National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 19 

1979 

BIOASSAY OF 

PROCARBAZINE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 366-70-1 

NCI-CG-TR-19 

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

Public Health Service 

National Institutes of Health 

Β10 AS SAY OF
 

PROCARBAZINE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20205
 

U.S. DEPARTMENT OF HEALTH, EDUCATION AND WELFARE
 

Public Health Service
 

National Institutes of Health
 

NIH Publication No. 79-819
 

BIOASSAY OF
 

PROCARBAZINE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

FOREWARD: This report presents the results of the bioassay of
 

procarbazine conducted for the Carcinogenesis Testing Program,
 

Division of Cancer Cause and Prevention, National Cancer
 

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

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

determine whether selected environmental chemicals have the
 

capacity to produce cancer in animals. A negative result, in
 

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

than control animals, does not necessarily mean that the test
 

chemical is not a carcinogen, inasmuch as the experiments are
 

conducted under a limited set of circumstances. A positive
 

result demonstrates 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 chemicals found to be
 

carcinogenic in animals requires a wider analysis.
 

CONTRIBUTORS; This bioassay of procarbazine was conducted by
 

Southern Research Institute (1 \ Birmingham, Alabama, initially
 

under direct contract to NCI and currently under a subcontract to
 

Tracor Jitco, Inc., Rockville, Maryland, prime contractor for the
 

NCI Carcinogenesis Testing Program.
 

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

Griswold (1), J. D· Prejean (1), E. K. Weisburger (2) and J. Η.
 

Weisburger (2,3). Ms. J. Belzer (1) and Mr. I. Brown fl) were
 

responsible for the care of the laboratory animals and the
 

administration of the test chemical. Data management and
 

retrieval were performed by Ms. C. A. Domini ck (1).
 

Histopathologic examinations were performed by Drs. S. D. Kosanke
 

(1) and J. C. Peckham (1), and the diagnoses included in this
 

report represent their interpretation.
 

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

Mason Research Institute (4). Statistical analyses were
 

performed by Dr. J. R. Joiner (5) and Ms. P. L. Yong (5), using
 

methods selected for the bioassay program by Dr. J. J. Gart (6).
 

iii
 
Chemicals used in this bioassay were analyzed under the direction
 

of Mr· C. Hewitt (7), and Dr. P. Lim (8), and analytical results
 

were reviewed by Dr. S. S. Olin (5).
 

This report was prepared at Tracor Jitco (5) under the direction
 

of NCI· Those responsible for the report at Tracor Jitco were
 

Dr. C. R. Angel, Acting Director of the Biossay Program; Dr. S.
 

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

toxicologist; Dr. G. L. Miller, Ms. L. A. Owen, and Mr. W. D.
 

Reichardt, bioscience writers; and Dr. Ε. W. Gunberg, technical
 

editor, assisted by Ms. Υ. Ε. Presley.
 

The following 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 (9), Dr. Richard A.
 

Griesemer, Dr. Harry A. Milman, Dr. Thomas W. Orme, Dr. Robert A.
 

Squire (10), and Dr. Jerrold M. Ward.
 

(1)	 Southern Research Institute, 2000 Ninth Avenue South,
 

Birmingham, Alabama.
 

(2)	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes
 

of Health, Bethesda, Maryland.
 

(3)	 Now with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

(4)	 EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

(5)	 Tracor Jitco, Inc., 1776 East Jefferson Street, Rockville,
 

Maryland.
 

(6)	 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.
 

(7) Drug Development Branch, Division of Cancer Treatment, 

National Cancer Institute, National Institutes of Health, 

Bethesda, Maryland. 

(8) Stanford Research Institute, Menlo Park, California.
 

iv
 
(9) Now with Clement	 Associates, Inc·, 1010 Wisconsin Avenue,
 

N.W., Suite 660, Washington D.C.
 

(10) Now with the Division of Comparative Medicine, Johns Hopkins
 

university, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of procarbazine for possible carcinogenicity was conducted
 

by administering the test chemical by intraperitoneal injection to
 

Sprague-Dawley rats and B6C3F1 mice.
 

Groups of 34 or 35 males and 35 or 36 females of both species were
 

administered procarbazine at one of two doses, either 15 or 30 mg/kg
 

for rats, and either 6 or 12 mg/kg for mice. Injections were made
 

three times per week for 26 weeks for the rats and 52 weeks for the
 

mice. Following the periods of injection, the dosed animals were
 

observed for a maximum period of 60 weeks for rats and 33 weeks for
 

mice, depending on survival. Vehicle controls, used for statistical
 

evaluation, consisted of 10 rats and 15 mice of each sex, administered saline solution on the same schedule as the test solution; the
 

same numbers of rats and mice served as untreated controls. Pooled
 

controls consisted of the vehicle controls from this bioassay
 

together with the vehicle controls from two other bioassays
 

similarly performed at the same laboratory. The pooled-control
 

groups consisted of 40 rats of each sex and 45 mice of each sex.
 

Surviving rats were killed at 86 weeks and surviving mice were
 

killed at 85 weeks.
 

Mean body weights of low- and high-dose rats and of high-dose female
 

mice were lower than those of the vehicle controls. Survival rates
 

of both rats and mice showed significant dose-related trends.
 

In rats, malignant lymphomas, adenocarcinomas of the mammary gland,
 

and the combination of olfactory neuroblastomas, adenocarcinomas, or
 

carcinomas of the brain, olfactory bulb, or cerebrum were induced in
 

statistically significant numbers.
 

In mice, malignant lymphomas or leukemias, olfactory neuroblastomas
 

or undifferentiated carcinomas, alveolar/bronchiolar adenomas, and
 

adenocarcinomas of the uterus were induced in statistically significant numbers.
 

It is concluded that under the conditions of this bioassay, procarbazine was carcinogenic for both Sprague-Dawley rats and B6C3F1 mice,
 

producing several types of tumors in both sexes of these two species.
 

vii
 

TABLE OF CONTENTS
 

Page
 

I· Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dosage Preparation 3
 

C. Animals 4
 

D. Animal Maintenance 4
 

E. Chronic Studies 8
 

F. Clinical and Pathologic Examinations 11
 

G. Data Recording and Statistical Analyses 12
 

III. Results - Rats 19
 

A. Body Weights and Clinical Signs (Rats) 19
 

B. Survival (Rats) 19
 

C. Pathology (Rats) 23
 

D. Statistical Analyses of Results (Rats) 26
 

IV. Results - Mice 31
 

A. Body Weights and Clinical Signs (Mice) 31
 

B. Survival (Mice) 31
 

C. Pathology (Mice) 35
 

D. Statistical Analyses of Results (Mice) 37
 

V. Discussion ······ 43
 

VI. Bibliography 49
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in Rats
 

Administered Procarbazine by Intraperitoneal
 

Inj ec ti on 51
 

Table Al Summary of the Incidence of Neoplasms in Male
 

Rats Administered Procarbazine by Intraperitoneal Injection 53
 

Table A2 Summary of the Incidence of Neoplasms in Female
 

Rats Administered Procarbazine by Intraperitoneal Injection 57
 

ix
 
Page 

Appendix Β Summary of the Incidence of Neoplasms in Mice 

Administered Procarbazine by Intraperitoneal Injection 61 

Table Bl Summary of the Incidence of Neoplasms in Male 

*Mice Administered Procarbazine by Intraperitoneal Injection 63 

Table B2 Summary of the Incidence of Neoplasms in Female 

Mice Administered Procarbazine by Intraperitoneal Injection 66 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Administered Procarbazine 

by Intraperitoneal Injection 71 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Administered 

Procarbazine by Intraperitoneal Injection .... 73 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Administered 

Procarbazine by Intraperitoneal Injection .... 78 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Administered Procarbazine 

by Intraperitoneal Injection 83 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Administered 

Procarbazine by Intraperitoneal Injection .... 85 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Administered 

Procarbazine by Intraperitoneal Injection .... 88 

Appendix Ε Analyses of the Incidence of Primary Tumors in 

Rats Administered Procarbazine by Intraperitoneal Injection 91 

Table El Analyses of the Incidence of Primary Tumors in 

Male Rats Administered Procarbazine by Intraperitoneal Injection 93 

Table E2 Analyses of the Incidence of Primary Tumors in 

Female Rats Administered Procarbazine by 

Intraperitoneal Injection 99 

χ 
Page 

Appendix F Analyses of Hie Incidence of Primary Tumors in 

Mice Administered Procarbazine by Intraperi­ toneal Injection 109 

Table Fl Analyses of the Incidence of Primary Tumors in 

Male Mice Administered Procarbazine by Intraperitoneal Injection . ·.. Ill 

Table F2 Analyses of the Incidence of Primary Tumors in 

Female Mice Administered Procarbazine by 

Intraperitoneal Injection 118 

TABLES 

Table 1 Design of the Chronic Studies of Procarbazine 

in Rats 9 

Table 2 Design of the Chronic Studies of Procarbazine 

in Mice. • • 10 

FIGURES 

Figure 1

Figure 2

 Growth Curves for Rats Treated with 

Procarbazine

 Survival Curves for Rats Treated with 

Procarbazine · ·. ·

 20 

 21 

Figure 3 Growth Curves for Mice Treated with 

Procarbazine · 32 

Figure 4 Survival Curves for Mice Treated with 

Procarbazine 33 

xi
 

I. INTRODUCTION
 

Procarbazine
 

Procarbazine (CAS 366-70-1; NCI C01810) is a methylhydrazine
 

derivative which has been shown to have effective antineoplastic
 

activity in advanced Hodgkin's disease and in oat-cell carcinoma
 

of the lung (Oliverio, 1973; Carter and Slavik, 1974). It has
 

also been shown to have carcinogenic activity in rats and mice
 

(Kelly et al., 1968; Kelly et al., 1969). The mechanism of the
 

cytotoxic action of this drug is not understood, although it is
 

clear that it leads to the inhibition of protein, RNA, and DNA
 

synthesis. Its oxidative metabolic products include formaldehyde, N-hydroxymethyl derivatives, and hydrogen peroxide, which
 

are capable of carcinostatic effects, and azomethine, which has
 

been shown to have carcinostatic and carcinogenic effects similar
 

to those of procarbazine (Oliverio, 1973). Methylation of
 

nucleic acids by the N-methyl group of procarbazine is also being
 

1 
studied (Oliverio, 1973; Carter and Slavik, 1974). Procarbazine
 

was selected for screening by the carcinogenesis bioassay program
 

in an attempt to evaluate the carcinogenic effects of certain
 

anticancer agents and other drugs which are used extensively and
 

for prolonged periods in humans.
 

2 
II. MATERIALS AND METHODS
 

A. Chemical
 

Procarbazine hydrochloride, which is the generic name for
 

N-iso-propyl-a-(2-methylhydrazino)-p-toluamide hydrochloride, was
 

purchased from Hoffman La Roche, Nutley, New Jersey, by the Drug
 

Development Branch, Division of Cancer Treatment, National Cancer
 

Institute. Elemental analyses (C, H, N, Cl) of the single batch
 

(Lot No. PP-9) gave a percentage composition equivalent to
 

theoretical values. In paper chromatographic analysis, the
 

sample showed two minor ultraviolet-absorbing impurities. No
 

attempt was made to identify or quantitate these impurities. The
 

infrared spectrum was comparable to spectra obtained from samples
 

known to be pure. Nuclear magnetic resonance and ultraviolet
 

spectra were as expected for procarbazine hydrochloride. The
 

chemical was stored at -20 C.
 

B. Dosage Preparation
 

Concentrations of 0.60 and 1.2% (w/v) procarbazine for rats and
 

0.06 and 0.12% (w/v) for mice were prepared in buffered saline
 

(pH 6.9) for intraperitoneal injection of the chemical. To
 

3 
minimize decomposition of the drug, fresh solutions were prepared
 

immediately before injection of the test animals.
 

C. Animals
 

Sprague-Dawley rats and B6C3F1 mice were obtained from the
 

Charles River Breeding Laboratories, Inc., Wilmington,
 

Massachusetts, through contracts with the Division of Cancer
 

Treatment, National Cancer Institute. Upon arrival at the
 

laboratory, the male rats were 28 days old, the females 35 days
 

old, and the male and female mice 28 days old. All animals were
 

housed within the test facility for 1 week. Animals with no
 

clinical signs of disease were assigned to dosed or control
 

groups and were earmarked for individual identification.
 

D. Animal Maintenance
 

All animals were housed in temperature-and humidity-controlled
 

rooms. Air was maintained at 20 to 24 C and 40 to 60% relative
 

humidity* Fresh air was filtered through fiberglass roughing
 

filters and was changed 15 times per hour. In addition to
 

natural light, illumination was provided by fluorescent light for
 

4 
9 hours per day. Wayne Sterilizable Lab Blox (Allied Mills,
 

Inc., Chicago, 111.) and water were supplied daily and were
 

available aci libitum*
 

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.). The bottoms of the rat cages were lined
 

with Iso-Dri hardwood chips (Carworth, Edison, N.J.), and cage
 

tops were covered with disposable filter bonnets beginning at
 

week 22; mouse cages were provided with Sterolit clay bedding
 

(Englehard Mineral and Chemical Co., New York, N.Y.). Bedding
 

was replaced one time per week; cages, water bottles, and feeders
 

were sanitized at 82 C one time per week; and racks were
 

cleaned one time per week.
 

