National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 57 

1978 

BIOASSAY OF 

b -TGdR 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 789-61-7 

NCI-CG-TR-57 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

BETA-2'-DEOXY-6-THIOGUANOSINE MONOHYDRATE
 

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) 78-1363
 

BIOASSAY OF
 

BETA-2'-DEOXY-6-THIOGUANOSINE MONOHYDRATE
 

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
 

beta-2'-deoxy-6-thioguanosine monohydrate 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 environmental
 

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 beta-2'-deoxy-6-thioguanosine
 

monohydrate was conducted by Southern Research Institute,
 

Birmingham, Alabama, initially under direct contract to NCI and
 

currently under a subcontract to Tracer Jitco, Inc., prime
 

contractor for the NCI Carcinogenesis Testing Program.
 

The experimental design and doses were determined by Drs. D. P.
 

Griswoldl, J. D. Prejean^, E. K. Weisburger-^, and J. H.
 

Weisburger^'^. MS. J. Belzer^- and Mr. I. Brown-"- were responsible
 

for the administration of the chemical and care of the animals.
 

Data management and retrieval were performed by Ms. C. A.
 

Dominick.1. Histopathologic examinations were performed by Drs.
 

S. D. Kosanke^ and R. B. Thompson^, and the diagnoses included in
 

this report represent their interpretation.
 

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

Mason Research Institute^. The statistical analyses were performed by Dr. J. R. Joiner-*, using methods selected for the
 

bioassay program by Dr. J. J. Gart6. Chemicals used in this
 

bioassay were obtained through Mr. C. Hewitt' and were analyzed
 

by Drs. R. H. Iwamoto^ and W. J. Haggerty^; the results of the
 

analyses were reviewed by Dr. S. S. Olin •
 

This report was prepared at Tracor Jitco-> under the direction of
 

NGI. Those responsible for the report at Tracor Jitco were
 

Dr. Marshall Steinberg, Director of the Bioassay Program; Dr. L.
 

A. Campbell, Deputy Director for Science; Drs. J. F. Robens and
 

R. W. Fogleman, toxicologists; Dr. R. L. Schueler, pathologist;
 

Dr. G. L. Miller, Ms. L. A. Waltz, 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 statistical analysis was reviewed by members of the
 

Mathematical Statistics and Applied Mathematics Section of NCI":
 

Dr. John J. Gart, Mr. Jun-mo Nam, Dr. Hugh M. Pettigrew, and Dr.
 

Robert E. Tarone.
 

The following other scientists at NCI were responsible for
 

evaluating the bioassay experiment, interpreting the results, and
 

reporting the findings:
 

Dr. Kenneth C. Chu
 

Dr. Cipriano Cueto, Jr.
 

Dr. J. Fielding Douglas
 

Dr. Dawn G. Goodman
 

Dr. Richard A. Griesemer
 

Dr. Harry A. Milman
 

Dr. Thomas W. Orme
 

Dr. Robert A. Squire10
 

Dr. Jerrold M. Ward
 

^Southern Research Institute, 2000 Ninth Avenue South,
 

Birmingham, Alabama.
 

iv
 
^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

%ow with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

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

Rockville, Maryland.
 

^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.
 

^Drug Development Branch, Division of Cancer Treatment,
 

National Cancer Institute, National Institutes of Health,
 

Bethesda, Maryland.
 

"Stanford Research Institute, Menlo Park, California.
 

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

Missouri.
 

l^Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

V 

SUMMARY
 

A bioassay of beta—2'-deoxy-6-thioguanosine monohydrate (ß-TGdR)
 

for possible carcinogenicity was conducted by administering the
 

test chemical by intraperitoneal injection to Sprague-Dawley rats
 

and B6C3F1 mice.
 

Groups of 35 rats of each sex were administered ß-TGdR in a
 

buffered saline and polysorbate 80 vehicle at one of two doses,
 

either 3.5 or 7 mg/kg body weight, three times per week for 52
 

weeks, then observed for an additional 26 weeks. Controls
 

consisted of groups of 10 rats of each sex, which were either
 

administered the vehicle alone (matched vehicle controls) or were
 

untreated (matched untreated controls). Pooled controls
 

consisted of the matched vehicle controls of each sex from the
 

current bioassay, combined with 20 corresponding vehicle controls
 

of each sex from similar bioassays of two other test chemicals.
 

All surviving rats were killed at 78 or 79 weeks.
 

Groups of 35 mice of each sex were administered the chemical in a
 

buffered saline and polysorbate 80 vehicle at one of two doses,
 

either 2 or 4 mg/kg, three times per week for 52 weeks, then
 

observed for periods of up to 27 weeks, depending on length of
 

survival. Because of severe toxicity at the high dose, resulting
 

in loss of all mice by week 12 (males) or week 25 (females),
 

additional groups of 35 mice of each sex were administered 1
 

mg/kg on the same schedule. Controls consisted of groups of 15
 

mice of each sex, which were either administered the vehicle or
 

were untreated. Pooled controls consisted of groups of 15
 

vehicle-control animals of each sex from studies using the doses
 

of 2 or 4 mg/kg, combined with corresponding groups of 15
 

vehicle-control animals of each sex from the study using the dose
 

of 1 mg/kg.
 

ß-TGdR was toxic to rats at the doses used in this study. Mean
 

body weights of the high- and low—dose rats of both sexes were
 

lower than those of the corresponding vehicle controls throughout
 

the study. There was also severe early mortality in the high-


dose groups of both sexes and positive dose—related trends in
 

mortality over the period of the bioassay. However, 661 of the
 

vii
 
low-dose males and 77% of the low-dose females survived until
 

termination of the study.
 

In mice, ß-TGdR was toxic at the doses originally selected. Mean
 

body weights were not consistently affected; however, at the high
 

dose only three males and seven females lived past week 7, and
 

all were dead by week 25. In the mid-dose group, only 14% of the
 

males and 6% of the females survived until termination of the
 

study at week 79; in the low-dose group, the survival rate was
 

31% for the males and 29% for the females.
 

Because of the high mortality, time-adjusted statistical analyses
 

were performed for both rats and mice.
 

In rats, the incidence of carcinomas of the ear canal (combined
 

carcinomas and squamous-cell carcinomas) was statistically
 

significant in both sexes. In males, the results of the test for
 

dose-related trend were significant using either matched vehicle
 

(P = 0.046) or pooled vehicle (P = 0.014) controls, but direct
 

comparisons of dosed male rats with matched vehicle or pooled
 

vehicle controls did not show significant differences (matched
 

vehicle controls 0/10, pooled vehicle controls 0/28, low-dose
 

1/31, high-dose 2/7). In females, the results of the test for
 

dose-related trend were significant using either matched vehicle
 

(P = 0.002) or pooled vehicle (P < 0.001) controls, and the
 

incidence in the high-dose group was significantly higher than
 

that in either the matched vehicle (P = 0.023) or pooled vehicle
 

(P < 0.001) controls (matched vehicle controls 0/9, pooled
 

vehicle controls 0/28, low-dose 2/32, high-dose 6/13). There
 

were no such ear canal tumors among 165 historical vehicle
 

controls of either sex or among 220 female untreated controls at
 

the laboratory, and only two such tumors occurred among 215 male
 

untreated controls.
 

In mice, no tumors appeared in statistically significant
 

incidences in the dosed groups compared with the matched vehicle
 

controls, and there was no significant evidence of dose-related
 

trend for any tumors. The incidences of the combination of
 

lymphoma and leukemia were significantly higher in the matched
 

viii
 
vehicle controls of each sex than in the corresponding matched
 

untreated controls (males: matched untreated controls 1/30,
 

matched vehicle controls 19/29; females: matched untreated
 

controls 2/30, matched vehicle controls 21/29). This high
 

incidence in the matched vehicle controls may have been due to a
 

systematic procedural problem associated with injection of the
 

drug.
 

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

low survival of the dosed and vehicle-control groups of mice, as
 

well as the possible procedural problem that may have affected
 

the incidences of tumors in these groups, does not allow a
 

determination to be made of the carcinogenic potential of ß-TGdR
 

in this species. ß-TGdR in the vehicle of 0.05% polysorbate 80
 

was, however, carcinogenic in rats, producing carcinomas of the
 

ear canal in the females and possibly also in the males.
 

ix
 

TABLE OF CONTENTS
 

Page
 

I. Introduction	 1
 

II. Materials and Methods	 3
 

A. Chemical	 3
 

B. Dosage Preparation	 4
 

C. Animals	 4
 

D. Animal Maintenance	 5
 

E. Subchronic Studies	 8
 

F. Designs of Chronic Studies	 9
 

G. Clinical and Pathologic Examinations	 13
 

H. Data Recording and Statistical Analyses	 15
 

III. Results - Rats	 21
 

A. Body Weights and Clinical Signs (Rats)	 21
 

B. Survival (Rats)	 21
 

C. Pathology (Rats)	 24
 

D. Statistical Analyses of Results (Rats)	 25
 

IV. Results - Mice	 31
 

A. Body Weights and Clinical Signs (Mice)	 31
 

B. Survival (Mice)	 31
 

C. Pathology (Mice)	 34
 

D. Statistical Analyses of Results (Mice)	 39
 

V. Discussion	 43
 

VI.	 Bibliography 47
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in
 

Table Al Summary of the Incidence of Neoplasms in
 

Rats Treated with ß-TGdR 49
 

Male Rats Treated with ß-TGdR 51
 

Table A2 Summary of the Incidence of Neoplasms in
 

Female Rats Treated with ß-TGdR 54
 

xi
 
Page 

Appendix B Summary of the Incidence of Neoplasms in 

Mice Treated with ß-TGdR 57 

Table Bl Summary of the Incidence of Neoplasms in 

Male Mice Treated with ß-TGdR (Control Groups) 59 

Table B2 Summary of the Incidence of Neoplasms in 

Female Mice Treated with ß-TGdR (Treated Groups)... 62 

Table B3 Summary of the Incidence of Neoplasms in 

Female Mice Treated with ß-TGdR (Control Groups)... 65 

Table B4 Summary of the Incidence of Neoplasms in 

Female Mice Treated with ß-TGdR (Treated Groups)... 68 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Treated with ß-TGdR 71 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Treated with ß-TGdR 73 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Treated with ß-TGdR 76 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Treated with ß-TGdR 79 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Treated with ß-TGdR 

(Control Groups) 81 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Treated with ß-TGdR 

(Treated Groups) 84 

Table D3 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Treated with ß-TGdR 

(Control Groups) 87 

Table D4 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Treated with ß-TGdR 

(Treated Groups) 90 

Appendix E Analyses of the Incidence of Primary Tumors 

in Rats Treated with ß-TGdR 93 

xii
 
Page 

Table El Analyses of the Incidence of Primary Tumors 

in Male Rats Treated with ß-TGdR 95 

Table E2 Analyses of the Incidence of Primary Tumors 

in Female Rats Treated with ß-TGdR 98 

Table E3 Analyses of the Incidence of Primary Tumors 

in Male Rats Treated with ß-TGdR, 

Using Pooled Controls 103 

Table E4 Analyses of the Incidence of Primary Tumors 

in Female Rats Treated with ß-TGdR, 

Using Pooled Controls 106 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Treated with ß-TGdR Ill 

Table Fl Analyses of the Incidence of Primary Tumors 

in Male Mice Treated with ß-TGdR 113 

Table F2 Analyses of the Incidence of Primary Tumors 

in Female Mice Treated with ß-TGdR 115 

Table F3 Time-adjusted Analyses of the Incidence of 

Primary Tumors in Male Mice Treated with ß-TGdR.... 117 

Table F4 Time-adjusted Analyses of the Incidence of 

Primary Tumors in Female Mice Treated with ß-TGdR.. 119 

TABLES 

Table 1 Design of

in Rats

 Chronic Studies of ß-TGdR 

 10 

Table 2 Design of Chronic Studies of ß-TGdR 

in Mice 11 

xiii
 
 1

 2

 3

 4

 5

 6

Page
 

FIGURES
 

Figure Growth Curves for Rats Treated with 0-TGdR 22
 

Figure Survival Curves for Female Mice
 

Figure Survival Curves for Rats Treated with 0-TGdR 23
 

Figure Growth Curves for Male Mice Treated with /3-TGdR.... 32
 

Figure Growth Curves for Female Mice Treated with /3-TGdR.. 33
 

Figure Survival Curves for Male Mice Treated with 0-TGdR.. 35
 

Treated with 0-TGdR 36
 

xiv
 
I. INTRODUCTION
 

Beta-2'-deoxy-6-thioguanosine monohydrate (ß-TGdR) (CAS 789-61-7;
 

NCI C01581) is an experimental anticancer drug and a derivative
 

of the anticancer drug 6-thioguanine (6-TG). Both drugs are
 

sulphur analogs of the purine base guanine, a similarity that
 

allows them to be incorporated into DNA. In tests using extracts
 

of murine and human tissues, ß—TGdR was phosphorylated to
 

nucleotide mono—, di-, and triphosphates; in further tests using
 

extracts of human tissues, the nucleotide triphosphate that was
 

formed was found to be incorporated into DNA (Peery and Lepage,
 

1969). Because of the carcinostatic effects of ß-TGdR in certain
 

mouse tumor systems that were resistant to 6-TG (Lepage et al. ,
 

1964), it was hoped that this compound could be used to bypass
 

the resistance to 6-TG that develops in man. At this time,
 

however, there are not sufficient data from clinical trials to
 

determine whether ß-TGdR has any real advantage over 6-TG (Loo et
 

al., 1975). ß-TGdR was selected for testing in the
 

carcinogenesis bioassay program as part of an attempt to evaluate
 

the carcinogenicity of drugs that may be used in humans for
 

prolonged periods.
 

1 

II. MATERIALS AND METHODS
 

A. Chemical
 

The ß-TGdR used in this study was obtained through the Drug
 

Development Branch, Division of Cancer Treatment, National Cancer
 

Institute (NCI). Lot No. RN C19052F was manufactured by Riker
 

Laboratories, Northbridge, California, and Lot No. AJ-31, by
 

Aerojet General, Sacramento, California. Both lots were used
 

during the chronic study.
 

The identity and purity of Lot No. RN C19052F were confirmed at
 

Stanford Research Institute by melting point, elemental analyses
 

(C, H, N) for CioH13N5°3s*H2°> and infrared and ultraviolet
 

spectra. Karl-Fischer analysis for water (6.0%) was correct for
 

one molecule of water of hydration. Paper chromatography showed
 

no detectable thioguanine or 2-deoxyribose.
 

The identity and purity of Lot No. AJ-31 were confirmed at
 

Midwest Research Institute by melting point (differential thermal
 

analysis); elemental analyses (C, H, N, 0, S); and infrared,
 

ultraviolet, and nuclear magnetic resonance spectra. Karl-


Fischer analysis for water (6.7 ± 0.4%) was slightly high (6.0%
 

theoretical). The absence of thioguanine, alpha anomer, and
 

2-deoxyribose was confirmed by thin-layer and paper
 

chromatography.
 

3 
In conclusion, no impurities were found in either lot. Both lots
 

appeared to be > 98% pure ß-2'-deoxy-6-thioguanosine monohydrate.
 

The different lots of bulk chemical were stored in their original
 

containers at 5°C.
 

