALIGNANT TOHORS
 

TOTAL HALIGNANT TUHORS
 

TOTAL ANIHALS WITH SECONDARY TOHORSt
 

TOTAL SECONDARY TUHORS
 

TOTAL ANIHALS WITH TUHORS UNCERTAIN­ BENIGN OR HAIIGNANT
 

TOTAL UNCERTAIN TUHORS
 

TOTAL ANIHALS WITH TUHORS UNCERTAIN­ PRIHARY OR HBTASTATIC
 

TOTAL UNCERTAIN TUHORS
 

(«6) (ill)
 

2 (5%)
 

50 •50
 

6 7
 

2 17
 

15 5
 

27 18
 

3
 

3H UO
 

61 112
 

33 38
 

55 68
 

5 26
 

5 35
 

1
 

H
 

1 9
 

1 9
 

* PRIHARY TUHORS: ALL TOHORS EXCEPT SECONDARY TOHORS
 

# SECONDARY TUHORS: HETASTATIC TOBORS OR TOHORS INVASIVE INTO AN ADJACENT ORGAN
 

58
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIHALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEH
 

*SKIN
 

SQOAHOnS CELL PAPILLOMA
 

SQUABOUS CELL CARCINOMA
 

BASAL-CELL CARCINOMA
 

ADENOMA, NOS
 

*SUBCUT TISSUE
 

SCUABOUS CELL CABCINOHA
 

ADENOMA. NOS
 

SARCOBA, NOS
 

FIBROMA
 

FIBROSARCOBA
 

RESFIBATOBY SYSTEB
 

fLONG
 

CARCINOMA, NOS, BETASTATIC
 

SQUAMODS CELL CARCINOMA, METASTA
 

ALVEOLAR/BRONCHIOLAR ADENOMA
 

ALVEOLAR/BRONCHIOLAR CARCINOMA
 

PHEOCHROMOCYTOMA, METASTATIC
 

HEBATCFOIETIC SYSTEM
 

*BOITIPLE ORGANS
 

MALIGNANT LYMPHOHA, NOS
 

LEUKEMIA, NOS
 

UNDIFFERENTIATED LEUKEMIA
 

MYELOMONOCYTIC LEUKEHIA
 

»LYMPH NODE
 

SC.UAMOUS CELL CARCINOMA^ METASTA
 

HIGH DOSE
 

CONTROL
 

a so
 

48
 

46
 

(48)
 

(48)
 

1 (2%)
 

3 <6X)
 

1 (2%)
 

(48)
 

•
 

1 (2%}
 

1 (2%)
 

(48)
 

1 (2%)
 

1 (2%)
 

4 (8*)
 

(44)
 

HIGH DOSE
 

50
 

48
 

48
 

(48)
 

2 (4X)
 

2 (4X)
 

1 (2%)
 

1 (2%)
 

(48)
 

1 (2%)
 

1 (2%)
 

5 (10X)
 

(48)
 

1 (2%)
 

1 (2%)


2 (4«)
 

3 (6X)
 

1 (2X)
 

(48)
 

1 (2%)
 

3 (6S)
 

(33)
 

1 (3%)
 

t NUMBER OF ANIMALS HITH TISSUE EXABINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

» 50 ANIHALS WERE INITIALLY IN THE STUDY, BUT ONE ANIMAL MAS
 

FOUND TO BE A FEMALE IN A MALE GROUP.
 

59
 
A2. MALE RATS (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

HIGH DOSE
 

CONTROL
 HIGH DOSE
 

CIRCULATOBY SYSTEM
 

•HEABT/VENTHICLE	 (48) (48)
 

PHEOCHROMOCYTOHA, METASTATIC 1 (2%)
 

DIGESTIVE SYSTEM
 

•SALIVARY GLAND	 (U7) (45)
 

ADENOCARCINOHA, NOS 1 (2X)
 

SARCOMA, NOS 1 (2X)
 

tLIVER (48) (48)
 

BILE DUCT ADENOMA 1 (2%)
 

NEOPLASTIC NODULE 14 (29X)
 

HEPATOCELLULAR CARCINOMA 1 (2X) 8 (17X)
 

*BILE DUCT (48) (48)
 

BILE DUCT CARCINOMA 1 (2X)
 

•STOMACH	 (48) (42)
 

SCnAMOOS CELL PAPILLOMA 1 (2X)
 

fJEJUNUM (46) (46)
 

CYSTADENOCARCINOMA, NOS 1 (2%)
 

*ILEUM (46) (46)
 

SARCOMA, NOS 1 (2X)
 

•COLON	 (46) (38)
 

ADENOCARCINOMA, NOS 1 (3%)
 

ADENOMATOOS POLYP, NOS 1 (3X)
 

UBINAB Y SYSTEM
 

•KIDNEY	 (48) (48)
 

TBANSITIONAL-CELL CARCINOMA 1 (2%)
 

TOBULAR-CELL ADENOMA 1 (2%)
 

•URINARY	 BLADDER (43) (<*2)
 

TBANSITIONAL-CELL PAPILLOMA 1 (2%)
 

ENECCEINE SYSTEM
 

•EITUITABY	 (38) (41)
 

ADENOMA^ NOS 9 I24S1 10 (24£)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

60
 
A2. MALE RATS (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

CHROHOPHOBE ADENOHA
 

IADRENAL
 

CORTICAL ADENOHA
 

PHEOCHROHOCYTOBA
 

PHEOCHROBOCYTOHA, HALIGNANT
 

ITHYROID
 

MEOPLASB, NOS
 

FOLLICOLAR-CELL CARCINOHA
 

C-CELL CARCINOMA
 

•PARATHYROID
 

ADENOHA, NOS
 

•PANCREATIC ISLETS
 

ISLET-CELL ADE80HA
 

REPRODUCTIVE SYSTEB
 

*PREPUTIAL GLAND
 

CARCINOHA.NOS
 

ADENOHA, NOS
 

ITESTIS
 

INTERSTITIAL-CELL TtJHOR
 

NERVOUS SYSTEH
 

tBRAIN
 

GLIOHA, NOS
 

ASTROCYTOBA
 

OLIGODENDROGLIOBA
 

SPECIAL SENSE ORGANS
 

*ZYBBAL'S GLAND
 

CARCINOHA.NOS
 

SQUAHODS CELL CARCINOBA
 

HUSCDIOSKELETAL SYSTEH
 

_. BONE
 

HIGH DOSE
 

CONTROL
 

(47)
 

7 (15X)
 

1 (2X)
 

(US)
 

1 (2X)
 

(28)
 

1 (45t)
 

(46)
 

(48)
 

(47)
 

42 (89X)
 

(48)
 

1 (256)
 

(48)
 

HIGH DOSE
 

1 (2*)
 

(48)
 

1 (2X)
 

9 (19%)
 

1 (2»)
 

(47)
 

1 (2X)
 

1 (2X)
 

2 (4»)
 

(23)
 

(43)
 

4 <9X)
 

(48)
 

1 (2X)
 

4 (8X)
 

(47)
 

40 (85S)
 

(47)
 

1 (2X)
 

1 (2X)
 

(48)
 

4 (8X)
 

3 (6X)
 

t SDHEER OF ANIHALS HITH TISSUE EXA HIKED HICROSCOPICALLY
 

* NDHBER OF ANIHALS NECROPSIED
 

61 
A2. MALE RATS (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

BODY CAVITIES
 

*BOD¥ CAVITIES 0*8) (48)
 

HESOTHELIOHA, NOS 3 (6X)
 

HESOTHELIOHA, HALIGNANT 2 (4X)
 

All CTHEB SYSTEHS
 

SITE UNKNOWN
 

CARCINOHA,NOS 1
 

ANIHAI DISPOSITION SOHHARY
 

ANIHALS INITIALLY IN STUDY 50 50
 

NATURAL DEATHS 6 6
 

MORIBUND SACRIFICE 8 13
 

SCHEDULED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 30 26
 

ANIF1AL HISSING
 

ANIHAL DELETED(HRONG SEX) 1
 

a INCLUDES AUTOLYZED ANIHALS
 

TUHOR SUHHARY
 

TOTAL ANIHALS WITH PRIHARY TUHORS* 44 47
 

TOTAL PRIMARY TUHORS 80 141
 

TOTAL ANIHALS WITH BENIGN TUHORS 43 43
 

TOTAL BENIGN TUHORS 62 85
 

TOTAL ANIHALS WITH HALIGNANT TUHORS 17 32
 

TOTAL HALIGNANT TUHORS 18 38
 

TOTAL ANIHALS WITH SECONDARY TUHORS* 1 3
 

TOTAL SECONDARY TUHOBS 1 5
 

TOTAL ABIHALS HITH TUHORS UNCERTAIN­ BENIGN OR HALIGNANT 18
 

TOTAL UNCERTAIN TUHORS 18
 

TOTAL ANIHALS HITH TOBORS UNCERTAIN­ PRIHARY OB HETASTATIC
 

TOTAL UNCERTAIN TUHORS
 

* PRIHABY TDHOBS: ALL TOHOBS EXCEPT SECONDARY TOHORS
 * SECONDARY TUHORS: HBTASTATIC TUHORS OB TOHORS INVASIVE INTO AH ADJACENT ORGAN
 

62
 
TABLE A3. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET(LOW DOSE AND CONTROL) 

AHIHALS INITIALLY IN STUDY
 

ANIHALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

SQUAMOUS CELL PAPILLOHA
 

SQUAMOUS CELL CARCINOMA
 

*SOBCUT TISSUE
 

SQUANOUS CELL CARCINOMA
 

SARCOMA, NOS
 

FIBROMA
 

FIBROSARCOHA
 

RESPIRATORY SYSTEM
 

#LUNG
 

CARCINOMA, NOS, METASTATIC
 

ADENOCARCINONA, NOS, METASTATIC
 

HEPATOCELLULAR CARCINOMA, MBTAST
 

ALVEOLAR/BFONCHIOLAR ADENOMA
 

ALVEOLAR/BRONCHIOLAR CARCINOMA
 

SARCOMA, NOS, METASTATIC
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS
 

MALIGNANT LYMPHOMA, NOS
 

MALIG.LYMPHOHA, LYMPHOCYTIC TYPE
 

MYELOMONOCYTIC LEUKEMIA
 

MONOCYTIC LEUKEMIA
 

#LYMPH NODE
 

SARCOMA, NOS, METASTATIC
 

*RENAL LYMPH NODE
 

ADENOQARCINOflAi NQSt_flETASIATIQ _.
 

LOW DOSE
 

CONTROL LOW DOSE
 

50 50
 

49 an
 

U9 U3
 

(«9) (i»i*)
 

1 (2*)
 

2 (5%)
 

(U9) (Hk)
 

1 (2«)
 

2 (5%)
 

2 (5%)
 

1 (2*)
 

(«9) (43)
 

1 (2*)
 

1 (2X) 1 (2%)
 

1 (2«)
 

1 (2*) 3 (7%)
 

1 (2^)
 

1 (21)
 

(U9) (HU)
 

1 (2«)
 

2 (*%)
 

1 (2%)
 

2 (U%)
 

(11) (36)
 

1 (3%)
 

(ai> (36)
 

1_12SL_...
 

t NUMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

63
 
A3. FEMALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

\
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

#LIVfiR
 

ADENOCARCINOMA, NOS, METASTATIC
 

HEPATOCEI.LULAR CARCINOMA
 

fPANCREAS
 

CARCINOMA, NOS, METASTATIC
 

fSTOMACH
 

CARCINOMA, NOS, METASTATIC
 

#SMALL INTESTINE
 

ADENOMATOUS POLYP, NOS
 

MUCINOUS CYSTADENOCARCINOMA
 

*ILEUH
 

ADENOCARCINOMA, NOS
 

ADENOMATOUS POLYP, NOS
 

tCOLON
 

ADENOCARCINOMA, NOS
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

tPITUITARY
 

ADENOMA, NOS
 

ADENOCARCINOMA, NOS
 

CHROM3PHOBF ADENOMA
 

#ADPENAL
 

PHEOCHROMOCYTOMA
 

fTHYROID
 

ADENOMA^ NCS
 

LOW DOSE
 

CONTROL
 

(49)
 

1 (2%)
 

2 (it*)
 

(46)
 

(«8)
 

(U7)
 

(47)
 

(43)
 

(«3)
 

3 (7%)
 

2 (5%)
 

15 (35%)
 

(06)
 

2 («%)
 

(«7)
 

1 1221 _ .
 

LOW DOSE
 

(43)
 

1 (2%)
 

(39)
 

1 (3«)
 

(41)
 

1 (2%)
 

(40)
 

1 (3S)
 

1 (3%)
 

(40)
 

1 (3«)
 

1 (3%)
 

HS)
 

1 (3%)
 

(^0,
 

2 (7T,)
 

13 (43«)
 

(41)
 

1 (2%)
 

(40)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NDMBER OF ANIMALS NECROPSIFD
 

64
 
A3. FEMALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

ADENOCARCINOMA , NO S 

C-CELL ADENOM A 

#THYROI D FOLLICLE 

PAPILLAR Y CYSTADENOMA , NO S 

tPANCREATIC ISLETS 

ISLET-CELL ADENOM A 

REPRODUCTIV E SYSTEM 

*MAHMAR Y GLAN D 

CARCINOMA , NOS 

ADENOMA , NO S 

ADENOCAHCINOMA , NOS 

PAPILLAR Y ADENOCARCINOM A 

PAPILLAR Y CYSTADENOMA , NO S 

PAPILLAR Y CYSTADENOCARCINOMA,NO S 

FIBROADENOM A 

*CLITORIS 

CARCINOMA , NOS 

*CLITORAL GLAN D 

SQOAMOOS CELL PAPILLOM A

 

SQUAMOU S CELL CARCINOM A 

ADENOMA, NCS

 

#UTEROS 

NEOPLASM , NOS 

ADENOCAHCINOMA , NOS 

LEIOHYOSARCON A 

ENDOBETRIA L STROMAL POLYP 

#OVAR Y 

CARCINOMA , NOS, METASTATIC 

NERVOO S SYSTEM 

NONE 

SPECIAL SENSF ORGAN S 

*EAR CANA L 

FIBROM A 

LOW DOSE 

CONTROL LOW DOSE 

2 (UX ) 1 (3%) 

1 (2%) 2 (S«) 

(47) (40) 

1 (3% ) 

(46) (39 ) 

1 (3X ) 

(49) (44 ) 

1 (2X) 

1 (2X) 

1 (2%) 4 (<)%) 

1 (2%) 

1 (2%) 

1 (2X) 2 (5*) 

4 (3%) 4 (9%) 

(49) (44 ) 

1 (2X) 

(49) (44 ) 

1 (2% ) 

1 (2X ) 

3 (1%) 

(48) (43) 

2 (5%) 

4 (8%) 11 (26X) 

1 (2*) 

10 (21%) 11 (26%) 

(47) (43 ) 

1 (2%) 

(49) (44)

 

1 (2%) 

#	 NUMBER OF ANIBALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 HOMBER OF ANIMALS S3CROPSIED
 

65
 
A3. FEMALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

*ZYMBAL'S GLAND
 

CARCINOMA, NOS
 

SQUAHOnS CELL CARCINOMA
 

BUSCULOSKELETAI SYSTEM
 

NONE
 

BODY CAVITIES
 

*BODY CAVITIES
 

MESOTHELIOBA, MALIGNANT
 

ALL OTHER SYSTEMS
 

SITE UNKNOWN
 

LEIOMYOSARCOBA
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

1 INCLUDES AH10LXZ£B_A.NIflAlS__
 

LOW DOSE
 

CONTROL
 

(<»9)
 

(<*9)
 

1 (2%)
 

50
 

5
 

7
 

15
 

23
 

LOW DOSE
 

(««)
 

5 (11%)
 

5 (11%)
 

(1»U)
 

1
 

50
 

15
 

15
 

5
 

15
 

* NUMBER OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS HECROPSIED
 

66
 
A3. FEMALE RATS (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

TDBOE SOHHAR7
 

TOTAL ANIMALS WITH PBIHABY TUMORS* 32 uo
 

TOTAL PRIHARY TOHORS 56 97
 

TOTAL AHIHALS WITH BENIGN TUHOKS 27 31
 

TOTAL BENIGN TOHORS 39 U8
 

TOTAL ANIMALS WITH MALIGNANT TONORS 15 3«
 TOTAL MALIGNANT TUMORS 17 l»7
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2 3
 

TOTAL SECONDARY TUMORS U 7
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT 2
 

TOTAL UNCERTAIN TUMORS 2
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUHORS EXCEPT SECONDARY TUMORS
 

t SECONDARY TUMORS: NETASTATIC TUBORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

67
 
TABLE A4.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

AHIHALS INITIALLY IN STUDY
 

ANIBAIS HECBCPSIED
 

ANIMALS EXAHINED HISTOPATHOLOGICALLY
 

INTEGUHENTABY SYSTEH
 

*SKIN
 

SQUAMOUS CELL PAPILLOHA
 

SQDAHOOS CELL CABCINOHA
 

BASAL-CELL CARCINOBA
 

SEBACEOUS ADENOCARCINOHA
 

*SOBCUT TISSOE
 

FIBBOHA
 

FIBBOSABCOMA
 

RESFISATOBY SYSTEM
 

#LUNG
 

SQOAHOUS CELL CARCINOMA, METASTA
 

ALVEOLAR/BRONCHIOLAR ADENOMA
 

ALVEOLAR/BRONCHIOLAR CARCINOMA
 

HEHATCPOIETIC SYSTEM
 

*MUITIPLE ORGANS
 

ONDIFFEBENTIATED LEDKEMIA
 

MYELOMONOCYTIC LEUKEMIA
 

LYHFHOCYTIC LEUKEMIA
 

#SPLEEN
 

UNDIFFERENTIATED LEUKEMIA
 

CIBCULATOEY SYSTEM
 

NONE
 

HIGH OOSE
 

CONTROL
 

50
 

50
 

50
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 

1 (2X)
 

