National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 58 

1978 

BIOASSAY OF 

THIO-TEPA 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 52-24-4 

NCI-CG-TR-58 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

THIO-TEPA
 

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) 78-1308
 

BIOASSAY OF
 

THIO-TEPA
 

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
 

thio-TEPA conducted for the Carcinogenesis Testing Program,
 

Division of Cancer Cause and Prevention, National Cancer
 

Institute (NCI), National Institutes of Health, Bethesda,
 

Maryland· This is one of a series of experiments designed to
 

determine whether selected chemicals have the capacity to produce
 

cancer in animals. Negative results, in which the test animals
 

do not have a greater incidence of cancer than control animals,
 

do not necessarily mean that the test chemical is not a carcinogen, inasmuch as the experiments are conducted under a limited
 

set of circumstances· Positive results demonstrate that the test
 

chemical is carcinogenic for animals under the conditions of the
 

test and indicate that exposure to the chemical is a potential
 

risk to man. The actual determination of the risk to man from
 

animal carcinogens requires a wider analysis.
 

CONTRIBUTORS: This bioassay of thio-TEPA was conducted by
 

Southern Research Institute, Birmingham, Alabama, initially under
 

direct contract to NCI and currently under a subcontract to
 

Tracor Jitco, Inc., prime contractor for the NCI Carcinogenesis
 

Testing Program.
 

Weisburger *^. Ms. J. Belzer* and Mr. I. Brown* were responsible
 

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

Griswold1 

, J. D. Prejean1 , E. K. Weisburger2 , and J. Η. 2

for the care of the laboratory animals and administration of the
 

test chemical. Data management and retrieval were performed by
 

Ms. C. A. Dominick*. Histopathologic examinations were performed
 

by Drs. S. D. Kosanke* and J. C. Peckham*, and the diagnoses
 

included in this report represent their interpretation.
 

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

Mason Research Institute^. The statistical analyses were performed by Dr. J. R· Joiner^ and Ms· P· L· Yong^, using methods
 

selected for the bioassay program by Dr. J. J. Gart". Chemicals
 

used in this bioassay were analyzed under the direction of Dr. E.
 

Murrill^, and the analytical results were reviewed by Dr. S. S.
 

01in-\ The chemical structure was supplied by NCI.
 

This report was prepared at Tracor Jitco^ under the direction of
 

NCI. Those responsible for the report at Tracor Jitco were
 

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

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

toxicologist; Dr. R. L. Schueler, pathologist; Dr. G. L. Miller,
 

Ms. L. A. Waitz, and Mr. W. D. Reichardt, bioscience writers; and
 

Dr. E. W. Gunberg, technical editor, assisted by Ms. Υ. Ε.
 

Presley.
 

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.
 

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

^Southern Research Institute, 2000 Ninth Avenue South,
 

Birmingham, Alabama.
 

^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

% o w with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

iv
 
^Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute, National
 

Institutes of Health, Bethesda, Maryland.
 

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

Missouri.
 

"Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of thio-TEPA for possible carcinogenicity was
 

conducted by administering the test chemical by intraperitoneal
 

injection to Sprague-Dawley rats and B6C3F1 mice.
 

Groups of 31-39 rats of each sex were administered thio-TEPA in
 

phosphate-buffered saline at one of three doses» either 0.7, 1.4,
 

or 2.8 mg/kg body weight, three times per week for a maximum of
 

52 weeks, then observed for additional periods of time. The 

maximum time on study (administration of chemical and 

observation) was 86 weeks. The groups at the low dose were 

started 69 weeks after those at the mid and high doses, because
 

of high mortalities observed in the groups at the higher doses.
 

Matched controls consisted of groups of 10 untreated rats and 10
 

vehicle-control rats of each sex. Pooled-control groups also
 

were used. Surviving control rats were killed at 82-87 weeks;
 

surviving dosed rats were killed at 81 or 82 weeks.
 

Groups of 35 mice of each sex were administered thio-TEPA at one
 

of two doses, either 1.15 or 2.3 mg/kg body weight, three times
 

per week for a maximum of 52 weeks, then observed for a maximum
 

additional period of 34 weeks. Matched controls consisted of
 

groups of 15 untreated mice and 15 vehicle-control mice of each
 

sex. Pooled controls also were used. Surviving control and
 

dosed mice were killed at 86 or 87 weeks.
 

Thio-TEPA was toxic to both rats and mice, causing decreased mean
 

body weight gains and early deaths in the mid- and high-dose rats
 

and in the high-dose mice. Because of the early deaths,
 

statistical analyses were based only on time-adjusted incidences
 

of tumors. Since all high-dose male and female rats had died by
 

21 weeks, microscopic evaluation of tissues was performed only on
 

the low- and mid-dose animals.
 

In rats, the incidence of combined neoplasms of the hematopoietic
 

system (lymphoma, lymphocytic leukemia, or granulocytic leukemia)
 

was significant in the males in both the low-dose (P • 0.020) and
 

mid-dose (P • 0.001) groups, using pooled controls (pooled
 

vii
 
controls 0/29, low-dose 6/34; pooled controls 0/30, mid-dose
 

6/16).
 

Squamous-cell carcinoma of the skin or ear canal occurred at a
 

significant incidence in the male rats in both the low-dose (P «
 

0.009) and mid-dose (P = 0.023) groups, using pooled controls
 

(pooled controls 0/29, low-dose 7/33; pooled controls 0/30,
 

mid-dose 3/13) and in the mid-dose females (P < 0.001), using
 

pooled controls (pooled controls 0/28, mid-dose 8/21); in 

addition, two low-dose females had such tumors, with none 

occurring in the corresponding low-dose controls. 

The incidence of adenocarcinoma of the uterus was significant in
 

the mid-dose female rats (P = 0.001), using pooled controls
 

(pooled controls 0/28, mid-dose 7/21); in addition, two low-dose
 

females had adenocarcinoma of the uterus, with no such tumor
 

occurring in the corresponding low-dose controls.
 

In rats, neuroepitheliomas (neuroblastomas) or nasal carcinomas
 

occurred in three low-dose males, two low-dose females, and two
 

mid-dose females. Although these are not statistically
 

significant incidences, these tumors did not occur among control
 

animals and no such tumors have occurred in 380 Sprague-Dawley
 

control rats of each sex in other bioassays at the same
 

laboratory. Thus, they may be associated with administration of
 

the chemical.
 

In the high-dose groups of both male and female mice, but not in
 

the low-dose groups, the incidences of lymphoma or lymphocytic
 

leukemia were significantly higher (P < 0.001) for each sex than
 

those of either the vehicle or pooled controls (males: vehicle
 

controls 1/8, pooled controls 1/18, low-dose 2/24, high-dose
 

26/28; females: vehicle controls 0/14, pooled controls 0/29,
 

low-dose 5/26, high-dose 32/32).
 

In the low-dose male mice squamous-cell carcinoma was found in
 

the skin of seven animals, in the preputial glands of six
 

viii
 
animals, and in the ear canal of two animals· A carcinoma of the
 

preputial gland was also found in a high-dose male· When the
 

incidences of the tumors at the different sites were combined,
 

the incidence in the low-dose group was statistically significant
 

using either the vehicle (P = 0.004) or the pooled (P < 0.001)
 

controls (vehicle controls 0/8, pooled controls 0/18, low-dose
 

14/24, high-dose 1/2).
 

It is concluded that under the conditions of this bioassay,
 

thio-TEPA was carcinogenic in both Sprague-Dawley rats and B6C3F1
 

mice. In the rats, the chemical induced squamous-cell carcinoma
 

of the skin or ear canal in both males and females, and
 

hematopoietic neoplasms in the males; in the mice, it induced
 

lymphoma or lymphocytic leukemia in both sexes and squamous-cell
 

carcinoma in the skin and associated glands of males.
 

ix
 

TABLE OF CONTENTS
 

Page
 

I. Introduction . . 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

Β. 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 14
 

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) 26
 

D. Statistical Analyses of Results (Rats) 33
 

IV. Results - Mice 37
 

A. Body Weights and Clinical Signs (Mice) 37
 

B. Survival (Mice) 37
 

C. Pathology (Mice) 40
 

D. Statistical Analyses of Results (Mice) 43
 

V. Discussion 45
 

VI. Bibliography. 49
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms
 

in Rats Given Intraperitoneal Injections
 

of Thio-TEPA 51
 

Table Al Summary of the Incidence of Neoplasms
 

in Male Rats Given Intraperitoneal
 

Injections of Thio-TEPA (Control Groups)... 53
 

xi
 
Page 

Table A2 Summary of the Incidence of Neoplasms 

in Male Rats Given Intraperitoneal 

Injections of Thio-TEPA (Treated Groups)... 56 

Table A3 Summary of the Incidence of Neoplasms 

in Female Rats Given Intraperitoneal 

Injections of Thio-TEPA (Control Groups)... 60 

Table A4 Summary of the Incidence of Neoplasms 

in Female Rats Given Intraperitoneal 

Injections of Thio-TEPA (Treated Groups)... 63 

Appendix Β Summary of the Incidence of Neoplasms 

in Mice Given Intraperitoneal Injections 

of Thio-TEPA 67 

Table Bl Summary of the Incidence of Neoplasms 

in Male Mice Given Intraperitoneal 

Injections of Thio-TEPA 69 

Table B2 Summary of the Incidence of Neoplasms 

in Female Mice Given Intraperitoneal 

Injections of Thio-TEPA 72 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Given Intraperitoneal 

Injections of Thio-TEPA 75 

Table CI Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Given Intraperitoneal 

Injections of Thio-TEPA (Control Groups)... 77 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Given Intraperitoneal 

Injections of Thio-TEPA (Treated Groups)... 80 

Table C3 Summary of the Incidence of Nonneoplastic 

Lesions of Female Rats Given Intraperitoneal 

Injections of Thio-TEPA (Control Groups)... 84 

Table C4 Summary of the Incidence of Nonneoplastic 

Lesions of Female Rats Given Intraperitoneal 

Injections of Thio-TEPA (Treated Groups)... 87 

xii
 
Page
 

Appendix D Summary of the Incidence of Nonneoplastic
 

Lesions in Mice Given Intraperitoneal
 

Injections of Thio-TEPA 91
 

Table Dl Summary of the Incidence of Nonneoplastic
 

Lesions in Male Mice Given Intraperitoneal
 

Injections of Thio-TEPA 93
 

Table D2 Summary of the Incidence of Nonneoplastic
 

Lesions in Female Mice Given Intraperitoneal
 

Injections of Thio-TEPA 96
 

Appendix Ε Analyses of the Incidence of Primary
 

Tumors in Rats Given Intraperitoneal
 

Injections of Thio-TEPA 99
 

Table El Analyses of the Incidence of Primary
 

Tumors in Low-Dose Male Rats Given
 

Intraperitoneal Injections of Thio-TEPA.... 101
 

Table E2 Analyses of the Incidence of Primary
 

Tumors in Mid-Dose Male Rats Given
 

Intraperitoneal Injections of Thio-TEPA.... 107
 

Table E3 Analyses of the Incidence of Primary
 

Tumors in Low-Dose Female Rats Given
 

Intraperitoneal Injections of Thio-TEPA.... 110
 

Table E4 Analyses of the Incidence of Primary
 

Tumors in Mid-Dose Female Rats Given
 

Intraperitoneal Injections of Thio-TEPA.... 116
 

Table E5 Time-adjusted Analyses of the Incidence
 

of Primary Tumors in Low-Dose Male Rats
 

Given Intraperitoneal Injections of
 

Thio-TEPA. :
 121
 

Table E6 Time-adjusted Analyses of the Incidence
 

of Primary Tumors in Mid-Dose Male Rats
 

Given Intraperitoneal Injections of
 

Thio-TEPA 127
 

xiii
 
Page 

Table E7 Time-adjusted Analyses of the Incidence 

of Primary Tumors in Low-Dose Female Rats 

Given Intraperitoneal Injections of 

Thio-TEPA 130 

Table E8 Time-adjusted Analyses of the Incidence 

of Primary Tumors in Mid-Dose Female Rats 

Given Intraperitoneal Injections of 

Thio-TEPA 136 

Appendix F Analyses of the Incidence of Primary 

Tumors in Mice Given Intraperitoneal 

Injections of Thio-TEPA 143 

Table Fl Analyses of the Incidence of Primary 

Tumors in Male Mice Given Intraperitoneal 

Injections of Thio-TEPA 145 

Table F2 Analyses of the Incidence of Primary 

Tumors in Female Mice Given Intraperitoneal 

Injections of Thio-TEPA 154 

Table F3 Time-adjusted Analyses of the Incidence 

of Primary Tumors in Male Mice Given 

Intraperitoneal Injections of Thio-TEPA.... 158 

Table F4 Time-adjusted Analyses of the Incidence 

of Primary Tumors in Female Mice Given 

Intraperitoneal Injections of Thio-TEPA.... 164 

TABLES 

Table 1 Design of Chronic Studies of 

Thio-TEPA in Rats 10 

Table 2 Design of Chronic Studies of 

Thio-TEPA in Mice 12 

FIGURES 

Figure 1

Figure 2

 Growth Curves for Male Rats Treated 

with Thio-TEPA....

 Growth Curves for Female Rats Treated 

with Thio-TEPA.

 22 

 23 

xiv
 
Page
 

Figure 3 Survival Curves for Male Rats Treated 

with Thio-TEPA 24 

Figure 4 Survival Curves for Female Rats Treated 

with Thio-TEPA 25 

Figure 5 Growth Curves for Mice Treated with 

Thio-TEPA. 38 

Figure 6 Survival Curves for Mice Treated with 

Thio-TEPA 39 

xv
 

I. INTRODUCTION
 

Thio-TEPA (CAS 52-24-4; NSC 6396; NCI C01649) is an ethyleneimine
 

alkylating agent that was introduced in 1953 for clinical use in
 

cancer chemotherapy· Following oxidative desulfurization of
 

thio-TEPA, the three ethylenimine groups that the chemical
 

contains become activated to e thy len imonium ions (Montgomery and
 

Struck, 1973; Lederle Laboratories, 1977). The presence of
 

several such groups raises the possibility that the drug will
 

bind to more than one site on the DNA molecule and will have
 

increased cytotoxic effects (Calabresi and Parks, 1975).
 

At one time thio-TEPA was an important therapeutic drug in the
 

management of ovarian carcinoma (Calabresi and Welch, 1962). It
 

has been used effectively in the treatment of Hodgkins disease,
 

bronchogenic carcinoma, bladder cancer, retinoblastoma, and
 

breast cancer and for the control of pleural, pericardial, and
 

peritoneal neoplastic effusions (Wheeler, 1973; Carter and
 

Slavik, 1974; Calabresi and Parks, 1975). In the pulmonary tumor
 

response test in strain A mice (Shimkin et al. , 1966), thio-TEPA
 

showed weak activity. On the basis of these results, the drug
 

was retested in a lifetime carcinogen bioassay. This drug is one
 

of a series of anticancer agents that were selected for testing
 

because they may be administered chronically in humans.
 

II. MATERIALS AND METHODS
 

A. Chemical
 

Thio-TEPA
 

Thio-TEPA is the common name for tris(l-aziridinyl)phosphine
 

sulfide, manufactured by Lederle Laboratories, Pearl River, New
 

York. Lederle Lot No. D9878, which conformed to USP
 

specifications, was used during the chronic study. The purity of
 

this lot was 98.0 + 1.0% as determined by titration of the
 

aziridine groups with tetrabutylammonium iodide and perchloric
 

acid. Elemental analysis for sulfur was slightly high (18.2 +;
 

0.6% vs. 17.0% theoretical), but may have been affected by
 

phosphorus; analysis of carbon, hydrogen, nitrogen, and
 

phosphorus verified the chemical composition of this product.
 

The melting point was 52.0-52.5°C and was comparable to that of
 

51.5°C reported elsewhere (U.S. Patent, 1954). Thin-layer
 

3 
chromatography showed one trace impurity which remained at the
 

origin. Vapor phase chromatography gave one homogeneous peak.
 

The infrared and nuclear magnetic resonance spectra were
 

consistent with the spectra given in the literature for this
 

compound.
 

B. Dosage Preparation
 

Solutions of thio-TEPA in phosphate-buffered saline were prepared
 

on the day that they were used by blending for 20 seconds in a
 

10-ml Potter-Elvehjem tissue grinder with a Teflon pestle; unused
 

solutions of the drug were discarded. Concentrations of
 

thio-TEPA ranged from 0.028% to 0.112% for the chronic studies
 

using rats and from 0.012% to 0.023% for the chronic studies
 

using mice. Volumes of solutions used for injection were 0.25
 

ml/100 g body weight for the rats and 1.0 ml/100 g body weight
 

for the mice.
 

C. Animals
 

For the subchronic studies, male Sprague-Dawley rats and male
 

Swiss mice were obtained from Charles River Breeding
 

Laboratories, Inc., Wilmington, Massachusetts. On arrival at the
 

laboratory, they were 30 days of age. The rats were quarantined
 

for 5 days and the mice for 20 days prior to being placed on
 

study.
 

4 
In the chronic studies, Sprague-Dawley rats and B6C3F1 mice of
 

each sex were obtained from Charles River Breeding Laboratories
 

under a contract with the Division of Cancer Treatment, National
 

Cancer Institute. Male rats were received at 29 days of age,
 

female rats at 36 days of age, and male and female mice at 32
 

days of age. All animals were quarantined for periods of 6 days
 

for rats and 10 days for mice. After these periods, animals with
 

no visible signs of disease were assigned to control or dosed
 

groups and earmarked for individual identification. Additional
 

groups of male and female Sprague-Dawley rats for the chronic
 

study were received at 30 days of age from Charles River Breeding
 

Laboratories and quarantined for 28 days weeks prior to being
 

placed on study.
 

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 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 meal (Allied Mills,, Inc., Chicago, 111.) and water
 

were supplied daily and were available an libitum.
 

5 
Rats were housed five per cage and mice seven 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.); mouse cages were
 

provided with Sterolit® clay bedding (Englehard Mineral and
 

Chemical Co., New York, N.Y.). For initial studies with rats,
 

cage tops were covered with disposable filter bonnets beginning
 

at week 24; for later studies with rats, cage tops were covered
 

with filter bonnets for the entire test. Filter bonnets were not
 

used for the mouse cages. Bedding was replaced once per week;
 

cages, water bottles, and feeders were sanitized at 82°C once per
 

week; and 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
 

thio-TEPA were maintained in the same rooms as animals of the
 

same species being 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 monohydrοchloride
 

(MAAM) (NSC 141549)
 

6 
acronycine (CAS 7008-42-6)
 

5-azacytidine (CAS 320-67-2)
 

beta-2'-deoxy-6-thioguanosine monohydrate (beta-TGdR)
 

(CAS 789-61-7)
 

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

emetine dihydrοchloride tetrahydrate (CAS 316-42-7)
 

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

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

(+)-4,4'-(1-methyl-1,2-ethanediyl)bis-2,6-piperazinedione
 

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

Ν,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) (CAS 63-92-3)
 

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

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

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

adriamycin (CAS 23214-92-8)
 

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-toluenesulfonyl)-N'-hexamethyleniminourea
 

(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)
 

7
 
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)
 

beta~2'-deoxy-6~thioguanosine monohydrate (beta-TGdR)
 

(CAS 789-61-7)
 

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)
 

(+)-4,4Wl-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) (CAS 63-92-3)
 

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

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

E. Subchronic Studies
 

Subchronic studies were conducted using male Sprague-Dawley rats
 

and male Swiss mice to estimate the maximum tolerated doses of
 

thio-TEPA, on the basis of which two concentrations (hereinafter
 

referred to as "low doses11 and "high doses11) were determined for
 

administration in the chronic studies. Rats were administered
 

the drug at doses of 0.14, 0.35, 0.7, 1.4, or 2.8 mg/kg, and mice
 

at doses of 0.23, 0.58, 1.15, 2.3, or 4.6 mg/kg. Dosed animals
 

were injected intraperitoneally with thio-TEPA three times per
 

week for 45 days and then observed for an additional 45 days.
 