B. Dosage Preparation
 

Test solutions of ß-TGdR were prepared by adding the drug to a
 

vehicle of 0.05% polysorbate 80 in saline (for prechronic
 

studies) or buffered saline (for chronic studies) and mixing the
 

final suspension in a Potter-Elvehjem tissue grinder with a
 

Teflon pestle for 20 seconds. The concentrations of ß-TGdR that
 

were prepared were 0.01, 0.02, 0.04, 0.14, and 0.28%. The
 

solutions were stored in glass bottles at 5°C for up to 1 week.
 

C. Animals
 

Female Sprague-Dawley rats and male Swiss mice were used in the
 

subchronic studies, Sprague-Dawley rats and B6C3F1 mice of both
 

sexes were used in the chronic studies. The Sprague-Dawley rats
 

and B6C3F1 mice were obtained from Charles River Breeding
 

Laboratories, Inc., Wilmington, Massachusetts, through contracts
 

of the Division of Cancer Treatment, NCI. The Swiss mice were
 

obtained from Charles River Laboratories and from Purina
 

Laboratories, Staatsburg, New York. Rats used in the chronic
 

4 
studies were 30 days of age when received at the laboratory, and
 

mice were 41 days of age. Groups of mice started later in the
 

study were 32 days of age when received. On arrival at the
 

laboratory, all animals were quarantined (rats for 8 days, mice
 

in original study for 7 days, mice in later studies for 17 days).
 

Following this period, animals with no clinical signs of disease
 

were assigned to control or dosed groups and earmarked for
 

individual identification.
 

D. Animal Maintenance
 

All animals were housed in temperature- and humidity-controlled
 

rooms. The temperature range was 20-24°C, and the relative
 

humidity was maintained at 40-60%. The room air was changed 15
 

times per hour and passed through both intake and exhaust
 

fiberglass roughing filters. In addition to natural light,
 

illumination was provided by fluorescent light for 9 hours per
 

day. Wayne® Lab Blox animal feed (Allied Mills, Inc., Chicago,
 

111.) and water were supplied daily and were available ad
 

libitum.
 

Rats and mice were housed five per cage in solid-bottom stainless
 

steel cages (Hahn Roofing and Sheet Metal Co., Birmingham, Ala.).
 

Rat cages were provided with Iso-Dri® hardwood chip bedding
 

(Carworth, Edison, N.J.), and the cage tops were covered with
 

5 
disposable filter bonnets throughout the study. Mouse cages were
 

provided with Sterolit® clay bedding (Englehard Mineral and
 

Chemical Co., New York, N.Y.), and the cage tops were covered
 

with disposable filter bonnets for mid- and high-dose mice
 

beginning at week 63 and for low-dose mice beginning at week 51.
 

Bedding was replaced once per week; cages, water bottles, and
 

feeders were sanitized at 82°C once per week; racks were cleaned
 

once per week.
 

The rats and mice were housed in separate rooms. Control animals
 

were housed with respective dosed animals. Animals administered
 

ß-TGdR were maintained in the same rooms as animals of the same
 

species administered the following chemicals:
 

RATS
 

Gavage Studies
 

cholesterol (p-(bis(2-chloroethyl)amino)phenyl)acetate
 

(phenesterin) (CAS 3546-10-9)
 

estradiol bis((p-(bis(2-chloroethyl)amino)phenyl)acetate)
 

(estradiol mustard) (CAS 22966-79-6)
 

Intraperitoneal Injection Studies
 

4'-(9-acridinylamino)methansulfon-m-aniside monohydrochloride
 

(MAAM) (NSC 141549)
 

acronycine (CAS 7008-42-6)
 

5-azacytidine (CAS 320-67-2)
 

1,4-butanediol dimethanesulfonate (busulfan) (CAS 55-98-1)
 

emetine dihydrochloride tetrahydrate (CAS 316-42-7)
 

3,3'-iminobis-l-propanol dimethanesulfonate (ester)
 

hydrochloride (IPD) (CAS 3458-22-8)
 

Of)-4,4'-(l-methyl-l,2-ethanediyl)bis-2,6-piperazinedione
 

(ICRF-159) (CAS 21416-87-5)
 

6 
N,3-bis(2-chloroethyl)tetrahydro-2H-l,3,2-oxazaphosphorin-2­

amine-2-oxide (isophosphamide) (CAS 3778-73-2)
 

N-(2-chloroethyl)-N-(l-methyl-2-phenoxyethyl)benzylamine
 

hydrochloride (phenoxybenzamine hydrochloride) (CAS 63-92-3)
 

N-(l-methylethyl)-4-((2-methylhydrazino)methyl)benzamide
 

monohydrochloride (procarbazine) (CAS 366-70-1)
 

tris(l-aziridinyl)phosphine sulfide (thio-TEPA) (CAS 52-24-4)
 

2,4,6-tris(dimethylamino)-s-triazine (CAS 645-05-6)
 

MICE
 

Feed Studies
 

4-acetyl-N-((cyclohexylamino)carbonyl)benzenesulfonamide
 

(acetohexamide) (CAS 968-81-0)
 

anthranilic acid (CAS 118-92-3)
 

l-butyl-3-(p-tolylsulfonyl)urea (tolbutamide) (CAS 64-77-7)
 

4-chloro-N-((propylamino)carbonyl)benzenesulfonamide
 

(chlorpropamide) (CAS 94-20-2)
 

5-(4-chlorophenyl)-6-ethyl-2,4-pyrimidinediamine
 

(pyrimethamine) (CAS 58-14-0)
 

2,6-diamino-3-(phenylazo)pyridine hydrochloride
 

(phenazopyridine hydrochloride) (CAS 136-40-3)
 

L-tryptophan (CAS 73-22-3)
 

N-9H-fluoren-2-ylacetamide (CAS 53-96-3)
 

N-(p-t oluenesulf ony1)-N'-hexame thyleniminourea
 

(tolazamide) (CAS 1156-19-0)
 

1-phenethylbiguanide hydrochloride (phenformin) (CAS 114-86-3)
 

pyrazinecarboxamide (pyrazinamide) (CAS 98-96-4)
 

4,4'-sulfonyldianiline (dapsone) (CAS 80-08-0)
 

4,4'-thiodianiline (CAS 139-65-1)
 

ethionamide (CAS 536-33-4)
 

Gavage Studies
 

cholesterol (p-(bis(2-chloroethyl)amino)phenyl)acetate
 

(phenesterin) (CAS 3546-10-9)
 

estradiol bis((p-(bis(2-chloroethyl)amino)phenyl)acetate)
 

(estradiol mustard) (CAS 22966-79-6)
 

Intraperitoneal Injection Studies
 

4'-(9-acridinylamino)methansulfon-m-aniside monohydrochloride
 

(MAAM) (NSC 141549)
 

acronycine (CAS 7008-42-6)
 

5-azacytidine (CAS 320-67-2)
 

1,4-butanediol dimethanesulfonate (busulfan) (CAS 55-98-1)
 

7 
emetine dihydrochloride tetrahydrate (CAS 316-42-7)
 

3,3'-iminobis-l-propanol dimethanesulfonate (ester)
 

hydrochloride (IPD) (CAS 3458-22-8)
 

(+)-4,4'-(l-methyl-l,2-ethanediyl)bis-2,6-piperazinedione
 

(ICRF-159) (CAS 21416-87-5)
 

N,3-bis(2-chloroethyl)tetrahydro-2H-l,3,2-oxazaphosphorin-2­

amine-2-oxide (isophosphamide) (CAS 3778-73-2)
 

N-(2-chloroethyl)-N-(l-methyl-2-phenoxyethyl)benzylamine
 

hydrochloride (phenoxybenzamine hydrochloride) (CAS 63-92-3)
 

N-(l-methylethyl)-4-((2-methylhydrazino)methyl)benzamide
 

monohydrochloride (procarbazine) (CAS 366-70-1)
 

tris(l-aziridinyl)phosphine sulfide (thio-TEPA) (CAS 52-24-4)
 

2,4,6-tris(dimethylamino)-s-triazine (CAS 645-05-6)
 

adriamycin (CAS 23214-92-8)
 

E.	 Subchronic Studies
 

»
 

In the subchronic studies, ß-TGdR in 0.05% polysorbate 80 in
 

saline was administered by intraperitoneal injection to female
 

Sprague-Dawley rats and male Swiss mice to estimate the maximum
 

tolerated doses (MTD) of ß-TGdR, on the basis of which "high"
 

(estimated MTD) and "low" (1/2 estimated MTD) doses were determined for administration in the chronic studies. Rats received
 

doses of 3, 7, 15, 30, or 60 mg/kg body weight. Doses of 5, 12,
 

25, 50, or 100 mg/kg were given to mice initially, and doses of
 

0.38, 0.75, 1.5, or 3 mg/kg were given in a second study. The
 

animals were injected three times per week for 45 days, then
 

observed for an additional 45 days. Five animals of each species
 

were tested at each dose, 10 animals of each species were
 

maintained as vehicle controls, and 10 as untreated controls.
 

In rats, death occurred in none of the animals administered 3 or
 

8 
7 mg/kg, in 3/5 administered 15 mg/kg, in 3/5 administered 30
 

mg/kg, and in 4/5 administered 60 mg/kg. Mean body weights of
 

the rats at doses of 3 or 7 mg/kg were essentially unaffected;
 

mean body weights of animals at 15, 30, or 60 mg/kg were markedly
 

depressed at the end of the period of administration,, but were
 

comparable to those of the controls by the termination of the
 

study. No gross abnormalities were seen in any of the animals at
 

necropsy. The low and high doses for the chronic studies using
 

rats were set at 3.5 and 7 mg/kg.
 

In mice, death occurred in all animals administered doses of 5 to
 

100 mg/kg and in 1/5 administered 3 mg/kg; there were no deaths
 

at doses below 3 mg/kg. No gross abnormalities were found at.
 

necropsy. Because mean body weights of the mice at 3 mg/kg or
 

lower were comparable to those of the controls, the high dose was
 

set midway between the lowest dose causing excessive death and
 

the highest dose at which no effects on body weight were
 

observed. Thus, the low and high doses for the chronic studies
 

using mice were set at 2 and 4 mg/kg.
 

F. Designs of Chronic Studies
 

The designs of the chronic studies are shown in tables 1 and 2.
 

Since the number of rats in the matched vehicle—control group was
 

small, a pooled vehicle-control group also was used for
 

9 
Table 1. Design of Chronic Studies of ß-TGdR in Rats
 

Sex and 

Test 

Initial 

No. of 

ß-TGdR 

Doseb 

Time on Study 

Dosed Observed 

Group Animals3 me /kg /day (weeks) (weeks ) 

Male 

Matched 

Untreated-Control 10 0 79 

Matched 

Vehicle-Control 10 Oc 52 27 

Low-Dose 35 3.5 52 26 

High-Dose 35 7 52 26 

Female 

Matched 

Untreated-Control 10 0 79 

Matched 

Vehicle-Control 10 QC 52 27 

Low-Dose 35 3.5 52 26 

High-Dose 35 7 52 26 

a

All animals were 38 days of age when placed on study.
 

b

The drug was administered intraperitoneally in a vehicle of
 

phosphate-buffered saline and polysorbate 80 at a volume of 0.25
 

ml/100 g body weight, three times per week during the period of
 

administration of the drug. Doses were based on individual
 

weights. The same needle for injection was used for each group of
 

animals within a cage.
 

c

Vehicle-control groups received only phosphate-buffered saline
 

and polysorbate 80 at a volume 0.25 ml/100 g body weight. The
 

same bottle of vehicle solution was used for all vehicle-control
 

animals on test at any given time.
 

10
 
Table 2. Design of Chronic Studies of ß-TGdR in Mice
 

Sex and
 

Test
 

Group
 

Male
 

Low-Dose Matched
 

Untreated-Control
 

Low-Dose Matched
 

Vehicle-Control
 

Low-Dose
 

Mid- and High-Dose
 

Matched Untreated-


Control
 

Mid- and High-Dose
 

Matched Vehicle-


Control
 

Mid-Dosee
 

High -Dose
 

Female
 

Low-Dose Matched
 

Untreated-Control
 

Low-Dose Matched
 

Vehicle-Control
 

Low-Dose
 

Mid- and High-Dose
 

Matched Untreated-


Control
 

Mid- and High-Dose
 

Matched Vehicle-


Control
 

Mid-Dosee
 

High-Dose
 

Initial
 

No. of
 

Animals3
 

15
 

15
 

35
 

15
 

15
 

35
 

35
 

15
 

15
 

35
 

15
 

15
 

35
 

35
 

ß-TGdR
 

Doseb
 

rag /kg /day
 

0
 

Oc
 

1
 

0
 

QC
 

2
 

4
 

0
 

Oc
 

1
 

0
 

QC
 

2
 

4
 

Time
 

Dosed
 

(weeks)
 

52
 

52
 

52
 

52
 

12f
 

52
 

52
 

52
 

52
 

25*
 

on Study
 

Observed
 

(weeks)
 

78
 

26
 

25
 

79
 

17d
 

27
 

78
 

26
 

25
 

79
 

l?d
 

27
 

a

All animals were 48-49 days of age when placed on study.
 

11
 
Table 2. Design of Chronic Studies of ß-TGdR in Mice
 

(continued)
 

"The drug was administered intraperitoneally in a vehicle of
 

phosphate-buffered saline and polysorbate 80 at a volume of 1
 

ml/100 g body weight, three times per week during the period of
 

administration of the drug. Doses were based on the mean weight
 

of the animals in each cage. The same needle for injection was
 

used for each group of animals within a cage.
 

c

Matched vehicle-control groups received only phosphate-buffered
 

saline and polysorbate 80 at a volume of 1 ml/100 g body weight.
 

The same bottle of vehicle solution was used for all vehicle-


control animals on test at any given time.
 

^Observation of animals terminated at the time indicated,
 

due to the death of all animals.
 

e

lhe mid-dose group was originally the low-dose group; however,
 

because of toxicity, a new low—dose group was started after 12
 

weeks of the study.
 

^Administration of ß-TGdR to the animals terminated at the time
 

indicated, due to the death of all animals.
 

12
 
statistical comparisons. The 10 matched vehicle-control rats of
 

each sex from the current studies on ß-TGdR were combined with 10
 

rerun vehicle-control rats of each sex from studies on thio-TEPA
 

and 10 rerun vehicle-control rats of each sex from studies on
 

3,3'-iminobis-l-propanol dimethanesulfonate (ester) hydrochloride
 

(IPD), to give pooled vehicle-control groups of 30 rats of each
 

sex. The vehicle-control rats that were used in the pooled
 

vehicle-control groups were of the same strain and from the same
 

supplier, and they were examined by the same pathologists;
 

further, the different vehicle-control groups of which the pooled
 

groups were composed were pla ed on study at starting times
 

differing by no more than 3 months.
 

For statistical tests using mice, the 15 untreated-control mice
 

of each sex used for the low-dose groups in the current bioassay
 

were combined with the 15 untreated-control mice of each sex used
 

for the mid- and high-dose groups in the current bioassay, to
 

give final groups of 30 untreated-control mice of each sex.
 

Groups of 30 vehicle-control mice of each sex were made up
 

similarly.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity, and
 

animals that were moribund were killed and necropsied, except for
 

those dying prior to day 100, due, presumably, to toxicity of the
 

13
 
test chemical. Rats were individually weighed every 2 weeks for
 

62 weeks and once per month thereafter; mice were individually
 

weighed every 2 weeks for 62 weeks in the original study, for 51
 

weeks in the rerun study, and once per month thereafter.
 