(50)
 

1 (2X)
 

1(2%)
 

(50)
 

1 (2«)
 

3 (6X)
 

(1*8)
 

1 (2%)
 

HIGH DOSE
 

50
 

49
 

49
 

(49)
 

1 (2X)
 

3 (6X)
 

1 (2X)
 

(49)
 

1 (2X)
 

(48)
 

4 (8%)
 

2 (4X)
 

(49)
 

4 (8X)
 

2 (4X)
 

(48)
 

# NUMBER OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* BUMBEB OF ANIHALS NECEOPSIED
 

68
 
A4. FEMALE RATS (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

DIGESTIVE SYSTEM
 

ILIVER
 

NEOPLASTIC NODDLE
 

HEPATOCELLULAR CARCINOMA
 

tILEUN
 

IEIOHYOSABCOHA
 

URINA6Y SYSTEM
 

tURINABY BLADDEB
 

TRANSITIONAL-CELL PAPILLOBA
 

ENECCBINE SYSTEM
 

•PITUITARY
 

ADENOMA, NOS
 

CHROMOPHOBE ADENOMA
 

IADBENAL
 

CORTICAL ADENOMA
 

PHEOCHHOMOCYTOHA
 

*ADRENAL MEDULLA
 

GANGLIONEDROMA
 

tTHYROID
 

FOLLICOLAH-CELL ADENOMA
 

FOLLICULAB-CELL CARCINOMA
 

C-CELL ADENOMA
 

C-CELL CARCINOMA
 

tPAHCHEATIC ISLETS
 

ISLET-CELL ADENOMA
 

BEFBOCUCTIVE SYSTEM
 

*MAHHAEY GLAND
 

iDENOCARCINOHA, NOS
 

FIBROADEBOBA
 

HIGH DOSE
 

CONTROL
 

(50)
 

(<*8)
 

1 (2%)
 

(46)
 

C*0)
 

17 (43X)
 

(49)
 

1 (2%)
 

3 (6%)
 

(49)
 

1 (2X)
 

(«5)
 

1 (2«)
 

1 (2%)
 

1 (2*)
 

(48)
 

2 (4X)
 

(50)
 

19 U8S1
 

HIGH DOSE
 

(48)
 

3 (6*)
 

3 (6*)
 

(46)
 

(43)
 

1 (2X)
 

(39)
 

19	 (49%)
 

1 (3«)
 

(47)
 

3 (6«)
 

4 (9X)
 

(47)
 

(44)
 

1 (2*)
 

1 (2X)
 

1 (2X)
 

(47)
 

(49)
 

3 (6X)
 

11 J22X1
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

69 
A4. FEMALE RATS (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

*CLITORIS
 

CARCINOMA. NOS
 

SQUAMOUS CELL CARCINOMA
 

AEENOHA, NOS
 

*CLITOfiAL GLAND
 

SQOAMOUS CELL PAPILLOMA
 

SQOAHOOS CELL CARCINOMA
 

AEENOMA, NOS
 

tUTEROS
 

ADENOCARCINOMA. NOS
 

ENDOMETHIAL STROMAL POLYP
 

ENDOHETRIAL STROBAL SARCOMA
 

tOTERUS/ENDONETRIUM
 

ADEHOCARCINOHA, NOS
 

#OVARY
 

GHANULOSA-CELL TUMOR
 

NERVOUS SYSTEM
 

*BRAIN
 

ASTROCYTOMA
 

SPECIAL SENSE ORGANS
 

*ZYMBAL'S GLAND
 

CARCINOMA, NOS
 

SQUAMOUS CELL CARCINOMA
 

MOSCOIOSKELETAL SYSTEM
 

NONE
 

BCEY CAVITIES
 

NONE
 

HIGH DOSE
 

CONTROL
 

(50)
 

(50)
 

1 (2X)
 

2 (4X)
 

(50)
 

1 (2%)
 

10 (20%)
 

1 (2%)
 

(50)
 

09)
 

1 (2%)
 

(50)
 

(50)
 

HIGH DOSE
 

("9)
 

1 (2X)
 

2 (>4%)
 

3 (6X)
 

(49)
 

1 (2X)
 

(U9)
 

10 (2 OX)
 

5 (10X)
 

(49)
 

1 (2X)
 

(47)
 

(48)
 

2 (4X)
 

(49)
 

3 (6X)
 

9 (18X)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

70
 
A4. FEMALE RATS (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

ALL OTHER SYSTEMS
 

* MULTIPLE ORGANS 

ADENOCARCINOMA, DOS, BETASTATIC 

(50) (i»9) 

1 (2X) 

SITE UNKNOWN 

SQUABOUS CELL CARCINOHA 1 

ANIBA1 DISPOSITION SUBMARY
 

ANIBALS INITIALLY IN STUDY 50 50
 

NATURAL DEATHS 5 7
 

HORIBUND SACRIFICE 3 15
 

SCHEDDLED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 

TERBINAL SACRIFICE 37 23
 

ANIHAL MISSING
 

8 INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH FEIBAEY TUMORS* 38 45
 

TOTAL PRIMARY TUMORS 73 106
 

TOTAL ANIMALS WITH BENIGN TUMORS 35 3U
 

TOTAL BENIGN TUMORS 59 55
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 12 35
 

TOTAL MALIGNANT TUMORS 13 US
 

TOTAL ANIBALS WITH SECONDARY TUMORS * 1 1
 

TOTAL SECONDARY TUBOHS 1 1
 

_
 TOTAL ANIMALS WITH TDMORS UNCERTAIN
 

BENIGN OR MALIGNANT 1 3
 

TOTAL UNCERTAIN TUMORS 1 3
 

TOTAL ANIBALS WITH TUMORS UNCERTAIN
 -
 PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PBIHARY TDMORS: ALL TUHORS EXCEPT SECONDARY TOHORS
 « SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

71 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE FED 3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

IN THE DIET
 

73
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

ANIMALS INITIAILY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECEOPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

HEMANGIOSARCONA
 

RESPIRATORY SYSTEM
 

*LONG
 

ALVEOLAR/BRONCHIOLAR ADENOMA
 

ALVEOLAR/BRONCHIOLAR CARCINOMA
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS
 

MALIGNANT LYMPHOMA, NOS
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

#SPLEEN
 

HEMANGIOMA
 

tLIVER
 

MALIG.LYMPHOBA, HISTIOCYTIC TYPE
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

OLIVER
 

HEPATOCELLULAR CARCINOMA
 

LOW DOSE
 

CONTROL


50
 

48
 

US
 

(48)
 

(48)
 

6 (13%)
 

(48)
 

2 (4%)
 

2 (4%)
 

(47)
 

1 (2%)
 

(48)
 

(48)
 

7 i15%L .
 

 LOW DOSE
 

50
 

1
 

44
 

44
 

(44)
 

1 (2%)
 

(42)
 

5 (12%)
 

1 (2*)
 

(44)
 

2 (5%)
 

(43)
 

1 (2%)
 

(44)
 

1 (2%)
 

(44)
 

32 173%L .
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

75 
B1. MALE MICE (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

HBBANGIOBA 

*GALLBLADDBR 

ADENOHATOUS POLYP, HOS 

tSTOHACH 

SQUAMOUS CELL CABCIHOHA 

URINAH Y SYSTEH 

NONE 

ENDOCRINE SYSTEH 

*THYROID 

FOLLICULAR-CELL ADENOHA 

REPFODOCTIVE SYSTEM 

NONE 

NERVOUS SYST5H 

NONE 

SPECIAL SENSE ORGANS 

NONE 

MUSCOLOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEHS 

NONE „ 

LOW DOSE 

CONTROL LOW DOSE 

1 (2%) 

(08) (40) 

1 (2X) 

(07) (02) 

1 (2X) 

(07) (35) 

1 (2S) 

#	 NUHBER OP ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSIED
 

76
 
B1. MALE MICE (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

1

LOW DOSE
 

CONTROL LOW DOSE
 

ANIMAL DISPOSITION SONHABY
 

ANIHALS INITIALLY IN STUDY 50 SO
 

NATURAL DEATHS 3
 7
 

MORIBUND SACRIFICE 1
 

SCHEDULED SACRIFICE 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE U2 in
 

ANIMAL HISSING 1
 

9 INCLUDES AUTOLYZED ANIHALS
 

TUMOR S U M M A RS U M M A RY
 

TOTAL ANIHALS WITH P R I M A RP R I M A RY TUMORS* 17 35
 

TOTAL PRIMARY TUMOR TUMORSS 21 <•IC«
 1

TOTAL ANIHALS WITH BENIGN TUMORS 2 7


TOTAL BENIGN TUMORS 3 7


TOTAL ANIHALS WITH MALIGNAN MALIGNANT TUMOR TUMORSS 15 333


TOTAL MALIGNAN MALIGNANT TUMORS 18 337


TOTAL ANIHALS WITH SECONDAR SECONDARY TUMORSt
 

TOTAL SECONDAR SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN UNCERTAIN-­ BENIGN OR MALIGNAN MALIGNANT
 

TOTAL UNCERTAI UNCERTAIN TUMORS
 

TOTAL ANIHALS WITH TUHORS UNCERTAIN UNCERTAIN-­ PRIHARY OR HETASTATI HETASTATIC
 

TOTAL UNCERTAI UNCERTAIN TUHORS
 

* PRIMARY TUHORS: ALL TUHORS EXCEPT SECONDARY TOHORS
 

# SECONDARY TUHOHS: HBTASTATIC TUMORS OR TUHORS INVASIVE INTO AH ADJACENT ORGAN
 

77
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE FED 

3-AMINO-g-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

ANIHALS INITIALLY IN STUDY
 

AHIHALS HISSING
 

ANIHALS NECROPSIED
 

ANIHALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESEIFATORY SYSTEM
 

#IONG
 

HEPATOCELLULAR CAECINOMA, METAST
 

ALVEOLAR/ERONCHIOLAR ADENOMA
 

ALVEOLAR/BRONCHIOLAR CARCINOHA
 

HIMATCPOIETIC SYSTEM
 

+BUITIPLE ORGANS
 

MALIG.LYBPHOHA, HISTIOCYTIC TYPE
 

UNDIFFERENTIATED LEUKEMIA
 

*SPLEEN
 

HEMANGIOSARCOMA
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

KLYMPH NODE
 

HEPATOCELLDLAR CARCINOMA, METAST
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

#PEYEBS PATCH
 

MALIG.LYMPHOHA, HISTIOCYTIC TYPE
 

#KIDNEY
 

MALIG.LYBPHOMA, HISTIOCYTIC TYPE
 

CIECUIATORY SYSTEM
 

NCNE
 

HIGH DOSE
 

CONTROL HIGH DOSE
 

50	 50
 

1
 

49 50
 

49 50
 

(49)	 (49)
 

1 (2%} 3 (6X)
 

5 (10*) 4 (8*)
 

5 (10*) 3 (6X)
 

(49)	 (50)
 

3	 (6X) 2 (4X)
 

1 (2%)
 

(49)	 (49)
 

1 (2%)
 

1 (2%)
 

(42)	 (39)
 

1 (3X)
 

1 (251) 1 (3X)
 

(49)	 (47)
 

1 (2%)
 

(49)	 (50)
 

1 (2X)
 

f	 NUMEER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBEE OF ANIMALS NECROPSIED
 

78
 
B2. MALE MICE (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

DIGESTIVE SYSTEM
 

ILIVER
 

HEPATOCELLULAR ADENOMA
 

HBPATOCELLUIAR CARCINOMA
 

HENANGIOSARCOHA, DNC PRIM OB MET
 

^GALLBLADDER
 

ADEHOMATOOS POLYP, NOS
 

*BECTOM
 

ADENOCABCINOMA, NOS
 

UBINA6Y SYSTEM
 

NONE
 

ENECCEINE SYSTEM
 

#ADRENAL
 

PHEOCHROMOCYTOMA
 

*THYROID
 

FOLLICOLAB-CELL ADENOMA
 

PAPILLARY CYSTADENOMA, NOS
 

BEEBODUCTIVE SYSTEM
 

ITESTIS
 

INTERSTITIAL-CELL TUMOR
 

*EPIDIDYMIS
 

HIBERNOHA
 

NEBVOOS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE_ _ ,_ ^^_ _ ____ _
 

HIGH DOSE
 

CONTROL
 

(<»8)
 

2 {UX)
 

6 (13X)
 

1 (2%)
 

C»9)
 

(49)
 

(«K»)
 

1 (2%)
 

(45)
 

(48)
 

(49)
 

HIGH DOSE
 

(i»9)
 

41 (84X)
 

(50)
 

2 (««)
 

(50)
 

1 (2*)
 

(47)
 

1 (2X)
 

(46)
 

3 (7*)
 

1 (2X)
 

(49)
 

1 (2*)
 

(50)
 

1 (2*)
 

* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

79
 
B2. MALE MICE (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

* NOMBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

80
 
B2. MALE MICE (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS ilTH PBIMABY TUMORS* 22 U5
 

TOTAL PHIHARY TOHORS 26 6U
 

TOTAL ANIMALS WITH BENIGN TUMOBS 8 13
 

TOTAL BENIGN TOMOHS 8 13
 

TOTAL ANIMALS WITH MALIGNANT TDMOBS 15 HH
 

TOTAL MALIGNANT TUMORS 17 51
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 1 4
 

TOTAL SECONDARY TUMORS 1 H
 

TOTAL ANIMALS WITH TUMOBS UNCERTAIN­ BENIGN OR MALIGNANT
 

TOTAL UNCEBTAIN TUMORS
 

TOTAL ANIMALS KITH TUMORS DNCERTAIN­ PBIHAHY OR METASTATIC 1
 

TOTAL UNCERTAIN TUMORS 1
 

* PRIHARY TUHORS: AIL TUMORS EXCEPT SECONDARY TUHOHS
 * SECONDARY TUMORS: HETASTATIC TDHORS OR TOHORS INVASIVE INTO AN ADJACENT ORGAN
 

81
 
TABLE 83.

 

SUMMARY OF TH INCIDENCE OF NEOPLASMS IN FEMALE MICE FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

ANIMALS INITIALLY IN STUDY
 

ANIHALS HISSING
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOP ATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SOBCOT TISSUE
 

FIBROSARCOMA
 

LEIOMYOSARCOMA
 

RESPIRATORY SYSTEM
 

tLUNG
 

HBPATOCELLULAR CARCINOMA, BETAST
 

ALVEOLAR/BRONCHIOLAR ADENTMA
 

ALVEOLAR/BRONCHIOLAR CARCINOMA
 

HEMATOPOI^TIC SYSTEM
 

*HULTIPLE ORGANS
 

MALIGNANT LYMPHOMA, NOS
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

*SPLEEN
 

HEMANGIOSARCOMA
 

BALIG. LYMPHOMA, HISTIOCYTIC TYPE
 

ILYMPH NODE
 

MALIGNANT LYMPHOMA, NOS
 

tTHYHOS
 

MALIGNANT LYMPHOMA, NOS
 

CIRCULATORY SYSTEM
 

NQNE _ _
 

LOW DOSE
 

CONTROL
 

50
 

48
 

U7
 

(«8)
 

1 (2X)
 

(1*6)
 

3 (7X)
 

1 (2%)
 

(<48)
 

1 (2%)
 

2 (<»%)
 

(<46)
 

1 (2%)
 

1 (2*)
 

(39)
 

(3D
 

1 (3%)
 

LOW DOSE
 

50
 

2
 

U5
 

143
 

(«5)
 

2 (KX)
 

(42)
 

2 (5?)
 