Five animals of each species were administered the chemical at
 

8
 
each dose, 10 animals of each species were maintained as vehicle
 

controls, and 10 animals of each species were maintained as
 

untreated controls·
 

There were no deaths in the rats at any dose tested· Mean body
 

weight gains at 45 days in animals at 0·14, 0.35, or 0.7 mg/kg
 

were unaffected in comparison with those of the vehicle controls;
 

at 1.4 mg/kg the mean body weights were 79% of those of the
 

controls, and at 2· 8 mg/kg they were 70% of those of the
 

controls. After 90 days, mean body weight gains were comparable
 

to those of the controls in all groups except those at 1.4 mg/kg
 

and 2.8 mg/kg, where they were approximately 85% of those of the
 

controls. The vehicle controls gained slightly less weight than
 

the untreated controls. The low and high doses for the chronic
 

studies using rats were set at 1.4 and 2.8 mg/kg·
 

In mice, 3/5 animals died at 4.6 mg/kg and 1/5 died at 0.58
 

mg/kg. At 45 days, mean body weight gains in dosed animals were
 

not greatly affected, except for slight losses in the group at
 

4.6 mg/kg. The low and high doses for the chronic studies using
 

mice were set at 1.15 and 2.3 mg/kg.
 

F· Designs of the Chronic Studies
 

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

9 
Table 1. Design of Chronic Studies of Thio-TEPA in Rats
 

Sex and 

Test 

Group 

Initial

No. of 

Animalsa 

 Thio-TEPA 

Doseb 

(mg/ka) 

Time 

Dosed 

(weeks) 

on Study 

Observed 

(weeks) 

Male 

Low-Dose 

Untreated-Controlc 

Low-Dose 

Vehicle-Control0 

Low-Dosec 

10 

10 

39 

0 

0^ 

0.7 

52 

52 

83 

30 

30 

Mid- and High-Dose 

Untreated-Control 

Mid- and High-Dose 

Vehicle-Control 

Mid-Dose 

High-Dose 

10 

10 

35 

35 

0 

0d 

1.4 

2.8 

34e 

34e 

198 

87 

52 

44f 

Female 

Low-Dose 

Untreated-Control0 

Low-Dose 

Vehicle-Control0 

Low-Dose° 

10 

10 

31 

0 

0d 

0.7 

52 

52 

82 

30 

30 

Mid- and High-Dose 

Untreated-Control 

Mid- and High-Dose 

Vehicle-Control 

Mid-Dose 

High-Dose 

10 

10 

35 

35 

0 

0d 

1.4 

2.8 

34e 

34e 

218 

87 

53 

47 

aAges of rats when placed on study: mid- and high-dose males,
 

35 days; mid- and high-dose females, 42 days; low-dose males and
 

females, 58 days.
 

"Thio-TEPA was administered intraperitoneally in phosphate-


buffered saline three times per week at a volume of 0.25 ml/100 g
 

body weight; doses were based on individual weights.
 

10
 
Table 1. Design of Chronic Studies of Thio-TEPA in Rats
 

(continued)
 

c

Because of deaths in the high- and mid-dose groups, new
 

dosed and control groups were started 69 weeks after the
 

start of the original study.
 

^Vehicle controls received phosphate-buffered saline at the same
 

volume as the dosed animals.
 

Administration of the chemical to mid-dose males and females and
 

to mid- and high-dose vehicle controls terminated at week 34, due
 

to toxicity in the dosed groups.
 

fObservation of mid-dose males terminated at week 78, due to
 

death of all animals.
 

^Administration of the chemical to high-dose males and females
 

terminated at times indicated, due to death of all animals.
 

11
 
Table~2. Design of Chronic Studies of Thio-TEPA in Mice
 

Sex and Initial Thio-TEPA Time on Study 

Test No. of Doseb Dosed Observed 

Group Animalsa (mg/kg) (weeks) (weeks) 

Male 

Untreated-Control 15 0 87 

Vehicle-Control 15 0c 52 35 

Low-Dose 35 1.15 52 34 

High-Dose 35 2.3 52 4d 

Female 

Untreated-Control 15 0 87 

Vehicle-Control 15 0c 52 35 

Low-Dose 35 1.15 52 34 

High-Dose 35 2.3 43e 

aAll animals were 42 days of age when placed on study.
 

^Thio-TEPA was administered intraperitoneally in phosphate-


buffered saline three times per week at a volume of 1.0 ml/100 g
 

body weight based on the mean weight of the animals in each cage.
 

c

Vehicle controls received only phosphate-buffered saline
 

solution, at the same volume as dosed mice.
 

^Observation of high-dose males terminated at week 56, due to
 

death of all animals.
 

e

Administration of the chemical to high-dose females terminated at
 

week 43, due to death of all animals.
 

12
 
Since the numbers of rats and mice in the control groups were
 

small, pooled-control groups also were used for statistical
 

comparisons. Pooled-control groups for the low-dose rats
 

consisted of vehicle-control groups of 10 animals of each sex
 

from the bioassay of thio-TEPA, combined with corresponding
 

vehicle-control groups of 10 animals of each sex from similar
 

bioassays of beta-2'-deoxy-6-thioguanosine monohydrate (3-TGDR)
 

and 3,3'-iminobis-l-propanol dimethanesulfonate (ester) [IPD];
 

pooled-control groups for the mid-dose rats consisted of vehicle-


control groups of 10 animals of each sex from the bioassay of
 

thio-TEPA, combined with corresponding vehicle-control groups of
 

10 animals of each sex from similar bioassays of procarbazine and
 

IPD. The total number of animals was 30 in both the low-dose
 

pooled-control group and the mid-dose pooled-control group. For
 

the mice, pooled-control groups consisted of vehicle-control
 

groups of 15 animals of each sex from the bioassay of thio-TEPA,
 

combined with corresponding vehicle-control groups of 15 animals
 

of each sex from a similar bioassay of IPD, to give a total of 30
 

animals per group. The bioassays of the chemicals other than
 

thio-TEPA were also conducted at Southern Research Institute and
 

were started no more than 3 months apart from those for thio-


TEPA· The vehicle-control groups of rats and mice used in the
 

pooled-control groups were of the same strain, obtained from the
 

same supplier, and examined by the same pathologists as in the
 

13
 
respective dosed groups; further, the same vehicle was used for
 

the different vehicle-control groups.
 

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
 

test chemical. Rats (mid- and high-dose) and mice were weighed
 

individually each week for 8 weeks and every 2 weeks thereafter;
 

low-dose rats were weighed once every 2 weeks for the period of
 

administration of the chemical 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
 

14
 
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
 

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
 

15
 
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
 

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 Ρ values have been reported for all tests except the
 

departure from linearity test, which is only reported when its
 

two-tailed Ρ 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,
 

16
 
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
 

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
 

 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 Ρ 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
 

17
 
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
 

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;
 

18
 
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
 

s tlle
 relative risk is defined as Pt/Pc where Pt i true binomial
 

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

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

spontaneous incidence of the same type of tumor in a control
 

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
 

19
 
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, Ρ < 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.
 

20
 
III. RESULTS - RATS
 

A· Body Weights and Clinical Signs (Rats)
 

Mean body weights of all dosed male rats, particularly those of
 

the mid- and high-dose groups, were depressed throughout the
 

study, when compared with either matched or vehicle controls;
 

those of the dosed females were less markedly depressed (figures
 

1 and 2). 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 wide variation·
 

There was clinical evidence of respiratory disease in the high-


and mid-dose groups of rats and their untreated and vehicle
 

controls· To control the respiratory disease, these animals
 

received oxytetracycline in drinking water at a dose of 0.6 mg/ml
 

during weeks 24-30 and 0.3 mg/ml during weeks 30-35. No other
 

signs of toxicity were reported for the rats.
 

B· Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for the male and female rats administered thio-TEPA at
 

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

are shown in figures 3 and 4.
 

For each sex, the results of the Tarone test were significant
 

21
 
TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves for Male Rats Treated with Thio-TEPA 

22 
40 50 60 70 

TIME ON STUDY (WEEKS) 

Figure 2. Growth Curves for Female Rats Treated with Thio-TEPA 

23 
40 50 60 70 

TIME ON STUDY (WEEKS) 

Figure 3. Survival Curves for Male Rats Treated with Thio-TEPA 
TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves for Female Rats Treated with Thio-TEPA 

25 
( < 0.001), indicating that the administration of thio-TEPA
 

decreased survival in a dose-related manner· At the high dose,
 

all animals were dead at week 19 (males) or week 21 (females)·
 

Survival at the mid dose was also low, with all males dead by
 

week 78, and with only 3/35 (8.6%) females surviving to
 

termination, while 6/39 (15.4%) low-dose males and 13/31 (42%)
 

low-dose females survived to the end of the study. Among the
 

male vehicle controls, 8/10 (80%) of the mid and high-dose
 

controls and 10/10 (100%) of the low-dose vehicle controls
 

survived to week 82; corresponding values for females were 7/10
 

(70%) and 9/10 (90%). Even using time-adjusted analyses,
 

eliminating those rats dying before week 52 on study, in some
 

dosed groups, insufficient numbers of animals were at risk for
 

the development of late-appearing tumors.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

summarized in Appendix C, tables C1-C4·
 

A variety of neoplasms were observed in the untreated controls,
 

vehicle controls (phosphate-buffered saline), and dosed groups.
 

Some types of neoplasms occurred only, or with a greater
 

frequency, in rats of dosed groups as compared with controls.
 

26
 
Most of these lesions, however, are not uncommon In this strain
 

of rat Independent of the administration of any chemical· The
 

Incidence of several tumors was higher in the dosed groups·
 

These tumors involved the skin, connective tissues, ear canal,
 

nasal cavity and brain, hematopoietic tissues, lymphoreticular
 

tissues, and uterus· The incidences of these tumors were as
 

follows:
 

Combineda Combined8 

Untreated Vehicle Low Mid 

MALES Controls Controls Dose Dose 

Number of rats necropsied (19) (20) (37) (30)
 

Integument
 

Squamous-cell papilloma 0 1 1 0
 

Squamous-cell carcinoma 0 0 5 3
 

Trichoepithelioma 0 0 1 0
 

Basal-cell carcinoma 0 0 0 1
 

Sarcoma, NOSb 0 0 5 0
 

Fibroma 0 2 2 0
 

Fibrosarcoma 0 0 1 0
 

aMid- and high-dose control group and low-dose group control were
 

combined·
 bNot otherwise specified
 

27
 
Number of rats necropsied
 

Ear Canal
 

Squamous-cell papilloma


Squamous-cell carcinoma


Nasal Cavity
 

Carcinoma, NOS


Hematopoietic and
 

Lymphoreticular Tissues
 

Malignant lymphoma,
 

histiocytic type
 

Leukemia, lymphocyt ic
 

Leukemia, granulocytic
 

Brainc
 

Sarcoma, NOS
 

Astrocytoma
 

Combined
 

Untreated
 

Controls
 

(19)
 

0


 0


 0


0
 

0
 

0
 

(18)
 

0
 

0
 

Neuroepithelioma (Neuroblastoma) 0
 

Other Sites
 

Sarcoma, NOS
 

FEMALES
 

Number of rats necropsied
 

Integument
 

Squamous-cell papilloma
 

Squamous-cell carcinoma
 

Sarcoma, NOS
 

Fibroma
 

Fibrosarcoma
 

(19)
 

0
 

(20)
 

0
 

0
 

0
 

1
 

0
 

Combined
 

Vehicle
 

Controls
 

(20)
 

0


 0


 0


0
 

0
 

0
 

(20)
 

0
 

0
 

0
 

(20)
 

0
 

(20)
 

0
 

0
 

0
 

0
 

0
 

Low
 

Dose
 

(37)
 

3


1
 

3
 

2
 

(36)
 

0
 

1
 

2
 

(37)
 

4
 

(30)
 

0
 

0
 

1
 

1
 

1
 

Mid
 

Dose
 

(30)
 

1 0
 

1
 

1 0
 

0
 

5
 

I
 

(28)
 

1
 

0
 

0
 

(30)
 

1
 

(33)
 

1
 

3
 

0
 

0
 

0
 

c

Number of rats with tissue examined microscopically
 

28
 
Combined Combined 

Untreated Vehicle Low Mid 

Controls Controls Dose Dose 

Number of rats necropsied (20) (20) (30) (33)
 

Ear Canal
 

Squamous-cell papilloma 0 0 0 1
 

Squamous-cell carcinoma 0 0 2 5
 

Nasal Cavity
 

Carcinoma, NOS 0 0 0 1
 

Brainc (20) (19) (30) (32)
 

Neuroepithelioma (Neuroblastoma) 0 0 2 1
 

Sarcoma, NOS 0 0 0 1
 

Uterusc (20) (20) (30) (32)
 

Adenocarcinoma, NOS 0 0 2 7
 

Squamous-cell carcinoma 0 0 1 0
 

Sarcoma, NOS 0 0 2 1
 

Stromal polyp 0 0 4 1
 

Other Sites0 (20) (20) (30) (33)
 

Sarcoma, NOS 0 0 0 1
 

c

Number of rats with tissue examined microscopically
 

The skin tumors were differentiated epithelial tumors with the
 

majority being keratinizing squamous-cell carcinomas located in
 

various regions of the head and neck. The squamous-cell tumors
 

varied from benign acanthotic, hyperkeratotic, and papillary
 

growths (papillomas) to less differentiated carcinomas that
 

invaded the dermis. These latter tumors contained nests of
 

29
 
squamous cells forming whorls of keratin and cords or small
 

groups of darkly staining anaplastic epithelial cells· The
 

predominant cells were stratified and polygonal-shaped, had ample
 

eosinophilic cytoplasm, and had large vesicular nuclei with
 

prominent nucleoli. Two tumors appeared to arise from the basal
 

cells of hair follicles, sebaceous glands, or other adnexal
 

structures· The basal-cell tumors were composed of closely
 

packed, small polygonal cells with scanty cytoplasm and darkly
 

staining purple or blue round to oval nuclei. The cells had a
 

tendency to form palisades· Various degrees of keratinization
 

and hair-follicle formation were present· These tumors were
 

classified as a trichoepithelioma, and a basal-cell carcinoma·
 

Several subcutaneous connective tissue tumors were observed in
 

both dosed and untreated groups. These were spindle-cell tumors
 

that varied from well-differentiated fibromas with extensive
 

collagenous formation to less differentiated fibrosarcomas with
 

less collagen and poorly differentiated sarcomas (sarcomas, NOS)
 

with little or no collagen and irregular cystic or vascular
 

spaces. The sarcomas occurred frequently around the head and
 

neck as well as other subcutaneous and visceral sites.
 

Squamous-cell tumors, both benign and malignant, were observed in
 

the ear canals (Zymbal's Gland). These tumors were similar in
 

morphology to the squamous-cell tumors observed in the skin·
 

30
 
Ten tumors involved the brain and nasal cavity. Seven of the
 

tumors were types which occur very rarely in untreated Sprague-


Dawley rats. These tumors were neuroepitheliomas (neuroblastomas) and nasal carcinomas, NOS.
 

Lymphocytic leukemias that involved multiple organs including
 

various lymph nodes, spleen, liver, lungs, ovaries, uterus,
 

kidneys, and bone marrow were the primary hematopoietic tumors
 

seen. A smaller number of animals had granulocytic leukemias·
 

Uterine adenocarcinomas were characterized by neoplastic
 

epithelial cells having large vesicular nuclei, prominent
 

eosinophilic nucleoli, and ample cytoplasm that were arranged
 

into glands. The glands were usually separated by fibrovascular
 

stroma. The neoplastic glandular tissue arose in the endometrium
 

and both projected into the uterine lumen and infiltrated the
 

underlying muscle layers. Frequently the neoplastic cells
 

penetrated the serosal surface of the uterus with transplantation
 

on the mesentery throughout the abdominal cavity. Pulmonary
 

metastases were also frequent. One low-dose female had a
 

cervical uterine squamous-cell carcinoma. A few animals had
 

poorly differentiated spindle-cell uterine sarcomas (sarcomas,
 

NOS) and endometrial stromal polyps.
 

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

31
 
proliferative, and inflammatory changes were encountered also in
 

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

these nonneoplastic lesions are commonly seen in aged
 

Sprague-Dawley rats.
 

None of the high-dose male or female rats were processed for
 

microscopic evaluation because of the short life spans, and
 

thereby, the lack of time for tumor development. These deaths
 

which occurred at less than 150 days were assumed to be the
 

result of chemical toxicity. Many mid-dose males had reduced
 

life spans that were associated with moderate to severe suppurative bronchopneumonia and bone-marrow atrophy. The lower
 

incidence of tumors in this group may have resulted in part from
 

the reduced life span. The female groups had fewer early deaths
 

associated with bone-marrow atrophy and bronchopneumonias. The
 

effects in the females of reduced life spans from toxicity were
 

not considered important. Chronic and interstitial nephritis
 

occurred frequently, but did not appear to be chemical related.
 

Administration of thio-TEPA resulted in an increase of epithelial
 

tumors in the skin, ear canal, nasal cavity, and uterus; of
 

hematopoietic tumors, primarily lymphocytic leukemias; of
 

spindle-cell sarcomas in the subcutis and various other
 

locations; and of neuroepithelial brain tumois.
 

32
 
Based on the histopathologic examination, thio-TEPA given
 

intraperitoneally to Sprague-Dawley rats at doses of 0.7 and 1.4
 

mg/kg appeared to be carcinogenic under the conditions of this
 

bioassay· The dose of 2.8 mg/kg proved to be too toxic for an
 

evaluation of carcinogenic activity·
 

D· Statistical Analyses of Results (Rats)
 

Tables E1-E8 in Appendix Ε contain the statistical analyses of
 

the incidences of those primary tumors that occurred in at least
 

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

one or more than one group· Analyses of the incidences in the
 

high-dose groups are not included in the tables, because, as a
 

result of the low survival, animals in these groups were not
 

examined histopathologically· The untreated controls also are
 

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

conditions of the vehicle controls more closely resemble those of
 

the dosed rats. Since the low-dose and mid-dose vehicle controls
 

were started at different dates, the low-dose and mid-dose groups
 

are analyzed separately with their respective controls. Due to
 

the low survivals of animals in the dosed groups, time-adjusted
 

analyses were performed, eliminating animals that died before
 

week 52 on study, unless a tumor was found at the anatomic site
 

of interest before week 52· When such an early tumor was found,
 

comparisons were based exclusively on animals that survived at
 

33
 
least as long as the animal in which the first tumor was found·
 

The statistical narrative in the following paragraphs is based on
 

time-adjusted data only·
 

In rats of each sex, when the vehicle controls are used, there is
 

no significant positive result in either the low-dose or mid-dose
 

groups when the Bonferroni probability value of 0.025 is used as
 

a significance level·
 

In male rats, when the pooled controls are used, a significant
 

incidence of squamous-cell carcinoma in either the skin or ear
 

canal is observed in both the low-dose and mid-dose groups (P =
 

0.009, low-dose; Ρ • 0.023, mid-dose). In female rats, a
 

significant incidence of squamous-cell carcinoma in either the
 

skin or ear canal is observed in the mid-dose group (P < 0.001);
 

however, the incidence was not significant in the low-dose group·
 

While the vehicles-control groups were too small for effective
 

analyses, the significant incidence of this tumor observed in
 

both male and female rats, using pooled controls, suggests a dose
 

association with the administration of thio-TEPA.
 