Palpation for masses was carried out at each weighing.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of major tissues, major organs, and all gross lesions
 

from killed animals and from animals found dead. The following
 

tissues were examined microscopically: skin, muscle, lungs and
 

bronchi, trachea, bone and bone marrow, spleen, lymph nodes,
 

thymus, heart, salivary gland, liver, gallbladder and bile duct
 

(mice), pancreas, esophagus, stomach, small intestine, large
 

intestine, kidney, urinary bladder, pituitary, adrenal, thyroid,
 

parathyroid, mammary gland, prostate or uterus, testis or ovary,
 

brain, and sensory organs. Peripheral blood smears were prepared
 

from each animal whenever possible. Occasionally, additional
 

tissues were also examined microscopically. 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 diagnosis.
 

A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals were
 

14
 
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 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.
 

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
 

15
 
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.
 

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
 

16
 
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, with
 

continuity correction (Armitage, 1971), was also used. Under the
 

assumption of a linear trend, this test determines if the slope
 

of the dose-response curve is different from zero at the one-


tailed 0.05 level of significance. Unless otherwise noted, the
 

direction of the significant trend is a positive dose relationship. This method also provides a two-tailed test of departure
 

from linear trend.
 

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
 

17
 
was found at the anatomic site of interest before week 52. When
 

such an early tumor was found, comparisons were based exclusively
 

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 (Fisher exact tests, Cochran-Armitage tests, etc.) 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 with 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
 

18
 
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
 

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.
 

19
 

III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights for the low- and high-dose rats of each sex
 

were lower than those for the corresponding matched vehicle
 

controls throughout the study (figure 1). 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.
 

Clinical signs, other than those of body weights, observed
 

sporadically in a few animals included alopecia, cutaneous
 

erythema and sores, tilting of the head, and exophthalmos.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered ß-TGdR at the
 

doses of this bioassay, together with those of the controls, are
 

shown in figure 2.
 

The result of the Tarone test for positive dose-related trend in
 

mortality is significant in either sex (P < 0.001), but an
 

indicated departure from linear trend is observed (P = 0.008 in
 

males, P = 0.003 in females), due to the steep increase in
 

mortality in the high-dose rats. In male rats, 6/35 (17%) of the
 

high-dose group, 23/35 (66%) of the low-dose group, 9/10 (90%) of
 

21
 
Figure 1. Growth Curves For Rats Treated With (ß -TGdR 

22 
Figure 2. Survival Curves For Rats Treated With ß-TGdR 

23 
the matched vehicle controls, and all 10 of the matched untreated
 

controls lived to the end of the study. In female rats, 7/35
 

(20%) of the high-dose group, 27/35 (77%) of the low-dose group,
 

9/10 (90%) of the matched vehicle controls, and 9/10 (90%) of the
 

matched untreated controls lived to the end of the study. In
 

spite of the early death in the high-dose groups, a sufficient
 

number of animals of each sex survived to exhibit a significant
 

increase in the incidence of tumors in the dosed groups.
 

•»
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

Appendix A, tables Al and A2; findings on nonneoplastic lesions
 

are summarized in Appendix C, tables Cl and C2.
 

A variety of tumors occurred both in the untreated controls and
 

the vehicle controls (phosphate-buffered saline and polysorbate
 

80) and in the dosed groups. Some types of neoplasms occurred
 

only in rats of dosed groups, or with greater frequency in dosed
 

groups when compared with controls. Most of these lesions are
 

not uncommon in this strain of rat independent of any treatment.
 

However, carcinomas or squamous-cell carcinomas of the ear canal
 

occurred in 1/35 low-dose males, 2/30 high-dose males, 2/32 low-


dose females, and 6/27 high-dose females. None were seen in the
 

controls.
 

24
 
In addition to the neoplastic lesions, a number of degenerative,
 

proliferative, and inflammatory changes were also encountered in
 

animals of the dosed and control groups (Appendix C). For the
 

most part, these nonneoplastic lesions are commonly seen in aged
 

rats.
 

The small number of tumors observed in the high-dose groups may
 

have been influenced by the decreased life span in these animals.
 

The lack of tumors in these animals is an indication of toxicity
 

and not of a lack of carcinogenic activity. The shortened life
 

spans of males and females in the high-dose groups, which were
 

frequently associated with bone-marrow atrophy and
 

bronchopneumonia, reduced the likelihood of detecting any
 

possible carcinogenic activity of the test chemical in these
 

groups.
 

In the judgment of the pathologists, ß-TGdR in phosphate-


buffered saline and polysorbate 80 induced squamous-cell
 

carcinomas of the ear canal in female Sprague-Dawley rats.
 

D. Statistical Analyses of Results (Rats)
 

Tables E1-E4 in Appendix E contain the statistical analyses of
 

the incidences of those primary tumors that occurred in at least
 

two animals and with an incidence of at least 5% in one or more
 

than one group. Two sets of controls are used in the analyses:
 

25
 
the matched vehicle controls and the pooled vehicle controls.
 

Untreated controls are not included in the tables and analyses,
 

because the test conditions of the matched vehicle controls more
 

closely resemble those of the dosed animals; however, the
 

incidences in the untreated and vehicle controls are not
 

statistically different.
 

In male rats, none of the incidences of tumors at any site are
 

significant in the positive direction. The analyses of the
 

incidences of squamous-cell carcinomas of the ear canal in female
 

rats show a significant trend by the Cochran-Armitage test when
 

the pooled vehicle controls are used (P = 0.010). The Fisher
 

exact test shows that the incidence in the high-dose group is
 

significantly greater than that in the pooled vehicle controls
 

(P = 0.019). The statistical conclusion suggests the possibility
 

of dose association. When the combined incidence of carcinomas
 

and squamous-cell carcinomas of the ear canal is considered,
 

increased significance is observed over that of squamous-cell
 

carcinomas alone.
 

Due to the severe early mortality in the high-dose rats of each
 

sex, time-adjusted analyses are performed, eliminating animals
 

that died before week 52 on study; however, when a tumor is found
 

at the body site of interest before week 52, comparisons are
 

based exclusively on animals that survived at least as long as
 

26
 
the animal in which the first tumor was found. These time-


adjusted analyses are shown in tables E3 and E4.
 

In male rats, there is a significant trend in the time-adjusted
 

incidence of the combined incidence of carcinomas and squamouscell carcinomas of the ear canal. These time-adjusted incidences
 

become 0/28, 0/10, 1/31 (3%), and 2/7 (29%) in the pooled
 

vehicle-control, matched vehicle-control, low-dose, and high-dose
 

groups, respectively. The Cochran-Armitage test result is
 

significant when either the pooled vehicle controls (P = 0.014)
 

or the matched vehicle controls (P = 0.046) are used. The Fisher
 

exact comparison of incidences between the high-dose and pooled
 

vehicle-control groups shows a probability level of 0.035, which
 

is above the 0.025 level required by the Bonferroni inequality
 

criterion when multiple comparison is considered. The time-


adjusted incidence of lymphoma in male rats shows a significant
 

trend (P = 0.025) by the Cochran-Armitage test when the pooled
 

vehicle-control group is used; however, the Fisher exact test
 

results are not significant.
 

In female rats, the only significant time-adjusted incidences of
 

tumors in the positive direction are those of the ear canal. The
 

time-adjusted incidences of squamous-cell carcinomas of the ear
 

canal in female rats become 0/30, 0/10, 2/31 (6%), and 5/12 (42%)
 

in the pooled vehicle-control, matched vehicle-control, low-dose,
 

27
 
and high-dose groups, respectively. The results of the Cochran-


Armitage test are significant (P < 0.005) when either control
 

group is used, but an indicated departure from linear trend is
 

observed (P = 0.043) when the pooled-control group is used,
 

because of the steep increase in incidence in the high-dose
 

group. The Fisher exact test shows that the incidence in the
 

high-dose group is significantly higher (P = 0.001) than that in
 

the pooled vehicle-control group. The Fisher exact comparison of
 

incidences between the high-dose and matched vehicle-control
 

groups indicates a probability level of 0.030, which is above the
 

0.025 level required by the Bonferroni inequality criterion when
 

multiple comparison is considered. When the combined time-


adjusted incidence of carcinomas and squamous-cell carcinomas of
 

the ear canal is considered, increased significance is observed
 

over that of the squamous-cell carcinomas alone.
 

The spontaneous rate of tumors compiled to date at this laboratory in the historical vehicle controls shows no incidence of
 

tumors of the ear canal in the 165 Sprague-Dawley rats of each
 

sex; however, 2/215 (1%) untreated male rats were observed to
 

have such tumors. No tumors of this type were seen in the 220
 

untreated female controls at this laboratory.
 

The time-adjusted analysis of female rats indicates a significant
 

(P = 0.022) negative result in the Fisher exact test when the
 

28
 
incidence of chromophobe adenomas of the pituitary gland in the
 

high-dose group of female rats is compared with that in the
 

matched vehicle controls. Except for this comparison, all of the
 

Fisher exact test results of comparisons of the incidences of
 

this tumor are above or equal to the 0.025 level required when
 

the multiple comparison criterion is considered. The additional
 

deaths which occurred in each of the dosed groups of female rats
 

compared with the controls may account for this negative result.
 

No other Fisher exact test result was significant in the negative
 

direction. The analysis of this tumor in male rats indicates no
 

significant results of the Fisher exact test when time-adjusted
 

data are used.
 

Due to the early deaths in the high-dose groups, several of the
 

results for the Cochran-Armitage test are in the direction of a
 

negative trend, as indicated in the tables.
 

In the non-adjusted tables, significant results in the negative
 

direction were obtained for the incidence of tumors of the
 

pituitary gland in both sexes, and of fibroadenomas of the
 

mammary gland in female rats.
 

In summary, the statistical conclusion suggests that the
 

incidence of neoplasms of the ear canal in dosed rats is
 

associated with the administration of ß-TGdR.
 

29
 

IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

In spite of the toxicity of ß-TGdR and the many early deaths in
 

the high-dose groups, mean body weights were not consistently
 

affected by administration of ß-TGdR (figures 3 and 4).
 

Fluctuations 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. Progressive weight loss occurred in a few animals.
 

No other clinical signs that could be related to administration
 

of the drug were recorded.
 

To control respiratory disease, the high- and mid-dose groups,
 

together with corresponding controls, received oxytetracycline in
 

the drinking water at 0.6 mg/ml for 5 days during week 42 and at
 

0.3 mg/ml for the following 5 days. The low-dose group, with
 

controls, received oxytetracyline in the same pattern beginning
 

at week 31. Also, propylene glycol was vaporized for about 2
 

months in the rooms housing the mice, beginning at week 42 for
 

the high- and mid-dose groups and at week 30 for the low-dose
 

groups.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

31
 
TIME ON STUDY (WEEKS) 

FigureS. Growth Curves For Male Mice Treated With ß-TGdR 

32 
TIME ON STUDY (WEEKS) 

Figure 4. Growth Curves For Female Mice Treated Withß-TGdR 

33 
survival for male and female mice administered ß-TGdR by intraperitoneal injection at the doses of this bioassay, together with
 

those of the controls, are shown in figures 5 and 6.
 

The results of the Tarone tests for positive dose-related trend
 

in mortality are significant (P < 0.001) in either sex. In male
 

mice, 0/35 (0%) of the high-dose group, 5/35 (14%) of the mid-


dose group, 11/35 (31%) of the low-dose group, 4/15 (27%) of the
 

low-dose matched vehicle-control group, 0/15 (0%) of the mid- and
 

high-dose matched vehicle-control group, 9/15 (60%) of the low-


dose matched untreated-control group, and 10/15 (67%) of the mid-


and high-dose matched untreated-control group lived to the end of
 

the study. In the females, 0/35 (0%) of the high-dose group,
 

2/35 (6%) of the mid-dose group, 10/35 (29%) of the low-dose
 

group, 3/15 (20%) of the low-dose matched vehicle controls, 0/1.5
 

(0%) of the mid- and high-dose matched vehicle controls, 12/15
 

(80%) of the low-dose untreated controls, and 13/15 (87%) of the
 

mid- and high-dose untreated controls survived to the end of the
 

study. The severe early mortality in the dosed male and female
 

mice may have prevented the observation of late-appearing tumors.
 

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.
 

34
 
TIME ON STUDY (WEEKS) 

Figure 5. Survival Curves For Male Mice Treated With ß-TGdR 

35 
TIME ON STUDY (WEEKS) 

Figure 6. Survival Curves For Female Mice Treated With /3-TGdR 

36 
There was a high incidence of lymphoreticular neoplasms,
 

primarily malignant lymphomas, in the matched vehicle controls
 

(phosphate-buffered saline and polysorbate 80) and the low- and
 

mid-dose groups. The incidences of animals bearing these
 

neoplasms were as follows:
 

Matched
 

Untreated
 

Controls
 

MALES
 

Number of animals
 

necropsied
 

Multiple organs (lymphore ­

ticular)
 

Malignant lymphoma, NOS*
 

Malignant lymphoma, undiff erentiated type
 

Malignant lymphoma,
 

lymphocytic type
 

Malignant lymphoma,
 

histiocytic type
 

Malignant lymphoma,
 

mixed type
 

Lymphocytic leukemia
 

Total number of animals
 

with neoplasms
 

Percent of animals
 

with neoplasms
 

*Not otherwise specified
 

30
 

0
 

0
 

1
 

0
 

0
 

0
 

1
 

3
 

Matched 

Vehicle Low Mid 

Controls Dose Dose 

29 35 32 

2 1 1 

0 2 6 

4 7 9 

12 4 0 

1 3 0 

0 0 2_ 

19 17 18 

66 49 56 

37
 
Matched Matched 

Untreated Vehicle Low Mid 

Controls Controls Dose Dose 

FEMALES
 

Number of animals
 

necropsied 30 29 33 29
 

Multiple Organs (lymphore
 

ticular)
 

Malignant lymphoma, NOS 1 4 2 2
 

Malignant lymphoma,
 i
 undiff erentiated type 0 4 17
 

Malignant lymphoma,
 

lymphocytic type 0 0 2 2
 

Malignant lymphoma,
 

histiocytic type 1 11 14 0
 

Lymphocytic leukemia 0 1 0 0_
 

Total number of animals
 

with neoplasms 2 20 19 21
 

Percent of animals
 

with neoplasms 7 69 58 72
 

Data for the high-dose groups are not presented, since all but
 

one of the animals in these groups died early, and they were not
 

examined.
 

Lymphocytic leukemia was diagnosed in one vehicle-control female
 

and two dosed males. This diagnosis was used because of the
 

presence of numerous neoplastic lymphoid cells in the vascular
 

sinusoids of the livers and spleens and peripheral blood smears.
 

Since there was extensive lymphoid cell infiltration in many
 

organs and tissues, these cases probably represented malignant
 

lymphomas with accompanying leukemia.
 

38
 
Malignant lymphomas appeared with approximately equal frequency
 

in the vehicle-control and dosed groups of both sexes. Thus, the
 

neoplasms do not appear to be chemical related.
 

The lack of tumors in the high-dose male and female groups of
 

mice could be related to marked decrease in life span, which is
 

an indication of the toxicity of ß-TGdR and does not necessarily
 

indicate a lack of carcinogenic activity.
 

The other neoplasms listed in Appendix B appeared with
 

approximately equal frequency in dosed and control mice, or
 

appeared in insignificant numbers.
 