1 (2X)
 

3 (7%)
 

(K5)
 

1 (2%)
 

(UO)
 

(")
 

1 (3X)
 

d7)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED HICROSCOPICRLLY
 

* NUMBER OF ANIMALS NECROPSIED
 

82
 
B3. FEMALE MICE (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

DIGESTIVE SYSTEM
 

#LIVER
 

HEPATOCELLULAR CARCINOMA
 

FIBROSARCOMA
 

*GALLBLADDER
 

PAPILLARY AD3NOMA
 

#STOMACH
 

SQUAMOOS CELL PAPILLOMA
 

*COLON
 

LEIOMYOSARCOMA
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

#PITUITARY
 

CARCINOB A,NOS
 

ADENOMA, NOS
 

#ADRENAL
 

CORTICAL ADENOMA
 

PHEOCHROMOCYTOMA
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND
 ADENOCAHCINOMA, NOS
 

tUTEHUS
 

LEIOMYOSABCOMA
 

ENDOMETRIAL STROMAL POLYP
 

#OVARY
 

TUBULAR ADENOMA
 

NFRVOUS SYSTEM
 

NONE
 

LOW DOSE
 

CONTROL
 

(47)
 

1 (2X)
 

1 (2%)
 

(U8)
 

(<*«)


1 (2%)
 

(10)
 

1 (3%)
 

(42)
 

1 (2%)
 

2 (5%)
 

C*5)
 

1 (2%)
 

(<*8)
 

(U5)
 

1 (2%)
 

3 (7«)
 

(<*5)
 

1 (2%)
 

LOW DOSE
 

(43)
 

37 (86»)
 

(45)
 

1 (2%)
 

(37)
 

(37)
 

(21)
 

1 (3%)
 

(40)
 

1 (3«)
 

3 (8«)
 

(45)
 

1 (2%)
 

(3«)
 

(39)
 

# SOMBER OF ANIMALS WITH TISSUE EXAMINED MICHOSCOPICA LLY
 * NUMBER OF ANIMALS NECHOPSIED
 

83
 
B3. FEMALE MICE (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

SPECIAL SENSE ORGAN S 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONE 

ANIHA L DISPOSITION SUMMAR Y 

ANIMALS INITIALLY IN STUDY 

NATURAL DEATHS 

MORIBUN D SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIMA L MISSING 

50 

6 

2 

5 

37 

50 

10 

3 

35 

2 

a iNciaess ifliaLizEE-Miaiis . 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

84
 
B3. FEMALE MICE (LOW DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

TUMOR SUMHARY
 

TOTAL ANIMALS HITH PRIMARY TUMORS* 20 38
 

TOTAL PRIMARY TUMORS 2U 52
 

TOTAL ANIMALS WITH BENIGN TUMORS 11 7
 

TOTAL BENIGN TUMORS 11 7
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 11 37
 

TOTAL MALIGNANT TUMORS 13 l»5
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2
 

TOTAL SECONDARY TUMORS 2
 

TOTAL ANIHALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS HITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 * SECONDARY TUMORS: HETASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

85
 
TABLE B4.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE FED 

3-AMINO-9 ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

J

 TM '" •" " ~'1: L. J- •" J- -nr ILH r ­ ­ _ r "• 

HIGH DOSE 

CONTROL HIGH DOSE 

ANIMALS INITIALLY IN STUDY 50 50 

ANIMALS HISSING 1 

ANIHALS NECBOPSIED 50 i»9 

ANIMALS EXAMINED HI STOP ATHOLOGIC ALLY 50 <»9 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE (50) (49)
 

SARCOMA, NOS 1 (2%)
 

RESEIBATOBY SYSTEM
 

#LUNG (50) (48)
 

HEPATOCELLOLAfi CARCINOMA, METAST 5 (10X)
 

ALVEOLAR/BBONCHIOLAR ADENOMA 2 (<»X) 3 (6%)
 

ALVEOLAR/BBONCHIOLAR CARCINOMA 1 (2%) 1 (2%)
 

HEMATCPOIETIC SYSTEM
 

*MDITIPLE ORGANS (50) <U9)
 

MALIGNANT LYMPHOMA. NOS 2 («%)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 2 (<»X)
 

*HEMATOPOIETIC SYSTEM (50) (t9)
 

NEOPLASM, NOS 1 (2X)
 

*LIVER (50) (49)
 

KUPFFER-CELL SARCOMA 3 (6X)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

tLIVEE (50) (49)
 

HEPATOCELLULAB CARCINOMA 1 12%) 43 188XJ_
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

86 
B4. FEMALE MICE (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

#STOBACH
 

SCUABOUS CELL PAPILLOBA
 

OBINABY SYSTEM
 

•KIDNEY
 

TUBOLAB-CELL ADENOHA
 

dUBINARY BLADDER
 

THANSITICHAL-CELL PAPILLOHA
 

ENCOCBINE SYSTEM
 

#PITUITARY
 

ADENOMA, NOS
 

CHEOMOPHOBE ADENOMA
 

REEBODUCTIVE SYSTEM
 

#OVARY/OVIDUCT
 

PAPILLARY ADENOMA
 

NEBVCUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

*HAEDEFIAN GLAND
 

PAPILLARY ADENOMA
 

MUSCOLOSKEIETAL SYSTEM
 

NONE
 

BOCY CAVITIES
 

*BODY CAVITIES
 

MESOTHELIOMA. MALIGNANT
 

HIGH DOSE
 

CONTROL
 

(49)
 

(50)
 

(48)
 

(42)
 

1 (2%)
 

2 (5X)
 

(47)
 

1 (2X)
 

(50)
 

1 (2%)
 

(50)
 

HIGH DOSE
 

(44)
 

1 (2X)
 

(45)
 

1 (2X)
 

C*3)
 

1 (2»)
 

(33)
 

(35)
 

(49)
 

(49)
 

1_- I1S1
 

t NUMBEB OF ANIMALS WITH TISSUE EXAMINED MICBOSCOPICALLY
 

* NUMBEB OF ANIMALS NECBOPSIED
 

87
 
B4. FEMALE MICE (HIGH DOSE AND CONTROL): NEOPLASMS (CONTINUED) 

HIGH DOSE
 

CONTROL
 

ALI OTHER SYSTEMS
 

NONE
 

ANIBA1 DISPOSITION SUMMARY 

ANIHALS INITIALLY IN STUDY 

NATURAL DEATHS 

MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIMAL HISSING 

50 

2 

10 

38 

9 INCLUDES AUTOLYZED ANIMALS 

TUMOR SUHBARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 10
 

TOTAL PRIMARY TUMORS 11
 

TOTAL ANIMALS WITH BENIGN TUMORS 7
 

TOTAL BENIGN TUMORS 7
 

TOTAL ANIMALS WITH MALIGNANT TUMORS tt
 

TOTAL MALIGNANT TDMORS 4
 

TOTAL ANIMALS WITH SECONDARY TUMORS* I
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIHALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIHALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUHORS
 

HIGH DOSE
 

50
 

11
 

2
 

5
 

31
 

1
 

46
 

58
 

6
 

6
 

44
 

51
 

5
 

5
 

1
 

1
 

* PRIMARY TUMORS: ALL TUHOHS EXCEPT SECONDARY TOflOBS
 * SECONDARY TUMORS: HETASTATIC TUHORS OB TUHOES INVASIVE INTO All ADJACENT ORGAN
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS FED 3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

IN THE DIET
 

89
 

TABLE C1. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

LOW

CON

 DOSE 

TROL LOW DOSE 

ANIMALS INITIALLY IN STUDY 

ANIMALS MISSING 

ANIMALS NECROPSIED 

ANIMALS EXAMINED HISTOP ATHOLOGICALLY 

50 

46 

46 

50 

3 

44 

43 

INTEGUMENTARY SYSTEM
 

*SKIN (46) (44)
 

INFLAMMATION, NOS 1 (2*)
 

DLCER, NOS 2 (5X)
 

*SUBCUT TISSUE (46) (44)
 

FIBROSIS 1 (2%)
 

RESPIRATORY SYSTEM
 

tTRACHEA (U5) (40)
 

INFLAMMATION, NOS 9 (20%) 0 (20%)
 

INFLAMMATION, CHRONIC 10 (22%)
 

ILUNG/BRONCHUS (t6) (43)
 

BBONCHIECTASIS 2 (5%)
 

INFLAMMATION, NOS 1 (2%)
 

INFLAMMATION, CHRONIC 8 (17%)
 

fBRONCHIAL MUCOUS GLA (46) (43)
 

ABSCESS, NOS 1 (2%)
 

NECROSIS, NOS 1 (2%)
 

HYPERPLASIA, ADBNOMATOUS 1 (2%)
 

tLUNG/BRONCHIOLE (46) (43)
 

IHFLAHHATION, NOS 1 (2%) 5 (12%)
 

INFLAMMATION, FOCAL 1 (2%)
 

tLUNG (46) (43)
 

ATELECTASIS 1 (2%)
 

CONGESTION, NOS 1 (2X)
 

EDEMA, NOS 1 (2%)
 

BBONCHOPNEUNONIAj. NOS 1_J2«L
 

t NUMBER OF ANIMALS HITH TISSDE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

91
 
C1. MALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

INFLAHHATICN, DOS
 

INFLAMMATION, FOCAL
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, SOPPURATIVE
 

INFLAMMATION, NECBOTIZING
 

PNEUMONIA, CHRONIC MURINE
 

INFLAMMATION, CHRONIC
 

PERIVASCOLITIS
 

HYPERPLASIA, NOS
 

HYPERPLASIA, EPITHELIAL
 

HEMATOPOIETIC SYSTEM
 

•SPLEEN
 

THROMBOSIS, NOS
 

FIBROSIS
 

FIBROSIS, FOCAL
 

INFARCT, HEALED
 

HEMOSIDEROSIS
 

RETICOLOCYTOSIS
 

HYPERPLASIA, HEMATOPOIETIC
 

HYPERPLASIA, ERYTHROID
 

HYPERPLASIA, RETICULUN CELL
 

fLYMPH NODE
 

INFLAMMATION, NOS
 

NECROSIS, CENTRAL
 

HYPERPLASIA, NOS
 

HYPERPLASIA, RETICULUM CELL
 

#MEDIASTINAL L.NODE
 

PLASMACYTOSIS
 

CIRCULATORY SYSTEM
 

"LYMPHATIC VESSELS
 

INFLAMMATION, NOS
 

tMYOCARDIOM
 

INFLAMMATION, NOS
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, CHRONIC FOCAL
 

FIBROSIS
 

LOW DOSE
 

CONTROL
 

1 (2%)
 

3 (7X)
 

1 (2S)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

5 (11X)
 

(U6)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2%)
 

12 (26X)
 

8 (17X)
 

(38)
 

1 (3*)
 

1 (3X)
 

3 (8%)
 

(38)
 

1 (3X)
 

(46)
 

1 (2%)
 

(<*6)
 

1 (2%)
 

22 (U8%)
 

3 (7«)
 

_ __ 2_.Q5Xi
 

LOW DOSE
 

1 (2*)
 

11 (268)
 

1 (2*)
 

1 (2*)
 

1 (2X)
 

(U2)
 

2 (5«)
 

4 (10S)
 

U (10S)
 

8 (19%)
 

(34)
 

1 (3«)
 

1 (3*)
 

3 (<5«)
 

(34)
 

(44)
 

(43)
 

12 (28S)
 

21 (49%)
 

17 J40*}_
 

# NUMBER 3F ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

92
 
C1. MALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

_. 1 .
 CONTROL LOW DOSE
 

* AORTA («6) (4U)
 

MINERALIZATION 1 (2%)
 

INFLAMMATION, NOS 1 (2%)
 

INFLAMMATION, CHRONIC FOCAL 1 (2%)
 

*PULHONARY ARTERY (U6) CKO
 

MINERALIZATION 5 (11%)
 

HYPERTROPHY, NOS 1 (2%)
 

DIGESTIVE SYSTEM
 

tSALIVARY GLAND (38) (38)
 

PERIVASCOLITIS 1 (3%)
 

HYPERPLASIA, FOCAL 1 (3S)
 

#LIVER (U6) (^2)
 

NECROSIS, FOCAL 3 (7%) U (10%)
 

NECROSIS, COAGOLATIVE 1 (2*)
 

METAMORPHOSIS FATTY 1 (2%) 8 (19%)
 

HYPEHPLASIA, FOCAL 23 (50%) 1« (45%)
 

HEBATOPOIESIS 1 (2%)
 

#LIVER/PERI PORTAL (46) (<*2)
 

FIBROSIS 1 (2%) 1 (2%)
 

*BILE DUCT (46) (!»4)
 

INFLAMMATION, NOS 6 (13%)
 

HYPSRPLASIA, NOS 32 (70%) 17 (39%)
 

HYPERPLASIA, FOCAL 1 (2%) 1 (2%)
 

#PANCREAS (42) (39)
 

INFLAMMATION, NOS 10 (24X) 18 (46%)
 

DBGENERATION, CYSTIC 1 (37-)
 

HYPERPLASIA, INTRADUCTAL 1 (2«) 1 (3%)
 

*PANCREATIC DUCT (U2) (39)
 

INFLAMMATION, NOS 1 (3*)
 

HYPERPLASIA, NOS 2 (5%)
 

IPANCREATIC ACINUS («2) (39)
 

ATROPHY, NOS t (10%) 3 (8%)
 

HYPERPLASIA, FOCAL 1 (3%)
 

tSTOMACH (U5) (t2)
 

EPIDERMAL INCLUSION CYST 1_12«1___
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

93
 
C1. MALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

INFLAMMATION, NOS 7 (17%)
 

ULCER, NOS 2 (4%)
 

HYPERPLASIA, NOS 6 (13%) 3 (7%)
 

HYPERPLASIA, FOCAL 1 (2%)
 

HYPERKERATCSIS 1 (2%) 2 (5%)
 

ACANTHOSIS 1 (2X) 3 (7X)
 

tSHALL INTESTINE (43) (41)
 

INFLAMMATION, FOCAL 1 (2X)
 

#PEYERS PATCH (43) (41)
 

HYPERPLASIA, NOS 7 (16*) 5 (12X) •
 

#COLON (43) (38)
 

NEMATODIASIS 3 (7X)
 

PARASITISM 1 (3%)
 

URINARY SYSTEM
 

#KIDNEY (46) (42)
 

GLOHEROLONEPHRITIS, NOS 33 (12%) 40 (95S)
 

INFLAMMATION, INTERSTITIAL 1 (2*)
 

ABSCESS, NOS 1 (2%)
 

FIBROSIS 1 (2%)
 

tORINARY BLADDER (42) (40)
 

INFLAMMATION, NOS 1 (2«) 4 (10X)
 

HYPERPLASIA, EPITHELIAL 3 (7%) 8 (20*)
 

ENDOCRINE SYSTEM 

*PITOITARY (41) (37) 

HYPERPLASIA, NOS 3 (7%) 

HYPERPLASIA, CHROMOPHOBE-CELL 2 (5*) 

tADRENAL CORTEX (43) (43) 

HYPERTROPHY, FOCAL 

HYPERPLASIA, NOS 

1

1

 (2X) 

 (2%) 1 (2X) 

HYPERPLASIA, FOCAL 1 (2X) 

tADRENAL MEDULLA 

NECROSIS, NOS 

_ CALCIFICATION^. NOS __ _ 

(43) 

1 (2X) 

1 I2S.1 . 

(43) 

_


t NOBBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * BOMBER OF ANIMALS NECROPSIED
 

94
 
C1. MALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HYPERPLASIA, NODULAB
 

HYPERPLASIA, 80S
 

*THYBOID
 

HYPEBPLASIA, ADENOHATOtJS
 

HYPERPLASIA, C-CELL
 

IPARATHYROID
 

HYPERPLASIA, DOS
 

IPANCRSATIC ISLETS
 

HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEB
 

*MABMARY GLANC
 

GALACTOCELE
 

HYPERPLASIA, NOS
 

*PREPOTIAL GLAND
 

ABSCESS, NOS
 

HYPERPLASIA, NOS
 

#PROSTATE
 

INFLAHHATION, NOS
 

INFLAMMATION, FOCAL
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

HYPEHPLASIA, PAPILLARY
 

METAPLASIA, SQHANOtJS
 

tTESTIS
 

HIHERALIZATION
 

ATROPHY, NOS
 

ASPERMATOGENESIS
 

HYPERPLASIA, INTERSTITIAL CELL
 

fTESTIS/TDBOLE
 

MINERALIZATION
 

DEGENERATION, NOS
 

NERVOUS SYSTEM
 

NONE _ __
 

LOW DOSE
 

CONTROL
 

1 (2*)
 

6 (14%)
 

(45)
 

1 (2%)
 

1 (2%)
 

(32)
 

(42)
 

2 (5%)
 

(46)
 

5 (11%)
 

(1*6)
 

1 (2%)
 

1 (2%)
 

(45)
 

21	 (47%)
 

3 (7%)
 

5 (11%)
 

2 (4%)
 

5 (11%)
 

(45)
 

2 (4%)
 

1 (2%)
 

19 (42 X)
 

(45)
 

6 (13%)
 

LOW DOSE
 

2 (5%)
 

1 (2%)
 

(40)
 

1 (3X)
 

1 (3%)
 

(24)
 

1 (4*)
 

(39)
 

3 (8%)
 

(44)
 

1 (2X)
 

9 (20%)
 

(44)
 

1 (2%)
 

(41)
 

16 (39%)
 

2 (5%)
 

1 (2%)
 

2 (5%)
 

3 (7%)
 

(43)
 

4 (9%)
 

11 (26%)
 

14 (33%)
 

(43)
 

1 (2%)
 

5 (12%)
 

* NUMBER OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

95
 
C1. MALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

SPECIAL SENSE ORGAN S 

*EYE 

CATARACT 

*EYE/RETINA 

ATROPHY, NOS 

*EAR CANA L 

NECROSIS, NOS 

KERATIN-PEAR L FORMATIO N 

*WA X GLAN D 

KERATIN-PEAR L FORMATION 

MUSCULOSKELETA l SYSTEM 

*CARTILAGE,NO S 

CYST, NOS 

BODY CAVITIES 

NON E 

A LL OTHER SYSTEMS 

NON E 

SPECIAL MORPHOLOG Y SUMMAR Y 

ANIMA L MISSING/NO NECROPS Y 

AUTO/NECROPSY/HISTO PERF 

MJTO/NECROPSY/NO HISTO 

AUTOLYSIS/NO NECROPSY 

# NUMBE R OF ANIMAL S WITH TISSUE

* NUMBER OF ANIMAL S NECROPSIED 

LOW DOSE 

CONTROL 

(46) 

(<*6) 

(46) 

(46) 

(46) 

1 (2%) 

1 

4 

LOW DOSE 

(44) 

(44) 

(44) 

1 (2%) 

1 (2%) 

(44 ) 

1 (2%) 

(44 ) 

3 

1 

1 

3 

 EXAMINE D MICROSCOPICALLY 

96 
TABLE C2. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

ANIMALS INITIALLY IN STUDY
 

ANIHAIS NECBOPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTi"
 

*SKIN
 

INFLAHMATION, NOS
 

INFLAMMATION, NECROTIZING
 

*SUECOT TISSUE
 

ABSCESS, NOS
 

FIBEOSIS
 

METAPLASIA, OSSEOUS
 

RESEIEATOBY SYSTEM
 

#TRACHEA
 

INFLAMMATION, NOS
 

#LONG/ERONCHUS
 

BBONCHIECTASIS
 

INFLAMMATION, NOS
 

METAPLASIA, SQUAMOUS
 

#LONG
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, NECROTIZING
 

PNEUMONIA, CHBONIC MURINE
 

GBANULOMA, FOREIGN BODY
 

HYPERPLASIA, EPITHELIAL
 

HEMATCPOIETIC SYSTEM
 

tBONE MABROW
 

MEGAKARYCCYTOSIS
 

#SPLEEN
 

INFLAHMlIONijgS
 

HIGH DOSE
 

CONTROL
 

850
 

48
 

1*8
 

(i»8)
 

C*8)
 

1 (2%)
 

(US)
 

2 (4%)
 

(48)
 

1 (2*)
 

7 (15%)
 

(48)
 

4 (8%)
 

1 (2%)
 

1 (2%)
 

1 (2*)
 

(47)
 

(48)
 

- -_
 

HIGH DOSE
 

50
 

48
 

48
 

(48)
 

1 (2%)
 

3 (6*)
 

(48)
 

1 (2*)
 

1 (2%)
 

(47)
 

2 (4%)
 

(48)
 

1 (2%)
 

3 (6%)
 

1 (2X)
 

(48)
 

16 (33%)
 

2 (4%)
 

1 (2%)
 

6 (13%)
 

(46)
 

2 (4%)
 

(«B)
 

2__[4%J___
 

* NUMBEP OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

S 50 ANIMALS WEBE INITIALLY IN THE STUDY, BUT ONE ANIMAL
 

MAS FOUND TO BE A FEMALE IN A MALE G80UP.
 