Leukemias were reported at a significant incidence (P = 0.001) in
 

mid-dose male rats when compared with the pooled controls. When
 

the incidences of leukemia or lymphoma are combined for analyses,
 

a significant incidence ( β 0.020) is present in the low-dose
 

34
 
male group when compared with that in the pooled controls· The
 

combination of leukemia or lymphoma did not appear at a significant incidence in the female groups· These results are based on
 

the use of the pooled controls and appeared in significant
 

incidences in males only; therefore, while suggestive of a dose
 

relationship, the statistical evidence is not as clear as for
 

squamous-cell carcinoma·
 

In female rats, adenocarcinoma was seen in a significant
 

incidence in the mammary gland ( 0.006) and in the uterus ( β
 

0·001) when the incidences in the mid-dose group were compared
 

with those in the pooled controls· The incidence of mammary
 

adenocarcinoma in the low-dose group also was higher than that in
 

the pooled controls (P « 0.033), while the incidence of uterine
 

carcinoma in the low-dose group was not significant· These data
 

suggest a positive association between the administration of the
 

test chemical and the formation of these tumors·
 

Significant results in the negative direction occurred in the
 

incidences of pituitary tumors in the low-dose and mid-dose
 

groups of female rats, and in the low-dose group of male rats·
 

In summary, the results of the statistical tests show that the
 

incidences of squamous-cell carcinoma in the skin or ear canal in
 

each sex, hematopoietic tumors in male rats, adenocarcinoma of
 

35
 
the mammary gland in female rats, and adenocarcinoma of the
 

uterus in female rats are associated with the administration of
 

thio-TEPA.
 

36
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of low-dose male and female mice were only
 

slightly lower than those of the controls throughout much of the
 

study (figure 5) . Those of the high-dose animals, particularly
 

those of the females, were lower throughout their period of
 

survival· 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 wide variation. No other signs of
 

toxicity were recorded.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered thio-TEPA at the
 

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

shown in figure 6.
 

For each sex, the Tarone test result is significant (P < 0.001),
 

indicating that thio-TEPA decreased survival in a dose-related
 

manner. At the high-dose, none of the males survived past week
 

56 and none of the females past week 43. At the low-dose, 15/35
 

(44%) of the males and 17/35 (49%) of the females survived to
 

termination of the study. Survival was 7/15 (47%) among vehicle37
 
TIME ON STUDY (WEEKS) 

Figure 5. Growth Curves for Mice Treated with Thio-TEPA 

38 
TIME ON STUDY (WEEKS) 

Figure 6. Survival Curves for Mice Treated with Thio-TEPA 

39 
control males and 12/15 (80%) among vehicle-control females in
 

the bioassay. Statistical analyses were time-adjusted, eliminating those mice dying before week 52; however, in some groups,
 

insufficient numbers of mice were at risk for the development of
 

late-appearing tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

Appendix B, tables Bl and B2; findings on nonneoplastic lesions
 

are summarized in Appendix D, tables Dl and D2.
 

Excluding the lymphoreticular and epithelial tumors, the
 

neoplasms listed in Appendix Β occurred with approximately equal,
 

or greater, frequency in the control mice than in the dosed mice,
 

or occurred in insufficient numbers for accurate evaluation·
 

These lesions are not uncommon in the B6C3F1 strain of mouse
 

independent of the administration of any chemical·
 

Administration of thio-TEPA increased the frequency of skin
 

tumors and lymphoreticular tumors. The distribution of these
 

tumors was as follows:
 

40
 
Vehicle Untreated Low High
 

MALE Control Control Dose Dose
 

Number of Mice Necropsied (14) (15) (30) (34)
 

Skin (including preputial glands
 

and subcutaneous tissues)
 

Squamous-cell or adnexal
 

carcinoma 0 0 14 1
 

Basal-cell carcinoma 1 0 0 0
 

Carcinoma, NOS 0 0 1 0
 

Multiple Organs,
 

Lymphoreticular
 

Malignant lymphoma 0 1 2 16
 

Leukemia, lymphocytic 1 0 0 10
 

Leukemia, granulocytic 0 0 1 0
 

FEMALE
 

Number of Mice Necropsied (15) (15) (30) (32)
 

Multiple Organs,
 

Lymphoreticular
 

Malignant lymphoma 0 1 2 20
 

Leukemia, lymphocytic 0 0 3 12
 

The morphology of the skin tumors was similar to that described
 

previously for the rats. The majority of the squamous-cell
 

tumors were malignant, occurred in the groin or pelvic area, and
 

were associated with preputial glands. Two squamous-cell
 

carcinomas metastasized to the lungs. One mouse had a poorly
 

differentiated spindle-cell carcinoma.
 

The malignant lymphomas were classified as lymphocytic,
 

histiocytic, or mixed types and were usually solid tumors
 

41
 
involving multiple organs including the spleen, lymph nodes, and
 

liver.
 

In addition to the malignant lymphomas, many female mice had
 

lymphocytic leukemias. These leukemias were composed of
 

neoplastic lymphocytes similar in appearance to those seen in the
 

malignant lymphomas, but they had a more generalized distribution
 

with sinusoidal involvement of the liver. One mouse had a
 

granulocytic leukemia that was composed of well-differentiated
 

neutrophils.
 

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

proliferative, and inflammatory changes were encountered also in
 

animals of the dosed and control groups (Appendix D) . These
 

nonneoplastic lesions are commonly seen in aged B6C3F1 mice. The
 

high-dose male and female mice had reduced life spans which were
 

associated with the high incidence of lymphoreticular neoplasms.
 

Based on the histopathologic examination, thio-TEPA given intraperitoneally to B6C3F1 mice was carcinogenic under conditions of
 

this bioassay. The tumors induced were malignant lymphomas and
 

lymphocytic leukemias in males and females and squamous-cell
 

carcinomas of the skin in males.
 

42
 
D· Statistical Analyses of Results (Mice)
 

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

the incidences of those primary tumors that occurred in at least
 

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

one or more than one group· The 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
 

mice. Due to the low survival of the high-dose mice, time-


adjusted analyses were performed, eliminating animals that died
 

before week 52 on study, unless a tumor was found at the anatomic
 

site of interest before week 52. When such an early tumor was
 

found, comparisons were based exclusively on animals that
 

survived at least as long as the animal in which the first tumor
 

was found. The statistical narrative in the following paragraphs
 

is based on time-adjusted data only.
 

In male mice, the results of the Cochran-Armitage test for dose-


related trend in the incidence of squamous-cell carcinoma at all
 

sites are significant (P = 0.018), and the Fisher exact test
 

shows that the incidence in the low-dose group is significantly
 

higher (P = 0.004) than that in the matched controls. There were
 

only two animals at risk in the high-dose group, and the
 

squamous-cell carcinoma occurred in one of these two animals.
 

This tumor was not present in significant incidences in female
 

43
 
mice; however, the statistical analysis suggests a positive dose
 

association in male mice·
 

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


related trend in the combined incidence of lymphoma or leukemia
 

in both male and female mice are significant (P < 0.001)· In the
 

female high-dose group, the incidence is 32/32 (100%) and in the
 

male high-dose group it is 26/28 (93%). The results of the
 

Fisher exact show that the incidences in the high-dose groups are
 

significantly higher (P < 0.001) than those in either type of
 

control groups. The statistical conclusion is that the
 

incidences of these hematopoietic tumors in mice are dose
 

associated.
 

In summary, the results of the statistical tests show that the
 

incidences of squamous-cell carcinoma at all sites in male mice
 

and of the hematopoietic tumors in each sex are associated with
 

the administration of thio-TEPA.
 

44
 
V. DISCUSSION
 

Under the conditions of this bioassay, thio-TEPA was toxic to
 

both rats and mice, as evidenced by reduced mean body weights and
 

by high mortality. Mean body weights of high-, mid-, and low-


dose male rats, high- and mid-dose female rats, and high-dose
 

male and female mice were all markedly lower than those of
 

corresponding controls· All high-dose male rats died by week 19,
 

all high-dose females by week 21, all mid-dose males by week 78,
 

91% of the mid-dose females by termination week 81, and 85% of
 

the low-dose males and 51% of the low-dose females by termination
 

week 82. Histopathologic evaluation of tissues was performed
 

only on the low- and mid-dose groups of rats, since all high-dose
 

males and females died before the occurrence of tumors could be
 

expected. All of the high-dose male mice died by week 56, all of
 

the high-dose females by week 43, and 7 7% of the low-dose males
 

and 51% of the low-dose females by termination week 86. Because
 

of this high mortality, time-adjusted analyses were performed on
 

incidences of tumors in each species and each sex.
 

In rats, the incidence of combined neoplasms of the hematopoietic
 

system (lymphoma, lymphocytic leukemia, or granulocytic leukemia)
 

was significant in the males in both the low-dose (P = 0.020) and
 

mid-dose (P = 0.001) groups, using pooled controls (pooled
 

45
 
controls 0/29, low-dose 6/34; pooled controls 0/30, mid-dose
 

6/16).
 

Squamous-cell carcinoma of the skin or ear canal occurred at a
 

significant incidence in the male rats in both the low-dose (P ­

0.009) and mid-dose (P « 0.023) groups, using pooled controls
 

(pooled controls 0/29, low-dose 7/33; pooled controls 0/30,
 

mid-dose 3/13) and in the mid-dose females (P < 0.001), using
 

pooled controls (pooled controls 0/28, mid-dose 8/21); in
 

addition, two low-dose females had such tumors, with none
 

occurring in the corresponding low-dose controls.
 

The incidence of adenocarcinoma of the uterus was significant in
 

the mid-dose female rats ( β 0.001), using pooled controls
 

(pooled controls 0/28, mid-dose 7/21); in addition, two low-dose
 

females had adenocarcinoma of the uterus, with no such tumor
 

occurring in the corresponding low-dose controls. Adenocarcinoma
 

of the mammary gland occurred at a significant incidence in the
 

mid-dose female rats (P = 0.006), using pooled controls (pooled
 

controls 1/28, mid-dose 8/24). In addition, seven low-dose
 

females had these tumors, but they were not significant, since
 

one of the low-dose pooled-control females had the tumor; and
 

3/10 low-dose untreated-control females had the tumor. Thus, the
 

occurrence of mammary adenocarcinoma in the female rats cannot be
 

clearly related to administration of the test chemical.
 

46
 
Also in rats, neuroepitheliomas or nasal carcinomas occurred in
 

three low-dose males, two low-dose females, and two mid-dose
 

females. Although these are not statistically significant
 

incidences, these tumors did not occur among control animals and
 

no such tumors have occurred in 380 or more Sprague-Dawley
 

control rats of each sex in other bioassays at the same
 

laboratory. Thus, they may be associated with administration of
 

the chemical.
 

In the high-dose groups of both male and female mice, but not in
 

the low-dose groups, the incidences of lymphoma or lymphocytic
 

leukemia were significantly higher (P < 0.001) for each sex than
 

those of either the vehicle or pooled controls (males: vehicle
 

controls 1/8, pooled controls 1/18, low-dose 2/24, high-dose
 

26/28; females: vehicle controls 0/14, pooled controls 0/29,
 

low-dose 5/26, high-dose 32/32). These tumors were observed even
 

though all of the high-dose males died as early as week 56 and
 

all of the high-dose females as early as week 43. In addition,
 

granulocytic leukemia occurred in one low-dose male mouse.
 

In the low-dose male mice, squamous-cell carcinoma was found in
 

the skin of seven animals, in the preputial glands of six
 

animals, and in the ear canal of two animals. A carcinoma of the
 

preputial gland was also found in a high-dose male. When the
 

incidences of the tumors at the different sites were combined,
 

47
 
the incidence in the low-dose group, but not in the high-dose
 

group was statistically significant using either the vehicle (P =
 

0.004) or the pooled (P < 0.001) controls (vehicle controls 0/8,
 

pooled controls 0/18, low-dose 14/24, high-dose 1/2).
 

In previous tests of thio-TEPA for carcinogenicity, Schmahl and
 

Osswald (1970) observed a variety of tumors in male rats of the
 

BR 46 strain that had been injected intravenously with 1 mg/kg
 

body weight weekly for 52 weeks. Shimkin et al. (1966), using a
 

pulmonary tumor test system, reported the induction of lung
 

tumors in A/J strain mice given intraperitoneal injections of
 

thio-TEPA three times per week for 4 weeks and observed for 39
 

weeks after the first injection. Stoner et al· (1973), using the
 

same test system, reported the induction of lung tumors in mice
 

of the A/He strain given total dosages of the chemical of 4.7 or
 

9.4 mg/kg by intraperitoneal injection three times per week for 4
 

weeks and observed 24 weeks after the first injection.
 

It is concluded that under the conditions of this bioassay,
 

thio-TEPA was carcinogenic in both Sprague-Dawley rats and B6C3F1
 

mice. In the rats, the chemical induced squamous-cell carcinoma
 

of the skin or ear canal in both males and females, and
 

hematopoietic neoplasms in the males; in the mice, it induced
 

lymphoma or lymphocytic leukemia in both sexes and squamous-cell
 

carcinoma in the skin and associated glands of males.
 

48
 
VI. BIBLIOGRAPHY
 

Armitage, Ρ·, Statistical Methods in Medical Research, John Wiley
 

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

Berenblum, I., ed., Carcinogenicity Testing, A Report on the
 

Panel on Carcinogeniity gf the Cancer Research Commission of
 

the UICC, Vol. 2, International Union Against Cancer,
 

Geneva, 1969.
 

Calabresi, P. and Parks, R. E., Jr., Alkylating agents,
 

antimetabolites, hormones, and other antiproliferative
 

agents. In: The Pharmacological Basis of Therapeutics,
 

Goodman, L. S., and Gilman, Α., eds., Macmillan Publishing
 

Co., Inc., New York, 1975, pp. 1254-1259 and 1265-1266.
 

Calabresi, P. and Welch, A. D·, Chemotherapy in neoplasms. Ann.
 

Rev. Med. 13:147-202, 1962.
 

Carter, S. K. and Slavik, M., Chemotherapy of cancer. Ann Rev.
 

Pharmacol. 14:157, 1974.
 

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

Soc. Β 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.
 

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

incomplete observations. J^ Amer. Statist. Assoc. 

^3:457-481, 1958. 

Lederle Laboratories. Physicians' Desk Reference, Baker, C. Ε.,
 

Jr., Medical Economics Co., Oradell, N. J., 1977, pp. 903
 

and 906.
 

Linhart, M. S·, Cooper, J., Martin, R. L. , Page, N. , and Peters,
 

J., Carcinogenesis bioassay data system. Comp. and Biomed.
 

2*230-248, 1974.
 

49
 
Miller, R. G., Jr., Simultaneous Statistical Inference,
 

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

Montgomery, J. A. and Struck, R. J., The relationship of the
 

metabolism of anticancer agents to their activity. Progress
 

±n Drug Research JJ: 363-364, 1973.
 

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.
 

Schmahl, V. D. and Osswald, H., Experimentelle Untersuchungen
 

uber carcinogene Wirkungen von Krebs-Chemotherapeutica und
 

Immunosuppressiva. Arzneimittel-Forschung 20 (10):1461­

1470, 1970.
 

Shimkin, M. B., Weisburger, J. Η., Weisburger, Ε. Κ., Guboreff,
 

Ν., and Suntzeff, V., Bioassay of 29 alkylating chemicals by
 

the pulmonary-tumor response in strain A mice. J^ Natl.
 

Cancer Inst. 36:915-935, 1966.
 

Stoner, G. D., Shimkin, M. B., Kniazeff, A. J., Weisburger, J.
 

Η., Weisburger, Ε. Κ., and Gori, G. B., Test for
 

carcinogenicity of food additives and chemotherapeutic
 

agents by the pulmonary tumors response in strain A mice.
 

Cancer Res. 13:3069-3085, 1973.
 

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

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

U.S. Patent 2,670,347 (February 23, 1954).
 

Wheeler, G. P., Alkylating agents. In: Cancer Medicine,
 

Holland, J. F., and Frei, E., Ill, eds., Lea & Febiger,
 

Philadelphia, 1973, pp. 791-805.
 

50
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

51
 

TABLE ΑΙ.
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA(CONTROL GROUPS)
 

MID AND HIGH
 

DOSE VEHICLE
 

CONTROL
 

10
 

10
 

10
 

(10)
 

1 (10%)
 

(10)
 

1 (10%)
 

(10)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

10
 

10
 

10
 

(10)
 

1 (10%)
 

(10)
 

(9)
 

3 /33%)
 

MID AND HIGH
 

DOSE UNTREATED
 

CONTROL 

ANIMALS INITIALLY

ANIMALS NECROPSIED 

ANIMALS EXAMINED

 IN STUD* 

 HISTOPATHOLOGICALLY

10 

10 10 

INTEGUMENTARY SYSTEM
 

•SKIN	 (10)
 

SQUAMOUS CELL PAPILLOMA
 

FIBROMA
 

•SUBCUT	 TISSUE (10)
 

FIBROMA
 

RESPIRATORY SYSTEM
 

NONE
 

HEMATOPOIETIC SYSTEM
 

NONE
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

NONE
 

UEINARY SYSTEM
 

ΝΟΝΞ
 

ENDOCRINE SYSTEM
 

#PITUITARY (10)
 

CHROMOPHOBE ADENOMA
 

LOW DOSE.
 

UNTREATED
 

CONTROL
 

10
 

9
 

9
 

(9)
 

(9)
 

(7)
 

1 (14%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

53
 
CHROMOPHOBE CARCINOMA 

REPRODUCTIVE SYSTEM 

•MAMMASY GLAND 

FIBROADENOMA 

NERVOUS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 

NATURAL DEATHS 

MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACfilFICE 

ANIMAL MISSING 

@ INCLUDES AUTOLYZED ANIMALS
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICBOSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

TABLE A1. MALE RATS (CONTROL GROUPS): NEOPLASMS (CONTINUEO) 

MID AND HIGH LOW DOSE 

DOSE UNTREATED UNTREATED 

CONTROL CONTROL 

(10) (9)

 

2 (20%)

 

10 10

 

2 2

 

8 8 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

(10) 

1 (10%) 

10 

1 

1 

8 

LOW DOSE

 

VEHICLE 

CONTROL 

1 (11*) 

(10) 

10 

10 

54
 
TABLE A1. MALE RATS (CONTROL GROUPS): NEOPLASMS (CONTINUED)

 

MID AND HIGH DOSE UNTREATED 

CONTROL 

LOW DOSE UNTREATED 

CONTROL 

MID AND HIGH DOSE VEHICLE 

CONTROL 

LOW DOSE VEHICLE 

CONTROL 

TUMOfi SUMMARY 

TOTAL ANIMALS WITH PRIMARY TUMORS* TOTAL PRIMARY TUMCRS 2 

2 

1 

1 

3 

3 

5 

5 

TOTAL ANIMALS WITH BENIGN TUMORS

TOTAL BENIGN TUMORS

 2 

2 

1 

1 

3 

3 

4 

4 

TOTAL ANIMALS WITH MALIGNANT TUMORS 

TOTAL MALIGNANT TUMORS 

1 

1 

TOTAL ANIMALS WITH SECONDARY TUMORS# 

TOTAL SECONDARY TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT TOTAL UNCERTAIN TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ EBIMARY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

• PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT OBGAH 

55
 
TABLE A2.
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA (TREATED GROUPS)
 

LOW DOSE MID DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY 

ANIMALS NECROPSIED 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 

39 

37 

37 

35 

30 

29 

35 

32 

0 

INTEGUMENTARY SYSTEM 

•SKIN 

SQUAMOUS CELL PAPILLOMA 

SQUAMOUS CELL CARCINOMA 

BASAL-CELL CARCINOMA 

TRICHOEPITHELIOMA 

(37) 

1 (3%) 

5 (14%) 

1 (3%) 

(30) 

3 (10%) 

1 (3%) 

(32) 

*SUBCUT TISSUE 

SARCOMA, NOS 

FIBROMA 

FIBROSARCOMA 

(37) 

5 (14%) 

2 (595) 

1 (3%) 

(30) (32) 

RESPIRATORY SYSTEM 

•NASAL CAVITY 

CARCINOMA,NOS 

(37) 

1 (3%) 

(30) (32) 

#LUNG 

SQUAMOUS CELL CARCINOMA, METASTA 

ALVEOLAR/BRONCHIOLAR ADENOMA 

(36) (29) 

1 (3%) 

1 (3%) 

HEMATOPOIETIC SYSTEM 

•MULTIPLE ORGANS 

MALIG.LYMPHOMA, HISTIOCYTIC

LYMPHOCYTIC LEUKEMIA 

GRANULOCYTIC LEUKEMIA 

 ΤΥΡΓ 

(37) 

1 (3%) 

3 [&%) 

2 (5%) 

(30) 

5 (17%) 

1 (3%) 

(32) 