In addition to the neoplastic lesions, a large number of
 

degenerative, proliferative, and inflammatory changes were
 

encountered in animals of the dosed and control groups (Appendix
 

D). For the most part, these nonneoplastic lesions are commonly
 

seen in aged mice and were not associated with increased
 

mortalities or decreased life spans.
 

In the judgment of the pathologists, this study failed to provide
 

conclusive evidence of carcinogenic activity of ß-TGdR in B6C3F1
 

mice.
 

D. Statistical Analyses of Results (Mice)
 

Tables F1-F4 in Appendix F contain the statistical analyses of
 

39
 
the incidences of those primary tumors that occurred in at least
 

two animals and with an incidence of at least 5% in one or more
 

than one group. The high-dose groups are not included in the
 

tables, since all but one of the animals in these groups died
 

early and they were not examined. The matched untreated controls
 

are not included in the tables and analyses, because the test
 

conditions of the vehicle controls more closely resemble those of
 

the dosed animals.
 

The combined incidences of lymphoma and leukemia in the matched
 

vehicle controls are significantly higher than those in the
 

matched untreated controls in both sexes. In male mice, they are
 

1/30 in the matched untreated controls and 19/29 in the matched
 

vehicle controls; in females they are 2/30 and 21/29 in the same
 

order. These differences may indicate a difficulty in the
 

execution of this bioassay.
 

Due to the severe early mortality in the dosed mice and in the
 

matched vehicle controls, time-adjusted analyses are performed,
 

eliminating animals that died before week 52 on study; however,
 

when a tumor is found at the body site of interest before week
 

52, comparisons are based exclusively on animals that survived at
 

least as long as the animal in which the first tumor was found.
 

These time-adjusted analyses are shown in tables F3 and F4.
 

40
 
The results of the Cochran-Armitage test for positive dose-


related trend in incidences and those of the Fisher exact test
 

for direct comparison of control and dosed groups are not
 

significant in either sex, before or after time adjustment.
 

In each of the 95% confidence intervals of relative risk, shown
 

in the tables, the value of one is included; this indicates the
 

absence of significant results. It should also be noted that
 

each of the intervals has an upper limit greater than one,
 

indicating the theoretical possibility of the induction of tumors
 

by this compound, which could not be detected under the
 

conditions of this test.
 

41
 

V. DISCUSSION
 

ß-TGdR administered intraperitoneally was toxic to rats at the
 

doses used in this study. Mean body weights of the high- and
 

low-dose rats of both sexes were lower than those of the
 

corresponding vehicle controls throughout the study. There was
 

also severe early mortality in the high-dose groups of both sexes
 

and positive dose-related trends in mortality over the period of
 

the bioassay. However, 66% of the low-dose males and 77% of the
 

low-dose females survived until termination of the study.
 

In mice, ß-TGdR was toxic at the doses originally selected. Mean
 

body weights were not consistently affected; however, at the high
 

dose, only three males and seven females lived past week 7, and
 

all were dead by week 25. In the mid-dose group, only 14% of the
 

males and 6% of the females survived until termination of the
 

study at week 79; in the low-dose group, the survival rate was
 

31% for the males and 29% for the females.
 

Because of the high mortality, time-adjusted statistical analyses
 

were performed for both rats and mice.
 

In rats, the incidence of carcinomas of the ear canal (combined
 

carcinoma and squamous-cell carcinoma) was statistically
 

significant in both sexes. In males, the results of the test for
 

dose-related trend were significant using either matched vehicle
 

43
 
(P = 0.046) or pooled vehicle (P = 0.014) controls, but direct
 

comparisons of dosed male rats with matched vehicle or pooled
 

vehicle controls did not show significant differences (matched
 

vehicle controls 0/10, pooled vehicle controls 0/28, low-dose
 

1/31, high-dose 2/7). In females, the results of the test for
 

dose-related trend were significant using either matched vehicle
 

(P = 0.002) or pooled vehicle (P < 0.001) controls, and the
 

incidence in the high-dose group was significantly higher than
 

that in either the matched vehicle (P = 0.023) or pooled vehicle
 

(P < 0.001) controls (matched vehicle controls 0/9, pooled
 

vehicle controls 0/28, low-dose 2/32, high-dose 6/13). There
 

were no such tumors of the ear canal among 165 historical vehicle
 

controls of either sex or 220 untreated female controls at the
 

laboratory, and only two such tumors among 215 untreated male
 

controls.
 

In mice, no tumors appeared in statistically significant
 

incidences in the dosed groups compared with the vehicle
 

controls, and there was no significant evidence of dose-related
 

trend for any tumors. The incidences of the combination of
 

lymphoma and leukemia were significantly higher in the matched
 

vehicle controls of either sex than in the corresponding
 

untreated controls (males: matched untreated controls 1/30,
 

matched vehicle controls 19/29; females: matched untreated
 

44
 
controls 2/30, matched vehicle controls 21/29). This high
 

incidence in the matched vehicle controls may have been due to a
 

systematic procedural problem associated with injection of the
 

drug, which made possible the transplantation of tumor cells.
 

The same needle for injection was used for each group of five
 

animals within a cage; furthermore, the same bottle of vehicle
 

was used for all vehicle-control animals.
 

The vehicle used for administering the ß-TGdR to all groups in
 

this bioassay contained polysorbate 80, which in itself has been
 

implicated as a carcinogen, but only in the production of local
 

sarcomas following subcutaneous injection (Grasso et al., 1971).
 

However, in these bioassays no local sarcomas were observed in
 

the vehicle-control animals administered polysorbate 80 by
 

intraperitoneal injection.
 

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

low survival of the dosed and vehicle-control groups of mice, as
 

well as the possible procedural problem that may have affected
 

the incidences of tumors in these groups, does not allow a
 

determination to be made of the carcinogenic potential of ß-TGdR
 

in this species. ß-TGdR in the vehicle of 0.05% polysorbate 80
 

was, however, carcinogenic in rats, producing carcinomas of the
 

ear canal in the females and possibly also in the males.
 

45
 

VI. BIBLIOGRAPHY
 

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 the 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.
 

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

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Statist. Inst. 39:148-169, 1971.
 

Grasso, P., Gangolli, S. D., Golberg, L., and Hooson, J.,
 

Physicochemical and other factors determining local sarcoma
 

production by food additives. J. Fd. Cosmet. Toxicol.
 

1:463-478, 1971.
 

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

incomplete observations. J. Amer. Statist. Assoc.
 

jv3:457-481, 1958.
 

Lepage, G. A., Junga, I. G. , and Bowman, B., Biochemical and
 

carcinostatic effects of 2'-deoxythioguanosine. Cancer Res.
 

14:835-840, 1964.
 

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.
 

Loo, T. L., Ho, D. H. W., Bodey, G. P., and Freireich, E. J. ,
 

Pharmacological and clinical studies of some nucleoside
 

analogs. Ann. N. Y. Acad. Sci. 255:252-260, 1975.
 

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

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

47
 
Peery, A. and Lepage, G. A., Nucleotide formation from a- and
 

/8-2'-deoxythioguanosine in extracts of murine and human
 

tissues. Cancer Res. 29:617-623, 1969.
 

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 oxide. Cancer Res. 32;1073-1081, 1972.
 

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

62(3):679-682, 1975.
 

48
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH ß-TGdR
 

49
 

TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH ß-TGdR 

HIGH DOSE
 

35
 

30
 

30
 

(3P)
 

1 (3X)
 

(30)
 

1 C7%)
 

(7)
 

ANIHALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEH
 

*SKIN
 

PAPILLOHA. DOS
 

SQUAHODS CELL CARCINOMA
 

TRICHOEPITHELIOHA
 

RESPIRATORY SYSTEH
 

NONE
 

HEMATOPOIETIC SYSTEH
 

•MULTIPLE ORGANS
 

HALIG.LYMPROHA, UNDIFPER-TYPE
 

HALIG.LYHPHOHA, LYHPHOCYTIC TYPE
 

tHANDIBDLAR L. NODE
 

SQOAMOtIS CELL CARCINOMA, PIETASTA
 

CIRCDLATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEH
 

NONE
 

URINARY SYSTEH
 

JJ2HE
 

UNTREATED VEHICLE* LOW DOSE 

CONTROL CONTROL 

10 10 35 

10 10 35 

10 10 35 

(10) (10) (35) 

1 (10*) 

1 <3«) 

(10) (10) (35) 

1 (3«) 

(10) (10) (17) 

1 (6%) 

* NOHBER OF ANIMALS WITH TISSUE EXMUNED MICROSCOPICALLY
 

* NDHBER OF HHIHias HECEOPSIED
 

51
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

UNTREATED VEHICLE
 

•	 CONTROL CONTROL
 

MDOCRIHB SYSTEH
 

tPITOITAPY (9) (9)
 CHROHOPHOBE ADESOBA 2 (22*1 4 (4UX)
 

•ADRENAL	 (1P) (10)
 PHEOCHROBOCYTOBA 1 (10X)
 

tPANCREATIC ISLETS (10) (10)
 ISLET-CELL ADENOHA 1 (10%)
 

REPPODDCTIVE SYSTEB
 

tTESTIS (10) (1*)
 

INTERSTITIAL-CELL TOHOR
 

NERVOUS SYSTEB
 

tBRAIN (10) (10)
 ASTROCYTOBA 1 (10*)
 

SPECIAL SENSE ORGANS
 

*EAR CANAL (10) (10)
 CARCINOHA.NOS
 

SQOABOUS CELL CARCINOBA
 

BOSCDLOSKELETAL SYSTEB
 

NONE
 

BODY CAVITIES
 

* PELVIS (10) (10)
 ADENOCARCINOBA, NOS 1 (10*)
 

ALL OTHER SYSTEBS
 

*BULTIPLE ORGANS (10) (10)
 

saOiBQS5_C2lt-C4ECIBQai
 

* NOBBER OF ANIBALS BITH TISSDE EXAHINED BICROSCOPICALLY
 

« NOBBEH OF ANIBALS NECROPSIED
 

52
 

LOW DOSE
 

(25)
 

3 (12X)
 

<3U)
 

(33)
 

1 (3*)
 

(3U)
 

1 (3*)
 

(33)
 

(35)
 

1 (3*)
 

(35)
 

(35)
 

.1.J2S1
 

HIGH DOSE
 

(20)
 

(29)
 

(27)
 

(28)
 

(27)
 

1 (!»*)
 

(30)
 

1 (3*)
 

1 (3*)
 

(30)
 

(30)
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

ANIHAL DISPOSITIOH SUMHARY
 

ANIHALS INITIALLY IN STODY
 

NATURAL DEATHS
 HORIBOND SACRIFICE
 

SCHEDULED SACRIFICE
 ACCIDENTALLY KILLED
 

TEHBINAL SACRIFICE
 

ANIHAL HISSING
 

i INCLUDES AUTOLTZED ANIBALS
 

TOHOF SOBHARY
 

TOTAL ANIBALS WITH PRIHART TOBORS*
 

TOTAL PRIBART TDHORS
 

TOTAL ANIHALS WITH BENIGN TDHORS
 TOTAL BENIGN TOHORS
 

TOTAL ANIHALS WITH BALIGMANT TOBORS
 

TOTAL BALIGHANT TUHOHS
 

TOTAL ANIHALS WITH SECONDARY TOHORS*
 

TOTAL SECONDAPI TOHORS
 

TOTAL ANIBALS WITH TOHORS UNCERTAINBENIGN OR HALIGNANT
 

TOTAL UNCERTAIN TOBORS
 

TOTAL ANIBALS WITH TUHORS ONCERTAINPRIHARY OH HETASTATIC
 TOTAL ONCEHTAIH TOHORS
 

UNTREATED
 

CONTROL
 

10
 

10
 

3
 

3
 

2
 

2
 

1
 

1
 

VEHICLE
 

CONTROL
 

10
 

1
 

9
 

7
 

8
 

6
 

7
 

1
 

1
 

LOW DOSE
 HIGH DOSE
 

35
 

6
 

6
 

23
 

9
 

9
 

6
 

6
 

3
 

3
 

1
 

1
 

* PRIBARY T0HORS: ALL TUMORS EXCEPT SECONDARY TOH3HS
 

* SECONDARY TOHORS: HETASTATIC TDHOBS OR TOHORS INVASIVE INTO AN ADJACENT ORGAN
 

53
 

35
 

13
 

16
 

6
 

5
 

6
 

5
 

6
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE 

RATS TREATED WITH ß-TGdR 

UNTREATED VEHICLE 

CONTROL CONTROL 

ANIMAL S INITIALLY IN STUDY 10 10 

ANIMAL S NECROPSIED 10 10 

ANIMALS EXAMINED HISTOPATHO LOGICALLY 10 10 

INTEGUMENTAR Y SYSTEM 

*SKIN (10) (10) 

SARCOMA , NOS 1 (10%) 

*SOBCUT TISSUE (10) (10) 

SARCOMA , NOS 

ANBLOBLASTOM A 

RESPIRATOR Y SYSTEM 

NONE 

RENATOPOIETIC SYSTEM 

•MULTIPLE ORGAN S (10) (10) 

MALIGNAN T LYMPHOHA , NOS 

HALIG.LYMPHOHA , UNDIPPER-TYPE 1 (10%) 

MALIG.LYMPHOMA , LYMPHOCYTIC TYPE 

tTHYHUS (8) (10) 

THYHOHA 

CIRCULATORY SYSTEM 

NONE 

DIGESTIVE SYSTEM 

NONE 

URINAR Y SYSTEM 

_ fiSfiE 

* NDMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS N3CROPSIED
 

LOW DOSE 

35 

32 

32 

(32) 

(32) 

1 (3X) 

1 (3%) 

(32) 

(31) 

1 (3%) 

HIGH DOSE 

35 

27 

27 

(27) 

(27) 

(27 ) 

1 («%) 

1 ("%> 1 (1%) 

(25) 

54
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

UNTREATED VEHICLE 

CONTROL CONTROL 

ENDOCRINE SYSTSM 

IPITUITAHY (10) (10 ) 

CHBOHOPHOBE ADENOM A 7 (70*) (3 (80*) 

REPRODUCTIVE SYSTEM 

«flAtlM4B Y GLAN D (10) (10 ) 

CARCINOMA , NOS 1 (10*) ADENOCARCINOMA . NOS 

FIBEOADEN0.1A 4 (!»0%) 2 (20%) 

IUTERU S (9) (10) 

ENDOHETRIA L STEOMAL POLY P 1 4 10J) 

NERVOU S SYSTEM 

NON E 

SPECIAL SENS E ORGAN S 

*EAB CANAL (10) (10)

 

CARCINOMA . NOS 

SQUAMOU S CELL CARCINOM A 

HUSCULOSKELETA L SYSTE M 

NON E 

BODY CAVITIES 

• MEDIASTINU M (10) (10)

 

SARCOMA , NOS 

A LL OTHER SYSTEM S 

THORACIC CAVITY 

ADENOCAHCINOMAj , NOS 

t NUMBER OF ANIMALS KITH TISSUE EXAMINED MICEOSCOP 1CA LLY 

* NUBBE E OF ANIMAL S NECEOPSIE D 

LOW DOSE 

M l)

12 (39%) 

(32) 

3 (9*) 

5 (16% ) 

CJ2 )

1 (3%) 

(32) 

2 (6*) 

(32) 

HIGH DOSE 

(23) 

2 (9*J 

(27) 