97
 
C2. MALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

I NUMBER OF ANIMALS WITH TISSUE EXAHINBD MICROSCOPICALLY
 

* NUMBER OF ANIMALS HECROPSIED
 

98
 
C2. MALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

•PANCREATIC ACINUS 

HYPERTROPHY, NOS 

HYPERTROPHY, FOCAL 

HYPEEPLASIA, FOCAL 

(46) 

1 (2%) 

(43) 

1

1

3

 (2X) 

 (2X) 

 (7%) 

tESOPHAGOS 

DYSPLASIA. NOS 

(<45) 

1 (2%) 

(38) 

•STOMACH 

INFLAMMATION, NOS 

INFLAMMATION, FOCAL 

HYPEEPLASIA, FOCAL 

HYPFRPLASIA, BASAL CELL 

HYPEBKERATOSIS 

ACANTHOSIS 

(48) 

1 (2%) 

1 (2*) 

2 (4%) 

2 (4%) 

(42) 

1

3

4

1

1 (2%) 

 (2X) 

 (7X) 

 (10%) 

 (2X) 

•PEYERS PATCH 

HYPERPLASIA, NOS 

(46) 

12 (26%) 

(46) 

12 (26%) 

tILEUM 

INFLAMMATION, NOS 

(46) 

2 (4») 

(46) 

•COLON 

PARASITISM 

(46) 

3 (7X) 

(38) 

1 (3%) 

URINARY SYSTEM
 

•KIDNEY	 (43) (<»8)
 

GLOHEHULONEPH8ITIS, NOS 47 (98X) 48 (100%)
 

INFLAMMATION, INTEBSTITIAL 1 (2%)
 

FIBBOSIS, DIFFUSE 6 (13X) 19 (40%)
 

GLOHERDLOSCLEROSIS, NOS 2 (4%)
 

HIPEBPLASIA, TDBDLAB CELL 1 (2%)
 

HYPERPLASIA, EPITHELIAL 1 (2%)
 

HYPERPLASIA, ADBHOHATOOS 1 (2%)
 

IORINARY BLADDER (43) C»2)
 

HYPERPLASIA, EPITHELIAL 1 (2X) 2 (5%)
 

E»rOCBINE SYSTEM
 

•PITUITARY	 (38) (41)
 

HYPERPLASI4, NOS .. 1 13X) .. 1 (2*L
 

* NOHBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

99
 
C2. MALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

HYPEFPLASIA, FOCAL	 2 (5X) 2 (5X)
 

•ADRENAL MEDULLA	 (47) (48)
 

HYPERPLASIA, NODULAR 1 (2%) 2 (4X)
 

HYPERPLASIA, FOCAL 4 (9X) 1 (2X)
 

•THYROID	 (48) (47)
 

LYMPHOCYTIC I NFLAHMATORY INFILT8 1 (2X)
 

HYPERPLASIA, C-CELL 3 (6X) 1 (2X)
 

•PARATHYROID	 (28) (23)
 

HYPERPLASIA, BOS 1 (4X) 3 (13X)
 

•PANCREATIC	 ISLETS (46) (43)
 

HYPERPLASIA, NOS 1 (2X)
 

HEEECCUCTIVE SYSTEM
 

*MAMMARY GLAND (48) (48)
 

GALACTOCELE 2 (4«)
 

HYPERPLASIA, NOS 4 (8X) 9 (19X)
 

*PREPOTIAL GLAND (48) (48)
 

NECROSIS, NOS 1 (2X)
 

•PROSTATE	 (44) (44)
 

INFLAMMATION, NOS 17 (39X) 20 (45X)
 

*SEMINAL VESICLE (48) (48)
 

HYPERPLASIA, PAPILLARY 1 (2X)
 

•TESTIS	 (47) (47)
 

MINERALIZATION 1 (2X) 1 (2X)
 

ATROPHY, NOS 6 (13X) 7 (15X)
 

HYPERPLASIA, NOS 2 (4X)
 

HYPERPLASIA, INTERSTITIAL CELL 3 (6X) 4 (9X)
 

•TES1IS/TOBULE	 (47) (47)
 

MINERALIZATION 2 (4%)
 

DEGENERATION, NOS 2 (4X)
 

NEBVCDS SYSTEM 

•ERAIN (48) (47) 

MINERALIZATION _ r _ ... . .. _T . 1_IZJL 

» NUMBER OF ANIMALS HITH TISSUE EXABINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

100
 
C2. MALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

SPECIAL SENSE ORGANS
 

NONE
 

NUSCULOSKELETAL SYSTEM
 

*BONE
 

OSTEOSCLEEOSIS
 

BODY CAVITIES
 

NONE
 

All OTHER SYSTEMS
 

CMENTUM
 

NECROSIS, NOS
 

NECROSIS, PAT
 

SPECIAL MORPHOLOGY SUMMARY
 

AUTOLYSIS/NO NECROPSY
 

HIGH DOSE
 

CONTROL HIGH DOSE
 

C»8) (U8)
 

1 (2%)
 

1
 

2
 

1 2
 

f NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OP ANIHALS NECROPSIED
 

101
 
TABLE C3. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

LOW DOSE
 

CONTROL LOW DOSE
 

ANIMALS INITIALLY IN STUDY 50 50
 

ANIMALS NECROPSIED 49 4U
 

ANIMALS EXAMINED HISTOPATHO LOGICALLY 49 1*3
 

INTEGUMENTARY SYSTEM
 

*SKIN (49) (HH)
 

INFLAMMATION, NOS 1 (2%)
 

ULCER, NOS 2 (5X)
 

RESPIRATORY SYSTEM 

tTRACHEA (48) («2) 

INFLAMMATION, NOS 9 (19%) 3 (7%) 

INFLAMMATION, ACUTE/CHRONIC 1 (2%) 

INFLAMMATION, CHRONIC 10 (21 %) 

POLYP, INFLAMMATORY 1 (2%) 

ILUNG/BRONCHUS (49) (43) 

BBONCKIECTASIS 

INFLAMMATION, NOS 

1

1

 (2%) 

 (2%) 

1

1

 (2%) 

 (2%) 

INFLAMMATION, CHRONIC 9 (18%) 

fLUNG/BRONCHIOLE (49) (43) 

INFLAMMATION, NOS 1 (2%) 

tLONG 

EDEMA, NOS 

(«9) (43) 

1 (2%) 

INFLAMMATION, NOS 1 (2%) 

INFLAMMATION, FOCAL 7 (14%) 

INFLAMMATION, INTERSTITIAL 2 (4%) 14 (33%) 

FIBROSIS, DIFFUSE 1 (2%) 

PEFIVASCULITIS 6 (12%) 

METAMORPHOSIS FATTY 1 (2%) 

HYPERPLASIA, NOS 1 (2%) 

HYPERPLASIA, EPITHELIAL 1 (2%) 

HYPERPLASIA, ALVEOLAR EPITHELIUM 2 (5%) 

tLUNG/ALVEOLI (U9) C*3) 

FIBROSISi_FOCAL 2_15%L 

t NUMBER OF ANIHALS HITH TISSUE EXAMINED HICROSCOPICALLY
 * NUMBER OF ANIHALS NECROPSIED
 

102 
C3. FEMALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

HENATOPOIETIC SYSTEM 

fSPLEBN (49) (43) 

INFLAMMATION, NOS 1 (2%) 

INFARCT, NOS 1 (2%) 

HEMOSIDEROSIS 3 (1%) 

HYPERPLASIA, NOS 

HYPERPLASIA. HEM ATOPOIETIC 

1

3

 (2%) 

 (6%) 19 (44*) 

HYPERPLASIA, ERYTHROID 17 (35%) 16 (31%) 

HYPERPLASIA, PLASMA CELL 1 (2%) 

HYPERPLASIA, RETICULUM CELL 11 (22%) 

EHYTHROPOIESIS 1 (2«) 

#LYMPH NODE 

INFLAHHATION, NOS 

CM)

3 (7%) 

1 1 f, 

h (17X) 

HYPERPLASIA, NOS 

PLASHACYTOSIS 

2

3

 (5*) 

 (7%) 

3

1

 (8%) 

 (3%) 

HYPERPLASIA, HEM ATOPOIETIC 2 (6%) 

HYPERPLASIA, PLASMA CELL 1 (2%) 1 (3%) 

HYPERPLASIA, LYMPHOID 2 (6«) 

tMEDIASTINAL L.NODE 

PLASMACYTOSIS 

I'M) (36) 

3 (8%) 

CIRCULATORY SYSTEM
 

JMYOCARDIOM («9) (43)
 

INFLAMMATION, NOS 1 (2%) 11 (26*)
 

INFLAMMATION, INTERSTITIAL 24 (49%) 16 (37%)
 

FIBROSIS 5 (10*) 4 (9%)
 

*POLHONARY ARTERY (49) (44)
 MINERALIZATION 1 (2%)
 

*PORTAL VEIN (49) (44)
 

THROMBUS, MORAL 1 (2%)
 

DIGESTIVE SYSTEM
 

tLIVER (49) (43)
 

INFLAHHATION, NOS 3 (7X)
 

_ F.IBBQSIS __ 1.J221 2_J5%L
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED HICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

103
 
C3. FEMALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

PERIVASCDLITIS 

DEGENERATION, NOS 

NECROSIS, FOCAL 

NECROSIS, COAGOtATIVE 

METAMORPHOSIS FATTY 

HYPERPLASIA, NODULAR 

HYPERPLASIA, FOCAL 

ANGIECTASIS 

HEMATOPOIESIS 

*BILE DUCT 

INFLAMMATION, NOS 

HYPERPLASIA , NOS 

tPANCREAS 

INFLAMMATION , NOS 

HYPERPLASIA , INTRADUCTA L 

tPANCREATIC DUCT 

HYPERPLASIA, NOS 

#PANCREATIC ACINUS 

ATROPHY , NOS 

tSTOHACH 

INFLAMMATION ,

INFLAMMATION ,

HYPERPLASIA ,

HYPERPLASIA,

HYPERPLASIA ,

ACANTHOSI S 

•GASTRIC MUCOS A 

HYPERPLASIA ,

fPEYERS PATCH 

HYPERPLASIA ,

*COLON 

NEBATODIASI S 

PARASITISM 

URINAR Y SYSTEM 

((KIDNEY 

 NOS 

 FOCAL 

 NOS 

 EPITHELIAL 

 FOCAL 

 NOS 

 NOS 

iSIERQNEPHROSIS 

LOW DOSE 

CONTROL LOW DOSE 

1 (2X) 

1 (2%) 

4 (8X) 3 (7«) 

2 (4X) 3 (7X) 

1 (2%) 5 (12X) 

1 (2X) 

22 (45X) 25 (58X) 

1 (2%) 1 (2%) 

1 (2%) 

(49) (44) 

5 (10X) 

27 (55X) 15 (34X) 

(146) (39) 

7 (15%) 14 (36X) 

2 (5X) 

(1*6) (39) 

1 (2X) 2 (5X) 

(46) (39) 

2 (UX ) 4 (10X ) 

(48) (41) 

2 (UX ) 2 (5X) 

2 («*) 

1 (2%) 

1 (2X) 

2 (555) 

2 (5%) 

(«8) (41 ) 

1 (2%) 

(U7) (40 ) 

6 (13X) 4 (10% ) 

(43) (35) 

3 (7X) 

2 (6%) 

(49) (43) 

1_12SL l.UZL 

f	 NUMBER 3F ANIHALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSIED
 

104 
C3. FEMALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE 

GLOMERULONEPHRITIS, NOS 

PYELONEPHRITIS, NOS 

INFLAHHATICN, INTERSTITIAL 

GLOMERULONEPHRITIS, HBMBRANOOS 

INFLAMHATION, CHRONIC 

HYPERPLASIA, HER ATOPOIETIC 

HYPERPLASIA, ERYTHROTD 

33

1

1

1

 (67%) 

 (2X) 

 (2%) 

 (2%) 

28

1

8

1

1

 (65%) 

 (2%) 

 (19%) 

 (2%) 

 (2%) 

tKIDNEY/CORTEX 

CYST, NOS 

(<»9) (43) 

1 (2%) 

#URINARY BLADDER 

INFLAMMATION, NOS 

HYPERPLASIA, EPITHELIAL 

(i»1) 

1 (2X) 

(39) 

U (10%) 

ENDOCRINE SYSTEM
 

*PITUITARY (<*3) (30)
 

HYPERPLASIA, NOS 2 (5%)
 

HYPERPLASIA, CHROMOPHOBE-CELL 1 (2«)
 

*ADRENAL («6) (U3)
 

METAMORPHOSIS FATTY 1 (2%)
 

HYPERPLASIA, FOCAL 1 (2%)
 

#ADRENAL CORTEX (U6) ("?)
 

FIBRIN BODY 1 (2%)
 

NODDLE 1 (2%)
 

HYPERPLASIA, NOS 7 (1?X) 1 (2*)
 

HYPERPLASIA, FOCAL 3 (7«)
 

fADRENAL MEDULLA (U6) C*3)
 

HYPERPLASIA, NOS U (9%)
 

*THYBOID («7) C*0)
 

HYPERPLASIA, FOLLICULAR-CELL 1 (2%)
 

fPANCREATIC ISLETS («6) (39)
 

HYPERPLASIA, NOS 1 (2%)
 

REPRODUCTIVE SYSTEM
 

*MAMHARY GLANC («9) (i*U)
 

GALACTOCELE . __5 110*1 . 3 (7%L
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

105
 
C3. FEMALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

HYPERPLASIA, NOS 17 (35*) 13 (30%)
 

HYPERPLASIA, PAPILLARY 1 (2%)
 

*CLITORAL GLAND (U9) (K14)
 

ABSCESS, NOS 1 (2%)
 

*VAGINA (U9) (UU)
 

NECROSIS, HEMORRHAGIC 1 (2%)
 

HYPERPLASIA, NOS 1 (2%)
 

fOTEHUS (08) («3)
 

HYDROBETRA 3 (6%)
 

HEMORRHAGE 3 (7%)
 

INFLAHHATION, SOPPORATIVE 1 (2%)
 

ABSCESS, NOS 2 (0%) 1 (2%)
 

HYPERPLASIA, FOCAL 1 (2%)
 

HYPERPLASIA, ADENOMATOOS 5 (10%) 1 (7«)
 

POLYP, INFLAHBATORY 1 (2X)
 

*OTEROS/ENDOHfTRIOB (U8) (43)
 

INFLABBATICN, NOS 1U (29%) 12 (28X)
 

IMFLAMHATION, FOCAL 1 (2%)
 

INFLABMATION, SOPPORATIVE 2 (HI) 7 (16%)
 

HIPERPLASIA, NOS 1 (2%)
 

HYPERPLASIA, FOCAL 1 (2%)
 

HYPERPLASIA, CYSTIC 2 <«%) 1 (2%)
 

HYPERPLASIA, ADENOMATOtJS 1 (2%)
 

tOVARY/OVIDOCT (U8) (43)
 

INFLABHATICN, NOS 1 (2«) 1 (2%)
 

#OVARY (i»7) (U3)
 

CYST, NOS U (9«) 1 (2%)
 

INFLAHHATION, FOCAL GRANULOBATOO 1 (2X)
 

HYPERPLASIA, INTERSTITIAL CELL 1 (2X)
 

NERVOUS SYSTEB
 

NONE
 

SPECIAL SENSE ORGANS
 

*EAR CANAL («9) (Hit)
 