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

were housed in the same room as their respective control
 

animals. Animals administered procarbazine were maintained in
 

the same rooms as animals of the same species administered the
 

following chemicals:
 

5 
RATS
 

Feeding Studies
 

(CAS 136-40-3)


Gavage Studies
 

(CAS 3546-10-9)


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

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

acetate) (phenesterin)
 

(CAS 22966-79-6) estradiol bis((p-(bis(2-chloroethyl)amino)phenyl)
 

acetate) (estradiol mustard)
 

Intraperitoneal Injection Studies
 

(CAS 3458-22-8) 3,3'-iminobis-l-propanol dimethanesulfonate 

(ester) hydrochloride (IPD) 

(CAS 320-67-2) 5-azacytidine 

(CAS 21416-87-5) (+)-4,4f-(l-methyl-l,2-ethanediyl)bis-2,6­

(CAS 789-61-7)


(CAS 55-98-1)


(CAS 7008-42-6)


(CAS 483-18-1)


(CAS 3778-73-2)


(CAS 52-24-4)


(CAS 63-92-3)


(NSC 141549)


MICE
 

Feeding Studies
 

(CAS 118-92-3)

(CAS 98-96-4)

(CAS 73-22-3)

piperazinedione (ICRF-159)
 

beta-2f-deoxy-6-thioguanosine (0-TGDR)
 

 1,4-butanediol dimethanesulfonate (busulfon)
 

 acronycine
 

 emetine dihydrochloride tetrahydrate
 

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

oxazaphosphorin-2-amine-2-oxide (isophosphamide)
 

 tris(l-aziridinyl)phosphine sulfide (thioTEPA)
 

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

benzylamine hydrochloride (phenoxybenzamine
 

hydrochloride)
 

4f-(9-acridinylamino)methansulfon-m-aniside
 

monohydrochloride (MAAM)
 

 anthranilic acid 

 pyrazinecarboxamide 

 L-tryptophan 

6 
(CAS 64-77-7)


(CAS 80-08-0)


(CAS 139-65-1)

(CAS 136-40-3)


(CAS 536-33-4)


(CAS 58-14-0)

(CAS 94-20-2)


(CAS 53-96-3)


(CAS 114-86-3)


(CAS 968-81-0)


(CAS 1156-19-0)


Gavage Studies
 

(CAS 3546-10-9)


 l-butyl-3-(p~tolylsulfonyl)urea (tolbutamide)
 

 4,4'-sulfonyldianiline
 

 4,4'-thiodianiline 

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

 2-ethyl-4-pyridinecarbothioamide (ethionamide)
 

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

(pyrimethamine)
 

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

 N-9H-fluorenyl-2-acetamide
 

 1-phenethylbiguanide hydrochloride(phenformin)
 

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

benzenesulfonamide (acetohexamide)
 

N-(p-toluenesulfonyl)-Nf

-hexamethyleniminourea
 

(tolazamide)
 

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

acetate (phenesterin)
 

(CAS 22966-79-6) estradiol bis((p-bis(2-chloroethyl)amino)phenyl)­

acetate) (estradiol mustard)
 

Intraperitoneal Injection Studies
 

(CAS 3458-22-8) 3,3'-iminobis-l-propanol dimethanesulfonate 

(ester) hydrochloride (IPD) 

(CAS 320-67-2) 5-azacytidine 

(CAS 21416-87-5) (+)-4,4'-(l-methyl-l,2-ethanediyl)bis-2,6-piper­

(CAS 789-61-7)


(CAS 55-98-1)


(CAS 7008-42-6)


(CAS-483-18-1)


(CAS 3778-73-2)


(CAS 52-24-4)


(CAS 63-92-3)


(NSC 141549)


(CAS 645-05-6)


azinedione (lCRF-159)
 

 beta-2'-dexy-6-thioguanosine (0-TGDR)
 

 1,4-butanediol dimethanesulfonate (busulfan)
 

 acronycine
 

 emetine dihydrochloride tetrahydrate
 

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

oxazaphosphorin-2-amine-2-oxide (isophosphamide)
 

 tris(l-aziridinyl)phosphine sulfide (thio-TEPA)
 

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

benzylamine hydrochloride (phenoxybenzamine
 

hydrochloride)
 

 4'-(9-acridinylamino)methanesulfon-m-aniside
 

monohydrochloride (MAAM)
 

 2,4,6-tris(dimethylamino)-s-triazine
 

7
 
. Chronic Studies
 

The test groups, doses administered, and durations of the chronic
 

studies are shown in tables 1 and 2.
 

Since the numbers of animals in the vehicle-control groups were
 

small, pooled-control groups also were used for statistical
 

comparisons. The pooled-control groups consisted of the vehicle
 

controls from the bioassay of procarbazine combined with the
 

vehicle controls from the bioassays of 3,3'-iminobis-1-propanol
 

dimethanesulfonate (ester) hydrochloride (CAS 3458-22-8) and
 

isophosphamide (CAS 3778-73-2), to give groups of 40 male or 40
 

female rats and 45 male or 45 female mice· The bioassays of the
 

two test chemicals other than procarbazine were also conducted at
 

Southern Research Institute and overlapped the bioassay of
 

procarbazine by at least 1 year using rats and at least 15 months
 

using mice· The vehicle-control groups of rats and the
 

vehicle-control groups of mice that were used in the respective
 

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

the same supplier, and their tissues were diagnosed by the same
 

pathologist.
 

The doses for the chronic studies were established on the basis
 

of results of an earlier study at Southern Research Institute in
 

8
 
Table 1. Procarbazine Chronic Studies in Rats
 

Sex and Initial Procarbazine Time on Study 

Test No. of Dosage (b) Dosed Observed 

Group Animals (a) (mg/kg) (weeks) (weeks) 

Male
 

Untreated-Control 10 0 86
 

Vehicle-Control(c ) 10 0 26 60
 

Low-Dose 34 15 26 34
 

High-Dose 35 30 26 17(d)
 

Female
 

Untreated-Control 10 0 86
 

Vehicle-Control(c ) 10 0 26 60(d)
 

Low-Dose 36 15 26 27(d)
 

High-Dose 35 30 26 5
 

(a)	 The males were 35 days of age, and the females were 42 days
 

of age when placed on study; however, all animals were
 

placed on study at the same time·
 

(b)	 Procarbazine was administered intraperitoneally three times
 

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

body weight during the period of administration·
 

(c)	 Vehicle controls were administered buffered saline (0.25
 

ml/100 g body weight).
 

(d)	 Observation of high-dose males and of low- and high-dose
 

females was terminated at the times indicated, due to the
 

deaths of all animals.
 

9 
Table 2. Procarbazine Chronic Studies in Mice
 

Sex and Initial Procarbazine Time on Study
 

Test No. of Dosage (b) Dosed Observed
 

Group Animals (a) (mg/kg) (weeks) (weeks)
 

Male
 

Untreated-Control 15 0 85
 

Vehicle-Control(c ) 15 0 52 33
 

Low-Dose 35 6 52 33
 

High-Dose 35 12 52 15(d)
 

Female
 

Untreated-Control 15 0 85 

Vehicle-Control(c) 15 0 52 33 

Low-Dose 35 6 52 33 

High-Dose 35 12 52 15(d) 

(a)	 All animals were 35 days of age when placed on study; all
 

animals were placed on study at the same time.
 

(b)	 Procarbazine was administered intraperitoneally three times
 

per week in buffered saline at a volume of 1.0 ml/100 g body
 

weight during the period of administration.
 

(c)	 Vehicle controls were administered buffered saline (1.0
 

ml/100 g body weight).
 

(d)	 Observation of the high-dose group was terminated, at the
 

times indicated, due to the deaths of all animals.
 

10
 
which procarbazine was administered three times per week for 6
 

months to Sprague-Dawley rats and Swiss Webster mice. The doses
 

selected (referred to in this report as "high doses11 and "low
 

doses") were 30 and 15 mg/kg for the Sprague-Dawley rats and 12
 

and 6 mg/kg for the B6C3F1 mice used in the present bioassay.
 

F · Clinical and Pathologic Examinations
 

All animals were observed twice daily. Observations to identify
 

sick, tumor-bearing, and moribund animals were made daily.
 

Animals were weighed individually each week for the first 8 weeks
 

and every 2 weeks thereafter, and palpated for masses at each
 

weighing. Animals that were moribund at the time of daily
 

examination were killed and necropsied.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of major tissues, major organs, and all gross
 

lesions. The tissues were preserved in 10% neutral buffered
 

formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin. Special staining techniques were utilized when
 

indicated. The following tissues were examined microscopically:
 

skin, muscle, lungs and bronchi, trachea, bone and bone marrow,
 

spleen, lymph nodes, thymus, heart, salivary gland, liver,
 

11
 
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. Occasionally, additional tissues were also examined microscopically.
 

Necropsies were also performed on all animals found dead, unless
 

precluded in whole or in part by autolysis or cannibalization.
 

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.
 

G. 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
 

12
 
tables were generated for verification of data transcription and
 

for statistical review.
 

These data were analyzed using the appropriate statistical
 

techniques described in this section. Those analyses of the
 

experimental results that bear on the possibility of carcinogenicity are discussed in the statistical narrative sections.
 

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 Taronef

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
 

13
 
in which that site is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic
 

examination was required to detect lesions prior to histologic
 

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

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

denominators consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

significantly higher proportion of tumors than did the control
 

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

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
 

14
 
continuity correction (Armitage, 1971), is also used. Under the
 

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

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

one-tailed 0.05 level of significance. Unless otherwise noted,
 

the direction of the significant trend is a positive dose
 

relationship. This method also provides a two-tailed test of
 

departure from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

first tumor was observed were excluded by basing the statistical
 

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

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).
 

15
 
The week during which an animal died naturally or was sacrificed
 

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

methods of comparing these curves were used for two groups;
 

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

groups. The statistical tests for the incidence of tumors which
 

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

noted, in the direction of a positive dose relationship.
 

Significant departures from linearity (P less than 0.05,
 

two-tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared to its control was calculated
 

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

relative risk is defined as ρ / where ρ is the true
 

L C C
 

binomial probability of the incidence of a specific type of tumor
 

in a dosed group of animals and ρ 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
 

16
 
relative risk have been included in the tables of statistical
 

analyses. The interpretation of the limits is that in
 

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

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

control group would be within the interval calculated from the
 

experiment· When the lower limit of the confidence interval is
 

greater than one, it can be inferred that a statistically
 

significant result (P less than 0.025 one-tailed test when the
 

control incidence is not zero, Ρ less than 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.
 

17
 

III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

The mean body weights of the high- and low-dose rats of each sex
 

were lower than those of the corresponding vehicle and untreated
 

controls from approximately week 10 on study to the end of the
 

survival periods (figure 1). Fluctuation in the growth curves
 

may be due to mortality; as the size of a group dimishes, the
 

mean body weight may be subject to wide variations. No records
 

were kept of specific clinical signs; however, as a part of a
 

colony treatment for control of an intercurrent respiratory
 

disease, the animals in this study were administered oxytetracycline in the drinking water during weeks 23 to 29 (0.6 mg/ml)
 

and weeks 29 to 34 (0.3 mg/ml).
 

B. Survival (Rats)
 

Estimates of the probabilities of survival for male and female
 

rats receiving procarbazine at the doses used in this bioassay,
 

together with those of the untreated and vehicle controls, are
 

shown by the Kaplan and Meier curves in figure 2.
 

19
 
TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves For Rats Treated With Procarbazine 

20 
TIME ON STUDY (WEEKS) 

Figure 2. Survival Curves For Rats Treated With Procarbazine 

21 
In both sexes, the Tarone test results for positive dose-related
 

trend in the proportions of dosed animals compared to vehicle
 

controls surviving over the test period are significant (P less
 

than 0.001).
 

In high-dose males, all of the animals died before week 44 of the
 

study, with a median time on study at death of 26 weeks; 30/33
 

high-dose males that were necropsied were observed to have some
 

kind of tumor. Mortality of the low-dose group was almost as
 

high as that of the high-dose group. All of the low-dose males
 

died before week 60, with a median time on study at death of 31
 

weeks; however, tumors were observed in 19/30 low-dose animals
 

that were necropsied. Mortality was low among the vehicle-control
 

males, with 80% of the animals living to the end of the study.
 

The dosed females also had poor survival, with no animal living
 

to the end of the study. Tumors were observed in 27/30 low-dose
 

females that were necropsied; the median time on study at death
 

for this group was 31 weeks. In high-dose females, 30/31 animals
 

that were necropsied had tumors, and the median time on study at
 

death for this group was 22 weeks. Survival was high among the
 

vehicle-control females, with 89% of the animals living to
 

termination of the study.
 

22
 
C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

are summarized in Appendix C, tables Cl and C2.
 

The frequency of animals developing tumors and the mean numbers
 

of tumors per animal increased in both the high- and low-dose
 

groups of males and females.
 

The primary tissues affected were neuroepithelial, epithelial,
 

and lymphoreticular or hematopoietic. The tumors observed
 

included 45 olfactory and nasal neuroblastomas, adenocarcinomas,
 

and carcinomas in 41 animals, 32 malignant lymphomas of
 

lymphocytic type, and 52 adenocarcinomas or cystadenocarcinomas
 

of the mammary gland. The neuroepithelial tumors, coded as
 

"olfactory neuroblastomas11

 were unusual. They have also been
 

termed "esthesioneuroepitheliomas" (Laskin et al., 1971) and
 

"olfactory neuroepithelial tumors11 (Herrold, 1964).
 

Histologic features which characterize the olfactory neuroblastomas (Obert et al., 1960) are:
 

23
 
(1)	 plexiform intercellular fibrils;
 

(2)	 poorly defined, almost nonexistent, cytoplasm;
 

(3)	 round to oval, usually oval, nuclei;
 

(4)	 chromatin usually distinct and sharply defined, but
 

either coarse or fine;
 

(5)	 compartmentation of sheets of neoplastic cells into
 

lobules by slender vascular fibrous septa; and
 

(6)	 true rosettes and pseudorosettes.
 

The tumors observed in this study met these criteria. In
 

addition, some tumors had areas of adenocarcinomatous tissue
 

which arose from the nasal epithelium and was mixed or associated
 

with the neoplastic neural tissues. The tumors extended
 

posteriorly into the adjacent brain. Lysis of the cranial bones
 

was observed. Olfactory neuroblastomas have not been seen in
 

untreated rats of this strain at Southern Research Institute.
 

The mammary glands of the dosed animals had an increased
 

proportion of malignant tumors when compared with the control
 

groups of this experiment and with untreated Sprague-Dawley rats
 

previously observed at Southern Research Institute (Prejean et
 

al., 1973). Malignant lymphoma, lymphocytic type (previously
 

termed "disseminated lymphosarcoma11

 or "lymphocytic leukemia")
 

and granulocytic leukemia occurred also in a greatly increased
 

number of animals.
 

24
 
The majority of nonneoplastic proliferative lesions involved the
 

lymphoreticular tissues and mammary glands of the dosed groups.
 

The hyperplasias observed in the mammary glands of some animals
 

not having tumors may have been associated with administration of
 

the test chemical.
 

In addition, a variety of other neoplasms that have been
 

encountered previously as spontaneous lesions in the rat occurred
 

in both dosed and control (untreated and vehicle) groups.
 

The dosed animals had shorter life spans than those in the
 

untreated- and vehicle-control groups. The high frequency of
 

neoplasia suggests that the reduction in life span is directly
 

related to carcinogenesis. However, many of the rats in both the
 

low-dose and high-dose groups had respiratory disease that was
 

believed to be chronic murine pneumonia.
 

Intraperitoneal administration of procarbazine to Sprague-Dawley
 

rats resulted in the occurrence of tumors associated with
 

administration of the test chemical. The principal tumors
 

observed were olfactory neuroblastomas, adenocarcinomas of the
 

mammary gland, and malignant lymphomas, lymphocytic type.
 

25
 
D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix Ε contain the statistical analyses
 

of the incidences of those specific primary tumors that were
 

observed in at least 5% of one or more dosed groups of either
 

sex. The untreated controls are not included in the tables,
 

since the test conditions of the vehicle controls were closer to
 

those of the dosed animals.
 

In male rats, the Cochran-Armitage test results for positive
 

dose-related trend in the incidence of lymphoma of the
 

hematopoietic system are highly significant (P = 0.002) when the
 

pooled-control group is used. The Fisher exact test shows that
 

the incidence in the high-dose males is significantly higher than
 

that in the pooled controls (P = 0·003). The lower limit of the
 

95% confidence interval of the relative risk shows a value
 

greater than one. In females, the Cochran—Armitage test results
 

are highly significant (P less than 0.001) when either the
 

pooled-control group or the vehicle-control group is used. The
 

departure from linear trend is highly significant (P less than
 

0.001) when the pooled-control group is used, and Ρ = 0·004 when
 

the vehicle-control group is used, since there is a steep rise in
 

incidence in the high-dose animals. The Fisher exact test shows
 

that the incidence in the high-dose females is significantly
 

26
 
higher ( less than 0.001) than that in either the pooled
 

controls or the vehicle controls. This positive finding is also
 

shown by the lower limits of the 95% confidence intervals of
 

these relative risks, which have values greater than one. The
 

statistical conclusion is that the incidence of lymphoma of the
 

hematopoietic system in rats is associated with procarbazine for
 

high-dose male and female rats in this experiment.
 

Leukemia was found exclusively in the high-dose males (3/33), and
 

a significant Cochran-Armitage test result (P = 0.031) is
 

observed when the pooled-control group is used; however, none of
 

the Fisher exact test results is significant; therefore, it is
 

inconclusive whether the incidence of leukemia in male rats is
 

related to administration of the chemical. No such tumor was
 

observed in females.
 