(26) 

(27) 

1 (IX) 

5 (19X) 

(27) 

1 C»XJ 

55
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

1
UNTREATED VEHICLE LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 10 10 35 35
 

NATURAL DEATHS 1 J 13
 

MORIBUND SACRIFICE 1 5 15
 

SCHEDULED SACRIFICE
 ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 9 9 27 7
 

ANIMAL HISSING
 

3) INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS BITH PRIMAR Y TUMORS* b j 17 10
 

TOTAL PRIMARY TUMOBS 12 1"4 2f> 12
 

TOTAL ANIMALS WITH BENIGN TUMORS d y 15 2
 

TOTAL BENIJN TUMORS 11 1 1 20 2
 

TOTAL ANIMALS KITH MALIJNAN T TUMORS 1 J h 10
 

TOTAL MALIGNANT TUMORi 1 3 D 10
 

TOTAL ANIMALS HITH SECONDARY TUMORS*
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS HITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY lUtiOKa
 * SECONDARY TUMORS: META5IAIIC TUMORS OR lUHOfJ INVASIV E INTO AN ADJACENT ORGftS
 

56
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH ß-TGdR
 

57
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

TREATED WITH ß-TGdR (CONTROL GROUPS) 

MIC 

U 

1 & HIGH DOSE 

NTREATED 

CONTROL 

LOW DOSE UNTREATED CONTROL MID & HIGH DOSE VEHICLE 

CONTROL 

ANIMALS INITIALLY IN STUDY 

ANIMALS NECROPSIED 

ANIMALS EXAMINED HISTOPAMOLOUICALLY 

15 

15 

15 

15 

15 

15 

15 

11* 

1« 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

• LUNG (1<O ('5) (lit)
 

ALVEOLAR/BRONCHIOLAR ADENOM A 2 (1U%) 1 (7X)
 

ALVEOLAR/BBONCH10LAB CARCINOMA 1 (7%)
 

HEMATOPOIETIC SYSTEM
 

'MULTIPLE ORGANS (15) (15) (It)
 MALI3NANT LYMPHOMA, NOS
 

MALI3. LYMP.10MA, LYMPKOCYTIC TYPE 1 i? X) J (2U)
 

MALIQ.LYMPHOMA, HISTIOCYTIC TYPE 6 It3*j
 

MALIGNANT LYMPHOMA, MIXED TYPE
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

tLIVER (15) (15) (1U)
 

HEPATOCELLULAR ADENOMA 1 <7X) 1 (7*>
 HEPAIOCELLULAR CARCINOMA 1 (7S)
 

*BIL£ DUCT (15) (15) (Hi)
 

BILE DUCT CARCINOMA 1 (7%)
 

U8INAHJ SYSTEM
 

IURINARY BLADDER (15) (15) (It)
 _ HEMANGIOMA 1 J7*l_
 

t NUMBER OP ANIMALS WITH TISSUE EXAMINED MICEOSCCFICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

59
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

15
 

15
 

15
 

(15)
 

(15)
 

2 CU%)
 

1 (7X)
 

b (UOX)
 

1 (7X)
 

(15)
 

1 (7S)
 

(15)
 

(15)
 
TABLE B1. MALE MICE (CONTROL GROUPS): NEOPLASMS (CONTINUED)

 

ENDOCRINE SYSTEM
 

HONE
 

REPRODUCTIVE SYSTEM
 

NONE
 

NEEVODS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

HUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

ANIHAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATH*
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

» IHEiHCES.AUTOLYZED.ABIHllS
 

MID & HIGH DOSE LOW DOSE MID & HIGH LOW DOSE UNTREATED UNTREATED DOSE VEHICLE VEHICLE CONTROL CONTROL CONTROL CONTROL 

15 15 15 15 

3 1 10 u 2 5 5 7 

IP 9 U 

t NUMBE R OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMAL S NECROPSIED
 

60 
TABLE B1. MALE MICE (CONTROL GROUPS): NEOPLASMS (CONTINUED) 

Ml D & HIGH DOSE UNTREATED CONTROL 

LOW DOSE UNTREATED CONTROL 

MID & HIGH DOSE VEHICLE CONTROL LOW DOSE VEHICLE CONTROL 

TOHOP SUMMAR Y 

TOTAL ANIMALS KITH PRIMAR Y TUNOPS* 

TOTAL PST MARY TUMORS 

<l 

5 

3 

U 

9 

10 

11 

11 

TOTAL ANIMAL S WITH BENIGN TUMORS 

TOTAL BENIGN TUMORS 

2 

2 2 3 

1 

1 

1 

1 

TOTAL ANIMALS WITH MALIGNANT TUMOPS 

TOTAL MALIGNANT THMOFS 

3 

3 

1 

1 

9 

9 

10 

10 

TOTAL ANIMALS WITH SECONDARY TUMORS* 

TOTAL SECONDAPY TUMOPS 

TOTAL ANIMAL S WITH TUMOHS UNCERTAINBENIGN OR MALIGNAN T •"•OTAL UNCERTAIN TUMOPS 

TOTAL ANIMALS WITH TUMOPS UNCEPTAIN­ PRIMARY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

* PRIMARY TUWORS: AIL TUMOHS EXCEPT SECONDARY TOMDPS * SECONDAPY TtJHOPS: METASTATIC TOHOPS OR TUMOPS INVASIVE INTO AN ADJACENT OP3AN 

61
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

TREATED WITH |3-TGdR (TREATED GROUPS)

 

LOW DOSE MID DOSE HIGH DOSE
 

MURALS INITIALLY IN STDDT 35 35 35
 ANIMALS NECPOPSIED 35 32 0
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 35 32 0
 

IHTEGONENTARY SYSTEM
 

NONE
 

RESPIBATOBY SYSTEH
 

NONE
 

HEBATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS (35) (32)
 

MALIGNANT LYMPHOHA, NOS 1 (3%) 1 (3%)
 HALIG.LYHPHOMA, ONDIFFEH-TYPE 2 <6X) 6 (19*)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 7 (20*) 9 (28X)
 

MALIG.LYHPHOHA, HISTIOCYTIC TYPE « (11%)
 MALIGNANT LYHPKONA, MIXED TYPE 3 (9«)
 

LYHPHOCYTIC LEUKEMIA 2 (6*)
 

CIBCULATORY SYSTEH
 

NONE
 

DIGESTIVE SYSTEM
 

tLIVER <3<») (3D
 

HSPATOCELLHLAR ADENOMA 1 (3%) 1 (3*)
 

HEHANGIOSABCOHA 1 (3*)
 

tJBINAHY SYSTEH
 

__fiQBB fc
 

t NUBBEP OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * BOBBER OF ANIMALS NECROPSIBD
 

62
 
TABLE B2. MALE MICE (TREATED GROUPS): NEOPLASMS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE 

ENDOCRINE SYSTEM 

*THYROID 

FOLLICULAS-CELL ADENOMA 

(31) (22) 

1 (5%) 

REPRODUCTIVE SYSTEM 

NONE 

NERVOUS SYSTEM 

NONE 

SPECIAL SENSE OPGANS 

NONE 

BUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 

NATURAL DEATH* 

HORIBDND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIMAL HISSING * 

35 

5 

19 

11 

35 

15 

15 

5 

35 

35 

a-!HCL.op.ES_ADTOL.!S!B ifilBHS 

t NUMBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

63
 
TABLE B2. MALE MICE (TREATED GROUPS): NEOPLASMS (CONTINUED) 

LOW DOSE
 MID DOSE HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS HITH PRIMARY TUMORS* 19 19
 

TOTAL PRIMARY TtlMORS 19 20
 

TOTAL ANIMALS WITH BENIGN TUMORS 1 1
 

TOTAL BENIGN TDMORS 1 2
 

TOTAL ANIMALS WITH MALIGNANT TUHOHS 18 18
 

TOTAL MALIGNANT TUMORS 18 18
 

TOTAL ANIMALS HITH SECONDARY TUHORS t
 

TOTAL SECONDARY TUHORS
 

TOTAL ANIMALS HITH TUMORS UNCERTAIN
 .
 

BENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TDMOPS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN
 -
 PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIHABY TUflORS: ALL TUHORS EXCEPT SECONDARY TUHOPS
 

* SECONDARY TUHORS: HETASTATIC TOHORS OR TOHORS INVASIVE INTO AN ADJACENT ORGAN
 

64
 
TABLE B3.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

TREATED WITH |3-TGdR (CONTROL GROUPS) 

MID & HIGH DOSE LOW DOSE MID & HIGH LOW DOSE 

UNTREATED UNTREATED DOSE VEHICLE VEHICLE 

CONTROL CONTROL CONTROL CONTROL 

ANIHAL S INITIALL Y IN STUDY 15 15 15 15 

ANIMALS NECROPSIED 15 15 15 m 

ANIHAL S EXAMINE D HISTOPATHO LOGICALLY 15 15 15 m 

INTEGOHENTAP Y SYSTEM 

NONE 

RESPIRATORY SYSTEM 

tLUNG (15) (15) (15) (I'M

 

ALVEOLAR/BRONCHIOLA P ADENOM A 1 f%)

 

OSTEOSARC08A, HFTASTATIC 1 <7«)

 

HBMATOPOIETIC SYSTEM 

*MnLTIPLE ORGAN S (15) (15) (15) (1U ) 

MALIGNAN T LYMPHOMA , NO S 1 (7*) 2 (13% ) 2 (W) 

MALIG.LYMPHOHA , UNDIFPER-TYPE 1 (7*) 3 (21X ) 

HALIG.LYHPHOMA , HISTIOCYTIC TYPE 1 (7X) 7 (U7X ) « (29*) 

LYMPHOCYTIC LEUKEMIA 1 (7K) 

tLIVER (15) (15) (15) (14)

 

HALIG.LYHPHOHA , ONDIFFER-TYPE 1 (7*)

 

CIRCULATORY SYSTEM 

HONE 

DIGESTIVE SYSTEM 

*LIVE8 (15) (15) (15) (1«)

 

HEPATOCELLULAP ADENOM A 1 (7*) 

DHINARY SYSTEM 

BQfiS __ __ __ _ . 

* NUMBER OF ANIHALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

« NUMBER OF ANIMALS NECROPSIED
 

65
 
ERDOCRIRE SYSTER
 

ROME
 

REPRODUCTIVE SYSTEH
 

NONE
 

NERVOUS SYSTEM
 

RONE
 

SPECIAL SENSE ORGANS
 

NONE
 

RDSCULOSKELETAL SYSTEM
 

•VERTEBRA
 

OSTEOSARCOMA
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEHS
 

RONE
 

ANIRAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL D EAT Hi
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE
 

ANIMAL MISSING
 

»_IBCIrSBES AUTOLYZED_4NI8aiS
 

i NUNBER OF ANIHALS KITH TISSUE EXAHINED HICROSCOPICALLY
 

» ROBBER OF ANIHALS NECEOPSIED
 

MID & HIGH DOSE LOW DOSE
 UNTREATED UNTREATED
 

CONTROL CONTROL
 

(15) (15) 

1 (7%) 

15 15 

1 1 

2 

1 

13 12 

MID & HIGH
 

DOSE VEHICLE
 

CONTROL
 

(15)
 

15
 

3
 

12
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

(1lt)
 

15
 

6
 

5
 

1
 

3
 

66 
TABLE B3. FEMALE MICE (CONTROL GROUPS): NEOPLASMS (CONTINUED) 

Ml D & HIGH DOSE 

UNTREATED 

CONTROL 

LOW DOSE 

UNTREATED CONTROL 

MID & HIGH 

DOSE VEHICLE 

CONTROL 

LOW DOSE 

VEHICLE 

CONTROL 

TOHOH SUHHARY 

TOTAL ANIHALS WITH PRIHARY TUHORS* 

TOTAL PRIHARY TUHORS 

1 

1 

i» U 

11 

11 

10 

10 

TOTAL ANIHALS WITH BENIGN TUHORS 

TOTAL BENIGN TUHORS 

1 

1 1 1 

TOTAL ANIHALS WITH HALIGNANT TUHORS 

TOTAL HALIGNANT TUHOPS 

3 

3 

11 

11 

10 

10 

TOTAL ANIHALS WITH SECONDARY TUHORS* 

TOTAL SECONDARY TUHORS 

1 

1 

TOTAL ANIHALS WITH TUHORS

BENIGN OR HALIGRANT TOTAL UNCERTAIN TUHORS 

 UNCERTAINTOTAL ANIHALS WITH TUHOHS UHCERTAIN­ PRIRARY OS HETASTATIC 

TOTAL UNCERTAIN TUHORS 

* PRIHART TOHORS: ALL TOHORS EXCEPT SECONDARY TOHORS * SECONDARY TOHORS: HETASTATIC TOHOBS OH TOHORS INVASIVE INTO AN ADJACBUT ORGAN 

67
 
TABLE B4.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

TREATED WITH ß-TGdR (CONTROL GROUPS) 

HIGH DOSE
 

35
 

1
 

1
 

(D
 

(D
 

1 (100X)
 

(1)
 

LOW DOSE
 

ANIMALS INITIALLY IN STUDY 35 

ANIHALS NECPOPSIED 33 ANIHALS EXAMINED HISTOPATHOLOGICALLY 33 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEH
 

«1DNG (33)
 

ALVEOLAR/BRONCHIOLAR ADENOMA
 

HEMATOPOIFTIC SYSTEM
 

*MOLTIPLE OPGANS (33)
 

MALIGNANT LYMPHOMA, HOS 2 (6X)
 

HiLIG. LYMPHOMA, UNDIPFER-TYPE 1 (3X)
 

BALIG.LYMPHOHA, LYMPHOCYTIC TYPE 2 (6*)
 

MALIG.LYHPHOMA, HISTIOCYTIC TYPE 1<t (i»2«)
 

CIPCULATOSY SYSTEH
 

NONE
 

DIGESTIVE SYSTEH
 

NONE
 

UBINA8Y SYSTEM
 

NORE
 

EMDOCBINE SYSTEM
 

tTHTROID (30)
 

FQLlICBLAErCSLI, ABEN.QHA „ _1.J3S1._.
 

MID DOSE
 

35
 

29
 

29
 

(28)
 

1 ("»*>
 

(29)
 

2 (7X)
 

17	 (59%)
 

2 (7%)
 

(25)
 

I NDHBER OF ANIMAL S WITH TISSUE EXAHISED HICROSCOPICALLY
 

* NUMBE R OF ANIHAL S NECROPSIED
 

68
 
TABLE B4. FEMALE MICE (TREATED GROUPS): NEOPLASMS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE
 

REPRODUCTIVE SYSTEM
 

*OVASY (32) (29) (D
 HEMANGIOSARCOMA 1 (3%)
 

NEPVOUS SYST5PI
 

NONS
 

SPFCIAL SENSE ORGAN S
 

NONE
 

HUSCULO SKELETAL SYSTEM
 

NONE
 

BODY CAVITIFS
 

NONE
 

ALL OTHER SYSTFfS
 

NONE
 

ANIMAL DISPOSITION SI1MPARY
 

ANIMALS INITIALLY IN STUDY 35 35 35
 

NATOPAL DEATHS 9 Ml 33
 

BORIBUND SACRIFICE 15 17 1
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED 1 2 1
 

TERMINAL SACRIFICE 10 2
 

ANIMAL MISSING
 

S_INCLUDES.AOTOLYZED_ ANIMALS
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECPOPSIED
 

69 
TABLE B4. FEMALE MICE (TREATED GROUPS): NEOPLASMS (CONTINUED) 

TtJHOR SUHNARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BSNIGN TUMORS
 

TOTAL BENIGN TDMOSS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUKOPS
 

TOTAL ANIMALS WITH SECONDARY TUMORS
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN
 

BENIGN OR MALIGNANT
 

TOTAL UNCPRTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN
 

PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN- TUMORS
 

LOW DOSE MID DOSE HIGH DOSE 

20 21 1 

21 22 1 

1 1 

1 1 

2P 21 1 

20 21 1 

* 

_ 

. 