KERATIN-PEARL FORHATION 1 I2XL_ __
 

* NUMBER OF ANIHALS WITH TISSUE EXAHINED MICROSCOPICALLY
 

* NOHBER OF ANIMALS NECHOPSIED
 

106
 
C3. FEMALE RATS (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

MUSCOLOSKELETAL SYSTEM 

HONii 

BODY CAVITIES 

NOME 

ALL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMMAR Y 

AUTO/N3CROPSY/NO HISTO 1 

AUTOLYSIS/HO NECROPSY 1 6 

f NDHBEB OF ANIHALS WITH TISSUE EXAHIHED HICBOSCOPICALLY
 

* ROBBER OF ANIHAtS HECROPSIED
 

107 
TABLE C4. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIOE IN THE DIET (HIGH DOSE AND CONTROL) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

ANIMALS INITIALLY IN STUDY 50 50
 

ANIBALS NECBOPSIBD 50 49
 

ANIHALS EXAHINED HISTOPATHOLOGIC ALLY 50 i»9
 

INTIGOHENTARY SYSTEM
 

*SKIN (50) (<I9)
 

EPIDERHAL INCLUSION CYST 2 (<»*)
 

INFLAMMATION, NOS 1 (2%)
 

*SUBCOT TISSOE (50) (49)
 

MINEBALIZATION 1 (2X)
 

ABSCESS, NOS 1 (2%)
 

BESFIFATOFY SYSTEM
 

ITPACHEA (149) (45)
 

INFLAHMATION, NOS 1 (2»)
 

ILONG/ERONCHOS (50) (48)
 

INFLAHMATION, NOS 3 (6*)
 

INFLAMMATION, FOCAL 2 (4X)
 

ILUNG (50) (48)
 

INFLAMMATION, INTERSTITIAL 6 (12X) 13 (27%)
 

FIBROSIS, FOCAL 1 (2%)
 

HYPERPLASIA, EPITHELIAL 1 (2%) 3 (6*)
 

METAPLASIA, SQUAMOUS 1 (2*)
 

HEMATCPOIETIC SYSTEM 

#EONE MARROW 

OSTEOSCLEBOSIS 

(i»6) 

1 (2X) 

(45) 

tSPLEEN 

INFLAHMATION, NOS 

FIBROSIS „__ 

(«8) (48) 

5

1

 (10H) 

 12») 

* NUMBEB OF ANIMALS WITH TISSUE EXAHINED MICROSCOPICALLY
 

* NOMBEB OF ANIHALS NECBOPSIED
 

108
 
C4. FEMALE RATSfHIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

HEMOSIDEEOSIS 12 (25X) 18 (38X)
 

HYPERPLASIA, HEHATOPOIETIC 25 (52X) 21 (4HX)
 

HYPERPLASIA, ERYTHROID 19 (40X) 28 (58X)
 

•SPLENIC CAPSOLE (48) (48)
 

HEHOEEHAGIC CYST 1 (2X)
 

#LYMPH NODE (47) (27)
 

INFLAMMATION, NOS 2 (7X)
 

HYPERPLASIA, NOS 3 (11X)
 

PLASMACYTOSIS 1 (2X) 1 C«X)
 

HYPERPLASIA, LYMPHOID » 0«) 3 (11X)
 

CIBCUIATOHY SYSTEM
 

•MYOCARDIUM	 (50) («8)
 

INFLAMMATION, NOS 1 (2X)
 

INFLAMMATION, INTERSTITIAL 23 (U6X) 22 (46X)
 

FIBROSIS 15 (30X) 13 (27X)
 

•ENDOCARDIUM	 (50) (US)
 

INFLAMMATION, NOS 1 (2X)
 

DIGESTIVE SYSTEM
 

ISALIVAEY GLAND (50) (HH)
 

HYPERPLASIA, FOCAL 1 (2X)
 

ILIVER (50) (US)
 

NECROSIS, FOCAL 2 (HX) 4 (8X)
 

NECROSIS, COAGULATIVE 1 (2X)
 

METAMORPHOSIS FATTY 6 (12X) 6 (13X)
 

CYTOPLASMIC VACUOLIZATION 3 <6X)
 

HYPERPLASIA, FOCAL 38 (76%) 16 (33X) 

HYPERPLASIA, ERYTHROID 

HEMATOPOIESIS 

1

2

 (2X) 

(>4%) 1 (2X) 

•LIVER/PEHIPOHTAL (50) (48) 

FIBROSIS 1 (2X) 

fLIVER/HEPATOCYTES (50) (48) 

DEGENERATION, NOS 1 (2X) 

*EILE DDCT (50) (49) 

INFLAMMATION. NOS l_I2Si__ 2 (<**) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

109
 
C4. FEMALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE 

CONTROL HIGH DOSE 

HYPEFPLASIA, NOS 

HYPERPLASIA, FOCAL 

32 (64X) 

1 (2X) 

27 (55X) 

•PANCREAS 

INFLAMMATION, NOS 

(48) 

6 (13X) 

(47) 

12(26%) 

IPANCREATIC DUCT 

HYPERPLASIA, NOS 

(48) (47) 

1 (2X) 

•STOMACH 

INFLAMMATION, NOS 

DEGENERATION, NOS 

HYPERPLASIA. NOS 

HYPEEPLASIA, FOCAL 

HYPEEPLASIA, BASAL CELL 

HYPEEKEBATOSIS 

ACANTHOSIS 

(48) 

1 (2X) 

2 (4X) 

(46) 

3 (7X) 

1 (2X) 

1 (2X) 

2 (4X) 

6 (13X) 

5 (11X) 

6 (13X) 

fPEYERS PATCH 

HYPERPLASIA, NOS 

(48) 

15 (31X) 

(46) 

6 (13X) 

tCOICN 

PARASITISM 

(46) 

2 (4X) 

(28) 

UBINAEY SYSTEM
 

IKIDNEY (50) (49)
 

GLOMEEOLONEPHRITIS, NOS 43 (86X) 45 (92X)
 

FIBBOSIS, DIFFUSE 1 (2X)
 

tDGINARY BLADDER (<»6) (43)
 

HYPERPLASIA, EPITHELIAL 6 (14X)
 

ENEOCRINE SYSTEH
 

fPITOITARY (40) (39)
 

PEfilVASCOLITIS 1 (3X)
 

HYPERPLASIA, FOCAL 3 (8X)
 

IADRBNAL (49) (47)
 

METAMORPHOSIS FATTY 1 (2X)
 

tADRENAL CORTEX (49) (47)
 

LIPOIDOSfS 1 (2XJ,.., . .
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS SECROPSIED
 

110
 
C4. FEMALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

HYPERTROPHY, FOCAL 1 (2%)
 

*ADRENAL MEDOLLA (19) (47)
 

HYPERPLASIA, NODULAR 3 (6%) 1 (2%)
 

HYPERPLASIA, FOCAL 3 (6%)
 

fTHYBOID (45) (44)
 

CYSTIC FOLLICLES 1 (2%)
 

HYPERPLASIA, PAPILLARY 2 (5%)
 

HYPEBPLASIA, C-CELL 1 (2%) 1 (2%)
 

tPANCREATIC ISLETS («»8) (47)
 

HYPERPLASIA, BOS 1 (2%)
 

REPRODUCTIVE SYSTEB 

*HAHBAHY GLAND 

GALACTOCELE 

HYPEEPLASIA, NOS 

(50) 

16

8

 (32%) 

 (16%) 

(49) 

6

13

 (12%) 

 (27%) 

tDTERDS 

HEHORRHAGE 

HYPERPLASIA, ADENOHATOOS 

(50) 

1 (2%) 

(49) 

1 (2%) 

2 (<•%) 

f OTERUS/ENDOBETHIOH 

INFLAMMATION, NOS 

INFLAMMATION, NECROTIZING 

HYPERPLASIA, NOS 

HYPEBPLASIA, CYSTIC 

HYPBBPLASIA, ADBNOMATOOS 

(50) 

22

6

1 (2X) 

 (44%) 

 (12%) 

(49) 

22 (45%) 

1 (2%) 

8 (16%) 

2 <t%) 

•OVARY/OVIDUCT 

INFLAMMATION, NOS 

INFLAMMATION, SOPPORATIVE 

HYPEBPLASIA, NOS 

(50) 

10

2

 (20%) 

 (U%) 

(49) 

2 («%> 

1 (2%) 

1 (2%) 

tOVARY 

CYST, NOS 

HYPEBPLASIA, NOS 

(49) 

8 (16%) 

(«7) 

6

3

 (13%) 

 (6%) 

SEBVCOS SYSTEM
 

NONE _ _
 

t NOHBBB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

111
 
C4. FEMALE RATS (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

SPECIAL SENSE ORGANS
 

*EYE
 

CATARACT
 

*EYE/RETINA
 

ATROPHY, NOS
 

*HARDEBIAN GLAND
 

HYPEBPLASIA, NOS
 

MOSCUIOSKEIETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL CTHIE SYSTEHS
 

OHENTUM
 

NECBOSIS, FAT
 

SPECIAL MORPHOLOGY SUMMARY
 

AOTO/NECROPSY/HISTO PERT
 

AOTOIYSIS/NO NECROPSY
 

HIGH DOSE
 

CONTROL HIGH DOSE
 

(50)	 (49)
 

1 (2*)
 

(50)	 (49)
 

1 (2«) 1 (2X)
 

(50)	 (49)
 

1 (2S) 1 (2%)
 

1	 2
 

1
 

1
 

*	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * SOMBER OF ANIMALS NECEOPSIED
 

112
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE FED 3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

IN THE DIET
 

113
 

TABLE 01. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE FED 

3 AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

LOW DOSE 

CONTROL LOW DOSE 

ANIMALS INITIAILY IN STUDY 50 51 

ANIMALS MISSING 1 

ANIMALS NECROPSIED 48 414 

ANIMALS EXAMINED HISTOP ATHOLOGICALLY 48 44 

INTEGUMENTARY SYSTEM
 

*SKIN
 

FIBROSIS
 

ALOPECIA
 

*SUBCUT TISSUE
 

ABSCESS, NOS
 

NECROSIS, NOS
 

RESPIRATORY SYSTEM
 

•LUNG/BRONCHUS
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

*LUNG
 

EDEMA, NOS
 

HEMORRHAGE
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

INFLAMMATION, INTERSTITIAL
 

HYPERPLASIA, EPITHELIAL
 

#LUNG/ALVEOLI
 

INFLAMMATION, FOCAL
 

FIBROSIS, FOCAL
 

HEMATOPOIETIC SYSTEM
 

•SPLEEN
 

INFLAMMATION, NOS
 

HYPERPLASIA. NOS
 

(US) 

1 (2X) 

1 (2%) 

(44) 

(48) 

1 (2X) 

(44) 

2 (SS) 

(K8) 

1 (2%) 

1 (2%) 

(42) 

2 (5%) 

(«8) 

1 (2%) 

14 (29%) 

2 (4%) 

(12) 

1 (2%) 

1 (2%) 

1 (2*) 

fl (19%) 

(48) 

2 (4%) 

1 (2%) 

(42) 

(47) 

1 (2%) 

2_J4S1 _. 

C*3) 

13 i30«J. 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIFD
 

115
 
01. MALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

HYPBRPLASIA. HEBATOPOIETIC 

HYPERPLASIA, ERYTHROID 

HYPEHPLASIA, LYHPH3ID 

2 (4%) 

2 (UX) 

2 (i*%) 

S

5

 (12*) 

 (12%) 

#LYMPH NODE 

HEMORRHAGIC CYST 

INFLAMMATION, NOS 

DEGENERATION, CYSTIC 

HYPERPLASIA. NOS 

HYPERPLASIA, HEMATOPOIETIC 

HYPERPLASIA. RETICULUM CELL 

HYPERPLASIA, LYHPHOID 

MYELOID METAPLASIA 

(UU) 

1

13

1

2

1

2

2

 (2%) 

 (30%) 

 (2*) 

 (5X) 

 (2X) 

 (5%) 

 (5X) 

(36) 

18

1

1

2

 (50%) 

 (3%) 

 (3%) 

 (6%) 

*MEDIASTINAL L.NODE 

NECROSIS, NOS 

(41*) 

1 (2%) 

(36) 

#PANCREATIC L.NODE 

INFLAMMATION, NOS 

(UH) 

1 (2%) 

(36) 

IMESENTERIC L. NODE 

HEMORRHAGE 

INFLAMMATION, NOS 

(«(») 

1

9

 (2X) 

 (20X) 

(36) 

#THYMUS 

NECROSIS, NOS 

(3U) 

1 (3X) 

(16) 

CIRCULATORY SYSTEM
 

tHEABT/VENTRICLE (48) («2)
 

MELANIN 2 (<»X)
 

tMYOCARDIOM («8) C*2)
 

INFLAMMATION, INTERSTITIAL 2 («X)
 

FIBBOSIS 5 (10X)
 

*BLOOD VESSEL («8) (144)
 

INFLAMMATION, NOS 2 (1%) 1 (2S)
 

* AORTA (18) (44)
 

INFLAMMATION, NOS 1 (2*)
 

*PULBONARY ARTERY (08) (44)
 

.MINERALIZATION 2_1UX1
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMAL S NECROPSIFD
 

116
 
01. MALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

DIGESTIVE SYSTEM
 

iSALIVARY GLAND (it?) (U1)
 

INFLAMMATION, NOS 2 (f»X)
 

PERIVASCULAR CUFFING 1 (2X)
 

#LIVER (U8) (UU)
 

DEGENERATION, NOS 1 (2%)
 

NECROSIS, FOCAL 13 (2756) 11 (25%)
 

METAMORPHOSIS FATTY 3 (6%)
 

HYPERPLASIA, NODULAR 2 («*)
 

HYPERPLASTIC NODULE 1 (2X)
 

HYPERPLASIA, FOCAL 1 (2X)
 

HYPERPLASIA, DIFFUSE 2 (5X)
 

ANGIECTASIS 1 (2%)
 

MYELOID METAPLASIA 1 (2X)
 

ILIVER/HEPATOCYTES («8) («i»)
 

DEGENERATION, NOS 1 (2%)
 

*GALLBLADDER (US) (UU)
 

INFLAMMATION, NOS 7 (16X)
 

INFLAMMATION, FOCAL 1 (2X)
 

HYPERPLASIA, PAPILLARY U (9X)
 

*BILE DUCT (U8) (HU)
 

HYPERPLASIA, NOS 2 (5X)
 

tPANCREAS (48) C>3)
 

INFLAMMATION, NOS 7 (15X) 1 (2X)
 

INFLAMMATION, FOCAL 1 (2X)
 

DEGENERATION, CYSTIC 1 12%)
 

METAMORPHOSIS FATTY 1 (2X)
 

tPANCREATIC DUCT (t8) <43)
 

HYPERPLASIA, NOS 1 (2X)
 

tPANCREATIC ACINUS («8) (i»3)
 

HYPERTROPHY, NOS 1 (2X)
 

HYPERTROPHY, FOCAL 1 (2X)
 

HYPERPLASIA, FOCAL 1 (2X)
 

tSTOBACH C»7) (42)
 

.ISFLAHfiATIOltt fiQS __ .. 13 12SXL 2 15S1__ .
 

f NUMBER OF ANIMALS WITH TISSUE BXAHIHED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

117
 
01. MALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

tJLCER, NOS 

INFLAMMATION, FOCAL 

INFLAMMATION, INTERSTITIAL 

HYPERPLASIA, NOS 

HYPEBPLASIA, FOCAL 

HYPEBKERATOSIS 

ACANTHOSIS 

1 (2X) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

3 (6%) 

3 (6%) 

3 (7X) 

i» (10%) 

5 (12%) 

tGASTRIC MUCOSA 

HYPERPLASIA, FOCAL 

(47) 

1 (2%) 

(42) 

#PEYERS PATCH 

HYPERPLASIA, NOS 

(14 S) 

2 («*) 

(i*3) 

5 (12*) 

ilLEOB 

HEMORRHAGE 

INFLAMMATION, NOS 

(148) 

1

2

 (2%) 

 (4%) 

(43) 

#COLON 

PARASITISM 

(<45) 

1 (2%) 

(36) 

URINARY SYSTEM
 

tKIDNEY (1*7) (I*U)
 

GLOMERULONEPHRITIS, NOS 6 (13%) 7 (16%)
 

INFLAMMATION, NOS 1 (2%)
 

INFLAMMATION, INTERSTITIAL 23 (49%) 18 (1*1%)
 

IKIDNEY/TOBT7LE (U7) (UK)
 

NECROSIS, FOCAL 1 (2%)
 

#ORINARY BLADDER (48) (42)
 

INFLAMMATION, NOS U (8%)
 

HYPERPLASIA, EPITHELIAL 9 (19%) 5 (12%)
 

ENDOCRINE SYSTEM
 

*PITUITARY (12) (3U)
 

HYPERPLASIA, NOS 3 (7%)
 

HYPERPLASIA, FOCAL 3 (7%)
 

tADRENAL (45) (43

)
 

NODULE _ 2 I5%L
 

t NUMBER 3F ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

118
 
01. MALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

-


tADRENAL/CAPSOLE
 

HYPERPLASIA, NOS
 

* ADRENAL CORTEX
 

NODULE
 

HYPERTROPHY, FOCAL
 

HYPERPLASIA, NOS
 

#ADRENAL MEDULLA
 

DEGENERATION, NOS
 

•THYROID
 

LYMPHOCYTIC INFLAHBATOR Y INFILTR
 

HYPERPLASIA, PAPILLARY
 

HYPERPLASIA, FOLLICULAR-CELL
 

IPANCREATIC ISLETS
 

HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEM
 

*PREPUTIAL GLAND
 

ABSCESS, NOS
 

ITESTIS
 

HYPERPLASIA, INTERSTITIAL CELL
 

#TESTIS/TUBULE
 

DEGENERATION, NOS
 

NERVODS SYSTEM
 

#BHAIN
 

MINERALIZATION
 

#CEREBRAL CORTEX
 

MINERALIZATION
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAI SYSTEM
 

NONE
 

LOW DOSE
 

CONTROL LOW DOSE
 

(15)	 (43)
 

6 (14X)
 

(45)	 (43)
 

1 (2*) 1 (2%)
 

1 (2*) 1 (2%)
 

1 (2X)
 

(45)	 (43)
 

1 (2*)
 

(47)	 (35)
 

1 (2%)
 

1 (2X)
 

1 (2X)
 

(48)	 (43)
 

2 (UX).
 