When these hematopoietic tumors (lymphoma and leukemia) are
 

grouped for analyses in male rats, the significance of the
 

Cochran-Armitage test becomes Ρ less than 0.001 and Ρ = 0.013
 

when the pooled controls and the vehicle controls are used,
 

respectively. The incidence of the combination of lymphoma and
 

leukemia in the high-dose groups compared with that in the pooled
 

controls is statistically significant due to the high incidence
 

of lymphoma.
 

27
 
In this study, adenocarcinomas, NOS (not otherwise specified), of
 

the mammary gland were found only in the dosed animals. In male
 

rats, the Cochran-Armitage test results are significant when the
 

pooled controls (P = 0.001) and the vehicle controls (P = 0.016)
 

are used. The Fisher exact test shows that the incidence in the
 

high-dose males is significantly higher than that in the pooled
 

controls (P = 0.003). This positive finding results in the value
 

of the lower limit of the 95% confidence interval of the relative
 

risk being greater than one. In females, the Cochran-Armitage
 

test results are significant (P less than 0.001) when either the
 

pooled-control group or the vehicle-control group is used. This
 

positive finding is confirmed by the Fisher exact test, where the
 

comparisons between the incidences in the dosed groups and the
 

control groups are highly significant (probability levels 0.003
 

or less) and the lower limits of the 95% confidence intervals of
 

these relative risks have values greater than one. The
 

statistical conclusion is that the occurrence of adenocarcinomas,
 

NOS, of the mammary gland in rats is related to administration of
 

procarbazine. One adenoma of the mammary gland was found in the
 

male high-dose group and another was observed in the female
 

low-dose group. The analyses of the incidence of the grouped
 

mammary gland tumors (adenomas and adenocarcinomas, NOS) show
 

increased significance over that of the adenocarcinomas, NOS,
 

alone.
 

28
 
There were three high-dose male rats with adenocarcinomas, NOS,
 

of the olfactory bulb, and the results of the Cochran-Armitage
 

test show in a probability level of 0.034 when the pooled-control
 

group is used. None of the Fisher exact test results are
 

significant. When the incidences of carcinomas, neuroblastomas
 

and adeno carcinomas of the brain and olfactory bulb are
 

combined, the Cochran-Armitage test results are significant (P =
 

0.003) using the pooled-control group. Departures from linear
 

trend are present (P = 0.001, pooled control; Ρ = 0.013, matched
 

control) because the incidences in the low-dose group exceeds
 

that in the high-dose. The results of the Fisher exact test
 

using the pooled controls are significant (P less than or equal
 

to 0.001) in both dosed groups. It is concluded that the
 

incidences of these tumors are related to administration of the
 

chemical. In females, although the results of the Cochran-


Armitage test on the incidence of olfactory neuroblastomas or
 

mucinous adenocarcinomas of the brain or cerebrum are not
 

significant, there is an indicated departure from linear trend (P
 

less than 0.001), since the proportion is higher in the low-dose
 

group than in the high-dose group. The Fisher exact test shows
 

that the incidence in the low-dose females is significantly
 

higher than that in either the vehicle or pooled-control groups
 

(P less than or equal to 0.007).
 

29
 
Brain, ear, and olfactory tumors were observed in both sexes as
 

early as 21 weeks. When groupings of the types of tumors are
 

made, as in adenomas and adenocarcinomas, NOS, of the mammary
 

gland, the incidences of the individual components are not
 

included in tables El and E2 unless the proportions are greater
 

than 5% in any of the dosed groups. However, a list of the
 

incidences of each type of tumor is provided in tables El and E2
 

of Appendix E. In summary, there are three types of tumors that
 

appear to be associated with this chemical and four other types
 

where the evidence is not statistically conclusive.
 

30
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

The mean body weights of the dosed mice were not consistently
 

different from those of the controls (figure 3)· Those high-dose
 

males still alive had low mean body weights starting at about
 

week 40. Weights of high-dose and vehicle-control females were
 

lower than those of low-dose and untreated-control females
 

starting at approximately week 30 on study. Fluctuation in the
 

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

diminishes, the mean body weight may be subject to wide variation.
 

B· Survival (Mice)
 

Estimates of the probabilities of survival of male and female
 

mice receiving procarbazine at the doses used in this bioassay,
 

together with those of the untreated and vehicle controls, are
 

shown by the Kaplan and Meier curves in figure 4.
 

In male mice, the Tarone test for positive dose-related trend in
 

the proportions of dosed animals compared with vehicle controls
 

surviving over the test period is highly significant (P less than
 

31
 
TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves For Mice Treated With Procarbazine 

32 
TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves For Mice Treated With Procarbazine 

33 
0.001), and a departure from linear trend (P = 0·012) is
 

observed, due to the sharp rise in mortality of the high-dose
 

group. Forty percent of the vehicle controls and 31% of the
 

low-dose group, but none of the high-dose group, lived to the end
 

of the study; their respective median times on study at death
 

were 62 weeks, 75 weeks, and 56 weeks. None of the vehicle
 

controls developed tumors. In the low-dose male mice, 18/30 that
 

were necropsied had developed tumors, and in the high-dose males,
 

20/31 animals that were necropsied had evidence of tumors;
 

therefore, there is a possibility that the early deaths were
 

associated with the administration of procarbazine.
 

In females, the Tarone test results are highly significant (P
 

less than 0.001), and a departure from linear trend (P = 0.049)
 

is observed, due to the steep increase in the mortality of the
 

dosed groups. Eighty percent of the vehicle controls and 21% of
 

the low-dose group, but none of the high-dose group, survived to
 

termination of the study. The median times on study at death
 

were 54 weeks and 66 weeks for the high-dose and low-dose
 

animals, respectively. In the low-dose group, 19/23 animals
 

necropsied had developed tumors, and in the high-dose group,
 

20/26 animals necropsied were observed to have tumors.
 

34
 
C. Pathology (Mice)
 

Histopathlogic 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.
 

Malignant tumors of the olfactory bulbs and mucosa were observed
 

in 23 high-dose male ancj female mice. The majority of these
 

tumors were classified as olfactory neuroblastomas and resembled
 

those observed in rats, as described previously. One of these
 

olfactory neuroblastomas had invaded the adjacent cranial and
 

facial bones. One mouse had a brain lesion which was classified
 

as an undifferentiated carcinoma. One tumor of the brain,
 

composed of reticulum cells or histiocytes, was coded as a
 

sarcoma, NOS.
 

Another organ frequently affected by neoplasia was the uterus in
 

both the high- and low-dose females. The most commonly observed
 

uterine tumors were adenocarcinomas (14 in the low-dose group, 8
 

in the high-dose group, but none in the controls). These
 

adenocarcinomas penetrated the serosa and invaded or proliferated
 

into adjacent tissues of the abdomen. Metastatic uterine
 

adenocarcinomas were observed in the lungs, heart, and liver.
 

35
 
Four dosed mice had uterine leiomyosarcomas and one had a
 

malignant lymphoma of histiocytic type. One mouse had an
 

undifferentiated spindle-cell sarcoma of the urinary bladder,
 

coded as a sarcoma, NOS, which also involved the uterus, oviduct,
 

ovaries, and mesenteric lymph nodes.
 

A variety of other neoplasms occurred in both dosed and control
 

(untreated and vehicle) groups, which- have been encountered
 

previously as spontaneous lesions in the mouse. Nonneoplastic,
 

proliferative lesions involved primarily the lymphoreticular
 

tissues. A few mice had suppurative lesions suggestive of
 

bacterial infections. These lesions were not associated with
 

increased mortality or decreased life spans.
 

The high-dose groups had slightly reduced life spans; the median
 

time of death was only slightly above a year. The high frequency
 

of neoplasia and the extensive involvement of the brain and
 

uterus could account for the majority of the deaths prior to
 

termination of the study. The cause of the mortalities in the
 

vehicle-control group could not be explained on the basis of
 

postmortem examination of lesions.
 

The results of this study indicate that olfactory neuroblastomas
 

in males and females and uterine adenocarcinomas in females were
 

36
 
related to administration of the chemical· Neither of these
 

tumors was observed in the control groups.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that were observed in
 

at least 5% of one or more dosed groups of either sex. The
 

untreated controls are not included in the tables, since the test
 

conditions of the vehicle controls were closer to those of the
 

dosed groups.
 

Alveolar/bronchiolar adenomas of the lungs occurred exclusively
 

in the dosed groups. For males, the Cochran-Armitage test for
 

positive dose-related trend in proportions has a probability
 

level of less than 0.001 when the pooled controls are used. The
 

Fisher exact test shows that the incidences in the dosed groups
 

are significantly higher than those in either the vehicle
 

controls (P = 0.020) or the pooled controls (P less than 0.001);
 

and this positive finding is accentuated by the values of the
 

lower limits of the 95% confidence interval of the relative risks
 

of the dosed group versus the control groups, which are greater
 

than one. In females, the incidence of alveolar /bronchiolar
 

37
 
adenomas in the high-dose group is six times as high as that in
 

the low-dose group; the Cochran-Armitage test results are significant when the vehicle controls (P = 0.016) and the pooled controls
 

(P = 0.001) are used. Moreover, the Fisher exact test shows that
 

the incidence in the high-dose females is significantly higher
 

than that in the pooled controls (P = 0.002), and the lower limit
 

of the 95% confidence interval of the relative risk shows a value
 

greater than one. The statistical conclusion is that the occurrence of alveolar/bronchiolar adenomas of the lung in male and
 

female mice is associated with procarbazine at the doses used in
 

this experiment. In male mice, alveolar/bronchiolar carcinomas
 

were observed in one animal of each dosed group and in one animal
 

in the untreated-control group. Alveolar/bronchiolar carcinomas
 

were not observed in female mice.
 

In females, since the incidence of lymphoma or leukemia of the
 

hematopoietic system in the low-dose animals (8/23 (35%)) is over
 

four times that in the high-dose animals (2/26 (8%)), the test
 

for linear trend indicates a departure from linear trend when
 

either the vehicle-control group (P = 0.002) or the pooled-


control group (P less than 0.001) is used. The Fisher exact test
 

shows that the incidence of lymphoma or leukemia in the low-dose
 

females is significantly higher than that in either the vehicle
 

controls (P = 0.013) or the pooled controls (P less than 0.001).
 

38
 
The lower limit of the 95% confidence interval of the relative
 

risk of the low-dose group versus the pooled controls shows a
 

value greater than one. The results of these statistical tests
 

suggest that the incidence of lymphoma or leukemia in female mice
 

may be related to administration of the test chemical.
 

The incidence of olfactory neuroblastomas or undifferentiated
 

carcinomas of the brain is observed exclusively in the high-dose
 

groups. In male mice, the Cochran-Armitage test results are
 

highly significant when the vehicle controls (P = 0.002) and the
 

pooled controls (P less than 0.001) are used. A significant
 

departure from linear trend (P = 0·020) is observed when the
 

pooled-control group is used, due to the sharp increase in the
 

incidence of tumors in the high-dose group. This positive
 

finding is confirmed by the Fisher exact test, which shows that
 

the incidence in the high-dose animals is significantly higher
 

than that in the pooled controls (P less than 0.001), and the
 

lower limit of the 95% confidence interval of this relative risk
 

shows a value greater than one. In females, the Cochran-Armitage
 

test results are highly significant (P less than 0.001) when
 

either of the control groups is used. An indicated departure
 

from linear trend is observed when the pooled-control group (P =
 

0.006) or the vehicle-control group (P = 0.030) is used, due to
 

39
 
the steep increase in the incidence of tumors in the high-dose
 

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

high-dose females is significantly higher than that in either the
 

vehicle controls (P = 0.003) or the pooled controls (P less than
 

0.001); because of this positive finding, values of the lower
 

limits of the 95% confidence intervals are greater than one. The
 

statistical conclusion is that the occurrence of olfactory neuroblastomas is related to administration of the test chemical.
 

The analyses of adenocarcinomas, NOS, of the uterus show that the
 

Cochran-Armitage test is highly significant (P less than 0.001)
 

when the pooled-control group is used. The departure from linear
 

trend is also significant (P less than 0.001) when either the
 

vehicle-control group or the pooled-control group is used, due to
 

the sharp increase of the incidence of tumors in the dosed groups
 

and the higher incidence of tumors in the low-dose group. The
 

Fisher exact test results are all significant. The statistical
 

conclusion is that the incidence of adenocarcinomas, NOS, of the
 

uterus in female mice is related to administration of the test
 

chemical.
 

Groupings of these types of tumors are made, as in fibromas and
 

fibrosarcomas of the subcutaneous tissue in male mice; however,
 

the incidences of the individual components are not listed in
 

40
 
tables Fl and F2 unless they are greater than 5% in any of the
 

dosed groups· One adenocarcinoma, NOS, of the salivary gland was
 

found a low-dose male mouse. Since this was the only tissue
 

examined, the incidence of tumors in the low-dose group becomes
 

1/1(100%); however, it is not listed in table Fl because of the
 

small sample size. A list of the incidences of each of tumor is
 

provided in Appendix D, tables Dl and D2.
 

41
 

V. DISCUSSION
 

The doses of procarbazine used in the bioassay were toxic, as
 

shown by the lowered body weights and/or rates of survival of the
 

dosed animals. Mean body weights of all groups of rats administered procarbazine were lower than those of untreated- and vehicle-


control groups. No consistent compound-related effect on weights
 

occurred in mice. Survival of both rats and mice was markedly
 

reduced and was dose related (P less than 0.001); however, high
 

incidences of tumors were found in both rats and mice.
 

In rats, 21/60 dosed males and 20/59 dosed females, but no
 

controls, had tumors involving epithelial and neuroepithelial
 

tissue of the region of the nasal turbinates and nasal cavity,
 

i.e., olfactory neuroblastomas, carcinomas NOS, adenocarcinomas
 

NOS, and mucinous adenocarcinomas. The most frequent tumor,
 

olfactory neuroblastoma, was present in a greater number of
 

low-dose than high-dose animals of each sex. Direct comparisons
 

of the combination of these tumors in the low-dose groups with
 

both vehicle-control groups were significant. In the male, the
 

incidence in the high-dose group was also significant, as was the
 

dose-related trend, using pooled controls.
 

Malignant lymphoma in both male and female dosed rats was
 

43
 
statistically significant by direct comparison of the high-dose
 

group with the pooled-control group. In female rats, direct
 

comparison of the high-dose group with the vehicle-control group
 

was also statistically significant. These lesions appeared as
 

early as 12 weeks in female rats. Additionally, one eosinophilic
 

leukemia and two granulocytic leukemias were found in high-dose
 

males.
 

Adenocarcinomas of the mammary gland occurred in the dosed rats
 

and were statistically significant for dose-related trend in both
 

sexes and for direct comparisons of the high-dose group in males
 

and both the high- and low-dose groups in females with vehicle
 

controls. Several other neoplastic and nonneoplastic lesions were
 

observed in the mammary glands of animals of each sex, including
 

cystadenocarcinomas, adenomas, fibroadenomas, hyperplasias, and
 

cysts.
 

Several types of squamous-cell tumors of the ear canal and
 

Zymbal's gland were found in increased numbers among the dosed
 

rats. These included keratocanthomas, squamous-cell papillomas,
 

and squamous-cell carcinomas in two high-dose male rats, one
 

low-dose, and one untreated-control male rat, and in four
 

high-dose, three low-dose, but no control female rats. Although
 

the incidence was low, tumors of the auditory canal were
 

44
 
previously reported in rats administered procarbazine by both
 

Kelly et al. (1968) and Deckers et al. (1969).
 