* PRIMARY TUMOF.S: ALL TUHORS EXCEPT SECONDARY TON3RS
 

* SECONDARY TOMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

70
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS TREATED WITH ß-TGdR
 

71
 

TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN MALE RATS TREATED WITH ß-TGdR 

UNTREATED CONTROL VEHICLE CONTROL LOW DOSE 

AMIBALS INITIALLY IN STUDY 

iHIBALS NECBOPSIED 

ANIBALS EXABINED HISTOPATHOLOGIC ALLY 

10 

10 

10 

10 

10 

10 

35 

35 

35 

INTEGUBENTART SYSTEM
 

•SKIN	 (10) (10) (35)
 INFLAMMATION, FOCAL
 

ULCEB, CHBONIC 1 (3*)
 

BESPIHATORY SYSTEM
 

HBACHEA (10) (10) (3UJ
 

LYMPHOCYTIC INFLAMMATORY INFILTR 2 (6%)
 

INFLABBATION, SUPPURAIIVE
 

*LDNG/BRO»CHUS (10) (10) (34)
 BSONCHIECTASIS
 

INFLABBAIIOS, SUPPBRATIVE
 

HYPEBPLASIA. LYMPHOID 1 (10X) 3 (9X)
 

tLUNG (10) (10) (34)
 

BBOICHOPNEUMONIA SUPPORATIVE 1 (10X)
 PNEOBONIA, CHRONIC BURINE 1 (10*) 1 (10X) 10 (29X)
 INFLAMBATION, CHBOSIC SUPPDRATIV
 

HEBATOPOIETIC SYSIEB
 

tBONE HABBOi (10) (9) (34)
 

ATROPHY, NOS
 

CIBCUltTORY SYSIEH
 

iBIOCABDIDfl (10) 110) (34)
 

INFLABBiTION, INTERSTITIAL 1 (3*)
 

*PULBONtBY ARTEBY	 (10) (10) (35)
 

..aYesftmoEHi<..»Qs . . .	 1 J3XJ.
 

t NUBBEB OP A11IBALS BUB TISSOE EXAMINED MICROSCOPICALLY 

* NUBBEB OF ANIMALS NECBOPSIED 

HIGH DOSE
 

35
 

30
 

30
 

(30)
 

1 (3X)
 

(29)
 

2 (7X)
 

2 (7X)
 

(27)
 

1 (4X)
 

1 I"*)
 

3 (11X)
 

(27)
 

2 (7X)
 

4 (15X)
 

1 (4X)
 

(25)
 

8 (32X)
 

(29)
 

1 (3X)
 

(30)
 

_
 

73 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

UNTREATED VEHICLE
 CONTROL CONTROL
 

DIGESTIVE SYSTEM
 

tLIVER (10) (10)
 THROMBOSIS, BOS
 

INFLAMMATION, P YOGR ANULOMATOUS
 

NECROSIS, COAGULATIVE
 

tHEPATIC CAPSULE (10) (10)
 

HEMORRHAGE
 

tLIVER/CENTRILOBULAR (10) (10)
 

NECROSIS, FOCAL
 

tPANCREAS (10) (10)
 

INFLAMMATION, CHRONIC
 

INFLAMMATION, CHRONIC DIFFUSE 1 (10*)
 

*STOMACH (9) (10)
 

ULCER, FOCAL 1 (10*)
 

tJEJUNUH (10) (10)
 

ULCER, FOCAL 1 (10X)
 

URINAR Y SYSTEM
 

IKIDNEY (10) (10)
 

INFLAMMATION, CHRONIC
 

INFLAMMATION, CHRONIC FOCAL
 

INFLAMMATION, CHRONIC DIFFUSE 1* (40*) 3 (30%)
 

ENDOCRINE SYSTEM
 

•ADRENAL CORTEX (10) (10)
 

CYTOLOGIC DEGENERATION
 

REPRODUCTIVE SYSTEM
 

NONE
 

NERVOUS SYSTEM
 

NONE
 

« NOHBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

» NUMBER OF ANIMALS NECROPSIED
 

74
 

LOW DOSE
 

(33)
 

1 (3*)
 

2 (6*)
 

(33)
 

1 (3*)
 

(33)
 

(33)
 

(33)
 

(33)
 

<3«)
 

1 (3*)
 1 <3X)
 

5 (15*)
 

(3^4)
 

HIGH DOSE
 

(28)
 

1 (4X)
 

1 («*)
 

(28)
 

(28)
 

1 («*)
 

(27)
 

1 (<4«)
 

(27)
 

(2U)
 

(28)
 

1 (<**)
 

(29)
 

1 (3%)
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

LOW DOSE HIGH DOSE 

NDSCOLOSKELETM, SYSTEM 

NONE 

BODY CAVITIES 

*PERITONEON 

INFLAMMATION, CHRONIC FOCAL 

(1C) (10) 

1 (10*) 

(35) (30) 

ALL OTHER SYSTEMS 

*HULTIPLE ORGANS 

HYPERPLASIA, LYMPHOID 

(1C) (10) (35) 

1 (3*) 

(30) 

SPECIAL MORPHOLOGY SDMMAPY 

NO LESION REPORTED 

NO NECROPSY PERFORMED 

1 6 7 

5 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECBOPSIED 

75
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN 

FEMALE RATS TREATED WITH ß-TGdR 

UNTREATED VEHICLE LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

ANIMALS INITIALLY IN STUDY 10 10 35 35
 ANIHALS NECPOPSIED	 10 10 32 27
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 10 10 32 27
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

*LDNG/BRONCHUS (10) (10) (32) (27)
 HYPEPPLASIA. LYHPHOID 1 (3*) 2 (7%)
 

»LUNG (10) (10) (32) (27)
 

BHONCHOPNEDMONIf. SUPPORATIVS 1 (U%)
 PNEUMONIA, CHRONIC MORINE 1 <3X) 2 (7X)
 

HEHATOPOIETIC SYSTEM
 

IBONE HARROW (8) (10) (32) (26)
 

ATROPHY. NOS 7 (27X)
 

tSPLEEN (10) (10) (32) (27)
 

HEMATOPOIESIS 1 (3«)
 

tNANDIBOLAR L. NODE (8) (8) (27) (10)
 

INFLAMMATION, CHPONIC DIFFUSE 1 (10*)
 

RYPERPLASIA, LYMPHOID 1 C**)
 

CIRCULATOR! SYSTEM
 

•MYOCARDIUM	 (10) (10) (32) (27)
 

INFLAMMATION, CHRONIC 1 (»%)
 

DIGESTIVE SYSTEM
 

*LIV«R (10) (10) (32) (27)
 

- —BBCEQSIS^ CQlSDUmE... ___ 1_I1SL-.
 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIHALS NECROPSIED
 

76
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

tLIVER/CENTBILOBULAR (10) (10)
 

NECROSIS, FOCAL
 

CYTOPLASMIC VACUOLIZATION
 

•BILE DUCT (1C) (10)
 

HYPERPLASIA, NOS
 

*PANC8EAS (1C) (10)
 

INFLAMMATION, CHRONIC
 

URINARY SYSTEM
 

*KIDNEY (10) (10)
 

INFLAMMATION, CHRONIC 1 (10%) 1 (10*)
 

INFLAMMATION, CHRONIC DIFFUSE 1 (10*) 1 (10*)
 

ENDOCRINE SYSTEM
 

IADPENAL (10) (10)
 

ANGIECTASIS 1 (IP*)
 

tADRENAL N5DOLLA (10) (10)
 

HEMORRHAGE
 

REPRODUCTIVE SYSTEM
 

tUTESUS/ENDOMETRIUM (9) (10)
 

INFLAMMATION, SDPPUFATIVE
 

*UTERUS/HYONETPTUM (9) (10)
 

THROMBOSIS, NOS
 

tOVARY (9) (10)
 

INFLAMMATION, SUPPORATIVE
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

__NQHS .
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECPOPSIED
 

77
 

LOW DOSE
 

(32)
 

(32)
 

1 (3*)
 

(32)
 

1 (3*)
 

(32)
 

(32)
 

it (13*)
 

(32)
 

1 (3*)
 

(32)
 

(32)
 

1 (3X)
 

(32)
 

1 (3*)
 

HIGH DOSE
 

(27)
 

1 («*)
 

1 (t«)
 

(27)
 

(27)
 

(27)
 

(26)
 

1 ("»*)
 

(26)
 

(26)
 

1 (U*)
 

(26)
 

(26)
 

5 (19*)
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

LOW DOSE HIGH DOSE 

MUSCULOSKELETA L SYSTE M 

NON E 

BODY CAVITIE S 

•MESENTER Y 

NECBOSIS, PA T 

(10) (10) (32) 

1 (3*) 

(27) 

ALL OTHEB SYSTEM S 

NON E 

SPECIAL MOBPHOLOG Y SUMMAR Y 

NO LESION BEPOBTED 

NO liECBOPSY PEBFOHME D 

AUTOLYSIS/N O NECROPS Y 

2 

• NUHBEH OP ANIMALS UITH TISSUE EXAHINED MICFCSCCPICALLY 

« NUMBEH OP ANIMALS NECROPSIED 

1 10 

2 

1 

14 

a 

78
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE TREATED WITH ß-TGdR
 

79
 

TABLE D1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE 

MICE TREATED WITH p-TGdR (CONTROL GROUPS)

 

MIC 

U 

>& HIGH DOSE 

NTREATED 

CONTROL 

LOW DOSE 

UNTREATED 

CONTROL 

MID & HIGH 

DOSE VEHICLE 

CONTROL 

LOW DOSE 

VEHICLE 

CONTROL 

ANIMALS INITIALLY IN STUDY 

ANIMALS NECBOPSIED 

ANIMALS EXAMINED HISTOPATHOLOGIC ALLY 

15 

15 

15 

15 

IS 

15 

15 

14 

1U 

15 

15 

15 

INTEGUMENTAR Y SYSTEM
 

NONE
 

RESPIRAT08Y SYSTEM
 

•TRACHEA	 (15) 05) (14) (14)
 

INFLAMMATION. SUPPUBATIVE 5 ij.iii) .. (13X)
 

•LUNG/BRONCHIOLE	 (14) (15) (1«) (15)
 

HYPERPLASIA. PLASMA CEIL 1 UOX)
 HYPEBPLASIA, LYMPHOID 1 i.T*1
 

•LUNG (1<t) (15) (14) (15)
 

ERONCHOPNEUMONIA JUPPU RATIV E 6 lljXJ 7 (U7S) 1 (7%)
 

HYPEHPLASIA, PLASMS CELL 1 (7*)
 

HEMATOPOIETIC SYSTEM
 

NONE
 

CIRCULATORY SYSTEM
 

•MYOCARDIUM	 (15) (15) (14) i15)
 

INFLAMMATION, SUPPU RATIV E 1 (7%) 1 (7%)
 

INFLAMMATION, CHRONIC DIFFUSE 1 i7f.)
 

INFLAMMATION, CHRONIC SUPPUBATIV 1 (7S)
 

•ENDOCARDIUM	 (15) (15) (14) (15)
 

INFLAMMATION. CHRONIC 1 (7*)
 

DIGESTIVE SYSTEM
 

•LIVER	 (15) (15) (14) (15)
 

INFLAMMATION^ SUPPUR&TIVE 1 17XJ.
 

* NUBBEB OF ANIHALS WITH TISSUE EXAHINED MlLFUSCC?1C ALLY
 * NUBBEB OF ANIMALS NECROPSIED
 

81
 
TABLE 01. MALE MICE (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

MID & HIGH
 

DOSE VEHICLE
 

CONTROL
 

1 (7X)
 2 (11%)
 

(1»)
 

1 (7%)
 

(TO
 

1 (7%)
 

(11»)
 

(14)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

1 (7%)
 2 (13%)
 

(15)
 

1 (7%)
 

(15)
 

(15)
 

(15)
 

1 (7%)
 

M
 

INFLAMMATION, CHBOHIC SOPPORATIV
 

NECROSIS, COAGULATIVE
 

HYPBRPLASIA, HODaLAE
 

URINARY SYSTEM
 

•KIDNEY
 

PYELONEPHRITIS SOPPORATIVE
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

•PROSTATE
 

INFLAMMATION, SOPPORATIVE
 

NERVOUS SYSTEH
 

NONE
 

SPECIAL SENSE ORGANS
 

•MIDDLE EAR
 

INFLAMMATION, SOPPORATITE
 

MUSCULOSKBLETAL SYSTEH
 

NONE
 

BODY CAVITIES
 

•PERITONEUM
 

INFLAMMATION, SUPPORATIVE
 

ALL OTHER SYSTEMS
 

NOJLB_ —
 

ID & HIGH DOSE
 

UNTREATED
 

CONTROL
 

2 (13%)
 

(15)
 

(15)
 

(15)
 

1 (7*)
 

(15)
 

LOW DOSE
 

UNTREATED
 

CONTROL
 

(15)
 

(15)
 

(15)
 

(15)
 

* HUHBER OF MIIHALS HITH TISSUE EXA8IHED HICROSCOPICALLY
 

* HDHBER OP AHIHM.S HECFOPSIED
 

82 
TABLE D1. MALE MICE (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

MID & HIGH DOSE LOW DOSE MID & HIGH LOW DOSE 

UNTREATED UNTREATED DOSE VEHICLE VEHICLE 

CONTROL CONTROL CONTROL CONTROL 

SPECIAL BOBPHOLOGY SOBBABY 

HO LESION SEPORTED U 6 1 3 

MJTO/HECHOPSY/NO HISTO 1 

AUTOLYSIS/NO NECROPSY 1 

* HOHBEH OP ANIHAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* HtUIBBR OF ANI8ALS NECPOPSIED 

83
 
TABLE 02.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE 

MICE TREATED WITH ß-TGdR (TREATED GROUPS) 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECBOPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGIC ALLY
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEH
 

*T8ACHEA
 

INFLAMMATION, SUPPU8ATIVE
 

*LUNG/BBONCHIOLE
 

HYPEHPLASIA, PLASMA CELL
 

ILUNG
 

BBONCHOPNEUMONIA SUPPURATIVE
 

HEMATOPOIETIC SYSTEM
 

NONE
 

CIBCULATGBY SYSTEM
 

NONE
 

DIGESTIVE SYSTEH
 

•LIVES
 

INFLAMMATION, HECROTI21NG
 

INFLAMMATION, CHRONIC NECROTIZIN
 

NECROSIS, COAGULATIVE
 

HYPE8PLASIA, NODULAR
 

IHEPATIC CAPSULE
 

HEMOBBHAGE
 

LOW DOSE MID DOSE HIGH DOSE 

35 35 35 

35 32 0 

35 32 0 

(35)
 

1


(35)
 

2


(35)
 

8


(34)
 

1


2


(31)
 

 13X)
 

 (6%)
 

(23%)
 

 (3»)
 

 (6X)
 

(32)
 

(31)
 

(31)
 

(3D
 

1


1


1


(31)
 

1


 (3X)
 

 (3X)
 

 (3S)
 

13*1_
 .
 