(i»8) (44)
 

2 (US)
 

(U7) (43)
 

4 (9T.)
 

(47)	 (43)
 

4 (9%)
 

(48)	 (43)
 

1 (2%)
 

(48)	 (41)
 

3 (6%)
 

#	 NUMBER OP ANIMALS WITH TISSDE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECBOPSIED
 

119
 
D1. MALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL HORPHOLOGY SOMHABY
 

ANIMAL MISSING/NO NECROPSY 1
 

AUTO/NECROFSY/HISTO PERF 1
 

AUTOLYSIS/NO NECROPSY 2 5
 

f NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECBOPSIED
 

120
 
TABLE D2. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

ANIMALS INITIALLY IN STUDY
 

ANIHAIS HISSING
 

ANIMALS NECBOPSIED
 

ANIHALS EXAHINED HISTOPATHOLOGICALLY
 

INTIGOMENTABY SYSTEM
 

*SKIN
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

INFLAMMATION, NECBOTIZING
 

BESEIFATOBY SYSTEM
 

•LONG/BBONCHOS
 

INFLAMMATION, FOCAL
 

ILONG/BRONCHIOLB
 

INFLAMMATION, FOCAL
 

• LONG
 

INFLAMMATION, INTEBSTITIAL
 

HYPERPLASIA, EPITHELIAL
 

HEMATCPOIETIC SYSTEM
 

•SPLEEN
 

HYPEBPLASIA, NOS
 

BBTICULOCYTOSIS
 

HYPEBPLASIA, HEMATOPOIETIC
 

HYPEBPLASIA, ERYTHBOID
 

HYPEBPLASIA, LYMPHOID
 

tLYHPH NODE
 

CONGESTION, NOS
 

INFLAMMATION, NOS
 

HYPEBPLASIA, NOS
 

BETICULOCYTOSIS
 

HIGH DOSE
 

CONTROL
 

50
 

1
 

49
 

49
 

C*9)
 

1 (2X)
 

3 (6*)
 

1 (2%)
 

(49)
 

1 (2%)
 

(49)
 

1 (2X)
 

(49)
 

10 (20%)
 

(49)
 

6 (12X)
 

1 (2%)
 

5 (10X)
 

1 (2%)
 

(42)
 

.


t NOMBEB OF ANIMALS BITH TISSUE EXAMINED


* NUHBEB OF ANIHALS NECROPSIED
 

10 (24%)
 

1 (2%)
 

2 (5*)
 

 MICROSCOPICALLY
 

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

(49)
 

(49)
 

(49)
 

6 (12X)
 

1 (2X)
 

(49)
 

1 (2«)
 

1 (2X)
 

3 (6X)
 

1 12%)
 

(39)
 

1 <3X)
 

3 (8X)
 

1 (3X1
 

121
 
02. MALE MICE (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HYPBBPLASIA, HEMATOPOIETIC
 

HYPERPLASIA, HETICDLUB CEIL
 

HYPERPLASIA, LYBPHOID
 

CIBCUIATOBY SYSTEM
 

tHEABT
 

MINEBALIZATION
 

IMYOCARDIUM
 

INFLAMMATION, NOS
 

DIGESTIVE SYSTEM
 

f LIVER
 

FIBBOSIS SEPTAL LIVER
 

NECBOSIS, FOCAL
 

METAMORPHOSIS FATTY
 

HYPERPLiSTIC NODULE
 

*GALLBLADDBR
 

INFLAMMATION, NOS
 

HYPEBPLASIA, PAPILLARY
 

*EILE DUCT
 

HYPEBPLASIA, NOS
 

#PANCREAS
 

INFLAMMATION, NOS
 

#STOMACH
 

INFLAMMATION, FOCAL
 

INFLAMMATION, NECBOTIZING
 

HYPEBPLASIA, FOCAL
 

HYPERKEBATOSIS
 

ACANTHOSIS
 

•PEYERS PATCH
 

HYPERPLASIA, NOS
 

tCOLCN
 

PARASITISM
 

HIGH DOSE
 

CONTROL
 

3 (7X)
 

(49)
 

1 (2X)
 

(49)
 

(18)
 

9 (19X)
 

1 (2*)
 

(49)
 

(U9)
 

(«7)
 

1 (2S)
 

(US)
 

2 (H%)
 

1 (2X)
 

1 (2«)
 

1 (2*)
 

1 (2%)
 

(49)
 

7 (14S)
 

(43)
 

_3_JIil
 

HIGH DOSE
 

4 (10*)
 

1 (3X)
 

2 (5X)
 

(49)
 

(49)
 

2 {««)
 

(49)
 

1 (2%)
 

3 (6X)
 

1 (2X)
 

3 (6X)
 

(50)
 

1 (2%)
 

2 (4X)
 

(50)
 

1 (2X)
 

(44)
 

(48)
 

(47)
 

2 (4%)
 

(39)
 

2 i55l)
 

I NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

122
 
02. MALE MICE (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

CONTROL HIGH DOSE
 

DRINARY SYSTEM 

•KIDNEY (49) (50) 

PCLYCYSTIC KIDNEY 1 (2*) 

GLOMERDLONEPHRITIS, NOS 

INFLAMMATION, INTERSTITIAL 

2

16

 (1*) 

 (33%) 

16

2

 (32%) 

 (4%) 

HYPERPLASIA, TUBDLAR CELL 1 (2%) 

tORINABY BLADDER 

HYPERPLASIA, EPITHELIAL 

(148)

4 (8%) (49) 

1 (2%) 

ENDOCRINE SYSTEM
 

*ADRENAL (44) (17)
 

HYPERPLASIA, NOS 3 (1%) 2 (»*)
 

IADRENAL/CAPSULE (44) (47)
 

HYPERPLASIA, NOS 3 (7%)
 

*ADRENAL CORTEX (HI) (47)
 

HYPERTROPHY, FOCAL 1 (2%)
 

#THYROID C»5) C»6)
 

HYPEBPLASIA, FOLLICULAR-CELL 2 (4%)
 

tPANCREATIC ISLETS (47) (44)
 

HYPERPLASIA, NOS 3 (7%)
 

REEECCUCTIVE SYSTEM
 

*PREPUTIAL GLAND (49) (50)
 

ABSCESS, NOS 1 (2%) 1 (2%)
 

#TESTIS (48) (49)
 

HYPERPLASIA, INTERSTITIAL CELL 1 (2%)
 

#TESTIS/TUBOLE (48) (49)
 

DEGENERATION, NOS 2 (4«)
 

*EPIDIDYMIS (49) (50)
 

INFLAMMATION, NOS 1 (2%)
 

NERVOUS SYSTEM
 

NONE
 

# NDBBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIHALS NECROPSIED
 

123
 
02. MALE MICE (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE 

CONTROL HIGH DOSE 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELE1AL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHIR SYSTEMS 

ADIEOSE TISSUE 

INFLAMMATION, ACUTE

OMENTOM 

NECBOSIS, FAT

 1 

1 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED

ANIMAL MISSING/NO NECROPSY

AOTO/NECROPSY/HISTO PERF

 5 

1 

2 

* NUMBER OF ANIHALS WITH TISSUE EXAMINED MICHOSCOPICALLY
 * NOHBEE OF ANIMALS NECROPSIED
 

124
 
TABLE D3. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE FED 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (LOW DOSE AND CONTROL) 

ANIHALS INITIALLY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SOBCUT TISSUE
 

MINERALIZATION
 

FIBROSIS
 

RESPIRATORY SYSTEM
 

#LUNG/BRONCHUS
 

INFLAMMATION, FOCAL
 

#LUNG
 

INFLAMMATION, INTERSTITIAL
 

HYPERPLASIA, EPITHELIAL
 

HEBATOPOIETIC SYSTEM
 

fBOHE MARROW
 

BYELOFIBROSIS
 

tSPLEEN
 

HYPEBPLASIA, NOS
 

HYPERPLASIA, HEBATOPOIETIC
 

HYPERPLASIA, ERYTHROID
 

HYPERPLASIA, LYMPHOID
 

HEBATOPOIESIS
 

MYELOPOIESIS
 

tLYMPH NODE
 

CYST, NOS
 

INFLAMMATION, NOS
 

HYPERPLASIA, NOS
 

_ IITICftI.QCI10S.IS __ ___
 

LOW DOSE
 

CONTROL
 

50
 

48
 

47
 

(H8)
 

1 (2X)
 

1 (2%)
 

(1*6)
 

1 (2%)
 

(1*6)
 

10 (22%)
 

3 (7X)
 

(1*5)
 

1 (2X)
 

(46)
 

16 (35X)
 

6 (13X)
 

10 (22%)
 

1 (2X)
 

1 (2%)
 

(39)
 

1 (3X)
 

15 (38X)
 

1 (3X)
 

1.J3X1
 

LOW DOSE
 

50
 

2
 

45
 

43
 

(45)
 

(42)
 

(42)
 

7 (17X)
 

(41)
 

(40)
 

14	 (35X)
 

6 (15X)
 

5 (13X)
 

(35)
 

18 (51X)
 

1 (3X)
 

t NUMBER OF AHIBALS WITH TISSUE EXABINBD BICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

125
 
03. FEMALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LYMPHOCYTOSIS
 

HIPERPLASIA, HEHATOPOIETIC
 

HYELOID HETAPLASIA
 

CIRCULATORY SYSTEH
 

tHEART/VEHTRICLE
 

HELANIN
 

DIGESTIVE SYSTEM
 

#SALIVARY GLAND
 

INFLAMMATION, NOS
 

PERIVASCOLAR CUFFING
 

#LIVER
 

INFLAMHATION, NOS
 

NECROSIS, FOCAL
 

NECROSIS, COAGULATIVE
 

BETAHORPHOSIS FATTY
 

HYPERPLASTIC NODULE
 

ANGIECTASIS
 

HEMATOPOIESIS
 

*GALLBLADDER
 

INFLAHHATION, NOS
 

HYPERPLASIA, PAPILLARY
 

*BILE DUCT
 

INFLAHMATICN, NOS
 

HYPERPLASIA, NOS
 

»PANCREAS
 

INFLAMMATION, NOS
 

PfiRIASTERITIS
 

fPANCREATIC DOCT
 

LYMPHOCYTIC INFLAHMATORY INFILTR
 

#STOMACH
 

INFLAMMATION, NOS
 

ULCER, NOS
 

INFLAMMATION^ FOCAL
 

LOW DOSE 

CONTROL LOW DOSE 

2 (516) 

1 OX) 

1 (3«) 

C*6) 

U (9*) 

(t2) 

(U5) 

2 C*«) 

U (9X) 

(37) 

(U7) 

1 (255) 

22 (<*7%) 

1 (2«) 

1 (2%) 

3 (6S) 

C*3) 

3 (7%) 

1 (2%) 

1 (2%) 

2 (5%) 

(US) 

3 (6X) 

(«5) 

2 (H%) 

(48) 

1 (2S) 

(45) 

1 (2*) 

(*»*») 

5 (11X) 

1 (2%) 

(11) 

C*1*) 

1 (2%) 

(U1) 

(«(*) 

7 (16«) 

1 (2%) 

1 i221_ . 

(") 

3_I8%1 _ 

f NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

126
 
03. FEMALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

HYPERPLASIA,

HYPEBPLASIA ,

HTPERPLASIA,

 NOS 

 EPITHELIAL 

 ADENOHATOO S 

1

1

1

 (2%) 

 (2«) 

(2%) 

HYPERKERATOSIS 1 (2%) 2 (5%) 

ACANTHOSIS 1 (2X) 2 (5«) 

fGASTRIC MUCOSA (i»4) (37) 

HYPERPLASIA , FOCAL 1 (2%) 

#PEYERS PATCH (HH) (40) 

HYPERPLASIA , NOS 1 (2*) 1 (3%) 

URINAR Y SYSTEM 

tKIDNE Y (l»6) (43)

GLOMEROLONEPHRITIS, NOS 1U (3P%) 6 (14%) 

INFLAMMATION, INTERSTITIAL 16 (35%) 9 (21%) 

tURINAR Y BLADDER (1*6) (34) 

INFLAMMATION, NOS H (9%) 3 (9%) 

HYPERPLASIA, EPITHELIAL 10 (22%) 2 (6%) 

ENDOCRINE SYSTE M 

*PITUITARY («2) (20) 

HYPERPLASIA , FOCAL 6 (14% ) 

tADRENAL/CAPSULE (us) (40) 

HYPERPLASIA, NOS 2 (5%) 

tADRENA L CORTEX (45) (40 ) 

NODOLE 3 (7%) 

ITHYROID (43 ) (37) 

FOLLICntAR CYST, NOS 1 (2%) 

INFLAMMATION , NOS 1 (2%) 

REPRODUCTIV E SYSTE M 

*MAMMAR Y GLAN D (48) (45) 

GALACTOCEL E 1 (2%)

JIYPERPLASIA^.NQS i_i8SL 3_i7%L__. 

# NUMBER OF RNINALS WITH TISSUE EXAMINED MICROSCOPIC*LLY
 

* NUMBFR OF ANIMALS NECROPSIED
 

127
 
D3. FEMALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE
 

CONTROL LOW DOSE
 

fUTERUS (45) (39)
 

HYDROMETPA 1 (2%) 3 (8%)
 

ABSCESS, NOS 3 (7%) 1 (3%)
 

FIBROSIS 1 (2X)
 

*DTERUS/ENDOHFTRIUM (i»5) (39)
 

INFLAMMATION, NOS 10 (22%) 1 (3%)
 

INFLAMMATION, SUPPURATIVE U (9%) 1 (3%)
 

HYPERPLASIA, NOS 4 (9%) u (10%)
 

HYPERPLASIA, CYSTIC 18 (flOX) m (49%)
 

HYPERPLASIA, ADENOMATOUS 1 (2*)
 

#OVAHY/OVIDtICT («5) (39)
 

INFLAMMATION, NOS 5 (11%)
 

#OVAKY (45) (39)
 

CYST, NOS 3 (7%) 2 (5%)
 

INFLAMMATION, NOS « (9X) 1 (3%)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 1 (2%)
 

INFLAMMATION, SUPPURATIVE 10 (22%)
 

ABSCESS, NOS 4 (9%)
 

DEGENERATION, CYSTIC 1 (2%)
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE CRGANS
 

*EYE (48) (45)
 

CATARACT 1 (2%)
 

*HARDERIAN GLAND (48) (45)
 

HYPERPLASIA, PAPILLARY 1 (2%)
 

MUSCULOSKELETAL SYSTEM
 

*BONE (48) (45)
 

INFLAMMATION, NOS 2 (US)
 

HESORPTION 3 (6X)
 

BODY CAVITIES
 

NONE
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

128
 
D3. FEMALE MICE (LOW DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

CONTROL LOW DOSE 

A LL OTHER SYSTEMS 

OMENTU M 

NECROSIS, FAT 1 1

 

SPECIAL HORPHOLOGY SUMMAR Y 

NO LESION REPORTED 1 

ANIMAL MISSING/NO NECROPSY 2 

AUTO/NECROFSY/HISTO PERF 2 1

 

AUTO/NECROPSY/NO HISTO 1 2

 

AUTOLYSIS/NO NECROPSY 2 3 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE8 OF ANIMALS NECROPSIED
 

129
 
TABLE D4. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE FED 

3-AMINO-g-ETHYLCARBAZOLE HYDROCHLORIDE IN THE DIET (HIGH DOSE AND CONTROL) 

ANIMALS INITIALLY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECBOPSIED
 

ANIMALS EXAMINED HI STOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SOBCUT TISSUE
 

ABSCESS, SOS
 

BESFIEATOEY SYSTEM
 

•LUNG/BRONCHUS
 

INFLAMMATION, FOCAL
 

#LUNG/BEONCHIOLE
 

HYPERPLASIA, NOS
 

#LONG
 

INFLAMMATION, INTERSTITIAL
 

HYPERPLASIA, EPITHELIAL
 

HEKATCPOIETIC SYS1EM
 

•SPLEEN
 

GROWTH, ALTERATION
 

HYPERPLASIA, NOS
 

HYPERPLASIA, HEMATOPOIETIC
 

HYPEPPLASIA, ERYTHROID
 

HYPERPLASIA, LYMPHOID
 

fHEMOLYMPH NODES
 

INFLAMMATION, NOS
 

HYPEHPLASIA, NOS
 

#LYMEH NODE
 

INFLAMMATION, NOS
 

HYPEPPLASIA, NOS
 

HIGH DOSE
 

CONTROL
 

50
 

50
 

50
 

(50)
 

1 (2X)
 

(50)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 

14 (28*)
 

(i»9)
 

9 (18*)
 

6 (12X)
 

2 (4X)
 

(49)
 

2 («*)
 

1 (2*)
 

(i»«)
 

9 (20X)
 

3 J7J1
 

HIGH DOSE
 

50
 

1
 

U9
 

U9
 

(K 9)
 