CIRCULATORY SYSTEM 

NONE 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED

• NUMBER OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

56
 
TABLE A2. MALE RATS (TREATED GROUPS): NEOPLASMS (CONTINUED)

 

DIGESTIVE SYSTEM
 

•LIVER
 

CARCINOMA,NOS
 

UEINABY SYSTEM
 

#KIDNEY
 

HAMAPTCMA
 

ENDOCRINE SYSTEM
 

tPITUITAFY
 

CHROMOPHOBE ADENOMA
 

REPBODUCTIVE SYSTEM
 

•MAMMAPY	 GLAND
 

ADENOCARCINOMA, NOS
 

NERVOUS SYSTEM
 

#BRAIN
 

SARCOFA, NOS
 

ASTROCYTOMA
 

NEUROBLASTOMA
 

OLFACTORY NEUROBLASTOMA
 

SPECIAL SENSE ORGANS
 

•EAR CANAL
 

SQUAMOUS CELL PAPILLOMA
 

SQUAMOUS CELL CARCINOMA
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

LOW DOSE MID DOSE HIGH DOSE 

(37) (29) 

1 (3%) 

(37) (29) 

1 (3%) 

(28) (24) 

1 (4%) 1 (4%) 

(37) (30) (32) 

1 (3%) 

(36) (28) 

1 (4%) 

1 <3X) 

1 (3%) 

1 (3%) 

(37) (30) (32) 

1 (3%) 

3 (8*) 1 (3%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

57 
TABLE A2. MALE RATS (TREATED GROUPS): NEOPLASMS (CONTINUED)

 

LOW DOSE MID DOSE HIGH DOSE
 

BODY CAVITIES
 

•MEDIASTINUM	 (37) (30) (32)
 

SARCOMA, NOS 1 (3%)
 

•ABDOMINAL	 CAVITY (37) (30) (32)
 

SARCOMA, NOS 1 (3%)
 

•MESENTERY	 (37) (30) (32)
 

SARCOMA, NOS 2 (5%)
 

ALL OTHER SYSTEMS
 

•MULTIPLE	 ORGANS (37) (30) (32)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (3%)
 

CRANIAL CAVITY
 

SARCOMA, NOS 1
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 39 35 35
 

NATURAL DEATH* 7 13 26
 

MORIBUND SACRIFICE 26 22 9
 

SCHEDULED SACRIFICE
 ACCIDENTALLY KILLED
 TERMINAL SACRIFICE 6
 

ANIMAL MISSING
 

9 INCLUDES AUTCLYZED ANIMALS
 

« NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

• NUMBER OF ANIMALS NECROPSIED 

58 
TABLE A2. MALE RATS (TREATED GROUPS): NEOPLASMS (CONTINUED)

 

LOW DOSE MID DOSE HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 27
 

TOTAL PRIMARY TUMORS 36 16
 

TOTAL ANIMALS WITH BENIGN TUMORS 6 2
 

TOTAL BENIGN TUMORS 7 2
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 26 12
 

TOTAL MALIGNANT TUMORS 29 14
 

TOTAL ANIMALS WITH SECONDARY TUMORS# 2
 

TOTAL SECONDARY TUMORS 2
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ FRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

• PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

59
 
TABLE A3.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA (CONTROL GROUPS) 

MID AND HIGH
 

DOSE VEHICLE
 

CONTROL
 

10
 

10
 

10
 

(10)
 

(10)
 

2 (20%)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

10
 

10
 

10
 

(10)
 

(8)
 

1 (13%)
 

MID AND HIGH
 

DOSE UNTREATED
 

CONTROL
 

ANIMALS INITIALLY IN STUDY 10
 ANIMALS NECROPSIED 10
 ANIMALS EXAMINED HISTOPATHOLOGICALLY 10
 

INTEGUMENTARY SYSTEM
 

•SUBCUT TISSUE	 (10)
 

FIBROMA 1 (10%)
 

RESPIRATORY SYSTEM
 

NONE
 

HEMATOPOIETIC SYSTEM
 

NONE
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

NONE
 

ORINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

•PITUITARY	 (10)
 

CHROMOPHOBE ADENOMA 5 (50%)
 

LOW DOSE
 

UNTREATED
 

CONTROL
 

10
 

10
 

10
 

(10)
 

(10)
 

6 (6 0%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

60 
TABLE A3. FEMALE RATS (CONTROL GROUPS): NEOPLASMS (CONTINUED)

 

CHROMOPHOBE CARCINOMA
 

REPRODUCTIVE SYSTEM
 

•MAMMARY GLAND
 

ADENOCARCINOMA, NOS
 

FIBROADENOMA
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

HUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHS)
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIHAL MISSING
 

a INCLUDES AUTOLYZED ANIMALS
 

MID AND HIGH
 

DOSE UNTREATED
 

CONTROL
 

(10)
 

2 (20%)
 

10
 

2
 

8
 

LOW DOSE
 

UNTREATED
 

CONTROL
 

1 (10%)
 

(10)
 

3 (30%)
 

6 (60%)
 

10
 

2
 

1
 

7
 

MID AND HIGH
 

DOSE VEHICLE
 

CONTROL
 

(10)
 

1


5


10
 

10
 

 (10%)
 

 (50%)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

(10)
 

2 (20%)
 

10
 

1
 

9
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

61
 
TABLE A3. FEMALE RATS (CONTROL GROUPS): NEOPLASMS (CONTINUED)

 

MID AND HIGH DOSE UNTREATED 

CONTROL 

LOW DOSE UNTREATED 

CONTROL 

MID AND HIGH DOSE VEHICLE CONTROL LOW DOSE VEHICLE CONTROL 

TOWQR SUMMARY 

TOTAL ANIMALS WITH PRIMARY TUMORS* TOTAL PRIMARY TUMORS 7 

8 

9 

16 

5 

8 

3 

3 

TOTAL ANIMALS WITH BENIGN TUMORS

TOTAL BENIGN TUMORS

 7 

8 

9 

12 

5 

7 

3 

3 

TOTAL ANIMALS WITH MALIGNANT TUMORS 

TOTAL MALIGNANT TUMORS 

4 

4 

1 

1 

TOTAL ANIMALS WITH SECONDARY TUMORSi 

TOTAL SECONDARY TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT TOTAL UNCERTAIN TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

• PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN 

62
 
TABLE A4.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA (TREATED GROUPS) 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECBOPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

• SKIN
 

SQUAMOUS CELL PAPILLOMA
 

SQUAMOUS CELL CARCINOMA
 

•SUBCUT TISSUE
 

SARCOMA, NOS
 

FIBROMA
 

FIBROSARCOMA
 

RESPIRATORY SYSTEM
 

•NASAL CAVITY
 

CARCINOMA,NOS
 

«LUNG
 

ADENOCARCINOMA, NOS, METASTATIC
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS
 

MALIG.LYMPHOMA, HISTIOCYTIC TYFE
 

LYMPHOCYTIC LEUKEMIA
 

GRANULOCYTIC LEUKEMIA
 

•MANDIBULAR L.	 NODE
 

MALIG.LYMPHOMA, UNDIFFER-TYPE
 

•THYMUS
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
 

CIRCULATORY SYSTEM
 

NONE
 

LOW DOSE MID DOSE
 

31 35
 

30 33
 

30 33
 

(3d) (33)
 

1 (3%)
 

3 (9%)
 

(30)	 (33)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

(30)	 (33)
 

1 (3%)
 

(30)	 (33)
 

2 (7%)
 

(30)	 (33)
 

1 (3%)
 1 (3%)
 

1 (3%)
 

(28) (1)
 

1	 m%)
 

(30)	 (33)
 

1 (3%)
 

HIGH DOSE
 

35
 

34
 

0
 

(34)
 

(34)
 

(34)
 

(34)
 

• NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIID
 

63
 
TABLE A4. FEMALE RATS (TREATED GROUPS): NEOPLASMS (CONTINUED)

 

LOW DOSE MID DOSE HIGH DOSE 

DIGESTIVE SYSTEM 

•SALIVARY GLAND (24) 

SARCOMA, NOS 1 (4%) 

URINARY SYSTEM 

•KIDNEY (30) 

HAMARTOMA 1 (3%) 

ENDOCRINE SYSTEM 

•PITUITARY (29) (32) 

CHROMOPHOBE ADENOMA 4 (14%) 

REPRODUCTIVE SYSTEM 

•MAMMARY GLAND (30) (33) (34) 

ADtNOMA, NOS 3 (9%) 

ADENOCARCINOMA, NOS 7 (23%) 8 (24%) 

FIBROADENOMA 8 (27%) 4 (12%) 

•UTERUS (30) (32) 

ADENOCARCINOMA, NOS 2 (7%) 7 (22%) 

SARCOMA, NOS 2 (7%) 1 (3%) 

ENDOMETRIAL STROMAL POLYP 4 (13%) 1 (3%) 

•CERVIX UTERI (30) (32) 

SQUAMOUS CELL CARCINOMA 1 (3%) 

•CVABY (30) (32) 

LUTEOMA 1 (3%) 

SARCOMA, NOS 1 (3%) 

NERVOUS SYSTEM 

•BRAIN (30) (32) 

SARCOMA, NOS 1 (3%) 

OLFACTORY NEUROBLASTOMA 2 (7%) 1 (3%) 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

64 
TABLE A4. FEMALE RATS (TREATED GROUPS): NEOPLASMS (CONTINUED)

 

SPECIAL SENSE ORGANS
 

*EAR CANAL
 

SQUAMOUS CELL PAPILLOMA
 

SQUAMOUS CELL CARCINOMA
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

•ABDOMINAL	 CAVITY
 

ADENOCARCINOMA, NOS, METASTATIC
 

ALL OTHER SYSTEMS
 

•MULTIPLE	 ORGANS
 

ADENOCARCINOMA. NOS
 

ADENOCARCINOMA, NOS, METASTATIC
 

ANIMAL DISPOSITION SUMMABY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TEBfllNAL SACRIFICE
 

ANIMAL MISSING
 

1 INCLUDES AUTOLYZED ANIMALS
 

LOW DOSE MID DOSE HIGH DOSE 

(30) (33) (34) 

1 (3%) 2 (7%) 5 (15%) 

(30) (33) (34) 

1 (3%) 1 (3%) 

(30) (33) (34) 

1 (3%) 

4 (12%) 

31 35 35 

9 10 15 

9 22 20 

13 3 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

65
 
TABLE A4. FEMALE RATS (TREATED GROUPS): NEOPLASMS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 26 26
 

TOTAL PRIMARY TUMORS 39 43
 

TOTAL ANIMALS WITH BENIGN TUMORS 14 9
 

TOTAL BENIGN TUMOES 18 11
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 17 22
 

TOTAL MALIGNANT TUMORS 21 32
 

TOTAL ANIMALS WITH SECONDARY TUMORSt 3 5
 

TOTAL SECONDARY TUMORS 3 5
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

* SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT OKGAN
 

66
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MICE
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

67
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

• SKIN
 

CARCINCMA#NOS
 

SQUAMOUS CELL CARCINOMA
 

BASAL-CELL CARCINOMA
 

•SUBCUT TISSUE
 

SQUAMOUS CELL CARCINOMA
 

SARCOMA, NOS
 

RESPIRATOFY SYSTEM
 

#LUNG
 

SQUAMOUS CELL CARCINOMA, METASTA
 ALVEOLAR/BRONCHIOLAR ADENOMA
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS
 

MALIGNANT LYMPHOMA, NO3
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
 MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 MALIGNANT LYMPHOMA, MIXED TYPE
 LYMPHOCYTIC LEUKEMIA
 

GRANULOCYTIC LEUKEMIA
 

•MESENTERIC	 L. NODE
 

MALIGNANT LYMPHOMA, NOS
 

CIRCULATORY SYSTEM
 

•MYOCARDIUM
 

HEMANGIOMA
 

VEHICLE UNTREATED
 

CONTROL CONTROL
 

15 15
 

14 15
 

14 15
 

(14) (15)
 

1 (7%)
 

(14) (15)
 

(15)
 

1 <7X) 4	 (21%)
 

(14) (15)
 

1 (7%)
 

(2)	 (5)
 

1 (20%)
 

(14) (15)
 

LOW DOSE
 

a)34
 

30
 

30
 

(30)
 

1 (3%)
 

7 (23%)
 

(30)
 

1 (3X)
 

1 (3%)
 

(30)
 

1 (3X)
 

5 (17X)
 

(30)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

(1)
 

(30)
 

1 i3X)
 

HIGH DOSE
 

35
 

34
 

33
 

(34)
 

(34)
 

(33)
 

1 <3X)
 

(34)
 

2 (6X)
 

14 (41X)
 

10	 (29X)
 

(15)
 

(32)
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

@NOTE: 35 ANIMALS WERE INITIALLY IN STUDY BUT ONE WAS FOUND TO BE A FEMALE IN A MALE GROUP AND
 

WAS DELETED.
 

69
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED)

 

LOW DOSE HIGH DOSE
 

(30) (33)
 

3 (10%)
 

1 (3%)
 

(30)	 (34)
 

6 (20%) 1 (3%)
 

(30) (34)
 

1 (3%)
 

2 (7%)
 

DIGESTIVE SYSTEM
 

•LIVER
 

HEPATOCELLULAR ADENOMA
 

HEPATOCELLULAR CARCINOMA
 

UHINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

•PREPUTIAL	 GLAND
 

SQUAMOUS CELL CARCINOMA
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

•EAR CANAL
 

SQUAMOUS CELL PAPILLOMA
 

SQUAMOUS CELL CARCINOMA
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OTHER	 SYSTEMS
 

NONE
 

VEHICLE
 

CONTROL
 

(14)
 

1 (7%)
 

1 (7%)
 

(14)
 

(14)
 

UNTREATED
 

CONTROL
 

(15)
 

2 (13%)
 

(15)
 

(15)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSIED
 

70
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE CONTROL UNTREATED CONTROL LOW DOSE HIGH DOSE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 

NATURAL DEATHS 

MORIBUND SACRIFICE SCHEDULED SACRIFICE ACCIDENTALLY KILLED TERMINAL SACRIFICE ANIMAL MISSING ANIMAL DELETED (WRONG SEX) 

15 

7 

1 

7 

15 

2 

13 

35 

13 

6 

15 

1 

35 

13 

22 

@ INCLUDES AUTOLYZED ANIMALS 

TUMOR SUMMARY 

TOTAL ANIMALS WITH PRIMARY TUMORS* 

TOTAL PRIMARY TUMORS 

4 

5 

6 

7 

19 

32 

27 

27 

TOT*L ANIMALS WITH BENIGN TUMORS 

TOTAL BENIGN TUMOFS 

2 

2 

5 

6 

8 

10 

TOTAL ANIMALS WITH MALIGNANT TUMORS 

TOTAL MALIGNANT TUMORS 

3 

3 

1 

1 

18 

22 

27 

27 

TOTAL ANIMALS WITH SECONDARY TUMORS# 

TOTAL SECONDARY TUMORS 

1 

1 

1 

1 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT TOTAL UNCERTAIN TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAINEfilMARY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

• PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

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

71
 
TABLE B2.
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

VEHICLE 

CONTROL 

UNTREATED 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY 

ANIMALS MISSING 

ANIMALS NECROPSILD 

ANIMALS EXAMINED HISTOPATHOLOGICALLΥ 

15 

15 

15 

15 

15 

15 

35 

1 

30 

30 

35 

2 

32 

32 

INTEGUMFNTARY SYSTEM 

• SKIN 

SQUAMOUS CELL PAPILLOMA 

(15) (15) (30) 

1 (3*) 

(32) 

RESPIRATORY SYSTEM 

#LUNG 

ADENOCARCINOMA, NCS, METASTATIC 

ALVEOLAR/BRCNCHIOLAR ADENOMA 

(15) (15) (29) 

1 (3%) 

5 (17%) 

(32) 

HEMATOPOIETIC SYSTEM 

•MULTIPLE ORGANS 

MALIGNANT LYMPHOMA, NOS 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 

MALIGNANT LYMPHOMA, MIXED TYPF 

LYMPHOCYTIC LEUKEMIA 

(15) (15) 

1 (7%) 

(30) 

2 (7%) 

3 (10%) 

(32) 

1 (3%) 

19 (59%) 

12 (38%) 

CIRCULATORY SYSTEM 

NONE 

DIGESTIVE SYSTEM 

* IVEK 

HEPATOCELLULAR ADENOMA 

(15) (15) (30) 

2 (7%) 

(32) 

URINARY SYSTEM 

NONE 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED

• NUMBER OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

72
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED)
 

ENDOCRINE SYSTEM 

•ADRENAL
 

CORTICAL CARCINOMA
 

PHEOCHROMOCYTOMA
 

REPRODUCTIVE SYSTEM 

«MAMMARY GLAND
 

ADENOCARCINOMA, NOS
 

•OVARY
 

CARCINOMA,NOS
 

MBRVOUS SYSTEM 

ΜΟΝΕ 

SPECIAL SENSE ORGANS 

•EAR	 CANAL 

SQUAMOUS CELL PAPILLOMA 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONE 

VEHICLE
 

CONTROL
 

(15 ) 

(15 ) 

(15 ) 

(15 ) 

UNTREATED 

CONTROL 

(15 ) 

(15 ) 

(15 ) 

(15) 

LOW DOSE 

(28) 

1 (M) 

1 (<•%) 

(30) 

1

(29) 

3

(30) 

1

 (3%) 

 (10X) 

 (3%) 

HIGH DOSE 

(32) 

(31) 

(32) 

4 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

73
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE CONTROL UNTREATED CONTROL LOW DOSE HIGH DOSE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY

NATURAL DEATHS

MORI3UND SACRIFICE SCHEDULED SACRIFICE ACCIDENTALLY KILLED TERMINAL SACPIFICE ANIMAL MISSING 

 15 

2 

1 

 12 

15 

1 

14 

35 

11 

4 

2 

17 

1 

35 

16 

17 

2 

d INCLUDES AUTCLYZED ANIMALS 

TUMOR SUMMARY 

TOTAL ANIMALS WITH ERIMARY TUMORS* 

TOTAL PRIMARY TUMORS 

1 

1 

17 

20 

32 

32 

TOTAL ANIMALS WITH BENIGN TUMORS 

TOTAL BENIGN TUMORS 

10 

10 

TOTAL ANIMALS WITH MALIGNANT TUMCRS 

TOTAL MALIGNANT TUMORS 

1 

1 

9 

10 

32 

32 

TOTAL ANIMALS WITH SECONDARY TUMORS» 

TOTAL SECONDARY TUMORS 

1 

1 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT TOTAL UNCERTAIN TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAINERIMABY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT OfiGAN 

74
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

75
 

TABLE CI. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIOTEPA (CONTROL GROUPS)
 

— •	 ••
 

MID AND HIGH LOW DOSE
 DOSE UNTREATED UNTREATED
 CONTROL CONTROL
 

ANIMALS INITIALLY IN STUDY 10 10
 ANIMALS NECROPSIED 10
 Q
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 10 9
 

INTEGUMENTARY SYSTEM
 

• SKIN	 (10) Ί9)
 ULCER, CHRONIC
 

RESPIRATORY SYSTEM
 

•TRACHEA	 (10) (9)
 INFLAMMATION, NOS 1 (10%)
 

INFLAMMATION, ACUTE/CHRONIC
 

INFLAMMATION, CHRONIC 1 (11%)
 

INFLAMMATION, CHRONIC SUPPORATIV
 

•LUNG/BRONCHUS	 (10) (9)
 BRONCHIECTASIS 1 (11%)
 

•LUNG/BRONCHIOLE	 (10) (9)
 HYPERPLASIA, LYMPHOID 1 (10%)
 

• LUNG	 (10) (9)
 BRONCHOPNEUMONIA SUPPURATIVE
 

HEMATOPOIETIC SYSTEM 

• BONE MARROW (10) (9) ATROPHY, NOS 2 (2055) 4 (44%) 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

•LIVER	 (10) (9)
 

HYPERPLASIA NODULAR 1 (11%)
 

• NUMBER OF ANIMALS WITH TISSUE EXAHINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