* NUMBER OF ABDULS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

84
 
TABLE D2. MALE MICE (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE
 

•LIVEB/CENTBILOBULAR 

NECROSIS, DOS 

<3U) (31) 

1 (3*J 

•PANCREAS 

INFLAMMATION. SUPPUHATIVE 

INFLAMMATION, CHRONIC 

(33) (32) 

1

2

 (3*) 

 (6X) 

•STOMACH 

INFLAMMATION, CHBOBIC SUPPUBATIV 

(3<t) (32) 

1 (3*) 

*ILEUM 

ULCEB, NOS 

INFLAMMATION, SUPPURATIVE 

(J3) (31) 

1

1

 (3X) 

 (3*) 

URINAR Y SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

•PROSTATE	 (35) (32)
 

INFLAMMATION, CHBONIC SUPPUHATIV 1 (3*)
 

*EPIDIDYMIS (35) (32)
 

INFLAMMATION, CHBONIC SUPPURATIV 2 (6%)
 

NEBVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCOLOSKBLETAL SYSTEM
 

NONE
 

* NUMBER OF ANIMALS HITH TISSUE EXAMINED HICKOSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

85
 
TABLE D2. MALE MICE (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE 

BODY CAVITIES 

*PERITONEUM 

INPLAHBATION, SUPPURATIVE 

INFLAMHATION, CHRONIC SUPPOPATIV 

(35) (32) 

2 <6X) 

3 (9») 

ALL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMBAPY 

NO LESION REPORTED 

NO NECROPSY PERFORMED 

8 6 

3 35 

* NUMBER OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECPOPSIED 

86
 
TABLE 03.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH (ß-TGdR (CONTROL GROUPS) 

MIID& HIGH DOSE 

UNTREATED 

CONTROL 

LOW DOSE 

UNTREATED 

CONTROL 

ARIHALS INITIALLY IH STUDY 

AHIHALS RECROPSIED 

AHIHALS EXAHINED HISTOPATHOLOGICALLY 

15 

15 

15 

15 

15 

15 

IHTEGOHBRTARY SYSTEH
 

HONE
 

HESPIBATORY SYSTEM
 

*TRACHEA (15) (15)
 

TRPLAHHATIOH, SOPPUBATIVE
 

tLDRG/BRORCHIOLE (15) (15)
 

HYPERPLASIA, PLASBA CELL 2 (13%) 1 (7%)
 

HYPERPLASIA, LYHPHOID 1 (7%)
 

*LURG (15) (15)
 

BHORCHOPREOHORIA SOPPDRATIVE 5 (33X)
 

BRORCHOPREURORIA CHBOMIC SOPPORA
 

HYPERPLASIA, PLASHA CELL 1 (7X)
 

HEHATOPOIETIC SYSTEH
 

MODE
 

CIRCULATORY SYSTEH
 

IRYOCARDIDH (15) (15)
 

IRPUHHATIOR, SOPPOFATIVE
 

IRFLADRATIOR, CHRONIC SOPPORATIV
 

tBRDOCARDIDH (15) (15)
 

IRFLARHATIOH, CHRONIC
 

DIGESTITE SYSTEH
 

ISU­

* ROBBER OP AHIHALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* RUBBER OP AHIHALS RECROPSIED
 

87
 

MID& HIGH
 

DOSE VEHICLE
 

CONTROL
 

15
 

15
 

15
 

(15)
 

2 (13%)
 

(15)
 

2 (13%)
 

(15)
 

« (27%)
 

(15)
 

1 (7%)
 

(15)
 

1 «7%)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

15
 

1U
 

1U
 

(13)
 

(1t)
 

<1»)
 

1 (7%)
 

1 (7%)
 

(1U)
 

1 (7%)
 

(11)
 

1 (7%)
 
TABLE 03. FEMALE MICE (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

URINARY SYSTEM
 

tORINARY BLADDER
 

ULCER, DOS
 

ENDOCRINF SYSTEM
 

NOHE
 

REPRODUCTIVE SYSTEM
 

* UTERUS/EN DOMETRIUN
 

INFLAMMATION, SUPPUPATIVE
 

tOVARY
 

INFLAMMATION, SUPPUEATIVE
 

NERVOUS SYSTEM
 

tBRAIN
 

HEMORRHAGE
 

SPECIAL SENSE ORGANS
 

*EYE/CORNEA
 

INFLAMMATION, CHRONIC
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*PERITONEUM
 

INFLAMMATION, SUPPUPATIVE
 

ALL OTHEB SYSTEMS
 

£ONE _ _
 

MID & HIGH DOSE
 

UNTREATED
 

CONTROL
 

(15)
 

(15)
 

1 (7*)
 

(15)
 

(15)
 

(15)
 

(15)
 

LOW DOSE
 

UNTREATED
 

CONTROL
 

(15)
 

1 (7X)
 

(15)
 

(15)
 

(15)
 

(15)
 

(15)
 

MID & HIGH
 

DOSE VEHICLE
 

CONTROL
 

(15)
 

(15)
 

(15)
 

(15)
 

(15)
 

1 (7*)
 

(15)
 

1 (7*)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

<1« >
 

(1t>
 

(14)
 

1 (7%)
 

(13)
 

1 (8%)
 

(1«)
 

(I'M
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

88 
TABLE 03. FEMALE MICE (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

________ _ __­

MID & HIGH DOSE LOW DOSE MID& HIGH LOW DOSE 

UNTREATED UNTREATED DOSE VEHICLE VEHICLE 

CONTROL CONTROL CONTROL CONTROL 

SPECIAL BORPHOLOGY SOHHAR Y 

NO LESION REPORTED 9 11 2

 

ACCIDENTAL DEATH 1

 

t NDHBEP OP MUHULS WITH TISSUE EXUHTNED HICROSCOPICHLI.Y 

* NUMBER OP MflHULS HECKOPSIED 

89 
TABLE D4. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH ß-TGdR (TREATED GROUPS) 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGOHENTAPY SYSTEM
 

NONE
 

RESPIRATOBY SYSTEM
 

tTRACHEA
 

INFLAMMATION, SDPPOEATIVE
 

•LUNG/BRONCHIOLE
 

HYPERPLASIA, PLASMA CELL
 

tLONG
 

BRONCHOPNEDHONIA SDPPORATIVE
 

HEHATOPOIETIC SYSTEM
 

tMESENTERIC L. NODE
 

INFLAMMATION, NECROTIZING
 

CIRCOLATOFY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

NONE
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

—fiQHS
 

LOW DOSE
 

35
 

33
 

33
 

(33)
 

5 (15X)
 

(33)
 

(33)
 

6 (18%)
 

(30)
 

MID DOSE HIGH DOSE
 

35 35
 

29 1
 

29 1
 

(29) 

1 (3%) 

(28) 

1 (U%) 

(28) 

l» (1«%) 

(1) 

(D 

(D 

(20) 

1 (5X) 

_ _ _» 

t NOHBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS HECROPSIED
 

90
 
TABLE D4. FEMALE MICE (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED) 

REPRODUCTIVE SYSTEM
 

•UTERDS/ENDOMETRIDM
 

INFLAMMATION, SOPPOPATIVE
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

*NIDDL5 EAR
 

INFLAMMATION, SUPPURATIVE
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

•PERITONEUM
 

INFLAMMATION, SOPP09ATIVE
 

INFLAMMATION, CHRONIC FOCAL
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 

ACCIDENTAL DEATH
 

NO NECROPSY PERFORMED
 

AUTOLYSIS/NO NECROPSY
 

LOW DOSE
 

(32)
 

tt (13*)
 

(33)
 

1 (3%)
 

(33)
 

8
 

1
 

1
 

MID DOSE HIGH DOSE
 

(29) (1)
 

(29) (1) 

(29) 

1 (3*) 

1 (3%) 

(1) 

3 

2 

3 

1 

1 

33 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICAILY
 

* NUMBER OF ANIHALS NECEOPSIED
 

91
 

APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS TREATED WITH ß-TGdR
 

93
 

__ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Topography: Morphology
 

Hematopoietic System
 

Lymphomab
 

P Values0>d
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe
 

Adenoma*5
 

P Values0>d
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Treated

Pooled
 

Vehicle
 

Control
 

0/30 (0)
 

U.S.
 

Control) f
 

Control)f
 

7/26 (27)
 

P = 0.008(N)
 

Control)^
 

Control)f
 

79
 

 with ß-TGdRa 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

—
 

4/9 (44)
 

P = 0.003(N)
 

79
 

Low
 

Dose
 

1/35 (3)
 

N.S.
 

Infinite
 

0.047
 

Infinite
 

Infinite
 

0.017
 

Infinite
 

66
 

3/25 (12)
 

N.S.
 

0.446
 

0.083
 

1.710
 

0.270
 

0.057
 

1.344
 

77
 

High
 

Dose
 

2/30 (7)
 

N.S.
 

Infinite
 

0.301
 

Infinite
 

Infinite
 

0.109
 

Infinite
 

43
 

0/20 (0)
 

P = 0.005*(N)
 

P = 0.012**(N)
 

0.000
 

0.000
 

0.637
 

0.000
 

0.000
 

0.449
 

—
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Treated with ß-TGdRa 

(continued) 

Pooled Matched 

Vehicle Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Ear Canal: Carcinoma, NOS, or
 

Squamous-cell Carcinoma^ 0/30 (0) 0/10 (0) 1/35 (3) 2/30 (7)
 

P Values0 »d N.S. N.S. N.S. N.S.
 

Relative Risk (Pooled Vehicle Control)f Infinite Infinite
 

Lower Limit 0.047 0.301
 

Upper Limit Infinite Infinite
 

Relative Risk (Matched Vehicl e Control)f Infinite Infinite
 

Lower Limit 0.017 0.109
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 76
 — —
 

a

Dosed groups received 3.5 or 7 mg/kg by intraperitoneal injection.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

G

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the matched vehicle-control group (*) or with the
 

pooled vehicle-control group (**) when P < 0.05 for either control group; otherwise, not
 

significant (N.S.) is indicated.
 

63 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Treated with ß-TGdRa
 

(continued)
 

^A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^The 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
 

Treated with ß-TGdRa
 

Pooled Matched 

Vehicle Vehicle 

Topography: Morphology Control Control 

Hematopoietic System:
 

Lymphoma 1/30 (3) 1/10 (10)
 

P Values

c»d N.S. N.S.
 

Relative Risk (Pooled Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 41 41
 

Pituitary: Chromophobe
 

Adenoma^ 14/28 (50) 8/10 (80)
 

P Values0 »d P = 0.002(N) P < O.OOl(N)
 

Relative Risk (Pooled Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 41 41
 

Low
 

Dose
 

0/32 (0)
 

N.S.
 

0.000
 

0.000
 

17.314
 

0.000
 

0.000
 

5.791
 

12/31 (39)
 

P = 0.027*(N)
 

0.774
 

0.407
 

1.482
 

0.484
 

0.342
 

1.077
 

68
 

High
 

Dose
 

3/27 (11)
 

N.S.
 

3.333
 

0.288
 

167.925
 

1.111
 

0.108
 

56.038
 

31
 

2/23 (9)
 

P < 0.001*(N)
 

P = 0.002**(N)
 

0.174
 

0.022
 

0.649
 

0.109
 

0.012
 

0.419
 

70
 
Treated

(continued) 

Pooled 

Vehicle 

Topography: Morphology Control 

Mammary Gland:
 

Adenocarcinoma, NOS° 1/30 (3)
 

P Values0 »d N.S.
 

Relative Risk (Pooled Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 80
 

Mammary Gland: Carcinoma or
 

Adenocarcinoma, NOS^ 2/30 (7)
 

P Values0 »d N.S.
 

Relative Risk (Pooled Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 79
 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats 

 with ß-TGdRa 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

—
 

1/10 (10)
 

N.S.
 

79
 

Low
 

Dose
 

3/32 (9)
 

N.S.
 

2.813
 

0.242
 

142.727
 

Infinite
 

0.209
 

Infinite
 

74
 

3/32 (9)
 

N.S.
 

1.406
 

0.173
 

15.880
 

0.938
 

0.091
 

47.629
 

74
 

High
 

Dose
 

0/27 (0)
 

N.S.
 

0.000
 

0.000
 

20.405
 

—
 

—
 

—
 

—
 

0/27 (0)
 

N.S.
 

0.000
 

0.000
 

3.673
 

0.000
 

0.000
 

6.825
 

—
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats 

Treated with ß-TGdRa 

(continued) 

Pooled
 

Vehicle
 

Control
 

6/30 (20)
 

P = 0.020(N)
 

Control)f
 

Control)f
 

79
 

0/30 (0)
 

P = 0.010
 

Control)f
 

Control)f
 

—
 

Topography: Morphology
 

Mammary Gland: Fibroadenoma^
 

P Values0 »d
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ear Canal: Squamous-cell
 

Carcinoma^
 

P Valuesc»d
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Vehicle
 

Control
 

2/10 (20)
 

P = 0.032(N)
 

79
 

0/10 (0)
 

N.S.
 

. —
 

Low
 

Dose
 

5/32 (16)
 

N.S.
 

0.781
 

0.211
 

2.750
 

0.781
 

0.165
 

7.520
 

74
 

2/32 (6)
 

N.S.
 

Infinite
 

0.282
 

Infinite
 

Infinite
 

0.102
 

Infinite
 

70
 

High
 

Dose
 

0/27 (0)
 

P = 0.016**(N)
 

0.000
 

0.000
 

0.675
 

0.000
 

0.000
 

1.210
 

—
 

5/27 (19)
 

P = 0.019**
 

Infinite
 

1.436
 

Infinite
 

Infinite
 

0.524
 

Infinite
 

63
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Treated with ß-TGdRa
 

(continued) 

Pooled Matched 

Vehicle Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Ear Canal: Carcinoma, NOS, or
 

Squamous-cell Carcinoma'3 0/30 (0) 0/10 (0) 2/32 (6) 6/27 (22)
 

P Valuesc»d P = 0.004 N.S. N.S. P = 0.008**
 

Relative Risk (Pooled Vehicle Control)f Infinite Infinite
 

Lower Limit 0.282 1.824
 

Upper Limit Infinite Infinite
 

Relative Risk (Matched Vehicle Control)f Infinite Infinite
 

Lower Limit 0.102 0.665
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tuiaor 70 48
 —
 —
 

a

Dosed groups received 3.5 or 7 mg/kg by intraperitoneal injection.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

G

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the matched vehicle-control group (*) or with the
 

pooled vehicle-control group (**) when P < 0.05 for either control group; otherwise, not
 

significant (N.S.) is indicated.
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Treated with ß-TGdRa
 

(continued)
 

"A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
__ __ 

Table E3. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Treated with ß-TGdRa
 

Topography: Morphology
 

Hematopoietic System:
 

Lymphoma (43)°
 

>d
 P Valuesc


Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe
 

Adenoma (52)b
 

P Values0 >d
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/29 (0)
 

P = 0.025
 

Control)f
 

Control)f
 

7/24 (29)
 

N.S.
 

Control)f
 

Control)f
 

79
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

4/9 (44)
 

P = 0.027(N)
 

79
 

Low
 

Dose
 

1/32 (3)
 

N.S.
 