In mice, tumors of the epithelium or neuroepithelium were also
 

found in the olfactory bulbs and mucosa in 10 high-dose males and
 

11 high-dose females. Olfactory neuroblastomas, the tumor occurring with the highest incidences, were found only in high-dose
 

males and high-dose females and were statistically significant by
 

direct comparisons with both pooled-control groups. These tumors
 

were also statistically significant by direct comparison in
 

high-dose females with the vehicle-control group.
 

In mice, malignant lymphomas or leukemias were observed in four
 

animals of each of the male dosed groups and in one untreated
 

control, but were not found among vehicle controls of either sex.
 

In females, the incidence in the low-dose group (6/23) was more
 

than three times that in the high-dose group (2/26). The incidence
 

of leukemia or lymphoma was significant in the low-dose group of
 

females when compared with vehicle controls. In the high-dose
 

female group, survival was markedly reduced, and none of the
 

animals lived to the end of the study.
 

Alveolar/bronchiolar adenomas were found in significant proportions in low- and high-dose male and high-dose female mice.
 

45
 
In female mice, adenocarcinomas of the uterus were found only in
 

dosed animals, and were significant in both dosed groups when
 

compared with either vehicle-control group. However, the incidence
 

was much higher in the low-dose group than in the high-dose group,
 

and there was a significant departure from linear trend·
 

The findings of this bioassay confirm previous studies in Osborne-


Mendel and Fischer 344 rats by Kelly et al. (1968), inbred R strain
 

rats of Wistar origin by Deckers et al. (1974), random-bred Charles
 

River CD strain rats by Grunberg and Prince (1969), (BALB/c χ
 

DBA/2)F1 mice by Kelly et al. (1969), and Swiss mice by Grunberg
 

and Prince (1969). These previously reported studies described
 

increased incidences of tumors of the lung, spleen, kidney, uterus,
 

mammary glands, sebaceous glands, and the ear duct. The olfactory
 

neuroblastomas observed in both rats and mice in the present
 

bioassay were not previously described. Studies on metabolites
 

and various degradation products of procarbazine in mice (Kelly et
 

al., 1969) suggest that tumor-associated activity of the parent
 

chemicaL is retained by the azo metabolite, N-isopropyl <*-(2­

methylazo)-p-toluamide, and also by the hydrazone metabolite, N-


isopropyl α -(2-methylhydrazone)-p-toluamide, but not by the
 

aldehyde oxidation product, N-isopropyl-p formylbenzamide.
 

It is concluded that under the conditions of this bioassay,
 

46
 
procarbazine was carcinogenic for both Sprague-Dawley rats and
 

B6C3F1 mice, producing several types of tumor in both sexes of
 

these two species·
 

47
 

VI. BIBLIOGRAPHY
 

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

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

Berenblum, I., ed., Care inogenic i ty Testing: A Report gf the
 

Panel on Carcinogenicity off the Cancer Research Commission of the
 

UICC, Vol. _2. International Union Against Cancer, Geneva, 1969.
 

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

Soc. Β 34:187-220, 1972.
 

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

1970, pp. 48-52.
 

Deckers, C , Deckers-Passau, L., Maisin, J., Gautheier, J. Μ.,
 

and Mace, F., Carcinogenicity of procarbazine. Z^. Krebsforsch.
 

81^:79-84, 1974.
 

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

significance tests, confidence limits and adjustments for
 

stratification. Rev. Irrt. Stat. Inst. 39^:148-169, 1971.
 

Grunberg, Ε. and Prince, H. N., Further studies on the
 

tumor-inhibitory and tumor-indueing properties of N-isopropyl-a­

(methylhydrazino)-p-toluamide hydrochloride (procarbazine
 

hydrochloride). Chemotherapy 14·:65-76, 1969.
 

Herrold, Κ. Μ., Induction of olfactory neruoepithelial tumors in
 

Syrian hamsters by diethylnitrosamine. Cancer Γ7:114-121, 1964.
 

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

incomplete observations. J^ Amer. Statist. Assoc. 53:457-481,
 

1958.
 

Kelly, M. G., O'Gara, R. W., Yancey, S. T., Gadekar, K., Botkin,
 

C , and Oliverio, V. T., Comparative carcinogenicity of
 

N-isopropyl-<*-(2-methylhydrazino)-p-toluamide HC1 (procarbazine
 

hydrochloride), its degradation products, other hydrazines, and
 

isonicotinic acid hydrazide. J. Natl Cancer Inst. 42(2):337-344,
 

1969.
 

Kelly, M. G., O'Gara, R. W., Yancey, S. T., and Botkin, C ,
 

Induction of tumors in rats with procarbazine hydrochloride. J.
 

Natl Cancer Inst. 40(5):1027-1035, 1968. ~
 

49
 
Laskin, S., Kuschner, M., Drew, R. T., Cappiello, V. P., and
 

Nelson, N., Tumors of the respiratory tract induced by inhalation
 

of bis(chloromethyl)ether. Arch. Environ* Health 23:135, 1971.
 

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

Peters, J. Α., Carcinogenesis bioassay data system. £. Cotnp.
 

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

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

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

Obert, G. J., Devine, K. D., and McDonald, J. R., Olfactory
 

neuroblastomas. Cancer 13:205-215, 1960.
 

Oliverio, V. T., Derivatives of triazenes and hydrazines. In:
 

Cancer Medicine, Holland, J. F. and Frei III, E., eds., Lea and
 

Febiger7~~Philadelphia, Pa., 1973, pp. 806-817.
 

Prejean, J. D., Peckham, J. C , Casey, A. E., Griswold, D. P.,
 

Weisburger, E. K., and Weisburger, J. Η., Spontaneous tumors in
 

Sprague-Dawley rats and Swiss mice. Cancer Res. 33:2768-2773,
 

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.
 

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

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

United States Pharmacopeial Convention, Sulfisoxazole. In: The
 

United States Pharmacopeia XIX, United States Pharmacopeial
 

Convention, Inc., Rockville, Md., pp. 480-482, 1975.
 

Weinstein, L., Antimicrobial agents. In: The Pharmacological
 

Basis of Therapeutics, Goodman, L. S. and Gilman, Α., eds·,
 

Macmillan Publishing Co., Inc., New York, pp. 1113-1124, 1975.
 

50
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS ADMINISTERED PROCARBAZINE
 

BY INTRAPERITONEAL INJECTION
 

51
 

TABLE ΑΙ. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS
 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION
 

UNTREATED CONTROL 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

A N I M A L S I N I T I A L L Y I N S T U D Y A N I M A L S N E C R O P S I E D ANIMALS EXAMINED HISTOPATHOLOGICALLY 

10 

9 

9 

10 

10 

10 

34 

31 

28 

35 

33 

33 

INTEGUMENTARY SYSTEM 

*SKIN 

BASAL-CELL CARCINOMA (9) ( 10) (31) (33) 

1 (3%) 

*SUBCUT TISSUE 

CARCINOMA,NOS SARCOMA, NOS 

(9) (10) (31) (33) 

1 (3%) 

1 (3%) 

RESPIRATORY SYSTEM 

«LUNG 

UNDIFFERENTIATED CARCINOMA METAS (9) (10) (28) (33) 

1 (3%) 

HEMATOPOIETIC SYSTEM 

^MULTIPLE ORGANS 

MALIG.LYMPHOMA, LYMPHOCYTIC

MALIG.LYMPHOMA, HISTIOCYTIC GRANULOCYTIC LEUKEMIA 

EOSINOPHILIC LEUKEMIA 

 TYPE 

 TYPE 

(9) ( 10 ) 

1 (10% ) 

(31) 

2 (6%) 

(33) 

9 (27X) 

1 (3%) 

1 (3%) 

SBONE MARRON GRANULOCYTIC LEUKEMIA (9) (10 ) (28) (29) 

1 (3%) 

«SMALL INTESTINE MALIG.LYMPHOMA, LYMPHOCYTIC TYPE (9) (9) (27) 

1 (4%) 

(32) 

CIRCULATORY SYSTEM 

N O N E 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

53
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)

 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

DIGESTIVE SYSTEM
 

fcPANXREAS (9) (9)
 ACINAR-CELL ADENOMA
 

«SMALL INTESTINE (9) (9)
 CYSTADENOCARCINOMA, NOS
 

ifCOLON (9) (9)
 ADENOCARCINOMA, NOS
 ADENOCA IN ADENOMATOUS POLYP
 

URINARY SYSTEM
 

SKIDNEY (9) (10)
 UNDIFFERENTIATED CARCINOMA
 

ENDOCRINE SYSTEM
 

iiPITUITARY (8) (9)
 CHROMOPHOBE ADENOMA 1 (13%) 2 (22%)
 CHROMOPHOBE CARCINOMA
 

«ADRENAL (9) (9)
 CORTICAL ADENOMA
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND (9) ( 10)
 ADENOMA, NOS
 ADENOCARCINOMA, NOS
 CYSTADENOCARCINOMA, NOS
 FIBROADENOMA 3 (33%)
 

NERVOUS SYSTEM
 

«CEREBRUM (9) (9)
 OLIGODENDROGLIOMA
 

itBRAIN (9) (9)
 

CARCINOMA,NOS
 

NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

(25)
 

(27)
 

(28)
 

1 (4%)
 

(28)
 

(26)
 

1 (4%)
 

(28)
 

3 ( 1 1%)
 

(31)
 

1 (3%)
 

(27)
 

(27)
 

HIGH DOSE
 

(31)
 

1 (3%)
 

(32)
 

1 (3%)
 

(31)
 

1 (3%)
 

(32)
 

1 (3%)
 

(31)
 

1 (3%)
 

(32)
 

2 (6%)
 

(33)
 

1 (3%)
 

7 (21%)
 

2 (6%)
 

(33)
 

1 (3%)
 

(33)
 

3 ( 9%)
 

54
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

SARCOMA, NOS 

ASTROCYTOMA OLFACTORY NEUROBLASTOMA 1 (11%) 

«OLFACTORY BULB ADENOCARCINOMA, NOS (9) (9) 

SPECIAL SENSE ORGANS
 

*EAR CANAL (9) (10)
 

SQUAMOUS CELL PAPILLOMA
 SQUAMOUS CELL CARCINOMA 1 (11%)
 KERATOACANTHOMA
 

MUSCULOSKELETAL SYSTEM
 

VMUSCLE OF HEAD (9) ( 10)
 SARCOMA, NOS 1 (10%)
 

BODY CAVITIES
 

XPERITONEUM (9) ( 10)
 FIBROSARCOMA
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 10 10
 NATURAL DEATHS) 2 2
 MORIBUND SACRIFICE
 SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE δ 8
 ANIMAL MISSING
 

3 INCLUDES AUTOLYZED ANIMALS
 

NUMBER OF ANIMALS UITH TISSUE EXAMINED MICROSCOPICALLY
 NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

1 (4%)
 

12 (44%)
 

(27)
 

(31)
 

1 (3%)
 

(31)
 

(3D
 

34
 

17
 

17
 

HIGH DOSE
 

7 (21%)
 

(33)
 

3 (9%)
 

(33)
 

2 (6%)
 

(33)
 

(33)
 

1 (3%)
 

35
 

15
 

20
 

55
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)

 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 TOTAL MALIGNANT TUMORS
 

UNTREATED VEHICLE 

CONTROL CONTROL LOW DOSE 

5 19 6 23 

2 

2 5 

2 2 18 2 2 18 

TOTAL ANIMALS WITH SECONDARY TUMORS»
 TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS LXCEPT SECONDARY TUMORS
 

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

HIGH DOSE
 

30
 

49
 

6
 

6
 

30
 

43
 

1
 

1
 

56
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION 

UNTREATED VEHICLE
 

CONTROL CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY ANIMALS NECROPSIED ANIMALS EXAMINED HISTOPATHOLOGICALLY

10 

10 10 

10 

10 

10 

36 

31 

30 

INTEGUMENTARY SYSTEM 

KSUBCUT TISSUE FIBROMA NEUROFIBROMA 

(10) ( 10) (3 1 ) 

1 (3% ) 

RESPIRATORY SYSTEM 

HUNG 

ADENOSQUAMOUS CARCINOMA ( 10) ( 10 ) (29) 

HEMATOPOIETIC SYSTEM 

^MULTIPLE ORGANS MALIG.LYMPHOMA, LYMPHOCYTIC TYP E ( 10) ( 10 ) (31) 

CIRCULATORY SYSTEM 

NONE 

DIGESTIVE SYSTEM 

«COLON ADENOMATOUS POLYP, NOS (9) ( 10) (27) 

URINARY SYSTEM 

NONE 

ENDOCRINE SYSTEM 

«PITUITARY CHROMOPHOBE ADENOMA (5) 

3

# NUMBE R O F ANIMAL S WIT H TISSU E EXAMINE D

* NUMBE R O F ANIMAL S NECROPSIE D 

 (60% ) 

(

 MICROSCOPICALL Y 

 10 ) 3 (30% ) 

(29) 

1 (3%) 

35
 

31
 

31
 

(3D
 

1 (3%)
 

(31)
 

1 (3%)
 

(31)
 2 0 (65% )
 

(30 )
 1 (3% )
 

(29 )
 

57 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED)

 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

^ADRENAL ( 10) ( 10)
 CORTICAL ADENOMA
 

REPRODUCTIVE SYSTEM
 

XMAMMARY GLAND ( 10) (10)
 ADENOMA, NOS
 ADENOCARCINOMA, NOS
 CYSTADENOMA, NOS
 CYSTADENOCARCINOMA, NOS
 FIBROMA
 FIBROADENOMA 2 (20%) 2 (20%)
 

SUTERUS (9) ( 10)
 LEIOMYOSARCOMA
 ENDOMETRIAL STROMAL POLYP 1 (11%)
 

SUTERUS/EHDOMETRIUM (9) ( 10)
 ADENOMA, NOS
 

NERVOUS SYSTEM
 

«CEREBRUM ( 10) (10)
 **MUCINOUS ADENOCARCINOMA
 

#BRAIN ( 10) ( 10)
 **OLFACTORY NEUROBLASTOMA
 

*CRANIAL NERVE ( 10) (10)
 NEUROFIBROMA
 

SPECIAL SENSE ORGANS
 

*EAR CANAL ( 10) (10)
 

SQUAMOUS CELL PAPILLOMA
 SQUAMOUS CELL CARCINOMA
 KERATOACANTHOMA
 

*ZYMBAL'S GLAND ( 10) ( 10)
 KERATOACANTHOMA
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

* * THESE TUMORS ARE OF NEUROEPITHELIAL ORIGIN
 

LOW DOSE
 

(30)
 

4 (13%)
 

(3D


1 (3%)
 16 (52%)
 

1 (3%)
 4 (13%)
 

(30)
 

1 (3%)
 

(30)
 

(28)
 

(28)
 17 (61%)
 

(31)
 

(3D
 

1 (3%)
 2 (6%)
 

(31)
 

HIGH DOSE
 

(31)
 

2 (6%)
 

(3D


1 (3%)
 25 (81%)
 1 (3%)
 1 (3%)
 

3 ( 10%)
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

(31)
 

1 (3%)
 

(31)
 

2 (6%)
 

(31)
 

1 (3%)
 

(31)
 

1 (3%)
 