MID AND HIGH
 

DOSE VEHICLE
 

CONTROL
 

10
 

10
 

10
 

(10)
 

(10,
 

1 (10%)
 

(10)
 

(10)
 

1 (10%)
 

(10)
 

1 (10%)
 

(10)
 

5 (50%)
 

(10)
 

LOW DOSE
 

VEHICLE
 

CONTROL
 

10
 

10
 

10
 

(10)
 

1 (10%)
 

(10)
 

1 (10%)
 

(10)
 

(10)
 

(10)
 

(10)
 

7 (70%)
 

(10)
 

77
 
TABLE CI. MALE RATS (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

•BILE DUCT
 

HYPERPLASIA, NOS
 

URINARY SYSTEM
 

«KIDNEY
 INFLAMMATION, INTERSTITIAL
 INFLAMMATION, SUPPURATIVE
 INFLAMMATION, CHRONIC
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

•MAMMARY	 GLAND
 

CYST, NOS
 

•PROSTATE
 

INFLAMMATION, CHRONIC SUPPURATIV
 

•TESTIS
 

ATROPHY, NOS
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

MID AND HIGH LOW DOSE MID AND HIGH LOW DOSE DOSE UNTREATED UNTREATED DOSE VEHICLE VEHICLE CONTROL CONTROL CONTROL CONTROL 

(10) (9) (10) (10) 

1 (10%> 

(10) (9) (10) (10) 

6 (60%> 4 (40%) 

1 (1095) 

6 (61%) 8 (80%) 

(10) (9) (10) (10) 

1 (10*) 

(10) (9) (10) (10) 

1 (11%) 1 (10%) 

(9) (9) (10) (10) 

1 (11%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

78
 
TABLE CI. MALE RATS (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

MID AND HIGH 

DOSE UNTREATED 

CONTROL 

LOW DOSE 

UNTREATED 

CONTROL 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

LOW DOSE 

VEHICLE 

CONTROL 

ALL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED 

NO NECROPSY PERFORMED 

2 

1 

2 

# NUMBER OF ANIMALS

• NUMBER OF ANIMALS

 WITH TISSUE EXAMINED

 NECROPSIED 

 MICROSCOPICALLY 

79
 
0 

TABLE C2. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA (TREATED GROUPS) 

LOW DOSE MID DOSE
 

ANIHALS INITIALLY IN STUDY 39 35
 AHIMALS NECROPSIED	 37 30
 ANIMALS EXAMINED HISTOPATHOLOGICALLY 37 29
 

INTEGUMENTARY SYSTEM
 

•SKIN	 (37) (30)
 

EPIDERMAL INCLUSION CYST 2 15%)
 

DERMAL INCLUSION CYST 1 (3%)
 

INFLAMMATION, SUPPURATIVE 1 (3%)
 

ABSCESS, CHRONIC 1 (3%)
 

HYPERKERATOSIS 1 (3%)
 

KERATIN-PEARL FORMATION 1 (3S) 1 (3%)
 

RESPIRATORY SYSTEM
 

#TRACHEA (37) (28)
 

INFLAMMATION, CHRONIC 2 (5%)
 

#LUNG/BRONCHIOLE (36) (29)
 

HYPERPLASIA, PLASMA CELL 2 (7%)
 

«LUNG (36) (29)
 

HEMORRHAGE 1 (356)
 

INFLAMMATION, SUPPURATIVE 1 (3%)
 BRONCHOPNEUMONIA SUPPURATIVE 2 (6%) 10 (34%)
 

BRONCHOPNEUMONIA CHRONIC SUPPURA 3 (8X) 5 (17%)
 

ABSCESS, CHRONIC 1 (3%)
 METAPLASIA, SQUAMOUS 1 (3%) 2 (7%)
 

HYPERPLASIA, PLASMA CELL 1 (3%)
 

HEHATOPOIETIC SYSTEM
 

#BONE MARROH (35) (29)
 

ATROPHY, NOS 17 (4 9%) 10 (34%)
 

•SPLEEN	 (37) (28)
 

INFLAMMATION. NECROTIZING 1 (3%)
 

# NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECBOPSIED
 

HIGH DOSE
 

35
 

32
 

(32)
 

80
 
TABLE C2. MALE RATS (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE MID DOSE
 

ATROPHY, NOS 1 (3%)
 

ANGIECTASIS 1 (3%)
 

HEMATOPOIESIS 4 (11%) 1 (4%)

 

•HANDIBULAB L.	 NODE (31) (6)

 

CONGESTION, NOS 1 (3%)
 

HYPERPLASIA, RETICULUM CELL 1 (3%)
 

•MESENTERIC L.	 NODE (31) (6) 

CONGESTION, NOS 1 (3%)
 

INFLAMMATION, GRANULOMATOUS 1 (3%)
 

CYTOMEGALY 1 (3%)
 

CIRCULATORY SYSTEM 

•MYOCARDIUM (37) (2 8)
 

INFLAMMATION, INTERSTITIAL 1 (3%)
 

CALCIFICATION, METASTATIC 1 (3%)
 

DIGESTIVE SYSTEM
 

•LIVER	 (37) (2 9)
 HEMORRHAGE 1 (3%)
 

ANGIECTASIS 1 (3%)
 

•PANCREAS	 (37) (29)
 

NECROSIS, FAT 1 (3X)
 

•ESOPHAGUS	 (36) (27)
 

ULCER, NOS 1 (3%)
 

INFLAMMATION, SUPPURATIVE 1 (3%)
 

•GASTRIC	 MUCOSA (37) (29)
 

CALCIFICATION, METASTATIC 1 (3%)
 

•COLON	 (36) (29)
 

HEMORRHAGE 1 (3%)
 

•CECUM	 (36) (29)
 

THROMBOSIS, NOS 1 (3%)
 

URINARY SYSTEM
 

•KIDNEY	 (37) (29)
 

CALCULUS, NOS LJ14J
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 

81
 
TABLE C2. MALE RATS (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, CHRONIC
 

CALCIFICATION, METASTATIC
 

ENDOCRINE SYSTEM
 

«ADRENAL
 

ANGIECTASIS
 

REPRODUCTIVE SYSTEM
 

•TESTIS
 

ATROPHY, NOS
 

MBfiVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

• EYE
 

HEMORRHAGE
 

INFLAMMATION, SUPPURATIVE
 

•EYE/CORNEA
 

ULCBR, CHRONIC
 

•EYE/CONJUNCTIVA
 

INFLAMMATION, CHRONIC SOPPURA1IV
 

«MIDDLE EAR
 

INFLAMMATION, SUPPURATIVE
 

INFLAMMATION, CHRONIC SOPPURATIV
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

«PERITONEUM
 

INFLAMMATION. CHRONIC
 

LOW DOSE: MID DOSE HIGH DOSE 

7 (24%) 

26 (10%) 

1 OX) 

(3 7) (29) 

1 (35S) 

(36) (27) 

15 (42K) 

(37) (30) (32) 

1 (3%) 

1 (3%) 

(37) (30) (32) 

1 <3X) 

(37) (30) (32) 

1 (3%) 

(37) (30) (32) 

1 (3X) 

1 (3%) 

(37) (30) (32) 

1 i3%\ 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* MUMBER OF ANIMALS NECROΡSIED
 

82
 
TABLE C2. MALE RATS (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE MID DOSE HIGH DOSE 

•PLEURA 

INFLAMMATION, SUPPURATIVE 

(37) 

1 (3%) 

(30) (32) 

•MESENTERY 

NECROSIS, FAT 

(37) 

1 (3%) 

(30) (32) 

ALL OTHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION, GRANULOMATOUS 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED

NECROPSY PERF/NO HISTO PERFORMED

NO NECROPSY PERFORMED

AUTO/NECROPSY/NO HISTO

AUTOLYSIS/NO NECROPSY

 1 

1

 1 

1

 5

 31 

1 

3 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED

* NUMBER OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

83
 
TABLE C3.
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA (CONTROL GROUPS)
 

MID AND HIGH 

DOSE UNTREATED 

CONTROL 

LOW DOSE 

UNTREATED 

CONTROL 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

LOW DOSE 

VEHICLE 

CONTROL 

ANIMALS INITIALLY IN STUDY 

ANIMALS NECROPSIED 

ANIMALS EXAMINED HISTOPATHOLOGICALL

10 

10 10 

10 

10 

10 

10 

10 

10 

10 

10 

10 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

tTRACHEA (10) (10) (10) (10)
 

INFLAMMATION, NOS 1 (10%)
 

INFLAMMATION, CHRONIC 1 (10%)
 

ILUNG (10) (10) (10) (10)
 

PNEUMONIA, LIPID 1 (10%)
 

PNEUMONIA INTERSTITIAL CHRONIC 1 (10%)
 

BRONCHOPNEUMONIA CHRONIC SUPPURA 1 (10%)
 

HEMATOPOIETIC SYSTEM
 

#BONE MARROW (10) (10) (10) (9)
 ATROPHY, NOS 4 (40*) 7 (7 0%) 7 (7 0%) 7 (78%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

•LIV2R/PEBIPORTAL	 (10) (10) (10) (10)
 

FIBROSIS 1 (10X)
 

•PANCREATIC ACINUS	 (10) (10) (10) (10)
 

ATROPHY. NOS 1 (10*)
 

# NUMBER OF ANIMALS ft IT TISSUE EXAMINED MICROSCOPICALLY 

•	 NUMBER OF ANIMALS NECROPSEED 

84
 
TABLE C3. FEMALE RATS (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

——
 MID AND HIGH LOW DOSE
 

DOSE UNTREATED UNTREATED
 CONTROL	 CONTROL
 

OEINARY SYSTEM
 

•KIDNEY	 iiO) (10)
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, CHRONIC 3 (3 0%)
 

GLOMERULONEPHRITIS, CHRONIC 1 (10%)
 

ENDOCRINE SYSTEM
 

•ADRENAL	 (10) (10)
 

ANGIECTASIS 2 (20%) 1 (10%)
 

REPRODUCTIVE SYSTEM
 

•MAMMARY	 GLAND (10) (10)
 

CYST, NOS 2 (20%) 1 (10%)
 

HEMORRHAGIC CYST 1 (10%)
 

•UTERUS	 (10) (10)
 

INFLAMMATION, SUPPURATIVE
 

•UTERUS/ENDOMETRIUM	 (10) (10)
 

INFLAMMATION, SUPPURATIVE 5 (50%) 1 (10%)
 

•OVARY	 (10) (10)
 

CYST, NOS 1 (10%)
 

INFLAMMATION, HEMORRHAGIC 1 (10%)
 

INFLAMMATION, CHRONIC SUPPURATIV 1 (10%)
 

NERVOUS SYSTEM
 

• BRAIN	 (10) (10)


HYDROCEPHALUS, NCS


SPECIAL SENSE ORGANS
 

NON5
 

MUSCULOSKELETAL	 SYSTEM
 

NONE :	 , .


• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSISD
 

MID AND HIGH LOW DOSE
 

DOSE VEHICLE VEHICLE
 

CONTROL CONTROL
 

(10)	 (10)
 

1	 (10%)
 

1 (10%)
 

(10)	 (10)
 

1 (10%) 3 (30%)
 

(10)	 (10)
 

5 (50%) 1 (10%)
 

(10)	 (10)
 

1 (10%)
 

(10)	 (10)
 

(9)	 (10)
 

b (60%)
 

 (10)	 (9)
 

 1 (10%)
 

.
 

85
 
TABLE C3. FEMALE RATS (CONTROL GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)
 

MID AND HIGH LOW DOSE MID AND HIGH LOW DOSE 

DOSE UNTREATED UNTREATED DOSE VEHICLE VEHICLE 

CONTROL. CONTROL CONTROL CONTROL 

BODY CAVITIES 

•MESENTERY

STEATITIS

 (10) (10) (10) (10) 

 1 (10%) 

ALL OTHER

ADIPOSE

 SYSTEMS 

 TISSUE 

INFLAMMATION, FOCAL GRANULOMATOU 1 

SPECIAL H0BPH0L06Y SUMMARY 

NO LESION REPORTED # NUMBER OP ANIMALS WITH TISSUE EXAMINED

* NUMBER OF ANIMALS NEC BO Ρ SI ED 

1 MICROSCOPICALLY 2 

86
 
TABLE C4.
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA (TREATED GROUPS)
 

ANIMALS INITIALLY IN STUDY
 

ANIHALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

•SKIN
 

INFLAMMATION, SUPPURATIVE
 

HYPERKERATOSIS
 

PARAKERATOSIS
 

RESPIRATORY SYSTEM
 

•TRACHEA
 

INFLAMMATION, NOS
 

LYMPHOCYTIC INFLAMMATORY INFILTR
 

INFLAMMATION, CHRONIC
 

#LUNG/BRONCHIOLE
 

METAPLASIA, SQUAMOUS
 

• LUNG
 

EDEMA, NOS
 

HEMORRHAGE
 

BRONCHOPNEUMONIA SUPPURATIVE
 

ABSCESS, NOS
 

BRONCHOPNEUMONIA CHRONIC SUPPURA
 

INFLAMMATION, GRANULOMATOUS
 

CYTOMEGALY
 

HEMATOPOIETIC SYSTEM
 

•BONE MARROW
 

ATROPHY, NOS
 

•SPLEEN
 

HEMORRHAGE
 

LOW DOSE
 

31
 

30
 

30
 

(30)
 

(30)
 

3 (10%)
 

(30)
 

(30)
 

1 (3%)
 

1 (3%)
 

1 (3X)
 

1 (3%)
 

3 (10%)
 

2 (7%)
 

(29)
 

13 (45%)
 

(30)
 

1 (3%)
 

MID DOSE
 

35
 

33
 

33
 

(33)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

(33)
 

2 (6%)
 

1 (3%)
 

(33)
 

1 (3%)
 

(33)
 

4 (12%)
 

2 (6%)
 

4 (12%)
 

(33)
 

 (24%)
 

(33)
 

HIGH DOSE
 

35
 

34
 

0
 

(34)
 

• NUMBER OF ANIHALS WITH TISSUE EXAMINED MICEOSCOP1CALL!
 

* NUMBER OF ANIMALS NECROPSIED
 

87
 
TABLE C4. FEMALE RATS (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)
 

ANGIECTASIS
 

HEMATOPOIESIS
 

•MESENTERIC L.	 NODE
 

CONGESTION NOS
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

•LIVER
 

HEMORRHAGE
 

INFLAMMATION, CHRONIC NECROTIZIN
 

INFLAMMATION, GRANULOMATOUS
 

INFLAMMATION, NECRO GRAN
 

NECROSIS, FOCAL
 

CYTOPLASMIC VACUOLIZATION
 

HEPATOCYTOMEGALY
 

HEMATOPOIESIS
 

•LIVER/CENTRILOBULAR
 

NECROSIS, NOS
 

NECROSIS, COAGULATIVE
 

«PANCREAS
 

ATROPHY, NOS
 

•COLON
 

DLCER, NOS
 

•RECTUM
 

HEMATOMA, NOS
 

URINARY SYSTEM
 

•KIDNEY
 

HYDRONEPHROSIS
 

INFLAMMATION, INTERSTITIAL
 

GLOMERULONEPHRITIS, MEMBRANOUS
 

INFLAMMATION. CHRONIC
 

• NUMBER OF ANIMALS MITH TISSUE EXAMINED MICBOSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

1 (3%)
 

5 (17%)
 

(28)
 

2 (7%)
 

(30)
 

2 (7%)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

1 (3*)
 

1 (3%)
 

2 (7%)
 

1 (3%)
 

(30)
 

1 (3%)
 

(30)
 

(29)
 

1 (3%)
 

(30)
 

(30)
 

1 (3«)
 

10 133%ϊ
 

MID DOSE HIGH DOSE 

7 (21%) 

(1) 

(33) 

(33) 

1 (3%) 

(33) 

1 (3%) 

(30) 

(33) (34) 

1 (3%) 

(33) 

1 (3X) 

1 13%) 

88
 
TABLE C4. FEMALE RATS (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE MID DOSE HIGH DOSE
 

ENDOCRINE SYSTEM
 

•ADRENAL	 (30) (33)
 

ANGIECTASIS 11 (37%) 4 (12%)
 

REPRODUCTIVE SYSTEM
 

•MAMMARY GLAND	 (30) (33) (34)
 

CYST, NOS 13 (43%) 3 (9%)
 

INFLAMMATION, VESICULAR 1 (3%)
 

•VAGINA	 (30) (33) (34)
 

EPIDERMAL INCLUSION CYST 1 (3%)
 

INFLAMMATION, CHRONIC SUPPURATIV 1 (3%)
 

•UTERUS	 (30) (32)
 

HEMORRHAGE 1 (3%)
 

ANGIECTASIS 1 (3%)
 

METAPLASIA, SQUAMOUS 1 (3%)
 

•UTERUS/ENDOMETRIUM	 (30) (32)
 

INFLAMMATION, SUPPURATIVE 4 (13%) 4 (13%)
 

ULCER, CHRONIC 1 (3%)
 

INFLAMMATION, CHRONIC SUPPURATIV 6 (20%) 2 (6%)
 

HYPERPLASIA, CYSTIC 2 (7%)
 

•OVARY	 (30) (32)
 

CYST, NOS 14 (47%)
 

INFLAMMATION, SUPPURATIVE 1 (3%)
 

INFLAMMATION, CHRONIC SUPPURATIV 1 (3%)
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

•EAR CANAL	 (30) {33) (34)
 

KERATIN-PEARL FORMATION 1 (3%)
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

• NUMBEfi OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

• NUMBER CF ANIMALS NECROPSIED 

89 
TABLE C4. FEMALE RATS (TREATED GROUPS): NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE MID DOSE HIGH DOSE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 1
 

NECROPSY PERF/NO HISTO PERFORMED 34
 

AUTOLYSIS/NO NECROPSY 1 2 1
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

90
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MICE
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

91
 

TABLE D1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

• SKIN
 

EPIDERMAL INCLUSION CYST
 

INFLAMMATION, CHRONIC SUPPURATIV
 

*SUBCUT TISSUE
 

EDEMA# NOS
 

INFLAMMATION, CHRONIC SUPPURATIV
 

METAPLASIA, SQUAMOUS
 

RESPIRATORY SYSTEM
 

#LUNG
 

INFLAMMATION, INTERSTITIAL
 

BRONCHOPNEUMONIA SUPPURATIVE
 

PERIARTERITIS
 

HEMATOPOIETIC SYSTEM
 

#BONE MARROW
 

ATROPHY, NOS
 

HYPERPLASIA, GRANULOCYTIC
 

•SPLEEN
 

HYPERPLASIA, GRANULOCYTIC
 

HYPERPLASIA, LYMPHOID
 

HEMATOPOIESIS
 

•MESENTERIC L.	 NODE
 

CONGESTION, NOS
 

HYPERPLASIA, LYMPHOID
 

VEHICLE UNTREATED LOW DOSE HIGH DOSE 

CONTROL CONTROL 

15 15 $34 35 

14 15 30 34 

14 15 30 33 

(14) (15) (30) (34) 

1 (3%) 

1 (7%) 1 (3%) 

(14) (15) (30) (34) 

2 (6%) 

1 0%) 1 (3X) 

(14) (15) (30) (33) 

1 OX) 1 (7%) 

1 (3%) 

(13) (14) (29) (32) 

4 (13%) 

1 (3%) 

(14) (15) (30) (31) 

1 (3%) 

2 (7%) 

1 (7%) 2 (13*) 18 (60%) 10 (32%) 

(2) (5) 

4 (30%) 

(1) (15) 

1 (50%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

dNOTE: 35 ANIMALS WERE INITIALLY IN STUDY BUT ONE WAS FOUND TO BE A FEMALE IN A MALE GROUP AHD
 

WAS DELETED.
 