Infinite
 

0.049
 

Infinite
 

Infinite
 

0.018
 

Infinite
 

66
 

3/22 (14)
 

N.S.
 

0.468
 

0.088
 

1.763
 

0.307
 

0.065
 

1.511
 

77
 

High
 

Dose
 

2/10 (20)
 

N.S.
 

Infinite
 

0.885
 

Infinite
 

Infinite
 

0.330
 

Infinite
 

0/6 (0)
 

N.S.
 

0.000
 

0.000
 

1.672
 

0.000
 

0.000
 

1.293
 

—
 

43 
Table E3. lime-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Treated with ß-TGdRa
 

(continued) 

Pooled Matched 

Vehicle Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Ear Canal: Carcinoma, NOS, or
 

Squamous-cell Carcinoma (52)^ 0/28 (0) 0/10 (0) 1/31 (3) 2/7 (29)
 

P Values0 »d P = 0.014 P = 0.046 N.S. P = 0.035**
 

Relative Risk (Pooled Vehicle Con trol)f Infinite Infinite
 

Lower Limit 0.049 1.233
 

Upper Limit Infinite Infinite
 

Relative Risk (Matched Vehicle Co ntrol)f Infinite Infinite
 

Lower Limit 0.019 0.475
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 76 63
 —
 —
 

a

Dosed groups received 3.5 or 7 mg/kg by intraperitoneal injection.
 

"Number of tumor-bearing animals/number of animals examined at site (percent), based on animals
 

that lived at least as long as the number of weeks shown in parentheses after the description
 

of morphology.
 

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran—
 

Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the matched vehicle-control group (*) or with the
 

pooled vehicle-control group (**) when P < 0.05 for either control group; otherwise, not
 

significant (N.S.) is indicated.
 
Table E3. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats 

Treated with ß-TGdRa 

(continued) 

"A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

e

lhe probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

•^The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
__ 

Table E4. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Treated with ß-TGdRa
 

Pooled Matched 

Vehicle Vehicle 

Topography: Morphology Control Control 

Hematopoietic System:
 

Lymphoma (31)° 1/30 (3) 1/10 (10)
 

P Valuesc'd N.S. N.S.
 

Departure from Linear Trend6 P = 0.026 P = 0.029
 

Relative Risk (Pooled Vehicle Con trol)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Co ntrol)£
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 41 41
 

Pituitary: Chromophobe
 

Adenoma (41)b 14/28 (50) 8/10 (80)
 

P Valuesc'd P = 0.023(N) P = 0.002(N)
 

Relative Risk (Pooled Vehicle Con trol)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Co ntrol)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 41 41
 

Low
 

Dose
 

0/32 (0)
 

N.S.
 

0.000
 

0.000
 

17.314
 

0.000
 

0.000
 

5.791
 

12/31 (39)
 

P = 0.027*(N)
 

0.774
 

0.407
 

1.482
 

0.484
 

0.342
 

1.012
 

68
 

High
 

Dose
 

3/15 (20)
 

N.S.
 

6.000
 

0.526
 

289.415
 

2.000
 

0.198
 

96.600
 

31
 

2/14 (14)
 

P = 0.002*(N)
 

P = 0.025**(N)
 

0.286
 

0.036
 

1.002
 

0.179
 

0.046
 

0.658
 

70
 
__ 

Table E4. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Treated with ß-TGdRa
 

(continued)
 

Topography: Morphology
 

Mammary Gland:
 

Adenocarcinoma, NOS (52)b
 

P Valuesc»d
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Carcinoma or
 

Adenocarcinoma, NOS (52)°
 

Pooled
 

Vehicle
 

Control
 

1/29 (3)
 

N.S.
 

Control)f
 

Control)^
 

80
 

2/29 (7)
 

P Values0>d N.S.
 

Relative Risk (Pooled Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 79
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

—
 

1/10 (10)
 

N.S.
 

79
 

Low
 

Dose
 

3/31 (10)
 

N.S.
 

2.806
 

0.242
 

142.233
 

Infinite
 

0.216
 

Infinite
 

74
 

3/31 (10)
 

N.S.
 

1.403
 

0.174
 

15.813
 

0.968
 

0.094
 

49.097
 

74
 

High
 

Dose
 

0/12 (0)
 

N.S.
 

0.000
 

0.000
 

42.376
 

—
 

—
 

—
 

0/12 (0)
 

N.S.
 

0.000
 

0.000
 

7.546
 

0.000
 

0.000
 

14.663
 

—
 
Table E4. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Treated with ß-TGdRa
 

(continued) 

Pooled 

Vehicle 

Topography: Morphology Control 

Mammary Gland:
 

Fibroadenoma (52)b 6/29 (21)
 

P Valuesc»d N.S.
 

Relative Risk (Pooled Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 79
 

Ear Canal: Squamous-cell
 

Carcinoma (52)b 0/30 (0)
 

>d
 P Valuesc P < 0.001
 

Departure from Linear Trend6 P = 0.043
 

Relative Risk (Pooled Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 —
 —
 

Matched
 

Vehicle
 

Control
 

2/10 (20)
 

N.S.
 

79
 

0/10 (0)
 

P = 0.004
 

Low
 

Dose
 

5/31 (16)
 

N.S.
 

0.780
 

0.211
 

2.733
 

0.806
 

0.170
 

7.748
 

74
 

2/31 (6)
 

N.S.
 

Infinite
 

0.291
 

Infinite
 

Infinite
 

0.105
 

Infinite
 

70
 

High
 

Dose
 

0/12 (0)
 

N.S.
 

0.000
 

0.000
 

1.378
 

0.000
 

0.000
 

2.576
 

«._
 

5/12 (42)
 

P = 0.030*
 

P = 0.001**
 

Infinite
 

3.311
 

Infinite
 

Infinite
 

1.201
 

Infinite
 

63
 
Table E4. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Treated with ß-TGdRa
 

(continued) 

Pooled Matched 

Vehicle Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Ear Canal: Carcinoma, NOS, or
 

Squamous-cell Carcinoma (48) b

 0/28 (0) 0/9 (0) 2/32 (6) 6/13 (46)
 

P Valuesc»d P < 0.001 P = 0.002 N.S.	 P = 0.023*
 

P < 0.001**
 

Departure from Linear Trend6 P = 0.024
 

Relative Risk (Pooled Vehicle Control)f Infinite Infinite
 

Lower Limit 0.264 3.636
 

Upper Limit Infinite Infinite
 

Relative Risk (Matched Vehicle Control)f Infinite Infinite
 

Lower Limit 0.093 1.286
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor	 70 48
 — —
 

a

Dosed groups received 3.5 or 7 mg/kg by intraperitoneal injection.
 

^Number of tumor-bearing animals/number of animals examined at site (percent), based on
 

animals that lived at least as long as the number of weeks shown in parentheses after the
 

description of morphology.
 
Table E4. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats 

Treated with ß-TGdRa 

(continued) 

G

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the matched vehicle—control group (*) or with the
 

pooled vehicle-control group (**) when P < 0.05 for either control group: 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 probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^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 TREATED WITH ß-TGdR
 

111
 

__ 

Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Treated with ß-TGdRa
 

Topography: Morphology
 

Hematopoietic System: Lymphomab
 

P Values0>d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia"
 

P Values0>d
 

Relative Risk (Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Vehicle
 

Control
 

19/29 (66)
 

N.S.
 

39
 

0/29 (0)
 

N.S.
 

—
 

Low
 

Dose
 

17/35 (49)
 

N.S.
 

0.741
 

0.473
 

1.208
 

51
 

0/35 (0)
 

N.S.
 

—
 

—
 

—
 

Mid
 

Dose
 

16/32 (50)
 

N.S.
 

0.763
 

0.482
 

1.242
 

28
 

2/32 (6)
 

N.S.
 

Infinite
 

0.272
 

Infinite
 

54 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice 

Treated with ß-TGdRa 

(continued) 

Vehicle Low Mid
 

Topography: Morphology Control Dose Dose
 

Hematopoietic System: Lymphoma
 

or Leukemia^3 19/29 (66) 17/35 (49) 18/32 (56)
 

P Values

c»d N.S. N.S. N.S.
 

Relative Risk (Vehicle Control)f 0.741 0.859
 

Lower Limit 0.473 0.558
 

Upper Limit 1.208 1.354
 

Weeks to First Observed Tumor 39 51 28
 

a

Dosed groups received 1 or 2 mg/kg by intraperitoneal injection.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the vehicle-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 probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^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
 

Treated with ß-TGdRa
 

Topography; Morphology
 

Hematopoietic System: Lymphoma^
 

P Values0 »d
 

Relative Risk (Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Lymphoma
 

or Leukemia**
 

P Values0 »d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Vehicle
 

Control
 

20/29 (69)
 

N.S.
 

33
 

21/29 (72)
 

N.S.
 

27
 

Low
 

Dose
 

19/33 (58)
 

N.S.
 

0.835
 

0.560
 

1.285
 

64
 

19/33 (58)
 

N.S.
 

0.795
 

0.547
 

1.207
 

64
 

Mid
 

Dose
 

21/29 (72)
 

N.S.
 

1.050
 

0.728
 

1.501
 

34
 

21/29 (72)
 

N.S.
 

1.000
 

0.708
 

1.412
 

34
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice 

Treated with j8-TGdRa 

(continued) 

a

Dosed groups received 1 or 2 rag/kg by intraperitoneal injection.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the vehicle-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 probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
__ 

Table F3. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Mice
 

Treated with ß-TGdRa
 

Topography; Morphology
 

Hematopoietic System: Lymphoma^
 

P Valuesd
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Hematopoietic System: Leukemia0
 

P Valuesd
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Vehicle
 

Control
 

19/29 (66)
 

N.S.
 

0/16 (0)
 

N.S.
 

Low
 

Dose
 

17/35 (49)
 

N.S.
 

0.741
 

0.473
 

1.206
 

0/31 (0)
 

N.S.
 

—
 

—
 

Mid
 

Dose
 

16/31 (52)
 

N.S.
 

0.788
 

0.498
 

1.276
 

2/18 (11)
 

N.S.
 

Infinite
 

0.279
 

Infinite
 
Table F3. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Mice
 

Treated with ß-TGdRa
 

(continued) 

Vehicle Low Mid 

Topography; Morphology Control Dose Dose 

Hematopoietic System: Lymphoma 

or Leukemia^ 19/29 (66) 17/35 (49) 18/31 (58) 

P Valuesd N.S. N.S. N.S. 

Relative Risk (Vehicle Control)f 0.741 0.886 

Lower Limit 0.473 0.578 

Upper Limit 1.208 1.387 

a

Dosed groups received 1 or 2 mg/kg by intraperitoneal injection.
 

"Number of tumor-bearing animals/number of animals examined at site (percent) which survived at
 

least 28 weeks of the study.
 

G

Number of tumor-bearing animals/number of animals examined at site (percent) which survived at
 

least 52 weeks of the study.
 

"Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the vehicle-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^The 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
Table F4. Time-adjusted Analyses of the Incidence of Primary

Treated with ß-TGdRa 

Vehicle Low
 

Topography: Morphology Control Dose
 

Hematopoietic System: Lymphoma^ 20/28 (71) 19/32


P Values6 U.S. N.S.
 

Relative Risk (Vehicle Control )8 0.831
 

Lower Limit 

Upper Limit 

0.568 

1.266 

Hematopoietic System:

or Leukemia0 

 Lymphoma 

21/29 (72) 19/33

P Values6 N.S. N.S. 

Relative Risk (Vehicle Control) 8 0.795
 

Lower Limit 0.547
 

Upper Limit 1.207
 

 Tumors in

 (59)
 

 (58)
 

 Female Mice 

Mid
 

Dose
 

21/28 (75)
 

N.S.
 

1.050
 

0.740
 

1.473
 

21/28 (75)
 

N.S.
 

1.036
 

0.736
 

1.440
 
Table F4. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Mice 

Treated with ß-TGdRa 

(continued) 

a

Dosed groups received 1 or 2 mg/kg by intraperitoneal injection.
 

^Number of tumor-bearing animals/number of animals examined at site (percent) which survived
 

at least 33 weeks of the study.
 

°Number of tumor-bearing animals/number of animals examined at site (percent) which survived
 

at least 27 weeks of the study.
 

^Number of tumor-bearing animals/number of animals examined at site (percent) which survived
 

at least 52 weeks of the study.
 

e

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. 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 the vehicle-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

^The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

Slhe 95% confidence interval of the relative risk between each dosed group and the specified
 

control group.
 
Review of the Bioassay of Beta-Deoxythioguansine* (ß-TGdR)
 

for Carcinogenlcity
 

by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

January 18, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976 under the authority of the National
 

Cancer Act of 1971 (P.L. 92-218). The purpose of the Clear­ inghouse is to advise on the National Cancer Institute's
 

bioassay program to identify and evaluate chemical carcino­ gens 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 organic chemistry, bio­ statistics, biochemistry, toxicology, pathology, and epide­ miology. Representatives of various Governmental agencies
 

participate as ad hoc members. The Data Evaluation/Risk
 

Assessment Subgroup of the Clearinghouse is charged with
 

the responsibility of providing a peer review of NCI bioassay
 

reports on chemicals studied for carcinogenicity. In this
 

context, below is the edited excerpt from the minutes of
 

the Subgroup's meeting at which Beta-Deoxythioguansine
 

(p-TGdR)was reviewed.
 

The primary reviewer noted the unusual occurrence of
 

ear canal carcinomas in the ß-TGdR treated rats. Despite
 

the high mortality due to toxicity, he considered the
 

results of the study to be valid. He said that the Incidence
 

of lymphomas and leukemias in the matched vehicle control mice
 

may be related to the mineral oil vehicle. The primary
 

reviewer opined that too much attention was given to
 

statistical analyses to the detriment of biological
 

considerations.
 

The secondary reviewer said he agreed that ß-TGdR
 

was carcinogenic in the treated rats. He questioned the
 

primary reviewer's notion that Polysorbate 80 was related
 

to the mouse lymphomas and leukemias, since it has not
 

produced a similar effect in other studies in which it
 

had been used.
 

121
 
A Program staff pathologist commented that the Polysorbate 80 may not act in the same way as mineral oil. He
 

suggested that granulomas also would have been found if the
 

reaction was similar to that of mineral oil. Another
 

Program staff member said that it was clear that the rat
 

ear canal tumors were related to 3-TGdR treatment. He
 

added that a question of the significance of the lymphomas
 

in mice arose because of possible procedural irregularities.
 

A motion was made that Beta-Deoxythioguansine was
 

carcinogenic in the rat and that the mouse study was uninter­ pretable because of its inadequacies. The motion was
 

seconded and passed five to two. The opposing votes were
 

cast by Mr. Garfinkel and Dr. Kensler.
 

Members Present Were:
 

Arnold Brown (Acting Chairman), Mayo Clinic
 

Lawrence Garfinkel, American Cancer Society
 

Joseph Highland, Environmental Defense Fund
 

Charles Kensler, Arthur D. Little Company
 

Verald K. Rowe, Dow Chemical, U.S.A.
 

Sheldon Samuels, Industrial Union Department, AFL-CIO
 

Louise Strong, University of Texas Health Sciences Center
 

Sidney Wolfe, Health Research Group
 

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: 1978 260-889/3042 1-3 122 

DHEW Publication No. (NIH) 78-1363