3 (10%)
 

(31)
 

1 (3%)
 

58
 
TABLE A2. FEMALE RATS: NEOPLASMS ( ONTINUED)
 

BODY CAVITIES
 

*MESENTERY
 LIPOSARCOMA
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 NATURAL DEATHS)
 MORIBUND SACRIFICE
 SCHEDULED SACRIFICE
 ACCIDENTALLY KILLED
 TERMINAL SACRIFICE
 ANIMAL MISSING
 

«> INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

UNTREATED
 

CONTROL
 

 (10)
 

 10
 

1
 

9
 

TOTAL ANIMALS WITH PRIMARY TUMORS* TOTAL PRIMARY TUMORS 4 6 

TOTAL ANIMALS WITH BENIGN TUMORS TOTAL BENIGN TUMORS 4 6 

TOTAL ANIMALS WITH MALIGNANT TUMORS TOTAL MALIGNANT TUMORS 

TOTAL ANIMALS WITH SECONDARY TUMORS* TOTAL SECONDARY TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT TOTAL UNCERTAIN TUMORS 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC TOTAL UNCERTAIN TUMORS 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

(10) (31) (31) 

1 (3%) 

10 

1 

1 

8 

36 

15 

21 

35 

14 

21 

4 

5 

4 

5 

27 

49 

1 1 

14 

26 

35 

30 

68 

13 

17 

30 

51 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECOND RY TUMORS
 

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

59
 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE ADMINISTERED PROCARBAZINE
 

BY INTRAPERITONEAL INJECTION
 

61
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION 

UNTREATED VEHICLE
 

CONTROL CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY ANIMALS NECROPSIED ANIMALS EXAMINED HISTOPATHOLOGICALLY 15 

15 

15 

15 

14 

12 

35 

30 

30 

35 

31 

30 

INTEGUMENTARY SYSTEM 

*SKIN 

UNDIFFERENTIATED CARCINOMA 

*SUBCUT TISSUE SPINDLE CELL MELANOMA FIBROMA FIBROSARCOMA 

(15) 

(15) 

(14) 

(14) 

(30) 

(30) 

1 (3%) 1 (3%) 1 (3%) 

(31) 

1 (3%) 

(3D 

1 (3%) 

RESPIRATORY SYSTEM 

tttUNG HEPATOCELLULAR CARCINOMA, METAST ALVEOLAR/BRONCHIOLAR ADENOMA ALVEOLAR/BRONCHIOLAR CARCINOMA 

(15) 

2 (13%) 1 (7%) 

( 12) (30) 

10 (33%) 1 (3%) 

(30) 

1 (3%) 10 (33%) 1 (3%) 

HEMATOPOIETIC SYSTEM 

^MULTIPLE ORGANS MALIG.LYMPHOMA, LYMPHOCYTIC TYPE MALIG.LYMPHOMA, HISTIOCYTIC TYPE GRANULOCYTIC LEUKEMIA 

(15) 

1 (7%) 

(14) (30) 

1 (3%) 

2 (7%) 1 (3%) 

(3D 

1 (3%) 2 (6%) 1 (3%) 

CIRCULATORY SYSTEM 

NONE 

DIGESTIVE SYSTEM 

«SALIVARY GLA^D ADENOCARCINOMA, NOS 

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

(2) 

1 (50%) 

63 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

#LIVER HEPATOCELLULAR HEPATOCELLULAR HEMANGIOMA 

 ADENOMA CARCINOMA 

(15) 

1 (7X) 

(11) (30) 

3 (10ΣΟ 

1 (3%) 

(29) 

2 (7%) 1 (3%) 

URINARY SYSTEM 

NONE 

ENDOCRINE SY5TEM 

NONE 

REPRODUCTIVE SYSTEM 

NONE 

NERVOUS SYSTEM 

ttBRAIN 

••UNDIFFERENTIATED CARCINOMA ••OLFACTORY NEUROBLASTOMA 

(15) (8) (24) (29) 

1 (3%) 9 (31%) 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONE 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

** THESE TUMOB.S ARE OF NEUROEPITHELIAL ORIGIN 

64
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

UNTREATED VEHICLE
 

CONTROL CONTROL LOW DOSE HIGH DOSE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 15 15 35 35
 NATURAL DEATHS 7 15 12
 MORIBUND SACRIFICE 1 2 9 23
 SCHEDULED SACRIFICE 2
 ACCIDENTALLY KILLED
 TERMINAL SACRIFICE 14 6 9
 ANIMAL MISSING
 

a INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 5 18 20
 TOTAL PRIMARY TUMORS 5 23 30
 

TOTAL ANIMALS WITH BENIGN TUMORS 3 13 10
 TOTAL BENIGN TUMORS 3 15 12
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 2 8 16
 TOTAL MALIGNANT TUMORS 2 8 18
 

TOTAL ANIMALS WITH SECONDARY TUMORS» 1
 

TOTAL SECONDARY TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

65
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION

 

UNTREATED VEHICLE
 

CONTROL CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY 15 15 ANIMALS MISSING 1 ANIMALS NECROPSIED 12 14 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 12 14 

INTEGUMENTARY SYSTEM
 

XSKIN (12) (14)
 ADENOCARCINOMA, NOS
 

*SUBCUT TISSUE ( 12) ( 14)
 BASAL-CELL CARCINOMA
 

RESPIRATORY SYSTEM
 

ttLUNG ( 12) ( 14)
 ADENOCARCINOMA, NOS, METASTATIC
 ALVEOLAR/BRONCHIOLAR ADENOMA
 

HEMATOPOIETIC SYSTEM
 

^MULTIPLE ORGANS (12) (14)
 MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
 MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 GRANULOCYTIC LEUKEMIA
 

#LYMPH NODE (1)
 ADENOCARCINOMA, NOS, METASTATIC
 

tPULMONARY LYMPH NODE ( 1)
 ADENOCARCINOMA, NOS, METASTATIC
 

^MESENTERIC L. NODE (1)
 MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

#UTERUS (12) (13)
 MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

ft NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 x NUMBER OF ANIMALS NECROPSIED
 

66
 

35
 

1
 

23
 

23
 

(23)
 

(23)
 

1 (4%)
 

(23)
 

4 (17%)
 

1 (4%)
 

(23)
 

3 (13%)
 

1 (4%)
 

2 (9%)
 

(4)
 

(4)
 

2 (50%)
 

(4)
 

1 (25%)
 

(23)
 

1 (4%)
 

35
 

26
 

26
 

(26)
 

1 (4%)
 

(26)
 

(26)
 

4 (15%)
 

6 (23%)
 

(26)
 

1 (4%)
 

(6)
 

1 (17%)
 

(6)
 

(6)
 

1 (17%)
 

(25)
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CIRCULATORY SYSTEM
 

«MYOCARDIUM
 ADENOCARCINOMA, NOS, METASTATIC
 

DIGESTIVE SYSTEM
 

«LIVER
 ADENOCARCINOMA, NOS, METASTATIC
 HEMANGIOMA
 

«SMALL INTESTINE
 ADENOCARCINOMA, NOS, METASTATIC
 

URINARY SYSTEM
 

«URINARY BLADDER
 SARCOMA, NOS
 

ENDOCRINE SYSTEM
 

«ADRENAL
 PHEOCHROMOCYTOMA
 

«THYROID
 FOLLICULAR-CELL ADENOMA
 

REPRODUCTIVE SYSTEM
 

«UTERUS
 ADENOCARCINOMA, NOS
 LEIOMYOSARCOMA
 

NERVOUS SYSTEM
 

«BRAIN
 SARCOMA, NOS
 **OLFACTORY NEUROBLASTOMA
 NEUROFIBROSARCOMA
 

SPECIAL SENSE ORGANS
 

NONE
 

UNTREATED VEHICLE 

CONTROL CONTROL LOW DOSE HIGH DOSE 

(12) (14) (22) (23)

1 (4%) 

( 12) ( 14) (23) (26) 1 (4%) 1 (4%) 

( 12) (14) (22) (25) 1 (5%) 

( 12) (13) ( 18) (26) 

1 (4%) 

(12) ( 14) (22) (26) 

1 (4%) 

(11) ( 13) (18) 

1 (6%) 

( 19) 

( 12) ( 13) (23)'c 

14 (61%) 2 (9%) 

(25) 

8 (32%) 2 (8%) 

(11) ( 14) (22) (25) 

1 (4%) 11 (44%) 1 (4%) 

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

** THESE TUMORS ARE OF NEUROEPITHELIAL ORIGIN
 

67
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

MUSCULOSKELETAL SYSTEM 

XCRANIAL AND FACIAL Β OLFACTORY NEUROBLASTOMA, INVASIV (12) (14) (23) (26) 

1 (4%) 

BODY CAVITIES 

^MESENTERY ADENOCARCINOMA, NOS, METASTATIC (12) (14) (23) 

3 (13%) 

(26) 

ALL OTHER SYSTEMS 

^MULTIPLE ORGANS SARCOMA, NOS (12) (14) (23) (26) 

1 (45Ο 

SITE UNKNOWN ADENOMA, NOS 1 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY NATURAL DEATHS) MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED TERMINAL SACRIFICE ANIMAL MISSING 

15 

3 

1 1 

1 

15 

1 

2 

1 

11 

35 

18 

9 

6 

1 

1 

35 

17 

18 

3 INCLUDES AUTOLYZED ANIMALS 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED

* NUMBER OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

68
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

UNTREATED VEHICLE
 

CONTROL CONTROL LOW DOSE HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 19 20
 

TOTAL PRIMARY TUMORS 27 37
 

TOTAL ANIMALS WITH BENIGN TUMORS 2 9
 

TOTAL BENIGN TUMORS 2 9
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 19 20
 

TOTAL MALIGNANT TUMORS 25 28
 

TOTAL ANIMALS WITH SECONDARY TUMORS# 5 6
 

TOTAL SECONDARY TUMORS 11 7
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

69
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS ADMINISTERED PROCARBAZINE
 

BY INTRAPERITONEAL INJECTION
 

71
 

TABLE CI.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION 

UNTREATED
 

CONTROL
 

ANIMALS INITIALLY IN STUDY
 ANIMALS NECROPSIED
 ANIMALS EXAMINED HISTOPATHOLOGICALLY


INTEGUMENTARY SYSTEM
 

KSKIN
 

EPIDERMAL INCLUSION CYST
 INFLAMMATION, CHRONIC
 

FIBROSIS
 

XSUBCUT TISSUE
 INFLAMMATION, CHRONIC
 

RESPIRATORY SYSTEM
 

«TRACHEA
 CYST, NOS
 

INFLAMMATION, NOS
 

LYMPHOCYTIC INFLAMMATORY INFILTR
 INFLAMMATION, SUPPURATIVE
 INFLAMMATION, ACUTE
 INFLAMMATION, ACUTE SUPPURATIVE
 INFLAMMATION, ACUTE/CHRONIC
 

INFLAMMATION, CHRONIC
 

«LUNG/BRONCHUS
 

BRONCHIECTASIS
 

INFLAMMATION, CHRONIC
 

«LUNG/BRONCHIOLE
 HYPERPLASIA, LYMPHOID
 

«LUNG
 

EMBOLISM, NOS
 

CONGESTION, NOS
 EDEMA, NOS
 

HEMORRHAGE
 

10
 

9
 

9
 

(9)
 

1 (11%)
 

(9)
 

1 (11%)
 

(9)
 

2 (22%)
 

(9)
 

(9)
 

1 (11%)
 

(9)
 

VEHICLE
 

CONTROL
 

10
 

10
 

10
 

( 10)
 

( 10)
 

(9)
 

( 10)
 

(10)
 

3 (30%)
 

( 10)
 

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

73
 

LOW DOSE
 

34
 

31
 

28
 

(31)
 

1 (3%)
 

(3D
 

(27)
 

2 (7%)
 

3 (11%)
 

1 (4%)
 

1 (4%)
 

13 (48%)
 

(28)
 

(28)
 

(28)
 

1 (4%)
 

4 ( 14%)
 

4 (14%)
 

1 (4%)
 

HIGH DOSE
 

35
 

33
 

33
 

(33)
 

1 (3%)
 

(33)
 

(29)
 

3 (10%)
 

5 (17%)
 

1 (3%)
 

5 (17%)
 

1 (3%)
 

(33)
 

1 (3%)
 

1 (3%)
 

(33)
 

(33)
 

1 (3%)
 

1 (3%)
 
TABLE CI. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUEO)

 

UNTREATEO 

CONTROL 

VEHICLE 

CONTROL 

BRONCHOPNEUMONIA, NOS INFLAMMATION, INTERSTITIAL BRONCHOPNEUMONIA SUPPURATIVE LEUKOCYTOSIS, NOS HYPERPLASIA, LYMPHOID 

3 (30%) 

HEMATOPOIETIC SYSTEM
 

«BONE MARROW (9) (10)
 ATROPHY, NOS 2 (22%) 7 (70%)
 DEPLETION
 

HYPERPLASIA, HEMATOPOIETIC
 

«SPLEEN (9) (9)
 ATROPHY, NOS
 HYPERPLASIA, NOS
 HYPERPLASIA, HEMATOPOIETIC
 HYPERPLASIA, RETICULUM CELL
 HYPERPLASIA, LYMPHOID
 

HEMATOPOIESIS 1 (11%)
 

«LYMPH NODE
 ATROPHY, NOS
 

«MEDIASTINAL L.NODE
 ATROPHY, NOS
 HYPERPLASIA, NOS
 

«PANCREATIC L.NODE
 ATROPHY, NOS
 

«MESENTERIC L. NODE
 ATROPHY, NOS
 HYPERPLASIA, NOS
 

«THYMUS (9) (9)
 HEMORRHAGE
 ATROPHY, NOS
 

CIRCULATORY SYSTEM
 

«MYOCARDIUM (9) (9)
 INFLAMMATION, INTERSTITIAL
 

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

74
 

LOW DOSE 

1 (4%)
 

(28)
 

1 (4%)
 

(26)
 

18 (69%)
 

2 (8%)
 

(20)
 

(20)


18 (90%)
 

(20)
 

(20)
 

1 (5%)
 

(23)
 

3 (13%)
 

19 (83%)
 

(27)
 

4 (15%)
 

HIGH DOSE 

1 (3%)
 

4 (12%)
 

5 (15%)
 

1 (3%)
 

2 (6%)
 

(29)
 

5 (17%)
 

1 (3%)
 

4 (14%)
 

(33)
 

13 (39%)
 

3 (9%)
 

2 (6%)
 

1 (3%)
 

1 (3%)
 

3 (9%)
 

(21)
 

5 (24%)
 

(21)
 

5 (24%)
 

1 (5%)
 

(21)
 

1 (5%)
 

(21)
 

1 (5%)
 

1 (5%)
 

(28)


1 (4%)
 

20 (71%)
 

(33)
 