93
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
 

VEHICLE UNTREATED
 CONTROL CONTROL
 

CIRCULATORY SYSTEM
 

•HYOCARDIUM	 (14) (15)
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, CHRONIC
 

DIGESTIVE SYSTEM
 

UIVER (14) (15)
 

MINERALIZATION
 

CONGESTION, NOS 1 (7%)
 

NECROSIS, NOS
 

NECROSIS, FOCAL
 

HYPERPLASIA, NODULAR 3 (20*)
 

•LIVEfi/CENTRILOBULAR	 (1*0
 (15)
 

NECROSIS, NOS
 

•LIVER/PEFIPOBTAL	 (14) (15)
 

FIBROSIS
 

URINARY SYSTEM
 

•KIDNEY	 (14) (15)
 

HYDRONEPHROSIS
 

INFLAMMATION, SUPPURATIVE
 

PYELONEPHRITIS SUPPURATIVE 1 (7%)
 

INFLAMMATION, CHRONIC
 

HYPERPLASIA, LYMPHOID
 

•URINARY	 BLADDER (14) (15)
 INFLAMMATION, ACUTE/CHRONIC 1 (7%)
 

INFLAMMATION, CHRONIC SUPPURATIV 1 (7%)
 

ENDOCRINE SYSTEM
 

•ADRENAL	 (14) (15)
 

ANGIECTASIS
 

BEPEODUCTIVE SYSTEM
 

•PROSTATE	 (14) (15)
 

INFLAMMATION SUPPURATIVE
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

(30)
 

1 (3%)
 

1	 0%)
 

(30)
 

1 (3%)
 

1 (3%)
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

(30)
 

(30)
 

1 (3X)
 

(30)
 

2 (7*)
 

HIGH DOSE
 

(32)
 

i^3)
 

1 (3%)
 

(3-»ϊ
 

(33)
 

(33)
 

1 (3%)
 

(33)
 

(33)
 

(33)
 

1 13%)
 

94
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE
 

1 (3%)
 

(30)
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

6
 

4
 

HIGH DOSE
 

(33)
 

1 (3%)
 

(33)
 

(34)
 

(34)
 

1
 

1
 

1
 

ABSCESS, NOS
 

«TESTIS
 

ATROPHY, NOS
 

tTONICA ALBUGINEA
 

MINERALIZATION
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE OBGANS
 

•EAB CANAL
 

KERATIN-PEARL FORMATION
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

•MULTIPLE	 ORGANS
 

HYPERPLASIA, LYMPHOID
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 

NECROPSY PERF/NO HISTO PERFORMED
 

AUTOLYSIS/NO NECROPSY
 

VEHICLE
 

CONTROL
 

(14)
 

(14)
 

(1*)
 

d<0
 

7
 

1
 

UNTREATED
 

CONTROL
 

(15)
 

(15)
 

(15)
 

(15)
 

5
 

i NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

95
 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA 

ANIMALS INITIALLY IN STUDY
 

ANIMALS MISSING
 

ANIHALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

•SUBCUT TISSUE
 

EDEMA, NOS
 

RESPIRATORY SYSTEM
 

#LUNG/BRONCHIOLE
 

HYPERPLASIA, LYMPHOID
 

«LUNG
 

INFLAMMATION, INTERSTITIAL
 

HYPERPLASIA, ALVEOLAE EPITHELIUM
 

HEMATOPOIETIC SYSTEM
 

iBONE MARROW
 

ATROPHY, NOS
 

ffSPLEEN
 

ANGIECTASIS
 

HEMATOPOIESIS
 

•MESENTERIC L.	 NODE
 

HYPERPLASIA, GRANULOCYTIC
 

HYPERPLASIA, RETICULUM CELL
 

HYPERPLASIA, LYMPHOID
 

•INGUINAL LYMPH NODE
 

CONGESTION, NOS
 

ATROPHY, NOS
 

CIRCULATORY SYSTEM
 

•MYOCARDIUM
 

INFLAMMATION. INTERSTITIAL
 

VEHICLE
 

CONTROL
 

15
 

15
 

15
 

(15)
 

(15)
 

1 (7%)
 

(15)
 

(15)
 

1 (7%)
 

(15)
 

3 (20%)
 

(2)
 

1 (50%)
 

1 (50%)
 

(2)
 

1 (50%)
 

1 (50%)
 

(15)
 

UNTREATED
 CONTROL
 

15
 

15
 

15
 

(15)
 

(15)
 

1 (7%)
 

(15)
 

(15)
 

(15)
 

(1)
 

(1)
 

(15)
 

LOW DOSE
 

35
 

1
 

30
 

30
 

(30)
 

(29)
 

1 (3%)
 

(29)
 

2 (7%)
 

1 (3%)
 

(29)
 

4 (14%)
 

(30)
 

1 (3%)
 

5 (17%)
 

(«0
 

1 (25%)
 

1 (25%)
 

(<*)
 

(29)
 

1 (3%)
 

HIGH DOSE
 

35
 

2
 

32
 

32
 

(32)
 

5 (16%)
 

(32)
 

(32)
 

(32)
 

(32)
 

2 (6%)
 

(24)
 

(24)
 

(32)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

96
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

VEHICLE UNTREATED LOW DOSE
 

CONTROL CONTROL
 

DIGESTIVE SYSTEM
 

•LIVER	 (15) (15) (30)
 

FIBROSIS, DIFFUSE 1 (3%)
 

NECROSIS, FOCAL 1 (7%)
 

INFARCT, NOS 1 (7%)
 

HYPERPLASIA, NODULAR 1 (3%)
 

ANGIECTASIS 1 (3%)
 

HEMATOPOIESIS 1 (3X)
 

«PANCREAS (15) (15) (28)
 

ATROPHY, NOS 1 (7%)
 

URINARY SYSTEM
 

#KIDNEY (15) (15) (30)
 

HYDRONEPHROSIS 1 (7%)
 

INFLAMMATION, INTERSTITIAL 1 (7%)
 

AMYLOIDOSIS 1 (3X)
 

HYPERPLASIA, LYMPHOID 1 (7%)
 

•UfilNARY BLADDER	 (15) (15) (29)
 

INFLAMMATION, CHBCNIC SUPPURATIV 1 <7«)
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

•UTERUS	 (15) (15) (29)
 

EDEMA, NOS 1 (7%)
 

•UTERUS/ENDOWETHIUM	 (15) (15) (29)
 

INFLAMMATION, CHRONIC SUPPURATIV 1 (7%)
 

HYPERPLASIA, CYSTIC 10 (67X) 10 (67%) 9 (31%)
 

•OVARY	 (15) (15) (29)
 

CYST, NOS 2 (7%)
 

HEMORRHAGE 1 (3X)
 

ATROPHY, NOS	 1 (3X)
 

MBBVOOS SYSTEM
 

NONE
 

* NUHBEB OF ANIMALS WITH TISSUE EXAMINED filCROSCOPICALLY 

* NOMBEfi OF ANIMALS NECROPSIED 

HIGH DOSE
 

(32)
 

(32)
 

(32)
 

(32)
 

(31)
 

(31)
 

(31)
 

1 (3X)
 

97 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

VEHICLE CONTROL UNTREATED CONTROL LOW DOSE HIGH DOSE 

SPECIAL SENSE ORGANS 

•EAR CANAL 

INFLAMMATION, CHRONIC SUPPURAIIV (15) (15) (30) 

1 OX) 

(32) 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

•PERITONEUM 

INFLAMMATION, NOS 

(15) 

1 (7%) 

(15) (30) (32) 

ALL OTHER SYSTEMS 

•MULTIPLE ORGANS 

PERIARTERITIS 

HYPERPLASIA, LYMPHOID 

(15) (15) 

1 (7%) 

(30) 

1 (3%) 

(32) 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED 

ANIMAL MISSING/NO NECROPSY 

AUTO/NECROPSY/HISTO PERF 

AUTOLYSIS/NO NECROPSY 

3 6 

1 

1 

4 

2 

1 

# NUMBER OF ANIMALS

• NUMBER OF ANIMALS

 WITH TISSUE EXAMINED

 NECROPSIED 

 MICROSCOPICALLY 

98
 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

99
 

__ 

Table El. Analyses

Male Rats Given

Topography: Morphology
 

Skin: Squamous-cell Carcinoma**
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Subcutaneous Tissue: Sarcoma, NOS^
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 of the Incidence of Primary Tumors in Low-Dose 

 Intraperitoneal Injections of Thio-TEPAa 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

0/30 (0) 0/10 (0)
 

0/30 (0) 0/10 (0)
 

Low
 

Dose
 

5/37 (14)
 

 - 0.045**
 

Infinite
 

1.044
 

Infinite
 

Infinite
 

0.381
 

Infinite
 

53
 

5/37 (14)
 

 - 0.045**
 

Infinite
 

1.044
 

Infinite
 

Infinite
 

0.381
 

Infinite
 

61
 
Table El, Analyses of the Incidence of Primary Tumors in Low-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Subcutaneous Tissue:
 

Sarcoma or Fibrosarcoma*5
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Subcutaneous Tissue: Fibroma**
 

 Values0

»d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

1/30 (3) 0/10 (0)
 

«... ....
 

0/30 (0) 0/10 (0)
 

—
 

Low
 

Dose
 

6/37 (16)
 

N.S.
 

4.865
 

0.642
 

216.767
 

Infinite
 

0.484
 

Infinite
 

61
 

2/37 (5)
 

N.S.
 

Infinite
 

0.244
 

Infinite
 

Infinite
 

0.088
 

Infinite
 

74
 
__ 

Table El· Analyses of the Incidence of Primary Tumors in Low-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Hematopoietic System:
 

Granulocytic Leukemia"
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Lymphoma
 

or Lymphocytic Leukemia*5
 

 Valuesc>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

0/30 (0) 0/10 (0)
 

0/30 (0) 0/10 (0)
 

Low
 

Dose
 

2/37 (5)
 

N.S.
 

Infinite
 

0.244
 

Infinite
 

Infinite
 

0.088
 

Infinite
 

55
 

4/37 (11)
 

N.S.
 

Infinite
 

0.767
 

Infinite
 

Infinite
 

0.279
 

Infinite
 

37
 
Table El· Analyses of the Incidence of Primary Tumors in Low-Dose
 

Male Rats Giv* Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Hematopoietic System:
 

Leukemia or Lymphoma­

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma*5
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

0/30 (0) 0/10 (0)
 

7/26 (27) 3/9 (33)
 

82
 

Low
 

Dose
 

6/37 (16)
 

 - 0.023**
 

Infinite
 

1.325
 

Infinite
 

Infinite
 

0.484
 

Infinite
 

37
 

1/28 (4)
 

 - 0.038*(Ν)
 

 - 0.019**(N
 

0.133
 

0.003
 

0.931
 

0.107
 

0.002
 

1.186
 

78
 
Table £1. Analyses of the Incidence of Primary Tumors in Low-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Ear Canal: Squamous-cell Carcinoma*5
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Skin and Ear Canal:
 

Squamous-cell Carcinoma*5
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

0/30 (0) 0/10 (0)
 

0/30 (0) 0/10 (0)
 

«...
 

Low
 

Dose
 

3/37 (8)
 

N.S.
 

Infinite
 

0.497
 

Infinite
 

Infinite
 

0.181
 

Infinite
 

74
 

7/37 (19)
 

 « 0.011**
 

Infinite
 

1.608
 

Infinite
 

Infinite
 

0.587
 

Infinite
 

53
 
Table El. Analyses of the Incidence of Primary Tumors in Low-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Low-Dose 

Pooled Vehicle Low 

Topography: Morphology Control Control Dose 

Mesentery: Sarcoma, NOS^ 0/30 (0) 0/10 (0) 2/37 (5)
 

 Values0>d N.S.
 

Relative Risk (Pooled Control)e Infinite
 

Lower Limit 0.244
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)e Infinite
 

Lower Limit 0.088
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 66
 

a

Treated group received dose of 0.7 mg/kg.
 

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

c

Beneath the incidence of tumors in a treated group is the probability level for the
 

Fisher exact test for the comparison of that treated group with the vehicle-control
 

group (*) or with the pooled-control group (**) when Ρ < 0.05 for either control group;
 

otherwise, not significant (N.S.) is indicated.
 

d

A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

e

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

control group.
 
Table E2. Analyses of the Incidence of Primary Tumors in Mid-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

Topography; Morphology
 

Skin: Squamous-cell Carcinoma^
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System:
 

Lymphocytic Leukemia^
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid-Dose 

Pooled Vehicle Mid 

Control Control Dose 

0/30 (0) 0/10 (0) 3/30 (10) 

N.S. 

Infinite 

0.614 

Infinite 

Infinite 

0.223 

Infinite 

44 

0/30 (0) 0/10 (0) 5/30 (17) 

 ­ 0.026** 

Infinite 

0.291 

Infinite 

Infinite 

0.471 

Infinite 

40 
Table Ε2· Analyses of the Incidence of Primary Tumors in Mid-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Mid-Dose
 

Pooled Vehicle
 

Topography: Morphology Control Control
 

Hematopoietic System: All Leukemia13 0/30 (0) 0/10 (0)
 

 Valuesc

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 ο
 

oo Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma** 2/29 (7) 0/10 (0)
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lowe Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid
 

Dose
 

6/30 (20)
 

 - 0.012**
 

Infinite
 

1.638
 

Infinite
 

Infinite
 

0.598
 

Infinite
 

40
 

1/24 (4)
 

N.S.
 

0.604
 

0.011
 

10.830
 

Infinite
 

0.024
 

Infinite
 

54
 
Table E2. Analyses of the Incidence of Primary Tumors in Mid-Dose
 

Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Mid-Dose
 

Pooled Vehicle Mid
 

Topography: Morphology Control Control Dose
 

Skin and Ear Canal: 

Squamous-cell Carcinoma'3 0/30 (0) 0/10 (0) 3/30 (10) 

 Values 0>d N.S. 

Relative Risk (Pooled Control)e Infinite
 

Lower Limit 0.614
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)e Infinite
 ο
 Lower Limit 0.223
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 44
 

a

Treated group received dose of 1.4 ing/kg·
 

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

c

Beneath the incidence of tumors in a treated group is the probability level for the
 

Fisher exact test for the comparison of that treated group with the vehicle-control
 

group (*) or with the pooled-control group (**) when Ρ < 0,05 for either control group;
 

otherwise, not significant (N.S.) is indicated,
 

d

A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

e

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

control group.
 
Table E3. Analyses of the Incidence of Primary Tumors in Low-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

Low-Dose
 

Pooled Vehicle
 

Topography: Morphology Control Control
 

Subcutaneous Tissue: Sarcoma, NOS,
 

or Fibrosarcoma^ 0/30 (0) 0/10 (0)
 

 Values0

»d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

 Relative Risk (Vehicle Control)e
 

 Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia
 

or Lymphoma** 1/30 (3) 0/10 (0)
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low
 

Dose
 

2/30 (7)
 

N.S.
 

Infinite
 

0.301
 

Infinite
 

Infinite
 

0.109
 

Infinite
 

79
 

2/30 (7)
 

N.S.
 

2.000
 

0.110
 

113.910
 

Infinite
 

0.109
 

Infinite
 

63
 
Table £3· Analyses of the Incidence of Primary Tumors in Low-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Low-Dose
 

Pooled Vehicle Low
 

Topography: Morphology Control Control Dose
 

Pituitary: Chromophobe Adenoma** 14/28 (50) 1/8 (13) 4/29 (14)
 

 Values0

>d  - 0.004**(N)
 

Relative Risk (Pooled Control)e 0.276
 

Lower Limit 0.078
 

Upper Limit 0.752
 

Relative Risk (Vehicle Control)e 1.103
 

Lower Limit 0.142
 

 Upper Limit 52.317
 

Weeks to First Observed Tumor 82 75
 

Ear Canal: Squamous-cell Carcinoma*5 0/30 (0) 0/10 (0) 2/30 (7)
 

 Values0

>d N.S.
 

Relative Risk (Pooled Control)e Infinite
 

Lower Limit 0.301
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)e Infinite
 

Lower Limit 0.109
 

Upper Limit Infinite
 

Weeks to First Observed Tumor
 ——
 74
 
Table £3· Analyses of the Incidence of Primary Tumors in Low-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Brain: Olfactory Neuroblastoma^
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma,
 

 Values0

>d
 

Relative Risk (Pooled Control)6
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

0/30 (0) 0/10 (0)
 

1/30 (3) 0/10 (0)
 

Low
 

Dose
 

2/30 (7)
 

N.S.
 

Infinite
 

0·301
 

Infinite
 

Infinite
 

0.109
 

Infinite
 

59
 

7/30 (23)
 

 - 0.026**
 

7.000
 

0.987
 

302.176
 

Infinite
 

0.726
 

Infinite
 

37
 
Table E3. Analyses of the Incidence of Primary Tumors in Low-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology 

Pooled 

Control 

Mammary Gland: Fibroadenoma^ 6/30 (20) 

 Values0 > d 

Relative Risk (Pooled Control)e 

Lower Limit 

Upper Limit 

Relative Risk (Vehicle Control)e 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Uterus: Adenocarcinoma, NOS^ 0/30 (0) 

 Valuesc > d 

Relative Risk (Pooled Control)e 

Lower Limit 

Upper Limit 

Relative Risk (Vehicle Control)e 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Low-Dose 

Vehicle 

Control 

Low 

Dose 

2/10 (20) 8/30 (27) 

N.S. 

1.333 

0.464 

4.085 

1.333 

0.348 

11.664 

82 66 

0/10 (0) 2/30 (7) 

N.S. 

Infinite 

0.301 

Infinite 

Infinite 

0.109 

Infinite 

— 70 
4 

Table E3. Analyses of the Incidence of Primary Tumors in Low-Dose
 

Female Rats Given Intraperitoneal Injections of Thio~TEPAa
 

(continued)
 

Topography: Morphology
 

uterus: Sarcoma, NOSb
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polypi
 

 Values0

»d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose 

Pooled Vehicle 

Control Control 

0/30 (0) 0/10 (0) 

1/30 (3) 0/10 (0) 

Low
 

Dose
 

2/30 (7)
 

N.S.
 

Infinite
 

0.301
 

Infinite
 

Infinite
 

0.109
 

Infinite
 

82
 

4/30 (13)
 

N.S.
 

4.000
 

0.428
 

189.625
 

Infinite
 

0.345
 

Infinite
 

63
 
Table E3. Analyses of the Incidence of Primary Tumors in Low-Dose 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

a

Treated group received dose of 0.7 mg/kg.
 

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

c

Beneath the incidence of tumors in a treated groups is the probability level for the
 

Fisher exact test for the comparison of that treated group with the vehicle-control
 

group (*) or with the pooled-control group (**) when Ρ < 0.05 for either control group;
 

otherwise, not significant (N.S.) is indicated.
 

^A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

e

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

 control group.
 
Table Ε4· Analyses of the Incidence of Primary Tumors in Mid-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

Mid-Dose
 

Pooled Vehicle Mid
 

Tocography; Morphology Control Control Dose
 

Skin: Squamous-cell Carcinoma^ 0/28 (0) 0/10 (0) 3/33 (9)
 

 Values0»d N.S.
 

Relative Risk (Pooled Control)6 Infinite
 

Lower Limit 0.522
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)e Infinite
 

Lower Limit 0.203
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 64
 

Hematopoietic System: Leukemia
 

or Lymphoma" 0/28 (0) 0/10 (0) 3/33 (9)
 

 Values0»d N.S.
 

Relative Risk (Pooled Control)e Infinite
 

Lower Limit 0.522
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)e Infinite
 

Lower Limit 0.203
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 25
 
Table E4. Analyses of the Incidence of Primary Tumors in Mid-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Pituitary: Chromophobe Adenoma*3
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenoma, NOS^
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid-Dose
 

Pooled Vehicle
 

Control Control
 

8/28 (29) 2/10 (20)
 

87
 

0/28 (0) 0/10 (0)
 

Mid
 

Dose
 

0/32 (0)
 

 - 0.001**(N)
 

0.000
 

0.000
 

0.374
 

0.000
 

0.000
 

1.028
 

3/33 (9)
 

N.S.
 