(48)
 

(48)
 

(48)
 

6 (13*)
 

1 (2%)
 

(46)
 

1 (2*)
 

3 (7*)
 

1 (2%)
 

4 (9*)
 

(46)
 

(35)
 

4 (11*)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

130
 
D4. FEMALE MICE (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

- ~- •—"---!- ­- ­- --T­ '-I- J_— ­ I— 'IT­ ­ 11 '- J • ­1 T 1 r JT J­ _ 

HIGH DOSE 

CONTROL HIGH DOSE 

BETICULOCYTOSIS 1 (2%) 1 (3%) 

HYPEBPLASIA, HEMATOPOIETIC 1 (2%) 

HYPEBPLASIA. LYHPHOID U (9«) 

CIBCUIATOBY SYSTEM
 

*CARDIOVASCULAB SYSTE (50) (49)
 

PEBIVASCULITIS 1 (2X)
 

IBYOCA8DIOH (50) (49)
 

IKFLAMHATION, FOCAL 1 (2%)
 

*AOHTA (50) (49)
 

INFLAMMATION, SOS 2 (4%)
 

DIGESTIVE SYSTEM
 

ISALIVABY GLAND (48) (46)
 

PERIVASCULAR CUFFING 3 (6%)
 

tLIVER (50) (49)
 

NECROSIS, FOCAL 7 (1M)
 

HYPEBPLASTIC NODULE 2 (4%)
 

tPANCREAS (48) (43)
 

INFLAHHATION, NOS 2 (H%) 1 (2%)
 

#STOMACH (i*9) (44)
 

INFLABHATION, NOS 1 (2%) 1 (2%)
 

INFLAMMATION, FOCAL 1 (2%) 1 (2«)
 

ACANTHOSIS 2 (4%) 3 (7%)
 

tPEYERS PATCH (48) (44)
 

HYPEEPLASIA, NOS 7 (15%) 2 (5%)
 

tCOLCN (38) (35)
 

EARASITISM 1 (3%)
 

UEINAFY SYSTEM
 

#KIDNEY (50) (45)
 

GIOMERULONEPHRITIS. NOS 4 (8&1 l_12g%l
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

131 
04. FEMALE MICE (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE 

CONTROL HIGH DOSE 

GLOBEBOLONEPH8ITIS, FOCAL 

INFLAMMATION, INTERSTITIAL 

1

12

 (2X) 

 (24X) 5 (11%) 

tKIDNEY/TUBOLE 

MINERALIZATION 

(50) 

1 (256) 

C*5) 

fURINARY BLADDEB 

INFLAHHATION, NOS 

HYPERPLASIA, EPITHELIAL 

HYPEEPLASIA, PAPILLARY 

(48) 

1 (2%) 

(U3) 

2

1

1

 (5*) 

 (2«) 

 (2X) 

ENCCCEINE SYSTEM
 

#ADRENAL (48) (U6)
 

HYPEEPLASIA, NOS 5 (11X)
 

#ADRENAL/CAPSULE (48) (46)
 

HYPEBPLASIA, NOS 5 (10X)
 

#ADRENAL CORTEX (48) (46)
 

NODULE 1 (2X)
 

HYPERPLASIA, NOS 1 (25E)
 

•THYROID	 («4) (<*2)
 

INFLAMMATION, FOCAL 1 (2%)
 

HYPERPLASIA, FOCAL 1 (2%)
 

HYPERPLASIA, PAPILLARY 2 <5S) 1 12%)
 

HYPEEPLASIA, ADENOMATOUS 1 (2%)
 

REEBCEOCTIVE SYSTEM
 

*MAMMARY GLAND (50) (49)
 

HYPERPLASIA, NOS 1 (2X)
 

*OTEBOS {1*7) (35)
 

HYDHOMETBA 13 (28X) 2 (6X)
 

•UTERUS/ENDOMETRIUM	 (47) (35)
 

INFLAMMATION, NOS 8 (17X) 1 (3X)
 

HYPERPLASIA, NOS 8 (17X) 2 (6X)
 

HYPERPLASIA, CYSTIC 6 (13)1) 2 (6X)
 

tOVARY/OVIDUCT (147) (35)
 

INFLAMMATION*. NOS
 <* J9?-l — _
 

t NOMBEB OP ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECBOPSIED
 

132
 
D4. FEMALE MICE (HIGH DOSE AND CONTROL): NONNEOPLASTIC LESIONS (CONTINUED) 

AESCSSS, NOS
 

#OVARY
 

CYST, NOS
 

HEKOPRHAGE
 

INFLAMMATION, NOS
 

PERIARTERITIS
 

DEGENERATION, CYSTIC
 

HYPERPLASIA, CYSTIC
 

NEEVCUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSC01CSKEIETAI SYSTEM
 

*EONE
 

INFLAMMATION, NOS
 

BOtY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SOBHARY
 

NO LESION REPORTED
 

ANIMAL MISSING/NO NECROPSY
 

AUTO/NECROPSY/HISTO PEEF
 

HIGH DOSE
 

CONTROL
 

1 (2%)
 

(U8)
 

10 (21%)
 

tt (8X)
 

1 (2X)
 

3 (6%)
 

(50)
 

3
 

1
 

HIGH DOSE
 

(HO)
 

2 (5%)
 

1 (3X)
 

1 (3«)
 

(49)
 

1 <2X)
 

1
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

133
 

APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS ADMINISTERED 3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

IN THE DIET
 

135
 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

Low-Dose High-Dose Low High
 

Topography: Morphology Control Control Dose Dose
 

Integumentary System:
 

Squamous-cell Carcinoma'3 0/36 (0) 0/48 (0) 3/44 (7) 3/48 (6)
 

P Values0 >d N.S. N.S.
 

Relative Risk6 Infinite Infinite
 

Lower Limit 0.497 0.602
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 78 86
 —
 —
 OJ
 

Integumentary System: Squamous-cell
 

Papilloma or Carcinoma'3 0/36 (0) 0/48 (0) 6/44 (14) 5/48 (10)
 

P Values0 »d P = 0.023 P = 0.028
 

Relative Risk6 Infinite Infinite
 

Lower limit 1.324 1.263
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 72 86
 —
 —
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Topography: Morphology
 

Integumentary System:
 

Basal-cell Carcinoma
 

of the Skinb
 

P Values0 »d
 

Relative Riske
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

U)


oo
 

Integumentary System: Basal-cell
 

or Squainous-cell Carcinoma or
 

Squamous-cell Papilloma"
 

V Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Uose
 

Control
 

0/36 (0)
 

—
 

0/36 (0)
 

—
 

High-Dose Low 

Control Dose 

0/48 (0) 4/44 (9) 

N.S. 

Infinite 

0.767 

Infinite 

__ 

86 

0/48 (0) 8/44 (18) 

P = 0.006 

Infinite 

1.892 

Infinite 

72 — 

High
 

Dose
 

1/48 (2)
 

N.S.
 

Infinite
 

0.053
 

Infinite
 

106
 

6/48 (13)
 

P = 0.013
 

Infinite
 

1.602
 

Infinite
 

86
 
__ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Low-Dose
 

Topography: Morphology Control
 

Integumentary System: Fibroma
 

in Subcutaneous Tissue*5 0/36 (0)
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 

Carcinoma" 0/36 (0)
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 —
 

High-Dose Low 

Control Dose 

3/48 (6) 1/44 (2) 

N.S. 

Infinite 

0.044 

Infinite 

95 78 

1/48 (2) 1/43 (2) 

N.S. 

Infinite 

0.045 

Infinite 

109 95 

High
 

Dose
 

5/48 (10)
 

N.S.
 

1.667
 

0.345
 

10.203
 

103
 

3/48 (6)
 

N.S.
 

3.000
 

0.252
 

154.112
 

106
 
__ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Low -Dose High-Dose Low High
 

Topography: Morphology Control Control Dose Dose
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma'5 0/36 (0) 1/48 (2) 3/43 (7) 5/48 (10)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 Infinite 5.000
 

Lower Limit 0.509 0.590
 

Upper Limit Infinite 231.143
 

Weeks to First Observed Tumor 109 95 91
 

Hematopoietic System: Lymphoma
 

or Leukemia'3 2/36 (4) 6/48 (13) 6/44 (14) 4/48 (8)
 

P Values0 >d N.S. N.S.
 

Relative Riske 2.455 0.667
 

Lower Limit 0.474 0.147
 

Upper Limit 23.746 2.628
 

Weeks to First Observed Tumor 79 93 94 100
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3 

(continued) 

Topography: Morphology 

Low-Dose 

Control 

High-Dose 

Control 

Low 

Dose 

Liver: Hepatocellular 

Carcinoma'5 0/36 (0) 1/48 (2) 6/42 (14) 

P Values0 »d P = 0.020 

Relative Risk6 

Lower Limit 

Upper Limit 

Infinite 

1.388 

Infinite 

Weeks to First Observed Tumor 

__ 

109 72 

Liver: Neoplastic Nodule or 

Hepatocellular Carcinoma'3 0/36 (0) 1/48 (2) 12/42 (29) 

P Values0 »d P < 0.001 

Relative Risk6 

Lower Limit 

Upper Limit 

Infinite 

3.185 

Infinite 

Weeks to First Observed Tumor — 109 72 

High
 

Dose
 

8/48 (17)
 

P = 0.015
 

8.000
 

1.138
 

346.323
 

78
 

22/48 (46)
 

P < 0.001
 

22.000
 

3.844
 

873.814
 

78
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued) 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Pituitary: Chromophobe Adenoma
 

or Adenoma, NOSb 12/32 (38) 9/38 (24) 12/37 (32) 11/41 (27)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 0.865 1.133
 

Lower Limit 0.420 0.483
 

Upper Limit 1.802 2.744
 

Weeks to First Observed Tumor 101 85 78 94
 

Adrenal: Pheochromocytoma or
 

Pheochromocytoma, Malignant*3 6/35 (17) 8/47 (17) 4/43 (9) 10/48 (31)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 0.543 1.224
 

Lower Limit 0.122 0.478
 

Upper Limit 2.109 3.257
 

Weeks to First Observed Tumor 107 107 78 101
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Topography: Morphology 

Low-Dose 

Control 

Thyroid: Follicular-cell 

Carcinoma'5 0/35 (0) 

P Values0>d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

__ 

Thyroid: Follicular-cell 

Adenoma or Carcinoma" 0/35 (0) 

P Values0 » d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

High -Dose 

Control 

Low 

Dose 

0/48 (0) 2/40 (5) 

N.S. 

__ 

Infinite 

0.262 

Infinite 

105 

0/48 (0) 3/40 (8) 

N.S. 

Infinite 

0.533 

Infinite 

— 103 

High
 

Dose
 

1/47 (2)
 

N.S.
 

Infinite
 

0.055
 

Infinite
 

106
 

1/47 (2)
 

N.S.
 

Infinite
 

0.055
 

Infinite
 

106
 
__ __ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9—Ethylcarbazole Hydrochloride in the Dieta
 

(continued)
 

Low-Dose High-Dose Low High
 

Topography: Morphology Control Control Dose Dose
 

Pancreatic Islets: Islet-cell
 

Adenoma'3 2/34 (6) 0/46 (0) 4/39 (10) 4/43 (9)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 1.744 Infinite
 

Lower Limit 0.269 0.994
 

Upper Limit 18.348 Infinite
 

Weeks to First Observed Tumor 105 97
 

Preputial Gland: Adenoma, NOS^ 0/36 (0) 0/48 (0) 1/44 (2) 4/48 (8)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 Infinite Infinite
 

Lower Limit 0.044 0.929
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 105 97
 — —
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Low-Dose High-Dose Low
 

Topography: Morphology Control Control Dose
 

Testis: Interstitial-cell
 

Tumorb 33/35 (94) 42/47 (89) 32/43 (74)
 

P Values0 »d P = 0.018(N)
 

Relative Risk6 0.789
 

Lower Limit 0.709
 

Upper Limit 0.986
 

Weeks to First Observed Tumor 78 78 72
 

Zymbal's Gland: Carcinoma, NOSb 0/36 (0) 0/48 (0) 1/44 (2)
 

P Values0 »d N.S.
 

Relative Risk6 Infinite
 

Lower Limit 0.044
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 104
 —
 —
 

High
 

Dose
 

40/47 (85)
 

N.S.
 

0.952
 

0.819
 

1.130
 

- 78
 

4/48, (8)
 

N.S.
 

Infinite
 

0.929
 

Infinite
 

48
 
__ __ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Dieta
 

(continued)
 

Low-Dose High-Dose Low High
 

Topography: Morphology Control Control Dose Dose
 

Zymbal's Gland: Squamous-cell
 

Carcinoma^ 0/36 (0) 0/48 (0) 4/44 (9) 3/48 (6)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 Infinite Infinite
 

Lower Limit 0.767 0.602
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 72 78
 

Zymbal's Gland: Carcinoma, NOS,
 

or Squamous— cell Carcinoma" 0/36 (0) 0/48 (0) 5/44 (11) 7/48 (15)
 

P Values0 »d P = 0.045 P = 0.006
 

Relative Risk6 Infinite Infinite
 

Lower Limit 1.044 1.944
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 72 48
 —
 —
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Low-Dose High-Dose Low High
 

Topography: Morphology Control Control Dose Dose
 

Body Cavities: Mesothelioma. NOS,
 

or Mesothelioma, Malignant 0/36 (0) 2/48 (4) 2/44 (5) 3/48 (6)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 Infinite 1.500
 

Lower Limit 0.244 0.180
 

Upper Limit Infinite 17.302
 

Weeks to First Observed Tumor 106 86 91
 —
 a

Dosed groups received 800 or 2,000 ppm.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

G

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher
 

exact test for the comparison of that dosed group with its matched control group when
 

P < 0.05; otherwise, not significant (N.S.) is indicated.
 

**A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

lhe 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
__ __ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole


Topography: Morphology
 

Integumentary System:
 

Squamous-cell Carcinoma'3
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Integumentary System: Squamous-cell
 

Papilloma or Carcinoma"
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Control
 

0/39 (0)
 

0/39 (0)
 

—
 

 Hydrochloride in the Diet3
 

High-Dose Low
 

Control Dose
 

0/50 (0) 3/44 (7)
 

N.S.
 

Infinite
 

0.537
 

Infinite
 

78
 

0/50 (0) 4/44 (9)
 

N.S.
 

Infinite
 

0.829
 

Infinite
 

78
 —
 

High
 

Dose
 

3/49 (6)
 

N.S.
 

Infinite
 

0.614
 

Infinite
 

98
 

4/49 (8)
 

N.S.
 

Infinite
 

0.946
 

Infinite
 

98
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-EthylcarbaEole Hydrochloride in the Diet3 

(continued) 

Low— Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Integumentary System: Fibroma of 

the Subcutaneous Tissueb 0/39 (0) 1/50 (2) 2/44 (5) 1/49 (2) 

P Valuesc » d N.S. N.S. 

Relative Riske Infinite 1.020 

Lower Limit 0.264 0.013 

Upper Limit Infinite 78.488 

Weeks to First Observed Tumor 102 78 107 

Integumentary System: 

Sarcoma, NOSb 0/39 (0) 0/50 (0) 2/44 (5) 0/49 (0) 

P Values0 » d N.S. N.S. 

__ 

Relative Risk6 Infinite 

Lower Limit 0.264 

Upper Limit Infinite — 

— 

Weeks to First Observed Tumor — 84 — 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Topography: Morphology 

Low-Dose 

Control 

High-Dose 

Control 

Low 

Dose 

High 

Dose 

Lung: Alveolar /Bronchiolar 

Adenoma or Carcinoma" 1/39 (3) 1/50 (2) 4/43 (9) 6/48 (13) 

P Values0 » d N.S. N.S. 

Relative Risk6 

Lower Limit 

Upper Limit 

3.628 

0.381 

174.220 

6.250 

0.801 

280.829 

Weeks to First Observed Tumor 107 110 93 97 

Hematopoietic System: Lymphoma 

or Leukemia" 4/39 (10) 5/50 (10) 2/44 (5) 6/49 (12) 

P Values0 >d N.S. N.S. 

Relative Risk6 

Lower Limit 

Upper Limit 

0.443 

0.042 

2.914 

1.224 

0.333 

4.751 

Weeks to First Observed Tumor 101 104 72 98 
__ 

Table E2. Analyses of the Incidence of Primary Tumors in
 Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in
 

(continued)
 

Low-Dose High-Dose
 

Topography: Morphology Control Control
 

Liver: Hepatocellular
 

Carcinoma'3 2/39 (5) 0/50 (0)
 

P Values0 «d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 97
 

Liver: Neoplastic Nodule or
 

Hepatocellular Carcinoma" 2/39 (5) 0/50 (0)
 

P Values0 »d
 

Relative Riske
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 97
 —
 

the Diet3
 

Low
 

Dose
 

1/43 (2)
 

N.S.
 

0.453
 

0.008
 

8.373
 

92
 

-


1/43 (2)
 

N.S.
 

0.453
 

0.008
 

8.373
 

92
 

High
 

Dose
 

3/48 (6)
 

N.S.
 

Infinite
 

0.626
 

Infinite
 

78
 

6/48 (13)
 

P = 0.012
 

Infinite
 

1.667
 

Infinite
 

78
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered 

3-Amino-9-Ethylcarbazole Hydrochloride in the Dieta 

(continued) 

Topography: Morphology 

Low-Dose 

Control 

Pituitary: Adenoma, NOS, or 

Chromophobe Adenoma'3 18/37 (49) 

P Values0 » d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 76 

Pituitary: Adenocarcinoma, NOS^ 2/37 (3) 

P Values0 » d 

Relative Risk8 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 107 

High-Dose 

Control 

Low 

Dose 

High 

Dose 

17/40 (43) 15/30 (50) 20/39 (51) 

N.S. N.S. 