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

UNTREATED VEHICLE
 

CONTROL CONTROL
 

«ENDOCARDIUM (9) (9)
 FIBROSIS
 

DIGESTIVE SYSTEM
 

«LIVER (9) (9)
 HEMORRHAGE
 LYMPHOCYTIC INFLAMMATORY INFILTR
 PERIARTERITIS
 NECROSIS, COAGULATIVE
 ANGIECTASIS
 

LEUKOCYTOSIS, NOS
 HEMATOPOIESIS
 

«LIVER/CENTRILOBULAR (9) (9)
 DEGENERATION, NOS
 NECROSIS, NOS
 

«PANCREAS (9) (9)
 INFLAMMATION, INTERSTITIAL
 GRANULOMA, NOS
 NECROSIS, FAT
 

«PANCREATIC ACINUS (9) (9)
 HYPERPLASIA, NODULAR
 

«STOMACH (9) (9)
 ULCER, NOS
 ULCER, FOCAL
 PERIARTERITIS
 

URINARY SYSTEM
 

«KIDNEY (9) ( 10)
 LYMPHOCYTIC INFLAMMATORY INFILTR
 INFLAMMATION, INTERSTITIAL 5 (56%) 5 (50%)
 NEPHROPATHY
 CALCIFICATION, NOS
 HYPERPLASIA, TUBULAR CELL
 LEUKOCYTOSIS, NOS
 

«URINARY BLADDER (9) (9)
 INFLAMMATION. NOS
 

NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

(27)
 

2 (7%)
 

(26)
 

1 (4%)
 

(26)
 

1 (4%)
 

2 (8%)
 

(25)
 

2 (8%)
 

1 (4%)
 

(25)
 

(28)
 

2 (7%)
 

2 (7%)
 

(28)
 

1 (4%)
 1 (4%)
 

 (<m


5 (18%)
 

(25)
 

HIGH DOSE
 

(33)
 

1 (3%)
 

(31)
 

1 (3%)
 1 (3%)
 2 (6%)
 1 (3%)
 

1 (3%)
 2 (6%)
 

(31)
 

(3D
 

1 (3%)
 

(31)
 

1 (3%)
 

(32)
 

2 (6%)
 

1 (3%)
 

(32)
 

1 (3%)
 

2 (6%)
 

1 (3%)
 

(33)
 

1 f "*5n
 

75
 
TABLE CI. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

INFLAMMATION, HEMORRHAGIC
 

ENDOCRINE SYSTEM
 

#ADRENAL (9) (9)
 

HEMATOPOIESIS
 

«ADRENAL CORTEX (9) (9)
 

DEGENERATION, NOS
 

HYPERPLASIA, NODULAR
 

«THYROID (9) (9)
 

ECTOPIA
 EPIDERMAL INCLUSION CYST
 HYPERPLASIA, CYSTIC
 

REPRODUCTIVE SYSTEM
 

XMAMMARY GLAND (9) ( 10)
 

CYST, NOS 1 (10%)
 

HYPERPLASIA, NOS
 

«PROSTATE (9) (9)
 

HEMORRHAGE
 INFLAMMATION, SUPPURATIVE
 

*SEMINAL VESICLE (9) ( 10)
 

HEMORRHAGE
 

«TESTIS (9) (9)
 

ADHESION, NOS
 

ATROPHY, NOS
 

^EPIDIDYMIS (9) ( 10)
 

INFLAMMATION, FOCAL GRANULOMATOU
 

NERVOUS SYSTEM
 

«CEREBRUM (9) (9)
 

HEMORRHAGE
 INFLAMMATION, NOS
 

GLIOSIS
 

« NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 < NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

1 (4%)
 

(28)
 

1 (4%)
 

(28)
 

(20)
 

1 (5%)
 

(31)
 

8 (26%)
 

(28)
 

1 (4%)
 

2 (7%)
 

(3D
 

1 (3%)
 

(25)
 

1 (4%)
 

6 (24%)
 

(31)
 

1 (3%)
 

(27)
 

1 (4%)
 

1 (4%)
 

HIGH DOSE
 

(32)
 

(32)
 

1 (3%)


3 (9%)
 

(28)
 

1 (4%)


1 (4%)
 

(33)
 

4 (12%)
 

(32)
 

(33)
 

(32)
 

20 (63%)
 

(33)
 

(33)
 

1 (3%)
 

76
 
TABLE CI. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)

 

UNTREATED VEHICLE 

CONTROL CONTROL 

MALACIA 

«OLFACTORY BULB (9) (9) HYPERPLASIA, NOS METAPLASIA, SQUAMOUS 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

^SKELETAL MUSCLE (9) (10)
 INFLAMMATION, CHRONIC
 

BODY CAVITIES
 

^PERITONEUM (9) ( 10)
 FIBROSIS
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 1
 

NECROPSY PERF/NO HISTO PERFORMED
 AUTO/NECROPSY/NO HISTO
 AUTOLYSIS/NO NECROPSY 1
 

41 NUMBER OF ANIMALS UITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

3 (11%)
 

(27)
 

(3D
 

(31)
 

1
 

2
 

3
 

HIGH DOSE
 

(33)
 

1 (3%)
 

1 (3%)
 

(33)
 

1 (3%)
 

(33)
 

1 (3%)
 

2
 

77
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION 

UNTREATED VEHICLE
 

CONTROL CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY ANIMALS NECROPSIED ANIMALS EXAMINED HISTOPATHOLOGICALLY 

10 

10 

10 

10 

10 

10 

36 

31 

30 

35 

31 

31 

INTEGUMENTARY SYSTEM 

*SUBCUT TISSUE INFLAMMATION, SUPPURATIVE (10) ( 10) (31) (31) 

1 (3%) 

RESPIRATORY SYSTEM 

ftTRACHEA LYMPHOCYTIC INFLAMMATORY INFILTR INFLAMMATION, SUPPURATIVE INFLAMMATION, ACUTE/CHRONIC 

(10) 

1 (10%) 

( 10) (29) 

2 (7%) 9 (31%) 

(29) 

6 (21%) 11 (38%) 

«LUNG/BRONCHUS BRONCHIECTASIS INFLAMMATION, NOS INFLAMMATION, CHRONIC 

(10) ( 10) (29) 

2 (7%) 

2 (7%) 

(31) 

1 (3%) 3 (10%) 

ftLUNG/BRONCHIOLE HYPERPLASIA, LYMPHOID ( 10) 

1 (10%) 

(10) 

2 (20%) 

(29) (31) 

«LUNG 

CONGESTION, NOS EDEMA, NOS BRONCHOPNEUMONIA, NOS INFLAMMATION, FOCAL BRONCHOPNEUMONIA DIFFUSE INFLAMMATION, INTERSTITIAL BRONCHOPNEUMONIA SUPPURATIVE ABSCESS, NOS BRONCHOPNEUMONIA CHRONIC SUPPURA INFLAMMATION, FOCAL GRANULOMATOU 

( 10) 

2 (20%) 2 (20%) 

( 10) 

1 (10%) 1 (10%) 

(29) 

1 (3%) 

1 (3%) 

1 (3%) 2 (7%) 

1 (3%) 1 (3%) 

(31) 

1 (3%) 1 (3%) 3 (10%) 

1 (3%) 1 (3%) 

HEMATOPOIETIC SYSTEM 

#BONE MARROW ATROPHY, NOS (8) 

5 (63%) 

( 10) 

5 (50%) 

(26) (31) 

2 (6%) 

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

78
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)

 

UNTREATED VEHICLE 

CONTROL CONTROL 

HYPERPLASIA, HEMATOPOIETIC 

«SPLEEN (9) (10) ATROPHY, NOS HYPERPLASIA, NOS HYPERPLASIA, HEMATOPOIETIC HEMATOPOIESIS 

«LYMPH NODE 

ATROPHY, NOS HYPERPLASIA, PLASMA CELL 

«MEDIASTINAL L.NODE ATROPHY, NOS HYPERPLASIA, NOS 

«THYMUS CONGESTION, NOS (9) (10) 

HEMORRHAGE ATROPHY, NOS HYPERPLASIA, NOS 

CIRCULATORY SYSTEM
 

«MYOCARDIUM (10) (10)
 INFLAMMATION, INTERSTITIAL
 

DIGESTIVE SYSTEM
 

«LIVER (9) (10)
 HEMORRHAGE
 GRANULOMA, NOS
 NECROSIS, NOS
 NECROSIS, FOCAL
 HEMATOPOIESIS
 

«LIVER/CENTRILOBULAR (9) (10)
 ATROPHY, NOS
 

«PANCREAS (9) (10)
 INFLAMMATION, INTERSTITIAL
 

«STOMACH (9) (10)
 ULCER. NOS
 

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

LOW DOSE
 

4 (15%)
 

(29)
 

14	 (48%)
 

1 (3%)
 

8 (28%)
 

3 (10%)
 

(26)
 

1 (4%)
 

1 (4%)
 

(26)


20 (77%)
 

(28)
 

23 (82%)
 

(29)
 

(29)
 

1 (3%)
 

1 (3%)
 

2 (7%)
 

(29)
 

1 (3%)
 

(27)
 

(27)
 

2 (7%)
 

HIGH DOSE
 

6 (19%)
 

(31)
 

8 (26%)
 

2 (6%)
 

(27)
 

(27)
 

9 (33%)
 

1 (4%)
 

(29)
 

1 (3%)
 

1 (3%)
 

12	 (41%)
 

2 (7%)
 

(31)
 

2 (6%)
 

(31)
 

1 (3%)
 

1 (3%)
 

2 (6%)
 

(31)
 

(28)
 

1 (4%)
 

(30)
 

79
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

URINARY SYSTEM
 

«KIDNEY (10) (10)
 

PYELONEPHRITIS, NOS
 

INFLAMMATION, INTERSTITIAL 2 (205O
 

NEPHROPATHY
 CALCIFICATION, FOCAL
 

«KIDNEY/CORTEX ( 10) (10)
 

FIBROSIS
 

INFARCT, HEALED
 CALCIFICATION, NOS
 

«KIDNEY/TUBULE (10) (10)
 

CALCIFICATION, NOS
 

«KIDNEY/PELVIS (10) (10)
 

HYPERPLASIA, EPITHELIAL
 

«URINARY BLADDER (9) (10)
 

INFLAMMATION, NOS
 

ENDOCRINE SYSTEM
 

«ADRENAL (10) (10)
 

THROMBOSIS, NOS
 

INFLAMMATION, GRANULOMATOUS
 

NECROSIS, FOCAL
 ANGIECTASIS 1 (10ΣΟ 2 (20%)
 

«ADRENAL CORTEX (10) (10)
 

HYPERPLASIA, NODULAR
 

REPRODUCTIVE SYSTEM
 

XMAMMARY GLAND (10) ( 10)
 

CYST, NOS 2 (20X) 3 (305O
 

HYPERPLASIA, NOS
 

HYPERPLASIA, CYSTIC
 

«UTERUS (9) ( 10)
 

CYST, NOS
 

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

LOW DOSE
 

(30)
 

(30)
 

1 (3%)
 

(30)
 

(30)
 

(29)
 

(30)
 

(30)
 

4 (135i)
 

(31)
 

1 (3·/.)
 

(30)
 

HIGH DOSE
 

(30)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

(30)
 

1 (3%)
 

1 (3%)
 

(30)
 

1 (35Ο
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

(31)
 

1 (3%)
 

1 (3%)
 

1 (3%)
 

(31)
 

(31)
 

2 (6%)
 

(30)
 

2 (7%)
 

80
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUEO)

 

PYOMETRA
 

«UTERUS/ENDOMETRIUM
 CYST, NOS
 INFLAMMATION, SUPPURATIVE
 FIBROSIS
 ATROPHY, NOS
 

«OVARY/OVIDUCT
 DILATATION, NOS
 HYPERPLASIA, NOS
 

«OVARY
 CYST, NOS
 

«OVARY/MEDULLA
 HYPERPLASIA, NOS
 

NERVOUS SYSTEM
 

«CEREBRUM
 DEGENERATION, CYSTIC
 

«BRAIN
 HEMORRHAGE
 MALACIA
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

*BONE
 

HYPERPLASIA, NOS
 

^BONE/PERIOSTEUM
 HYPERPLASIA, NOS
 

*BONE/ENDOSTEUM
 HYPERPLASIA, NOS
 

UNTREATED
 

CONTROL
 

(9)
 

1 (11% )
 

(9)
 

(9)
 

(9)
 

( 10 )
 

(10 )
 

( 10)
 

( 10)
 

(10)
 

VEHICLE
 

CONTROL
 

( 10)
 

2 (20%)
 

(10)
 

(10)
 

(10)
 

( 10)
 

(10)
 

(10)
 

(10)
 

( 10)
 

LOW DOSE
 

5 (17%)
 

(30)
 

27 (90%)
 

3 (10%)
 

(30)
 

1 (3%)
 

1 (3%)
 

(25)
 

1 (4%)
 

(25)
 

(28)
 

1 (4%)
 

(28)
 

1 (4%)
 

1 (4%)
 

(31)
 

(31)


1 (3%)
 

(31)
 

1 (3%)
 

HIGH DOSE
 

1 (3%)
 

(30)
 

1 (3%)
 

5 (17%)
 

1 (3%)
 

(30)
 

(23)
 

(23)
 

1 (4%)
 

(31)
 

(31)
 

(31)


1 (3%)
 

(3D
 

(31)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

81
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

BODY CAVITIES 

^PERITONEUM INFLAMMATION, NOS INFLAMMATION, SUPPURATIVE GRANULOMA, NOS 

(10) (10) (31) 

1 (33Ο 

(31) 

2 (6%) 

1 (3%) 

ALL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED AUTO/NECROPSY/NO HISTO 

AUTOLYSIS/NO NECROPSY 

2 1 1 

1 

5 

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

82
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE ADMINISTERED PROCARBAZINE
 

BY INTRAPERITONEAL INJECTION
 

83
 

TABLE D1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION 

ANIMALS INITIALLY IN STUDY
 ANIMALS NECROPSIED
 ANIMALS EXAMINED HISTOPATHOLOGICALLY


INTEGUMENTARY SYSTEM
 

HSUBCUT TISSUE
 EPIDERMAL INCLUSION CYST
 INFLAMMATION, NECRO GRAN
 

RESPIRATORY SYSTEM
 

«LUNG
 

INFLAMMATION, INTERSTITIAL
 BRONCHOPNEUMONIA SUPPURATIVE
 BRONCHOPNEUMONIA, CHRONIC


HYPERPLASIA, LYMPHOID
 

HEMATOPOIETIC SYSTEM
 

«BONE MARROW
 HYPERPLASIA, HEMATOPOIETIC
 

«SPLEEN
 

HYPERPLASIA, HEMATOPOIETIC
 

HYPERPLASIA, RETICULUM CELL
 HYPERPLASIA, LYMPHOID
 HEMATOPOIESIS
 

«PULMONARY LYMPH NODE
 HYPERPLASIA, LYMPHOID
 

«PANCREATIC L.NODE
 HYPERPLASIA, LYMPHOID
 

«MESENTERIC L. NODE
 INFLAMMATION, HEMORRHAGIC
 

UNTREATED
 

CONTROL
 

15
 

15
 

 15
 

(15)
 

1 (7%)
 

(15)
 

1 (7%)
 

(15)
 

(15)
 

2 (13%)
 

(1)
 

(1)
 

(1)
 

VEHICLE
 CONTROL
 

15
 

1*
 

12
 

(14)
 

(12)
 

(12)
 

(12)
 

1 (8%)
 

(1)
 

(1)
 

(1)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

85
 

LOW DOSE
 

35
 

30
 

30
 

(30)
 

1 (3%)
 

(30)
 

1 (3%)
 

1 (3%)
 

(28)
 

(30)
 

1 (3%)
 2 (7%)
 2 (7%)
 

(6)
 

1 C 17%)
 

(6)
 

(6)
 

1 (17%)
 

HIGH DOSE
 

35
 

31
 

30
 

(31)
 

(30)
 

2 (7%)
 

(28)
 

1 (4%)
 

(28)
 

1 (4%)
 