Infinite
 

0.522
 

Infinite
 

Infinite
 

0.203
 

Infinite
 

43
 
00 

Table E4. Analyses of the Incidence of Primary Tumors in Mid-Dose
 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Adenocarcinoma, NOS^
 

 Values0>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma^
 

 Values0

>d
 

Relative Risk (Pooled Control)e
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)e
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid-Dose 

Pooled Vehicle Mid 

Control Control Dose 

1/28 (4) 1/10 (10) 8/33 (24) 

 « 0.025** 

6.788 

1.004 

289.508 

2.424 

0.409 

103.495 

87 40 

10/28 (36) 5/10 (50) 4/33 (12) 

Ρ 0.020* (Ν) 

Ρ 0.030**(N) 

0.339 

0.088 

1.036 

0.242 

0.071 

0.948 

87 56 
Table £4· Analyses of the Incidence of Primary Tumors in Mid-Dose 

Female Rats Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

Toppferaphy; Morphology 

Pooled 

Control 

Mid-Dose 

Vehicle 

Control 

Mid 

Dose 

Uterus: Adenocarcinoma» 0/28 (0) 0/10 (0) 7/32 (22) 

 Values0 > d 

 ­ 0.009** 

Relative Risk (Pooled Control)e 

Lower Limit 

Upper Limit 

Infinite 

1.745 

Infinite 

VO 

Relative Risk (Vehicle Control)e 

Lower Limit 

Upper Limit 

Infinite 

0.680 

Infinite 

Weeks to First Observed Tumor 50 

Ear Canal: Squamous-cell Carcinoma** 0/28 (0) 0/10 (0) 5/33 (15) 

 Values0 

» d 

 ­ 0.040** 

Relative Risk (Pooled Control)e 

Lower Limit 

Upper Limit 

Infinite 

1.097 

Infinite 

Relative Risk (Vehicle Control)e 

Lower Limit 

Upper Limit 

Infinite 

0.427 

Infinite 

Weeks to First Observed Tumor 71 
Table E4. Analyses of the Incidence of Primary Tumors in Mid-Dose 

Female Rats Given Intraperitoneal Injections of Thio-TEFAa 

(continued)
 

Mid-Dose
 

Pooled Vehicle Mid
 

Toppgraphy; Morphology Control Control Dose
 

Skin and Ear Canal: 

Squamous-cell Carcinoma^ 0/28 (0) 0/10 (0) 8/33 (24) 

 Values 0>d 

 = 0.005** 

Relative Risk (Pooled Control)e Infinite
 

Lower Limit 1.991
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)e Infinite
 

ο
 Lower Limit 0.776
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 64
 

a

Treated group received dose of 1.4 mg/kg.
 

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

c

Beneath the incidence of tumors in a treated group is the probability level for the
 

Fisher exact test for the comparison of that treated group with the vehicle-control
 

group (*) or with the pooled-control group (**) when Ρ < 0.05 for either control group;
 

otherwise, not significant (N.S.) is indicated.
 

d

A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

e

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

control group.
 
Table £5. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

Low-Dose
 

Pooled Vehicle Low
 

Topography: Morphology Control Control Dose
 

Skin: Squamous-cell Carcinoma0 0/29 (0) 0/10 (0) 5/33 (15)
 

 Valuesd»e - 0.036**
 

Relative Risk (Pooled Control)* Infinite
 

Lower Limit 1.134
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control) Infinite
 

Lower Limit 0.427
 

N> Upper Limit Infinite
 

Weeks to First Observed Tumor 53
 

Subcutaneous Tissue: Sarcoma, NOSC 0/29 (0) 0/10 (0) 5/33 (15)
 

 Valuesd>e - 0.036**
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 1.134
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)* Infinite
 

Lower Limit 0.427
 

Upper Limit Infinite
 

Weeks to First Observed Tumor
 61
 
Table E5. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Subcutaneous Tissue: Sarcoma
 

or Fibrosarcoma0
 

e
 Valuesd

>
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Subcutaneous Tissue: Fibroma0
 

e
 Valuesd

»
 

Relative Risk (Pooled Control)*
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose
 

Pooled Vehicle
 

Control Control
 

1/30 (3) 0/10 (0)
 

0/29 (0) 0/10 (0)
 

Low
 

Dose
 

6/33 (18)
 

N.S.
 

5.455
 

0.722
 

241.878
 

Infinte
 

0.543
 

Infinite
 

61
 

2/33 (6)
 

N.S.
 

Infinite
 

0.265
 

Infinite
 

Infinite
 

0.099
 

Infinite
 

74
 
Table E5. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Low-Dose 

Pooled Vehicle Low 

Topography: Morphology Control Control Dose 

Hematopoietic System:
 

Granulocytic Leukemia0 0/29 (0) 0/10 (0) 2/34 (6)
 

 Valuesd>e N.S.
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.257
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.096
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 55
 

Hematopoietic System: Lymphoma or
 

Lymphocytic Leukemia^ 0/29 (0) 0/10 (0) 4/34 (12)
 

 Valuesd>e N.S.
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.088
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)^ Infinite
 

Lower Limit 0.304
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 37
 
Table E5. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Hematopoietic System: Leukemia
 

or Lymphoma**
 

e
 Valuesd

>
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)^
 

)
 Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma0
 

e
 Valuesd

»
 

Relative Risk (Pooled Control)^
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose 

Pooled Vehicle 

Control Control 

0/29 (0) 0/10 (0) 

—— 

7/25 (28) 3/9 (33) 

82
 

Low
 

Dose
 

6/34 (18)
 

 = 0.020**
 

Infinite
 

1.397
 

Infinite
 

Infinite
 

0.527
 

Infinite
 

37
 

1/27 (4)
 

 = 0.041*(N)
 

 - 0.019**(N)
 

0.132
 

0.003
 

0.925
 

0.111
 

0.003
 

1.227
 

78
 
Table E5. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Hale Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Low-Dose
 

Pooled Vehicle Low
 

Topography: Morphology Control Control Dose
 

£ar Canal: Squamous-cell Carcinoma0 0/29 (0) 0/10 (0) 3/33 (9)
 

 Valuesd>e N.S.
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.540
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.203
 

ho Upper Limit Infinite
 

Weeks to First Observed Tumor 74
 

Skin or Ear Canal: 

Squamous-cell Carcinomac 0/29 (0) 0/10 (0) 7/33 (21) 

e

 Valuesd» - 0.009** 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 1.748
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.659
 

Upper Limit Infinite
 

Weeks to First Observed Tumor
 53
 
Table E5. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Low-Dose 

Pooled Vehicle Low 

Τopography: Morphology Control Control Dose 

Mesentery: Sarcoma, NOSC 0/29 (0) 0/10 (0) 2/33 (6) 

Ρ Valuesd ,e N.S. 

Relative Risk (Pooled Control)f Infinite 

Lower Limit 0.265 

Upper Limit Infinite 

Relative Risk (Vehicle Control)f Infinite 

Lower Limit 0.099 

Upper Limit Infinite 

-­ Weeks to First Observed Tumor 66 

126
 

a
Ireated group received dose of 0.7 mg/kg.
 

b

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

at least 37 weeks of the study.
 

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

at least 52 weeks of the study.
 

d

Beneath the incidence of tumors in a treated goup is the probability level for the Fisher
 

exact test for the comparison of that treated group with the vehicle-control group (*) or with the
 

pooled-control group (**) when Ρ < 0.05; otherwise, not significant (N.S.) is indicated.
 

e
A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

f

The 95% confidence interval of the relative risk between each treated group and the control
 

group.
 
Table E6. Time-adjusted Analyses of the Incidence of Primary Tumors 

in Mid-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa 

Topography: Morphology 

Hematopoietic System: 

Lymphocytic Leukemia·3 

 Valuese>f 

Relative Risk (Pooled Control^
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: All Leukemia^
 

 Valuese»f
 

Relative Risk (Pooled Control)S
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)S
 

Lower Limit
 

Upper Limit
 

Mid-Dose 

Pooled Vehicle Mid 

Control Control Dose 

0/30 (0) 0/10 (0) 5/16 (31) 

 = 0.003** 

Infinite 

2.453 

Infinite 

Infinite 

0.892 

Infinite 

40 

0/30 (0) 0/10 (0) 6/16 (38) 

 = 0.035* 

 = 0.001** 

Infinite 

3.123 

Infinite 

Infinite 

1.135 

Infinite 

Weeks to First Observed Tumor
 
Table E6. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Mid-Dose
 

Pooled Vehicle Mid
 

Topography: Morphology Control Control Dose
 

Pituitary: Chromophobe Adenoma** 2/29 (7) 0/10 (0) 1/6 (17)
 

 Valuese>f N.S.
 

Relative Risk (Pooled Control)^ 2.417
 

Lower Limit 0.042
 

Upper Limit 34.031
 

Relative Risk (Vehicle Control)S Infinite
 

Lower Limit 0.096
 

ro Upper Limit Infinite
 

oo
 

Weeks to First Observed Tumor 54
 

Skin or Ear Canal: 

Squamous-cell Carcinoma0 0/30 (0) 0/10 (0) 3/13 (23) 

 Valuese»f = 0.023** 

Relative Risk (Pooled Control)S Infinite
 

Lower Limit 1.435
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)S Infinite
 

Lower Limit 0.521
 

Upper Limit Infinite
 

Weeks to First Observed Tumor
 44
 
Table E6. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Male Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

a

Treated group received dose of 1.4 mg/kg.
 

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

at least 40 weeks of the study.
 

c

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

at least 44 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 treated group is the probability level for the Fisher
 

exact test for the comparison of that treated group with the vehicle-control group (*) or with the
 

pooled-control group (**) when Ρ < 0.05; otherwise, not significant (N.S.) is indicated.
 

^A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

95% confidence interval of the relative risk between each treated group and the control
 

group.
 
Table £7· Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

Topography: Morphology
 

Subcutaneous Tissue: Sarcoma, NOS,
 or Fibrosarcoma0
 

e
 Values d

»
 

Relative Risk (Pooled Control) f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control) f
 

u>


ο
 Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia
 

or Lymphoma0
 

e
 Valuesd

»
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose 

Pooled Vehicle 

Control Control 

0/28 (0) 0/9 (0) 

1/28 (4) 0/9 (0) 

Low
 

Dose
 

2/29 (7)
 

N.S.
 

Infinite
 

0.292
 

Infinite
 

Infinite
 

0·103
 

Infinite
 

79
 

2/29 (7)
 

N.S·
 

1.931
 

0.107
 

109.859
 

Infinite
 

0.103
 

Infinite
 

63
 
Table E7. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Topography: Morphology 

Pooled 

Control 

Low-Dose 

Vehicle 

Control 

Pituitary: Chromophobe Adenomac 14/28 (50) 1/8 (13) 

 Valuesd > e 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

u> 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 82 

Ear Canal: Squamous-cell Carcinoma0 0/28 (0) 0/9 (0) 

 Valuesd > e 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Low
 

Dose
 

4/28 (14)
 

 = 0.005**(N)
 

0.286
 

0.087
 

0.777
 

1.143
 

0.147
 

54.086
 

75
 

2/29 (7)
 

N.S.
 

Infinite
 

0.292
 

Infinite
 

Infinite
 

0.103
 

Infinite
 

74
 
Table E7. Time-adjusted Analyses of the Incidence of Priinary Tumors
 

in Low-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Low-Dose 

Pooled Vehicle Low 

Topography: Morphology Control Control Dose 

Brain: Olfactory Neuroblastoma0 0/28 (0) 0/9 (0) 2/29 (7)
 

 Valuesd>e N.S.
 

Pvelative Risk (Pooled Control)^ Infinite
 

Lower Limit 0.292
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.103
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 59
 

Mammary Gland: Adenocarcinoma, NOS^ 1/28 (4) 0/9 (0) 7/30 (23)
 

 Valuesd>e = 0.033**
 

Relative Risk (Pooled Control)f 6.533
 

Lower Limit 0.926
 

Upper Limit 282.066
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.663
 

Upper Limit Infinite
 

Weeks to First Observed Tumor
 37
 
133 

Table E7. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Low-Dose 

Pooled Vehicle Low 

Topography: Morphology Control Control Dose 

Mammary Gland: Fibroadenoma0 6/28 (21) 2/9 (22) 8/29 (28)
 

 Valuesd>e N.S.
 

Relative Risk (Pooled Control)^ 1.287
 

Lower Limit 0.452
 

Upper Limit 3.917
 

Relative Risk (Vehicle Control)f 1.241
 

Lower Limit 0.335
 

Upper Limit 10.781
 

Weeks to First Observed Tumor 82 66
 

Uterus: Adenocarcinoma, NOSC 0/28 (0) 0/9 (0) 2/29 (7)
 

 Valuesd>e N.S.
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.292
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)^ Infinite
 

Lower Limit 0.103
 

Upper Limit Infinite
 

Weeks to First Observed Tumor ....
 70
 
Table Ε7· Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Low-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Uterus: Sarcoma, NOSC
 

e
 Valuesd

>
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

u> Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polypc
 

e
 Valuesd

»
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose 

Pooled Vehicle Low 

Control Control Dose 

0/28 (0) 0/9 (0) 2/29 (7) 

N.S. 

Infinite 

0-292 

Infinite 

Infinite 

0.103 

Infinite 

82 

1/28 (4) 0/9 (0) 4/29 (14) 

N.S. 

3.862 

0.416 

182.833 

Infinite 

0.326 

Infinite 

63 
Table E7. Time-adjusted Analyses of the Incidence of Primary Tumors 

in Low-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

a

Treated group received dose of 0.7 mg/kg.
 

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

at least 3 7 weeks of the study.
 

c

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 treated group is the probability level for the Fisher
 

exact test for the comparison of that treated group with the vehicle-control group (*) or
 

with the pooled-control group (**) when Ρ < 0.05; otherwise, not significant (N.S.) is indicated.
 

€A negative value (N) indicates a lower incidence in a treated group than in a control group.
 

u> 

Ln 95% confidence interval of the relative risk between each treated group and the control
 

group.
 
Table E8. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Female Ra?s Given Intraperitoneal Injections of Thio-TEPAa
 

Mid-Dose
 

Pooled Vehicle Mid
 

Topography; Morphology Control Control Dose
 

Skin: Squamous-cell Carcinoma* 0/28 (0) 0/10 (0) 3/21 (14)
 

 Values8>h N.S.
 

Relative Risk (Pooled Control)± Infinite
 

Lower Limit 0.824
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control) * Infinite
 

Lower Limit 0.320
 LO Upper Limit Infinite
 

Weeks to First Observed Tumor 64
 

Hematopoietic System: Lymphocytic
 

Leukemia or Lymphoma** 0/28 (0) 0/10 (0) 3/22 (14)
 

 Values8»h N.S.
 

Relative Risk (Pooled Control)i Infinite
 

Lower Limit 0.786
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)1 Infinite
 

Lower Limit 0.305
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 25
 
Table E8. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology 

Pooled 

Control 

Mid-Dose 

Vehicle 

Control 

Mid 

Dose 

Pituitary: Chromophobe Adenoma ^ 8/28 (29) 2/10 (20) 0/20 (0) 

 ValuesS>h 

 » 0.008**(N) 

Relative Risk (Pooled Control)1 

Lower Limit 

Upper Limit 

0.000 

0.000 

0.583 

Relative Risk (Vehicle Control)1 

Lower Limit 

Upper Limit 

0.000 

0.000 

1.609 

Weeks to First Observed Tumor 87 

Mammary Gland: Adenoma^ 0/28 (0) 0/10 (0) 3/22 (14) 

 ValuesS>h N.S. 

Relative Risk (Pooled Control)1 

Lower Limit 

Upper Limit 

Infinite 

0.786 

Infinite 

Relative Risk (Vehicle Control)1 

Lower Limit 

Upper Limit 

Infinite 

0.305 

Infinite 

Weeks to First Observed Tumor 43 
Table £8. Time-adjusted Analyses of the Incidence of Primary Tumors 

in Mid-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

Topography: Morphology
 

Mammary Gland: Adenocarcinoma, NOSC
 

 Values8>h
 

Relative Risk (Pooled Control)1
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)1
 

Lower Limit
 

Upper Limit
 00 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenomaf
 

 Values8»h
 

Relative Risk (Pooled Control)1
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)1
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid-Dose 

Pooled Vehicle Mid 

Control Control Dose 

1/28 (4) 1/10 (10) 8/24 (33) 

 - 0.006** 

9.333 

1.398 

388.857 

3.333 

0.567 

139.059 

87 40 

10/28 (36) 5/10 (50) 4/21 (19) 

N.S. 

0.533 

0.141 

1.555 

0.381 

0.113 

1.434 

87 56 
Table E8. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Mid-Dose
 

Pooled Vehicle Mid
 

Topography; Morphology Control Control Dose
 

Uterus: Adenocarcinoma, NOSe 0/28 (0) 0/10 (0)	 7/21 (33)
 

 Values8>h	 * 0.044*
 

 - 0.001**
 

Relative Risk (Pooled Control)1 Infinite
 

Lower Limit 2.684
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)1 Infinite
 

u>
 Lower Limit 1.044
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 50
 

Ear Canal: Squamous-cell Carcinoma^ 0/28 (0) 0/10 (0) 5/21 (24)
 

 Values8»h 

 - 0.011**
 

Relative Risk (Pooled Control)1 Infinite
 

Lower Limit 1.737
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)1 Infinite
 

Lower Limit 0.676
 

Upper Limit Infinite
 

Weeks to First Observed Tumor
 71
 
Table E8. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Mid-Dose 

Pooled Vehicle Mid 

Topography; Morphology Control Control Dose 

Skin and Ear Canal:
 

Squamous-cell Carcinoma^ 0/28 (0) 0/10 (0) 8/21 (38)
 

 Values8>h	 = 0.026*
 

 < 0.001**
 

Relative Risk (Pooled Control)* Infinite
 

Lower Limit 3.164
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control) Infinite
 

Lower Limit 1.230
 

Upper Limit Infinite
 

Weeks to First Observed Tumor	 64
 

a

Treated group received dose of 1.4 mg/kg.
 

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

at least 25 weeks of the study.
 

c

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

at least 40 weeks of the study.
 

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

at least 43 weeks of the study.
 
Table E8. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Mid-Dose Female Rats Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

e

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

at least 50 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·
 

^Beneath the incidence of tumors in a treated group is the probability level for the Fisher
 

exact test for the comparison of that treated group with the vehicle-control group (*) or with
 

the pooled-control group (**) when Ρ < 0.05; otherwise, not significant (N.S.) is indicated.
 

^A negative value (N) indicates a lower incidence in a treated group than in a control group·
 

95% confidence interval of the relative risk between each treated group and the control
 

group.
 

APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN MICE
 

GIVEN INTRAPERITONEAL INJECTIONS OF THIO-TEPA
 

143
 

Table Fl. Analyses of the Incidence of Primary Tumors in 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa 

Topography: Morphology 

Pooled 

Control 

Vehicle 

Control 

Low 

Dose 

High 

Dose 

Skin: Squamous-ceil 

Carcinoma*5 0/28 (0) 0/14 (0) 7/30 (23) 0/34 (0) 

 Values0 > d N.S. N.S. « 0.007** N.S. 

Departure from Linear Trend6 ? < 0.001 < 0.001 

Relative Risk (Pooled Control)^ 

Lower Limit 

Upper Limit 

Infinite 

1.863 

Infinite — 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

Infinite 

0.978 

Infinite 

—— 

— 

Weeks to First Observed Tumor ... — 67 
Table Fl. Analyses of the Incidence of Primary Tumors in 

Hale Mice Given Intraperitoneal Injections of Thio-TEPAa 

Low
 

Dose
 

5/30 (17)
 

N.S.
 

4.667
 

0.571
 

212.161
 

2.333
 

0.306
 

106.152
 

83
 

(continued) 

Topography: Morphology
 

Lung: Alveolar/Bronchiolar
 

Adenomab
 

 Values0>d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)f
 

Lover Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Control
 

1/28 (4)
 

N.S.
 

 » 0.007
 

——
 

Vehicle
 

Control
 

1/14 (7)
 

N.S.
 

 - 0.043
 

87
 

High
 

Dose
 

0/33 (0)
 

N.S.
 