1.028 

0.589 

1.741 

1.207 

0.717 

2.034 

78 64 77 

0/40 (0) "0/30 (0) 0/39 (0) 

N.S. 

0.000 

0.000 

4.099 

N.S. 

H_ 

— 

— 

— — — 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Topography: Morphology
 

Adrenal: Cortical Adenoma"
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma'3
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose High-Dose Low High 

Control Control Dose Dose 

0/37 (0) 1/49 (2) 0/43 (0) 3/47 (6) 

N.S. N.S. 

3.128 — 0.262 — 160.605 — 

110 88 — — 

2/37 (5) 3/49 (6) 1/43 (2) 4/47 (9) 

N.S. N.S. 

0.430 1.390 

0.007 0.248 

7.940 9.029 

108 109 105 107 
__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

 Hydrochloride in the Diet3
 

High-Dose Low
 

Control Dose
 

0/50 (0) 4/44 (9)
 

N.S.
 

3.545
 

0.372
 

170.387
 

64
 

19/50 (38) 4/44 (9)
 

N.S.
 

0.886
 

0.177
 

4.460
 

107 68
 

High
 

Dose
 

3/49 (6)
 

N.S.
 

Infinite
 

0.614
 

Infinite
 

74
 

11/49 (22)
 

N.S.
 

0.591
 

0.286
 

1.160
 

92
 

(continued)
 

Topography: Morphology
 

Mammary Gland:
 

Adenocarcinoma, NOS^
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 101 

Mammary Gland: Fibroadenoma" 4/39 (10) 

P Values0 » d 

Relative Riske 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 101 

3-Amino-9-Ethylcarbazole


Low-Dose
 

Control
 

1/39 (3)
 
__ 

__ 

__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued) 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Mammary Gland: Papillary
 

Cystadenocarcinoma, NOS^ 1/39 (3) 0/50 (0) 2/44 (5) 0/49 (0)
 

P Values0>d N.S. N.S.
 

Relative Risk6 1.773
 

Lower Limit 0.096
 

Upper Limit 102.188
 —
 

—
 

Weeks to First Observed Tumor 107 78
 

Mammary Gland: Carcinoma, NOS,
 

Adenocarcinoma, NOS, Papillary
 

Adenocarcinoma, or Papillary
 

Cystadenocarcinoma, NOS° 2/39 (5) 0/50 (0) 8/44 (18) 3/49 (6)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 3.545 Infinite
 

Lower Limit 0.765 0.614
 

Upper Limit 32.681 Infinite
 

Weeks to First Observed Tumor 101 64 74
 —
 
__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued) 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Clitoris or Clitoral Gland:
 

Adenoma, NOS*> 0/39 (0) 2/50 (4) 3/44 (7) 3/49 (6)
 

P Valuesc»d N.S. N.S.
 

Relative Risk6 Infinite 1.531
 

Lower Limit 0.537 0.183
 

Upper Limit Infinite 17.671
 

Weeks to First Observed Tumor 104 78
 

Uterus: Neoplasm, NOS^ 0/38 (0) 0/50 (0) 2/43 (5) 0/49 (0)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 Infinite «•«.
 

Lower Limit 0.264
 

Upper Limit Infinite
 —
 

—
 

Weeks to First Observed Tumor 103
 — — —
 

99 
__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole
 Hydrochloride in the Dieta
 

High-Dose Low
 

Control Dose
 

0/50 (0) 2/44 (5)
 

N.S.
 

Infinite
 

0.264
 

Infinite
 •
 

_-.
 99
 

1/50 (2) 11/43 (26)
 

N.S.
 

2.430
 

0.796
 

9.650
 

109 74
 

(continued)
 

Topography: Morphology
 

Clitoris or Clitoral Gland:
 

Carcinoma, NOS, or Squamous-cell
 

Carcinoma*5
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus or Endometrium:
 

Adenocarcinoma, NOS^
 

P Values0 »d
 

Relative Risk6
 

Lower Li-.it
 

Upper L: .nit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Control
 

0/39 (0)
 

4/38 (11)
 

95
 

High
 

Dose
 

4/49 (8)
 

N.S.
 

Infinite
 

0.946
 

Infinite
 

77
 

11/49 (22)
 

P = 0.002
 

11.224
 

1.736
 

470.753
 

92
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Topography: Morphology
 

Uterus: Endometrial Stromal
 

Polypb
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal
 

Polyp or Sarcoma"
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose High-Dose 

Control Control 

10/38 (26) 10/50 (20) 

78 78 

10/38 (26) 11/50 (22) 

78 78 

Low 

Dose 

High 

Dose 

11/43 (26) 5/49 (10) 

N.S. N.S. 

0.972 

0.425 

2.267 

0.510 

0.147 

1.511 

72 68 

11/43 (26) 5/49 (10) 

N.S. N.S. 

0.972 

0.425 

2.267 

0.464 

0.135 

1.331 

72 68 
__ __ 

Table E2. Analyses of the Incidence of Primary Tumors
 in Female Rats Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride
 

(continued)
 

Low-Dose High-Dose
 

Topography: Morphology Control Control
 

Zymbal's Gland: Carcinoma, NOSb 0/39 (0) 0/50 (0)
 

P Values0 >d
 

Relative Riske
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Zymbal's Gland: Squamous-cell
 

Carcinoma*5 0/39 (0) 0/50 (0)
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 — —
 

in the Dieta
 

Low
 

Dose
 

5/44 (11)
 

P = 0.037
 

Infinite
 

1.127
 

Infinite
 

68
 

5/44 (11)
 

P = 0.037
 

Infinite
 

1.127
 

Infinite
 

72
 

High
 

Dose
 

3/49 (6)
 

N.S.
 

Infinite
 

0.614
 

Infinite
 

88
 

9/49 (18)
 

P = 0.001
 

Infinite
 

2.684
 

Infinite
 

47
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats Administered 

3-Amino-9-Ethylcarbazole Hydrochloride in the Dieta 

(continued) 

Low-Dose High-Dose Low High
 

Topography: Morphology Control Control Dose Dose
 

Zymbal's Gland: Carcinoma, NOS,
 

or Squamous-cell Carcinoma'3 0/39 (0) 0/50 (0) 10/44 (23) 12/49 (24)
 

P Values0 »d P = 0.001 P < 0.001
 

Relative Risk6 Infinite Infinite
 

Lower Limit 2.661 3.742
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 68 47
 —
 —
 

a

Dosed groups received 800 or 2,000 ppm.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher
 

exact test for the comparison of that dosed group with its matched control group when
 

? < 0.05; otherwise, not significant (N.S.) is indicated.
 

^A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN MICE ADMINISTERED 3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

IN THE DIET
 

161
 

Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Lung: Alveolar/Bronchiolar 

Carcinoma^ 6/48 (13) 5/44 (11) 1/42 (2) 3/49 (6) 

P Valuesc>d N.S. N.S. 

Relative Risk6 0.190 0.539 

Lower Limit 0.004 0.088 

Upper Limit 1.477 2.605 

Weeks to First Observed Tumor 96 96 94 78 

Lune: Alveolar/Bronchiolar ° i 

Adenoma or Carcinoma0 6/48 (13) 10/44 (23) 6/42 (14) 7/49 (14) 

P Valuesc » d N.S. N.S. 

Relative Risk6 1.143 0.629 

Lower Limit 0.329 0.223 

Upper Limit 3.946 1.669 

Weeks to First Observed Tumor 96 96 94 78 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

Topography: Morphology
 

Hematopoietic System:
 

Lymphomab
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Low-Dose
 

Control
 

4/48 (8)
 

Weeks to First Observed Tumor 96
 

Hematopoietic System:
 

Lymphoma or Leukemia*5 4/48 (8)
 

P Values0 «d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 96
 

High-Dose Low 

Control Dose 

5/44 (11) 3/44 (7) 

N.S. 

0.818 

0.126 

4.560 

96 94 

5/44 (11) 3/44 (7) 

N.S. 

0.818 

0.126 

4.558 

96 94 

High
 

Dose
 

5/50 (10)
 

N.S.
 

0.880
 

0.217
 

3.582
 

95
 

6/50 (12)
 

N.S.
 

1.056
 

0.289
 

4.090
 

89
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Dieta
 

(continued) 

Topography: Morphology 

Low- Dose 

Control 

Liver: Hepatocellular 

Carcinoma^ 7/48 (15) 

P Values0 » d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 78 

Liver: Hepatocellular 

Adenoma or Carcinoma^3 7/48 (15) 

P Values0 » d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 78 

High-Dose 

Control 

Low 

Dose 

6/44 (14) 32/44 (73) 

P < 0.001 

4.987 

2.527 

10.814 

78 68 

8/44 (18) 32/44 (73) 

P < 0.001 

4.987 

2.527 

10.814 

78 68 

High
 

Dose
 

41/49 (84)
 

P < 0.001
 

6.136
 

3.102
 

13.256
 

78
 

41/49 (84)
 

P < 0.001
 

4.602
 

2.569
 

8.447
 

78
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued) 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Thyroid: Follicular-cell
 

Adenoma'' 1/47 (2) 0/40 (0) 0/35 (0) 3/46 (7)
 

P Values0>d N.S. N.S.
 

Relative Risk6 0.000 Infinite
 

Lower Limit 0.000 0.527
 

Upper Limit 24.848 Infinite
 

Weeks to First Observed Tumor 96 95
 — —
 

Thyroid: Follicular-cell Adenoma
 

or Papillary Cystadenoma, NOS" 1/47 (2) 0/40 (0) 0/35 (0) 4/46 (9)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 0.000 Infinite
 

Lower Limit 0.000 0.812
 

Upper Limit 24.848 Infinite
 

Weeks to First Observed Tumor 96 78
 — —
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued)
 

a

Dosed groups received doses of 800 or 1,200 ppm.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

G

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher
 

exact test for the comparison of that dosed group with its matched-control group when
 

P < 0.05; otherwise, not significant (N.S. is indicated.
 

"A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice Administered
 

3-Amino-9-Ethylcarbazole
 Hydrochlorid ie n the Diet3
 

High-Dose Low
 

Control Dose
 

1/45 (2) 3/42 (7)
 

N.S.
 

3.286
 

0.276
 

168.212
 

78 88
 

3/45 (7) 4/42 (10)
 

N.S.
 

1.095
 

0.217
 

5.515
 

78 86
 

High
 

Dose
 

1/48 (2)
 

N.S.
 

0.938
 

0.012
 

72.085
 

95
 

4/48 (8)
 

N.S.
 

1.250
 

0.224
 

8.117
 

75
 

Topography: Morphology
 

Lung: Alveolar/Bronchiolar
 

Carcinoma^
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma"
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low- Dose
 

Control
 

1/46 (2)
 

97
 

4/46 (9)
 

97
 
__ 

Table F2. Analyses of the Incidence of Primary Tumors in Female Mice Administered
 

3-Amino-9-Ethylcarbazole


(continued) 

Low- Dose 

Topography: Morphology Control 

Hematopoietic System:
 

Lymphomab 5/48 (10)
 

P Values0 »d
 

Relative Risk8
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 97
 

Hematopoietic System:
 

Kupffer-cell Sarcoma** 0/48 (0)
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 —
 

 Hydrochloride in the Diet3
 

High-Dose Low
 

Control Dose
 

2/45 (4) 2/45 (4)
 

N.S.
 

0.427
 

0.042
 

2.455
 

96 89
 

0/45 (0) 0/45 (0)
 

N.S.
 

—
 

—
 

—
 —
 

High
 

Dose
 

2/49 (4)
 

N.S.
 

0.918
 

0.069
 

12.222
 

65
 

3/49 (6)
 

N.S.
 

Infinite
 

0.554
 

Infinite
 

78
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice Administered 

3-Amino-9-Ethylcarbazole Hydrochloride in the Dieta 

(continued) 

Topography: Morphology 

Low-Dose 

Control 

High-Dose 

Control 

Low 

Dose 

High 

Dose 

Liver: Hepatocellular 

Carcinoma*5 1/47 (2) 1/45 (2) 37/43 (86) 43/49 (88) 

P Value sc»d P < 0.001 P < 0.001 

Relative Risk6 

Lower Limit 

Upper Limit 

40.442 

7.935 

1373.351 

39.490 

7.832 

1336.021 

Weeks to First Observed Tumor 97 96 79 58 

Pituitary: Adenoma, NOSb 2/42 (5) 1/38 (3) 1/29 (3) 0/33 (0) 

P Values0 >d N.S. N.S. 

Relative Risk6 

Lower Limit 

Upper Limit 

0.724 

0.013 

13.163 

0.000 

0.000 

21.280 

Weeks to First Observed Tumor 97 96 90 — 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3 

(continued) 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Pituitary: Chromophobe Adenoma^ 0/42 (0) 2/28 (5) 0/29 (0) 0/33 (0) 

P Values0>d N.S. N.S. 

Relative Risk6 0.000 — Lower Limit 0.000 — Upper Limit — 3.841 

Weeks to First Observed Tumor 96 — — — 

Pituitary: Adenoma, NOS, or 

Chromophobe Adenoma" 2/42 (5) 3/38 (8) 1/29 (3) 0/33 (0) 

P Values0 » d N.S. N.S. 

Relative Risk6 0.724 0.000 

Lower Limit 0.013 0.000 

Upper Limit 13.163 1.887 

Weeks to First Observed Tumor 97 96 90 — 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice Administered
 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3
 

(continued) 

Low-Dose High-Dose Low High 

Topography: Morphology Control Control Dose Dose 

Adrenal: Pheochromocytoma^5 1/45 (2) 0/43 (0) 3/40 (8) 0/46 (0) 

P Value s c » d N.S. N.S. 

__ 

Relative Risk6 3.375 

Lower Limit 0.284 

Upper Limit 172.561 — 

— 

Weeks to First Observed Tumor 97 88 — — 

Uterus: Endometrial Stromal 

Polypb 3/45 (7) 0/43 (0) 0/39 (0) 0/35 (0) 

P Values0 >d N.S. N.S.
 

Relative Risk6 0.000
 —
 Lower Limit 0.000
 —
 Upper Limit 1.905
 —
 

Weeks to First Observed Tumor 53
 — — —
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice Administered 

3-Amino-9-Ethylcarbazole Hydrochloride in the Diet3 

(continued) 

a

Dosed groups received doses of 800 or 1,200 ppm.
 

''Number of tumor-bearing animals/number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher
 

exact test for the comparison of theat dosed group with its matched-control group when
 

P < 0.05; otherwise, not significant (N.S.) is indicated.
 

^A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 

APPENDIX G
 

ANALYSIS OF FORMULATED DIETS FOR
 

3-AMINO-9-ETHYLCARBAZOLE HYDROCHLORIDE
 

AND ITS HYDROCHLORIDE SALT
 

175
 

APPENDIX G
 

Analysis of Formulated Diets for
 

3-Amino-9-Ethylcarbazole and Its Hydrochloride Salt
 

Duplicate 2-g samples of the diet mixture were each shaken with
 

50 ml 95% ethanol for 15 minutes. The mixture was allowed to
 

settle overnight, and the absorbance of the supernatant, after
 

appropriate dilution, was measured at 306 nra against a "blank"
 

extracted from 2 g of feed from the same lot used to prepare the
 

diet mixture. Concentrations were determined by comparison with
 

standard solutions. Recoveries were determined from duplicate
 

spiked feed samples worked up simultaneously with each set of
 

diet mixtures. Typical recoveries from spiked feed samples
 

(1,000 ppm) were 98% for the free amine and 88% for the hydrochloride.
 

Theoretical No. of Sample Coefficient of Range (ppm) 

Concentrations Samples Analytical Variation (%) 

in Diet (ppm) Mean (ppm) 

300 6 340 12.98 280-400 

600 6 710 6.61 660-800 

400* 4 350 17.89 300-410 

800* 4 730 11.89 650-800 

Hydrochloride salt 

177 

Review of the Bloassay of 3-Amino-9-Ethylcarbazole (Hydrochlori.de)*
 

for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be exposed,
 The members of the Clearinghouse have been drawn from
 

academia, industry, organized labor, public interest groups,
 State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

Subgroup of the Clearinghouse is charged with the responsi­ bility of providing a peer review of reports prepared on
 

NCI-sponsored bioassays of chemicals studied for carcinogenic­ ity. It is in this context that the below critique is given
 

on the bioassay of 3-Amino-9-Ethylcarbazole (Hydrochloride)
 

for carcinogenicity.
 

The reviewer noted that some carbazole compounds have
 

been shown to be carcinogens. Besides a statistically
 

significant incidence of liver tumors induced in both
 

treated rats and mice, several other tumor types were
 

found at increased rates in rats, including lung tumors.
 The reviewer said that the experimental design and study
 

appeared to be adequate to define the carcinogenicity of
 

the compound. In view of the results, the reviewer stated
 

that the compound should be considered to pose a potential
 

carcinogenic risk to man. He moved that the report on the
 

179
 
bioassay of 3-Amino-9-Ethylcarbazole (Hydrochloride) be
 

accepted as written. The motion was approved without
 

objection.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical U.S.A.
 Michael B. Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

*	 Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

«U.S. GOVERNMENT PRINTING OFFICE:B78 260-899/3206 1-3 180 
DHEW Publication No. (NIH) 78-1337