1 (4%)
 3 (11%)
 

(6)
 

1 ( 17%)
 

(6)
 

1 (17%)
 

(6)
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

HYPERPLASIA, PLASMA CELL
 HYPERPLASIA, RETICULUM CELL
 HYPERPLASIA, LYMPHOID
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

«LIVER
 CYST, NOS
 FIBROSIS, DIFFUSE
 HYPERPLASIA, NODULAR
 ANGIECTASIS
 HYPERPLASIA, LYMPHOID
 

#PEYERS PATCH
 HYPERPLASIA, RETICULUM CELL
 

URINARY SYSTEM
 

«KIDNEY
 INFLAMMATION, INTERSTITIAI
 INFLAMMATION, SUPPURATIVE
 PYELONEPHRITIS SUPPURATIVE
 INFARCT, NOS
 

«URINARY BLADDER
 CALCULUS, NOS
 INFLAMMATION, CHRONIC
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

«PROSTATE
 INFLAMMATION, SUPPURATIVE
 

UNTREATED
 

CONTROL
 

1 (100%)
 

( 15)
 

1 (7%)
 

( 15)
 

( 15)
 

( 15)
 

(15)
 

VEHICLE
 

CONTROL
 

(11)
 

1 (9%)
 

(11)
 

( 12)
 

1 (8%)
 

( 12)
 

1 (8%)
 

( 12)
 

LOW DOSE
 

1 (17%)
 3 (50%)
 

(30)
 

1 (3%)
 

1 (3%)
 1 (3%)
 

(28)
 

1 (4%)
 

(30)
 

2 (7%)
 

1 (3%)
 

(30)
 

(30)
 

2 (7%)
 

HIGH DOSE
 

4 (67%)
 

(29)
 

(30)
 

(30)
 

1 (3%)
 1 (3%)
 

(30)
 

1 (3%)
 

(30)
 

3 (10%)
 

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

86
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

INFLAMMATION, CHRONIC SUPPURATIV 1 (3%) 1 (3%) 

NERVOUS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

^PERITONEUM INFLAMMATION, CHRONIC 

^MESENTERY HEMATOMA, ORGANIZED 

( 15) 

(15) 

( 14) 

(14) 

(30) 

(30) 

1 (3%) 

(31) 

1 (3%) 

(31) 

ALL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED NECROPSY PERF/NO HISTO PERFORMED AUTO/NECROPSY/NO HISTO

AUTOLYSIS/NO NECROPSY 

6 

# NUMBER OF ANIMALS UITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

9 

1 

1 

1 

6 

5 

8 

1 

4 

87
 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

ADMINISTERED PROCARBAZINE BY INTRAPERITONEAL INJECTION

 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

ANIMALS INITIALLY IN STUDY 15 15
 ANIMALS MISSING 1
 ANIMALS NECROPSIED 12 14
 ANIMALS EXAMINED HISTOPATHOLOGICALLY 12 14
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

XNASAL MUCOSA (12) (14)
 FIBROSIS
 

«LUNG (12) (14)
 HYPERPLASIA, LYMPHOID 1 (8%)
 

HEMATOPOIETIC SYSTEM
 

«SPLEEN ( 12) (14)
 HEMATOMA, NOS
 HYPERPLASIA, RETICULUM CELL
 HEMATOPOIESIS 1 (8%) 1 (7%)
 

^MESENTERIC L. NODE ( 1)
 HYPERPLASIA, RETICULUM CELL
 HYPERPLASIA, LYMPHOID
 

CIRCULATORY SrSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

ttLIVER (12) (14)
 HYPERPLASIA, NODULAR 1 (8%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

35
 

1
 

23
 

23
 

(23)
 

(23)
 

(22)
 

2 (9%)
 4 (18%)
 

(4)
 

1 (25%)
 

(23)
 

HIGH DOSE
 

35
 

26
 

26
 

(26)
 

1 (4%)
 

(26)
 

(24)
 

1 (4%)
 1 (4%)
 4 (17%)
 

(6)
 

1 (17%)
 3 (50%)
 

(26)
 

88
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

ANGIECTASIS 1 (7%) 

«PANCREAS INFLAMMATION, SUPPURATIVE (11) (14) 

«COLON (12) ( 14) INFLAMMATION, NOS INFLAMMATION, SUPPURATIVE 

URINARY SYSTEM
 

«KIDNEY (12) (14)
 

INFLAMMATION, INTERSTITIAL 1 (7%)
 GLOMERULONEPHRITIS, CHRONIC
 AMYLOIDOSIS
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

«UTERUS (12) (13)
 PYOMETRA
 FIBROSIS
 HYPERPLASIA, ADENOMATOUS
 

«UTERUS/ENDOMETRIUM (12) (13)
 INFLAMMATION, SUPPURATIVE
 HYPERPLASIA, NOS
 HYPERPLASIA, CYSTIC 10 (83*) 12 (92%)
 

«OVARY (12) (13)
 INFLAMMATION, CHRONIC SUPPURATIV
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE
 

(21)
 

(21)
 

(22)
 

2 (9%)
 1 (5%)
 

(23)
 

(23)
 

2 (9%)
 

2 (9%)
 

(23)
 

1 (4%)
 

HIGH DOSE
 

1 (4%)
 

(23)
 

2 C 9%)
 

(24)
 

1 (4%)
 1 (4%)
 

(26)
 

(25)


1 (4%)
 1 (4%)
 4 C 16%)
 

(25)
 

2 (8%)
 1 (4%)
 

5 (20%)
 

(25)
 

89
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

MUSCULOSKElETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

^PERITONEUM
 INFLAMMATION, CHRONIC
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 ANIMAL MISSING/NO NECROPSY
 AUTOLYSIS/NO NECROPSY
 

"" • • • • • • • ­ — ' ­ "*­ ' 

UNTREATED VEHICLE 

CONTROL CONTROL LOW DOSE HIGH DOSE 

( 12) ( 14) (23) (26) 

1 (4%) 

2 1 1 2 1 1 

2 1 1 1 9 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

90
 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS ADMINISTERED PROCARBAZINE
 

BY INTRAPERITONEAL INJECTION
 

91
 

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

Administered Procarbazine by Intraperitoneal Injection (a)
 

Topography: Morphology
 

Hematopoietic System:
 

Malignant Lymphoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System:
 

Leukemia (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled 

Vehicle 

Control 

1/39 (3) 

 = 0.002 

Control)(f) 

Control)(f) 

0/39 (0) 

 = 0.031 

ControD(f) 

Control)(f) 

Matched
 

Vehicle
 

Control
 

1/10 (10)
 

N.S.
 

86
 

0/10 (0)
 

N.S.
 

Low
 

Dose
 

3/31 (10)
 

N.S.
 

3.774
 

0.321
 

191.295
 

0.968
 

0.094
 

49.097
 

32
 

0/31 (0)
 

—
 

—
 

—
 

—
 

—
 

High
 

Dose
 

9/33 (27)
 

 = 0.003**
 

10.636
 

1.599
 

447.256
 

2.727
 

0.475
 

114.803
 

21
 

3/33 (9)
 

N.S.
 

Infinite
 

0.718
 

Infinite
 

Infinite
 

0.203
 

Infinite
 

20
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued) 

Topography: Morphology
 

Hematopoietic System: Leukemia
 

or Malignant Lymphoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Cortical Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

1/39 (3)
 

 less than
 

0.001
 

0/36 (0)
 

N.S.
 

Matched
 

Vehicle
 

Control
 

1/10 (10)
 

 = 0.013
 

86
 

0/9 (0)
 

N.S.
 

Low
 

Dose
 

3/31 (10)
 

N.S.
 

3.774
 

0.321
 

191.295
 

0.968
 

0.094
 

49.097
 

32
 

3/28 (11)
 

N.S.
 

Infinite
 

0.784
 

Infinite
 

Infinite
 

0.219
 

Infinite
 

27
 

High
 

Dose
 

12/33 (36)
 

 less than
 

0.001**
 

14.182
 

2.294
 

578.099
 

3.636
 

0.685
 

148.380
 

20
 

2/32 (6)
 

N.S.
 

Infinite
 

0.336
 

Infinite
 

Infinite
 

0.093
 

Infinite
 

29
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Adenocarcinoma, NOS (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle ControlXf)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenoma, NOS
 

or Adenocarcinoma, NOS (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/39 (0)
 

 = 0.001
 

0/39 (0)
 

 = 0.001
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

 = 0.016
 

—
 

0/10 (0)
 

 = 0.009
 

Low
 

Dose
 

1/31 (3)
 

N.S.
 

Infinite
 

0.068
 

Infinite
 

Infinite
 

0.019
 

Infinite
 

28
 

1/31 (3)
 

N.S.
 

Infinite
 

0.068
 

Infinite
 

Infinite
 

0.019
 

Infinite
 

28
 

High
 

Dose
 

7/33 (21)
 

 = 0.003**
 

Infinite
 

2.323
 

Infinite
 

Infinite
 

0.659
 

Infinite
 

17
 

8/33 (24)
 

 = 0.001**
 

Infinite
 

2.736
 

Infinite
 

Infinite
 

0.776
 

Infinite
 

21
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Cystadenocarcinoma, NOS (b)
 

 Values (c?d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Olfactory Bulb: Adenocarcinoma, NOS (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/39 (0)
 

N.S.
 

0/38 (0)
 

 = 0.034
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

—
 

0/9 (0)
 

N.S.
 

Low
 

Dose
 

0/31 (0)
 

—
 

—
 

—
 

—
 

—
 

—
 

0/27 (0)
 

—
 

—
 

—
 

—
 

—
 

High
 

Dose
 

2/33 (6)
 

N.S.
 

Infinite
 

0.353
 

Infinite
 

Infinite
 

0.099
 

Infinite
 

17
 

3/33 (9)
 

N.S.
 

Infinite
 

0.702
 

Infinite
 

Infinite
 

0.185
 

Infinite
 

29
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Brain/Olfactory Bulb: Carcinoma, NOS,
 

Neuroblastoma or Adenocarcinoma (b)
 

 Values (c,d)
 

Departure from Linear Trend
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

VO Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ear Canal: Keratoacanthoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/38 (0)
 

 = 0·003
 

 = 0.001
 

—
 

0/39 (0)
 

N.S·
 

Matched
 

Vehicle
 

Control
 

0/9 (0)
 

N.S.
 

 = 0.013
 

—
 

0/10 (0)
 

N.S.
 

Low
 

Dose
 

12/27 (44)
 

 = 0.014*
 

 less than
 

0.001**
 

Infinite
 

5.280
 

Infinite
 

Infinite
 

1.401
 

Infinite
 

23
 

0/31 (0)
 

—
 

—
 

—
 

—
 

—
 

High
 

Dose
 

9/33 (27)
 

 = 0.001**
 

Infinite
 

3.072
 

Infinite
 

Infinite
 

0.817
 

Infinite
 

21
 

2/33 (6)
 

N.S.
 

Infinite
 

0.353
 

Infinite
 

Infinite
 

0.099
 

Infinite
 

26
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

(a) Dosed groups received doses of 15 or 30 mg/kg.
 

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

(c) Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when Ρ is less than 0.05; otherwise, not significant (N.S.) is
 

indicated. Beneath the incidence of tumors in a dosed group is the probability level for the
 

Fisher exact test for the comparison of that dosed group with the vehicle-control group (*)
 

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

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

(d) A negative trend (N) indicates a lower incidence in a dosed group than in the vehicle-control
 

group.
 

(e) The probability level for departure from linear trend is given when Ρ is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the
 

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

Administered Procarbazine by Intraperitoneal Injection (a)
 

Topography: Morphology
 

Hematopoietic System:
 

Malignant Lymphoma (b)
 

 Values (c,d)
 

Departure from Linear Trend (e)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/40 (0)
 

 less than
 

0.001
 

 less than
 

0.001
 

Matched
 

Vehicle


Control


0/10 (0)


 less than
 

0.001
 

 = 0.004
 

 Low
 

 Dose
 

 0/31 (0)
 

—
 

—
 

—
 

—
 

—
 

High
 

Dose
 

20/31 (65)
 

 less than
 

0.001*
 

 less than
 

0.001**
 

Infinite
 

8.567
 

Infinite
 

Infinite
 

2.363
 

Infinite
 

12
 
Table Ε2· Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Adrenal: Cortical Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

1/38 (3)
 

N.S.
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

—
 

Low
 

Dose
 

4/30 (13)
 

N.S.
 

5.067
 

0.536
 

240.212
 

Infinite
 

0.345
 

Infinite
 

21
 

High
 

Dose
 

2/31 (6)
 

N.S.
 

2.452
 

0.134
 

139.793
 

Infinite
 

0.105
 

Infinite
 

21
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Adenocarcinoma, NOS (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

2/38 (5)
 

 less than
 

0.001
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

 less than
 

0.001
 

Low
 

Dose
 

16/31 (52)
 

 = 0.003*
 

 less than
 

0.001**
 

9.806
 

2.594
 

79.193
 

Infinite
 

1.842
 

Infinite
 

22
 

High
 

Dose
 

25/31 (81)
 

 less than
 

0.001*
 

 less than
 

0.001**
 

15.323
 

4.543
 

102.409
 

Infinite
 

3.051
 

Infinite
 

16
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued) 

Topography: Morphology 

Mammary Gland: Adenoma, NOS 

Adenocarcinoma, NOS (b) 

 Values (c,d) 

ο 

ho 

Relative Risk (Pooled Vehicle Control)(f) 

Lower Limit 

Upper Limit 

Relative Risk (Matched Vehicle Control)(f) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Pooled
 

Vehicle
 

Control
 

3/38 (8)
 

 less than
 

0.001
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

 less than
 

0.001
 

Low
 

Dose
 

17/31 (55)
 

 - 0.002*
 

 less than
 

0.001**
 

6.946
 

2.290
 

32.191
 

Infinite
 

1.971
 

Infinite
 

21
 

High
 

Dose
 

25/31 (81)
 

 less than
 

0.001*
 

 less than
 

0.001**
 

10.215
 

3.746
 

38.520
 

Infinite
 

3.051
 

Infinite
 

16
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Cystadenocarcinoma, NOS (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Cystadenoma, NOS
 

or Cystadenocarcinoma, NOS (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/38 (0)
 

N.S.
 

0/38 (0)
 

N.S.
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

—
 

0/10 (0)
 

N.S.
 

„
 

Low
 

Dose
 

0/31 (0)
 

N.S.
 

—
 

—
 

—
 

—
 

—
 

0/31 (0)
 

—
 

—
 

—
 

—
 

—
 

High
 

Dose
 

1/31 (3)
 

N.S.
 

Infinite
 

0.066
 

Infinite
 

Infinite
 

0.019
 

Infinite
 

17
 

2/31 (6)
 

N.S.
 

Infinite
 

^0.366
 

Infinite
 

Infinite
 

0.105
 

Infinite
 

17
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Fibroadenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle ControlHf)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

10/38 (26)
 

 = 0·044 (Ν)
 

Matched
 

Vehicle
 

Control
 

2/10 (20)
 

N.S.
 

84
 

Low
 

Dose
 

4/31 (13)
 

N.S.
 

0.490
 

0.123
 

1.511
 

0.645
 

0.117
 

6.542
 

26
 

High
 

Dose
 

3/31 (10)
 

N.S.
 