0.000
 

0.000
 

15.686
 

0.000
 

0.000
 

7.858
 

——
 
Table Fl. Analyses of the Incidence of Primary Tumors in 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

Pooled Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Hematopoietic System: Lymphoma 

or Lymphocytic Leukemia** 1/28 (4) 1/14 (7) 2/30 (7) 26/34 (76) 

 Values0 > d < 0.001 < 0.001 N.S. < 0.001* 

 < 0.001** 

Departure from Linear Trende 

 = 0.001 » 0.003 

Relative Risk (Pooled Control)f 1.867 21.412 

Lower Limit 0.103 4.109 

Upper Limit 106.333 770.826 

Relative Risk (Vehicle Control)f 0.933 10.706 

Lower Limit 0.055 2.187 

Upper Limit 53.204 385.659 

Weeks to First Observed Tumor 78 61 26 
Table Fl. Analyses of the Incidence of Primary Tumors in
 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology 

Pooled 

Control 

Vehicle 

Control 

Low 

Dose 

High 

Dose 

Hematopoietic System: 

Leukemia or Lymphoma** 1/28 (4) 1/14 (7) 3/30 (10) 26/34 (76) 

 Values0 » d 

 < 0.001 < 0.001 N.S. < 0.001* 

 < 0.001** 

Departure from Linear Trende 

 ­ 0.003 - 0.006 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

2.800 

0.242 

141.729 

21.412 

4.109 

770.721 

GO Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

1.400 

0.129 

70.914 

10.706 

2.187 

385.659 

Weeks to First Observed Tumor 78 61 26 
Table Fl· Analyses of the Incidence of Primary Tumors in
 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology 

Liver: Hepatocellular 

Adenoma or Carcinoma^ 

 Valuesc>d 

Departure from Linear Trend8
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled Vehicle Low 

Control Control Dose 

1/28 (4) 1/14 (7) 4/30 (13) 

N.S. N.S. N.S. 

 = 0.023 

3.733 

0.401 

177.009 

1.867 

0.215 

88.558 

87 67 

High
 

Dose
 

0/33 (0)
 

N.S.
 

0.000
 

0.000
 

15.686
 

0.000
 

0.000
 

7.858
 
Table Fl. Analyses of the Incidence of Primary Tumors in
 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Pooled Vehicle Low
 

Topography; Morphology Control Control Dose
 

Ear Canal: Squamous-cell
 

Carcinoma** 0/28 (0) 0/14 (0) 2/30 (7)
 

 Values0

>d N.S. N.S. N.S.
 

Departure from Linear Trende 

 - 0.040
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.282
 

Upper Limit Infinite
 

Relative Risk (Vehicle Control)f Infinite
 Ο
 

Lower Limit 0.147
 

Upper Limit Infinite
 

Weeks to First Observed Tumor 83
 

High
 

Dose
 

0/34 (0)
 

N.S.
 

«...
 

—
 

——
 

mmmm
 
Table Fl. Analyses of the Incidence of Primary Tumors in 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

Pooled Vehicle Low High 

Topography; Morphology Control Control Dose Dose 

Preputial Gland: Squamous-cell 

Carcinoma*5 0/28 (0) 0/14 (0) 6/30 (20) 1/34 (3) 

 Values0 > d N.S. N.S. - 0.014** N.S. 

Departure from Linear Trende « 0.002 = 0.007 

Relative Risk (Pooled Control)f Infinite Infinite 

Lower Limit 1.534 0.045 

Upper Limit Infinite Infinite 

Relative Risk (Vehicle Control)f Infinite Infinite 

Lower Limit 0.805 0.023 

Upper Limit Infinite Infinite 

Weeks to First Observed Tumor 72 56 
Table Fl. Analyses of the Incidence of Primary Tumors in
 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

All Sites: Squamous-cell
 

Carcinoma**
 

 Values0

>d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 Ln
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Control
 

0/28 (0)
 

N.S.
 

 < 0.001
 

Vehicle
 

Control
 

0/14 (0)
 

N.S.
 

 < 0.001
 

Low
 

Dose
 

14/30 (47)
 

 < 0.001**
 

 = 0.001*
 

Infinite
 

4.211
 

Infinite
 

Infinite
 

2.209
 

Infinite
 

67
 

High
 

Dose
 

1/34 (3)
 

N.S.
 

Infinite
 

0.045
 

Infinite
 

Infinite
 

0.023
 

Infinite
 

56 
Table Fl. Analyses of the Incidence of Primary Tumors in
 

Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

a

Treated groups received doses of 1.15 or 2·3 mg/kg.
 

^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 Ρ < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in a treated group is the probability level for the Fisher exact test for
 

the comparison of that treated group with the vehicle-control group (*) or with the pooled-


control group (**) when Ρ < 0.05 for either control group; otherwise, not significant (N.S.)
 

is indicated.
 

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

Kjn
 

e

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

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

control group.
 
Table F2· Analyses of the Incidence of Primary Tumors in
 

Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

Topography: Morphology
 

Lung: Alveolar/Bronchiolar
 

Adenoma**
 

 Values0

>d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)*
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Control
 

0/30 (0)
 

N.S.
 

 - 0.001
 

—
 

Vehicle
 

Control
 

0/15 (0)
 

N.S.
 

 « 0.005
 

——
 

Low
 

Dose
 

5/29 (17)
 

 - 0.024**
 

Infinite
 

1.336
 

Infinite
 

Infinite
 

0.698
 

Infinite
 

86
 

High
 

Dose
 

0/32 (0)
 

N.S.
 

—
 

«...
 

——
 

—
 
Table F2· Analyses of the Incidence of Primary Tumors in
 

Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Pooled Vehicle Low
 

Topography: Morphology Control Control Dose
 

Hematopoietic System: Lymphocytic
 

Leukemia or Lymphoma^ 1/30 (3) 0/15 (0) 5/30 (17)
 

 Values0

»d < 0.001 < 0.001 N.S.
 

Departure from Linear Trende « 0.001 - 0.038
 

Relative Risk (Pooled Control)f 5.000
 

Lower Limit 0.609
 

Upper Limit 227.307
 

Ln
 Relative Risk (Vehicle Control)* Infinite
 

Lower Limit 0.675
 

Upper Limit Infinite
 

Weeks to First Observed Tumor —
 31
 

High
 

Dose
 

32/32 (100)
 

 < 0.001*
 

 < 0.001**
 

30.030
 

8.208
 

Infinite
 

Infinite
 

6.468
 

Infinite
 

26
 
Table F2. Analyses of the Incidence of Primary Tumors in
 

Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology
 

Liver: Hepatocellular
 

Adenoma*5
 

 Values0>d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)*
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)* 

Ui Lower Limit 

Upper Limit 

Weeks to First Observed Tumor
 

Ovary: Carcinoma, NOS^
 

 Values0

>d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)*
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)*
 

Lower Limit
 

Upper Limit
 

Weeks to First o1

 jerved Tumor
 —
 

Pooled
 

Control
 

0/30 (0)
 

N.S.
 

 * 0.040
 

0/29 (0)
 

N.S.
 

 = 0.011
 

Vehicle
 

Control
 

0/15 (0)
 

N.S.
 

...
 

0/15 (0)
 

•N.S.
 

 = 0.030
 

Low
 

Dose
 

2/30 (7)
 

N.S.
 

Infinite
 

0.301
 

Infinite
 

Infinite
 

0.157
 

Infinite
 

86
 

3/29 (10)
 

N.S.
 

Infinite
 

0.615
 

Infinite
 

Infinite
 

0.332
 

Infinite
 

78
 

High
 

Dose
 

0/32 (0)
 

N.S.
 

—
 

—
 

—
 

0/31 (0)
 

N.S.
 

—
 

—
 

—
 
Table F2. Analyses of the Incidence of Primary Tumors in
 

Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

a

Treated groups received doses of 1.15 or 2.3 mg/kg.
 

^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 Ρ < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in a treated group is the probability level for the Fisher exact test for
 

the comparison of that treated group with the vehicle-control group (*) or with the pooled-


control group (**) when Ρ < 0.05 for either control group; otherwise, not significant (N.S.)
 

is indicated.
 

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

e

The probability level for departure from linear trend is given when Ρ < 0.05 for any comparison,
 

f

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

control group·
 
Table F3. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

Topography: Morphology
 

Skin: Squamous-cell Carcinoma*1
 

 Valuese>f
 

Relative Risk (Pooled Control)11
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)11
 

Lower Limit
 

 ι Upper Limit
 r^
 

Ln
 00 Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 

Adenoma**
 

 Valuese

>f
 

Relative Risk (Pooled Control)11
 

Lower Limit
 

Upper Limit
 

Relative Risk (Hatched Control)h
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Control
 

0/18 (0)
 

N.S.
 

1/18 (6)
 

N.S.
 

Vehicle
 

Control
 

0/8 (0)
 

N.S.
 

1/8 (13)
 

N.S.
 

87
 

Low
 

Dose
 

7/24 (29)
 

 - 0.013**
 

Infinite
 

1.544
 

Infinite
 

Infinite
 

0.754
 

Infinite
 

67
 

5/24 (21)
 

N.S.
 

3.750
 

0.479
 

168.323
 

1.667
 

0.246
 

74.916
 

83
 

High
 

Dose
 

0/2 (0)
 

N.S.
 

——
 

0/2 (0)
 

N.S.
 

0.000
 

0.000
 

93.466
 

0.000
 

0.000
 

41.780
 
Table F3· Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology 

Pooled 

Control 

Vehicle 

Control 

Low 

Dose 

High 

Dose 

Hematopoietic System: Lymphoma 

or Lymphocytic Leukemia^ 1/18 (6) 1/8 (13) 2/24 (8) 26/28 (93) 

 Valuese > f < 0.001 < 0.001 N.S. < 0.001* 

 < 0.001** 

Departure from Linear TrendS < 0.001 = 0.002 

Relative Risk (Pooled Control)*1 

Lower Limit 

Upper Limit 

1.500 

0.086 

84.637 

16.714 

3.601 

358.473 

Relative Risk (Matched Control)h 

Lower Limit 

Upper Limit 

0.667 

0.043 

37.663 

7.429 

1.771 

159.766 

Weeks to First Observed Tumor 78 61 26 
Table F3· Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

Topography: Morphology 

Pooled 

Control 

Vehicle 

Control 

Low 

Dose 

High 

Dose 

Hematopoietic System:

or Lymphoma*3 

 Leukemia 

1/18 (6) 1/8 (13) 3/24 (13) 26/28 (93) 

 Valuese 

» f < 0.001 < 0.001 N.S. < 0.001* 

 < 0.001** 

Departure from Linear TrendS = 0.003 = 0.004 

ο 

Relative Risk (Pooled Control)*1 

Lower Limit 

Upper Limit 

Relative Risk (Matched Control)h 

Lower Limit 

Upper Limit 

2.250 

0.202 

112.716 

1.000 

0.103 

50.152 

16.714 

3.601 

358.473 

7.429 

1.771 

159.766 

Weeks to First Observed Tumor 78 61 26 
--

--

--

--

--

--

--

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

in Male Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Pooled Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Liver: Hepatocellular 

Adenoma or Carcinomad 1/18 (6) 1/8 (13) 4/24 (17) 0/2 (0) 

Ρ Valuese,f N.S. N.S. N.S. N.S. 

Relative Risk (Pooled Control)h 3.000 0.000 

Lower Limit 0.336 0.000 

Upper Limit 140.603 93.466 

Relative Risk (Matched Control)h 1.333 0.000 

Lower Limit 0.173 0.000 

Upper Limit 62.559 41.780 

-­ Weeks to First Observed Tumor 87 67 

161 

Ear Canal: Squamous-cell 

Carcinomad 0/18 (0) 0/8 (0) 2/24 (8) 

Ρ Valuese,f N.S. N.S. N.S. 

Relative Risk (Pooled Control)h Infinite 

Lower Limit 0.232 

Upper Limit Infinite 

Relative Risk (Matched Control)h Infinite 

Lower Limit 0.113 

Upper Limit Infinite 

Weeks to First Observed Tumor -­ 83 

0/2 (0)
 

N.S.
 
Table F3· Time-adjusted Analyses of the Incidence of
 

in Male Mice Given Intraperitoneal Injections of
 

(continued)
 

Pooled Vehicle
 

Topography: Morphology Control Control
 

Preputial Gland: Squamous-cell
 

Carcinoma** 0/18 (0) 0/8 (0)
 

 Valuese>f 

 - 0.014 N.S.
 

Relative Risk (Pooled Control)*1
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)11
 

Lower Limit
 

) Upper Limit
 

Weeks to First Observed Tumor
 

All Sites: Squamous-cell
 

Carcinoma** 0/18 (0) 0/8 (0)
 

 Valuese»f 

 - 0.001 - 0.018
 

Relative Risk (Pooled Control)11
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)h
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor -—
 

Primary Tumors
 

Thio-TEPAa
 

Low
 

Dose
 

6/24 (25)
 

 - 0.026**
 

Infinite
 

1.271
 

Infinite
 

Infinite
 

0.620
 

Infinite
 

72
 

14/24 (58)
 

 - 0.004*
 

 < 0.001**
 

Infinite
 

3.513
 

Infinite
 

Infinite
 

1.709
 

Infinite
 

67
 

High
 

Dose
 

1/2 (50)
 

N.S.
 

Infinite
 

1.487
 

Infinite
 

Infinite
 

0.232
 

Infinite
 

1/2 (50)
 

N.S.
 

Infinite
 

0.487
 

Infinite
 

Infinite
 

0.232
 

Infinite
 

56 

56 
Table F3. Time-adjusted Analyses of the Incidence of Primary Tumors 

in Male Mice Given Intraperitoneal Injections of Thio-TEPAa 

(continued) 

a

Treated groups received doses of 1.15 or 2·3 mg/kg·
 

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

at least 26 weeks of the study·
 

c

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

at least 32 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 the control group is the probability level for the Cochran-


M Arraitage test when Ρ < 0.05; otherwise, not significant (N.S·) is indicated. Beneath the
 

w incidence of tumors in a treated group is the probability level for the Fisher exact test for
 

the comparison of that treated group with the vehicle-control group (*) or with the pooled-


control group (**) when Ρ < 0.05; otherwise, not significant (N.S.) is indicated.
 

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

SThe probability level for departure from linear trend is given when Ρ < 0.05 for any comparison.
 

95% confidence interval of the relative risk between each treated group and the control
 

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

in Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

Topography: Morphology 

Pooled 

Control 

Vehicle 

Control 

Lung: Alveolar/Bronchiolar 

Adenoma^ 0/29 (0) 0/14 (0) 

 Valuese > f = 0.034 N.S. 

Relative Risk (Pooled Control)h 

Lower Limit 

Upper Limit 

Relative Risk (Matched Control)h 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor —— 

Hematopoietic System: Lymphoma or 

Lymphocytic Leukemia^ 0/29 (0) 0/14 (0) 

 Valuese , f < 0.001 < 0.001 

Departure from Linear TrendS « 0.006 = 0.011 

Relative Risk (Pooled Control)h 

Lower Limit 

Upper Limit 

Relative Risk (Matched Control)h 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor -_ 

Low
 

Dose
 

5/24 (21)
 

 = 0.015**
 

Infinite
 

1.567
 

Infinite
 

Infinite
 

0.795
 

Infinite
 

86
 

5/26 (19)
 

 = 0.019**
 

Infinite
 

1.445
 

Infinite
 

Infinite
 

0.733
 

Infinite
 

31
 

High
 

Dose
 

0/0 (-)
 

N.S.
 

«...
 

—
 

——
 

——
 

32/32 (100)
 

 < 0.001*
 

 < 0.001**
 

Infinite
 

12.195
 

Infinite
 

Infinite
 

6.060
 

Infinite
 

26
 
Table F4. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued) 

Topography: Morphology 

Pooled 

Control 

Liver: Hepatocellular Adenoma^ 0/29 (0) 

 Valuese > f N.S. 

Relative Risk (Pooled Control)** 

Lower Limit 

Upper Limit 

Relative Risk (Matched Control)h 

Lower Limit 

Upper Limit 

Ln 

Weeks to First Observed Tumor 

Ovary: Carcinoma, NOSd 0/28 (0) 

 Valuese > f N.S. 

Relative Risk (Pooled Control)h 

Lower Limit 

Upper Limit 

Relative Risk (Matched Control)*1 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Vehicle
 

Control
 

0/14 (0)
 

N.S.
 

0/14 (0)
 

N.S.
 

——
 

Low
 

Dose
 

2/25 (8)
 

N.S.
 

Infinite
 

0.350
 

Infinite
 

Infinite
 

0.177
 

Infinite
 

86
 

3/24 (13)
 

N.S.
 

Infinite
 

0.720
 

Infinite
 

Infinite
 

0.377
 

Infinite
 

78
 

High
 

Dose
 

0/0 (-)
 

N.S.
 

—
 

—
 

—
 

—
 

0/0 (-)
 

N.S.
 
Table F4. Time-adjusted Analyses of the Incidence of Primary Tumors
 

in Female Mice Given Intraperitoneal Injections of Thio-TEPAa
 

(continued)
 

a

Treated groups received doses of 1·15 or 2.3 mg/kg·
 

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

at least 26 weeks of the study·
 

c

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

at least 31 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 the control group is the probability level for the Cochran-


Armitage test when Ρ < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in a treated group is the probability level for the Fisher exact test for
 

the comparison of that treated group with the vehicle-control group (*) or with the pooled-


control group (**) when Ρ < 0.05; otherwise, not significant (N.S.) is indicated.
 

^A negative trend (N) indicates a lower incidence in a treated group than in a control group·
 

SThe probability level for departure from linear trend is given when Ρ < 0.05 for any comparison.
 

95% confidence interval of the relative risk between each treated group and the control
 

group.
 
Review of the Bioassay of Thio-TEPA* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

April 26, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be
 

exposed· The members of the Clearinghouse have been drawn
 

from academia, industry, organized labor, public interest
 

groups, State health officials, and quasi-public health and
 

research organizations. Members have been selected on the
 

basis of their experience in carcinogenesis or related
 

fields and, collectively, provide expertise in chemistry,
 

biochemistry, biostatistics, toxicology, pathology, and
 

epidemiology. Representatives of various Governmental
 

agencies participate as ad hoc members. The Data Evaluation/
 

Risk Assessment Subgroup of the Clearinghouse is charged
 

with the responsibility of providing a peer review of
 

reports prepared on NCI-sponsored bioassays of chemicals
 

studied for carcinogenicity. It is in this context that the
 

below critique is given on the bioassay of Thio-TEPA for
 

carcinogenicity.
 

The primary reviewer said that, under the conditions
 

of test, Thio-TEPA induced squamous-cell carcinomas of the
 

skin and ear canal in both sexes of treated rats and
 

hematopoetic neoplasms in the males. Lymphomas or lymphocytic
 

leukemia were induced in both sexes of mice and squamous-cell
 

carcinomas in the skin and associated glands of males. After
 

a brief description of the experimental design and conditions
 

of test, he noted the following points in his critique: the
 

poor survival among the high dose treated animals resulting
 

from excessive toxicity; the higher percentage of bone marrow
 

atrophy reported in control animals than in treated ones;
 

the high mortality among control animals necessitated the
 

use of pooled controls for statistical analysis; the large
 

number of other chemicals tested in the same room and at
 

the same time; and the negative trend for pituitary tumors
 

in treated animals as compared to controls. Despite the
 

experimental shortcomings, the primary reviewer said that
 

the conclusion on the carcinogenicity of Thio-TEPA was
 

still valid. He added that Thio-TEPA would appear to pose
 

a carcinogenic risk to man.
 

167
 
The secondary reviewer agreed with the primary reviewer's
 

critique of the bioassay of Thio-TEPA.
 

It was moved that the report on the bioassay of Thio-
 TEPA be accepted as written. The motion was seconded and
 

approved unanimously.
 

Members present were:
 

Michael Shimkin (Acting Chairman), University of California
 

at San Diego
 

Joseph Highland, Environmental Defense Fund
 

George Roush, Jr., Monsanto Company
 

Louise Strong, University of Texas Health Sciences Center
 

John Weisburger, American Health Foundation
 

*	 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.
 

168 

U.S. GOVERNMENT PRINTING OFFICE: 1978 260-899/3173 1-3 

DHEW Publication (NIH) No. 78-1308