0.368
 

0.071
 

1.279
 

0.484
 

0.069
 

5.328
 

20 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology


Brain: Olfactory Neuroblastoma (b)


 Values (c,d)


Departure from Linear Trend(e)


Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

 Control
 

 0/38 (0)*
 

 N.S.
 

 less than
 

0.001
 

Matched
 

Vehicle
 

Control
 

0/10 (0)
 

N.S.
 

 less than
 

0.001
 

—
 

Low
 

Dose
 

17/28 (61)
 

 = 0.001*
 

 less than
 

0.001**
 

Infinite
 

7.563
 

Infinite
 

Infinite
 

2.192
 

Infinite
 

20
 

High
 

Dose
 

2/31 (6)
 

N.S.
 

Infinite
 

0.366
 

Infinite
 

Infinite
 

0.105
 

Infinite
 

21
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued) 

Pooled Matched 

Vehicle Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Brain/Cerebrum: Olfactory Neuroblastoma
 

or Mucinous Adenocarcinoma (b) 0/38 (0) 0/10 (0) 17/28 (61) 3/31 (10)
 

 Values (c,d) N.5. N.S. = 0.007* N.S.
 

 less than
 

0.001**
 

Departure from Linear Trend(e) less than less than
 

0.001 0.001
 

Relative Risk (Pooled Vehicle ControD(f) Infinite Infinite
 

Lower Limit 7.564 0.746
 

Upper Limit Infinite Infinite
 

Relative Risk (Matched Vehicle ControlKf) Infinite Infinite
 

Lower Limit 1.598 0.216
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 20
 21 
Table Ε2· Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued) 

Topography: Morphology 

Pooled 

Vehicle 

Control 

Matched 

Vehicle 

Control 

Low 

Dose 

High 

Dose 

Ear Canal: Keratoacanthoma (b) 0/38 (0) 0/10 (0) 2/31 (6) 3/31 (10) 

 Values (c,d) N.S. N.S. N.S. N.S. 

Relative Risk (Pooled Vehicle Control)(f) 

Lower Limit 

Upper Limit 

Infinite 

0.366 

Infinite 

Infinite 

0.746 

Infinite 

Relative Risk (Matched Vehicle Control)(f) 

Lower Limit 

Upper Limit 

Infinite 

0.105 

Infinite 

Infinite 

0.216 

Infinite 

Weeks to First Observed Tumor 27 22 

(a) Dosed groups received doses of 15 or 30 mg/kg.
 

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

(c) Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when Ρ is less than 0.05; otherwise, not significant (N.S.) is
 

indicated. Beneath the incidence of tumors in a dosed group is the probability level for the
 

Fisher exact test for the comparison of that dosed group with the vehicle-control group (*)
 

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

otherwise, not significant (N.S·) is indicated.
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

(d) A negative	 trend (N) indicates a lower incidence in a dosed group than in the vehicle-control
 

group.
 

(e) The probability level for departure from linear	 trend is given when Ρ is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the
 

specified control group.
 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN
 

MICE ADMINISTERED PROCARBAZINE
 

BY INTRAPERITONEAL INJECTION
 

109
 

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

Administered Procarbazine by Intraperitoneal Injection (a)
 

Topography: Morphology
 

Subcutaneous Tissue: Fibroma or
 

Fibrosarcoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled-Vehicle Control)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched-Vehicle Control)
 

Lower Limit
 

Upper Limit
 

Weeks to Observed Tumor
 

Lung: Alveolar/Bronchiolar Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled-Vehicle Control)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched-Vehicle Control)
 

Lower Limit
 

Upper Limit
 

Weeks to Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/42 (0)
 

N.S.
 

(f)
 

(f)
 

0/39 (0)
 

 less than
 

0.001
 

(f)
 

(f)
 

Matched
 

Vehicle
 

Control
 

0/14 (0)
 

N.S.
 

—
 

0/12 (0)
 

N.S.
 

Low
 

Dose
 

2/30 (7)
 

N.S.
 

Infinite
 

0.417
 

Infinite
 

Infinite
 

0.147
 

Infinite
 

75
 

10/30 (33)
 

 = 0.020*
 

 less than
 

0.001**
 

Infinite
 

3.931
 

Infinite
 

Infinite
 

1.309
 

Infinite
 

60
 

High
 

Dose
 

1/31 (3)
 

N.S.
 

Infinite
 

0.073
 

Infinite
 

Infinite
 

0.025
 

Infinite
 

52
 

10/30 (33)
 

 = 0.020*
 

 less than
 

0.001**
 

Infinite
 

3.931
 

Infinite
 

Infinite
 

1.309
 

Infinite
 

38
 
Table Fl· Analyses of the Incidence of Primary Tumors in Male Mice 

Administered Procarbazine by Intraperitoneal Injection (a) 

(continued) 

Topography: Morphology
 

Lung: Alveolar/Bronchiolar Adenoma
 

or Carcinoma (b)
 

 Values (c,d)
 

Departure from Linea Trend (e)
 

Relative Risk (Pooled-Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched-Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Weeks to Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/39 (0)
 

 less than
 

0.001
 

 = 0.024
 

Matched
 

Vehicle
 

Control
 

0/12 (0)
 

N.S.
 

Low
 

Dose
 

11/30 (37)
 

 = 0.013*
 

 less than
 

0.001**
 

Infinite
 

4.392
 

Infinite
 

Infinite
 

1.461
 

Infinite
 

60
 

High
 

Dose
 

10/30 (33)
 

 = 0.020*
 

 less than
 

0.001**
 

Infinite
 

3.931
 

Infinite
 

Infinite
 

1.308
 

Infinite
 

38
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Hematopoietic System: Lymphoma or
 

Leukemia (b)
 

 Values (c,d)
 

Relative Risk (Pooled-Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched-Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Weeks to Observed Tumor
 

Liver: Hepatocellular Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/42 (0>
 

 = 0.028
 

0/39 (0)
 

N.S.
 

Matched
 

Vehicle
 

Control
 

0/14 (0)
 

N.S.
 

—
 

0/11 (0)
 

N.S.
 

——
 

Low
 

Dose
 

4/30 (13)
 

 = 0.027*
 

Infinite
 

1.310
 

Infinite
 

Infinite
 

0.465
 

Infinite
 

66
 

3/30 (10)
 

N.S.
 

Infinite
 

0.790
 

Infinite
 

Infinite
 

0.243
 

Infinite
 

75
 

High
 

Dose
 

4/31 (13)
 

 = 0.029*
 

Infinite
 

1.268
 

Infinite
 

Infinite
 

0.450
 

Infinite
 

37
 

3/29 (10)
 

N.S.
 

Infinite
 

0.818
 

Infinite
 

Infinite
 

0.123
 

Infinite
 

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

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Hematopoietic System: Malignant Lymphoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/42 (0)
 

N.S.
 

0/39 (0)
 

N.S.
 

Matched
 

Vehicle
 

Control
 

0/14 (0)
 

N.S.
 

—
 

0/11 (0)
 

N.S.
 

„
 

Low
 

Dose
 

3/30 (10)
 

N.S.
 

Infinite
 

0.849
 

Infinite
 

Infinite
 

0.301
 

Infinite
 

66
 

3/30 (10)
 

N.S.
 

Infinite
 

0.790
 

Infinite
 

Infinite
 

0.243
 

Infinite
 

75
 

High
 

Dose
 

3/31 (10)
 

N.S.
 

Infinite
 

0.822
 

Infinite
 

Infinite
 

0.291
 

Infinite
 

37
 

2/29 (7)
 

N.S.
 

Infinite
 

0.401
 

Infinite
 

Infinite
 

0.123
 

Infinite
 

56
 
__ 

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

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Liver: Hepatocellular
 

Adenoma or Carcinoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Weeks to Observed Tumor
 

Brain: Olfactory Neuroblastoma (b)
 

 Values (c,d)
 

Departure from Linear Trend (e)
 

Relative Risk (Pooled Vehicle ControlHf)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/39 (0)
 

N.S.
 

0/36 (0)
 

 less than
 

0.001
 

 = 0.028
 

Matched
 

Vehicle
 

Control
 

0/11 (0)
 

N.S.
 

—
 

0/8 (0)
 

 = 0.003
 

Low
 

Dose
 

3/30 (10)
 

N.S.
 

Infinite
 

0.790
 

Infinite
 

Infinite
 

0.243
 

Infinite
 

75
 

0/24 (0)
 

N.S.
 

—
 

—
 

—
 

—
 

High
 

Dose
 

3/29 (10)
 

N.S.
 

Infinite
 

0.818
 

Infinite
 

Infinite
 

0.251
 

Infinite
 

56
 

9/29 (31)
 

 less tli
 

0.001**
 

Infinite
 

3.327
 

Infinite
 

Infinite
 

0.844
 

Infinite
 

47
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued) 

Pooled Matched 

Vehicle Vehicle Low High 

Topography: Morphology Control Control Dose Dose 

Brain: Olfactory Neuroblastoma
 

or Undifferentiated Carcinoma (b) 0/36 (0) 0/8 (0) 0/24 (0) 10/29 (34)
 

 Values (c,d)	 less than = 0.002 less than
 

0.001	 0.001**
 

Departure from Linear Trend (e)	 = 0.020
 

Relative Risk (Pooled Vehicle Control) (f) —
 Infinite
 

Lower Limit — 3.768
 

Upper Limit — Infinite
 

Relative Risk (Matched Vehicle Control) (f) Infinite
 

Lower Limit — 0.955
 

Upper Limit — Infinite
 

Weeks to Observed Tumor	 mmm 47
 

(a) Dosed groups received doses of 6 or 12 mg/kg.
 

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

(c) Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when Ρ is less than 0.05; otherwise, not significant (N.S.) is
 

indicated. Beneath the incidence of tumors in a dosed group is the probability level for the
 

Fisher exact test for the comparison of that dosed group with the vehicle-control group (*)
 

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

otherwise, not significant (N.S.) is indicated.
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

(d) A negative	 trend (N) indicates a lower incidence in a dosed group than in the vehicle-control
 

group.
 

(e) The probability level for departure from linear trend is given when Ρ is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the
 

specified control group.
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

Topography: Morpho1ogy
 

Lung: Alveolar/Bronchiolar Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

oo
 

Hematopoietic System:
 

Malignant Lymphoma (b)
 

 Values (c,d)
 

Departure from Linear Trend (e)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/43 (0)
 

 = 0.001
 

0/43 (0)
 

N.S.
 

 = 0.002
 

Matched
 

Vehicle
 

Control
 

0/14 (0)
 

 = 0.016
 

—
 

0/14 (0)
 

N.S.
 

 - 0.012
 

—
 

Low
 

Dose
 

1/23 (4)
 

N.S.
 

Infinite
 

0.100
 

Infinite
 

Infinite
 

0.034
 

Infinite
 

77
 

6/23 (26)
 

 = 0.043*
 

 = 0.001**
 

Infinite
 

3.038
 

Infinite
 

Infinite
 

1.054
 

Infinite
 

55
 

High
 

Dose
 

6/26 (23)
 

 = 0.002**
 

Infinite
 

2.681
 

Infinite
 

Infinite
 

0.931
 

Infinite
 

50
 

2/26 (8)
 

N.S.
 

Infinite
 

0.493
 

Infinite
 

Infinite
 

0.170
 

Infinite
 

53
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Hematopoietic System: Lymphoma
 

or Leukemia (b)
 

 Values (c,d)
 

Departure from Linear Trend (e)
 

Relative Risk (Pooled Vehicle ControlXf)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/43 (0)
 

N.S.
 

 less than
 

0.001
 

Matched
 

Vehicle
 

Control
 

0/14 (0)
 

N.S.
 

 = 0.002
 

—
 

Low
 

Dose
 

8/23 (35)
 

 = 0.013*
 

P less than
 

0.001**
 

Infinite
 

4.351
 

Infinite
 

Infinite
 

1.510
 

Infinite
 

55
 

High
 

Dose
 

2/26 (8)
 

N.S.
 

Infinite
 

0.493
 

Infinite
 

Infinite
 

0.170
 

Infinite
 

53
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Thyroid: Follicular-cell Adenoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control)(f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle ControD(f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled Matched 

Vehicle Vehicle 

Control Control 

0/42 (0) 0/13 (0) 

N.S. * N.S. 

„ 

Low
 

Dose
 

1/18 (6)
 

N.S.
 

Infinite
 

0.125
 

Infinite
 

Infinite
 

0.041
 

Infinite
 

81
 

High
 

Dose
 

0/19 (0)
 

—
 

—
 

—
 

——
 

—
 

—
 

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

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Uterus: Adenocarcinoma, NOS (b)
 

 Values (c,d)
 

Departure from Linear Trend (e)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/42 (0)
 

 less than
 

0.001
 

 less than
 

0.001
 

Matched
 

Vehicle
 

Control
 

0/13 (0)
 

N.S.
 

 less than
 

0.001
 

—
 

Low
 

Dose
 

14/23 (61)
 

 less than
 

0.001*
 

 less than
 

0.001**
 

Infinite
 

8.237
 

Infinite
 

Infinite
 

2.731
 

Infinite
 

64
 

High
 

Dose
 

8/25 (32)
 

 = 0.022*
 

 less than
 

0.001**
 

Infinite
 

3.903
 

Infinite
 

Infinite
 

1.297
 

Infinite
 

53
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

Topography: Morphology
 

Uterus: Leiomyosarcoma (b)
 

 Values (c,d)
 

Relative Risk (Pooled Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Vehicle Control) (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Vehicle
 

Control
 

0/42 (0)
 

N.S.
 

Matched
 

Vehicle
 

Control
 

0/13 (0)
 

N.S.
 

——
 

Low
 

Dose
 

2/23 (9)
 

N.S.
 

Infinite
 

0.544
 

Infinite
 

Infinite
 

0.180
 

Infinite
 

55
 

High
 

Dose
 

2/25 (8)
 

N.S.
 

Infinite
 

0.501
 

Infinite
 

Infinite
 

0.165
 

Infinite
 

44
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued) 

Topography: Morphology 

Pooled 

Vehicle 

Control 

Matched 

Vehicle

Control

 Low 

 Dose 

High 

Dose 

Brain: Olfactory Neuroblastoma (b) 0/43 (0) 0/14 (0) 0/22 (0) 11/25 (44) 

 Values (c,d) less than 

0.001 

 less than 

0.001 

 - 0.003* 

 less than 

0.001** 

Departure from Linear Trend (e) « 0·006 - 0.030 

Relative Risk (Pooled Vehicle Control)(f) Infinite
 

Lower Limit — 5.829
 

Upper Limit — Infinite
 

Relative Risk (Matched Vehicle Control)(f) —
 Infinite
 

Lower Limit — 2.023
 

Upper Limit — Infinite
 

Weeks to First Observed Tumor —
 44
 

(a) Dosed groups received doses of 6 or 12 mg/kg.
 

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

(c) Beneath the incidence of tumors in the vehicle-control group is the probability level for the
 

Cochran-Armitage test when Ρ is less than 0.05; otherwise, not significant (N.S.) is
 

indicated. Beneath the incidence of tumors in a dosed group is the probability level for the
 

Fisher exact test for the comparison of that dosed group with the vehicle-control group (*)
 

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

otherwise, not significant (N.S.) is indicated.
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered Procarbazine by Intraperitoneal Injection (a)
 

(continued)
 

(d) A negative trend (N) indicates a lower incidence in a dosed group than in the vehicle-control
 

group.
 

(e) The probability level for departure from linear trend is given when Ρ is less than 0.05 for any
 

comparison.
 

(f) The 95% confidence interval of the relative risk between each dosed group and the
 

specified control group.
 

NIH Publication No. 79-819