1978 

/if 

OFFICIAL NTP
 

FILE COPY
 

DO NOT REMOVE FROM THIS OFFICE
 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 95
 

BIOASSAY OF 

3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 6959-48-4 

NCI-CG-TR-95 

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

Public Health Service 

National Institutes of Health 

• ,'3-v , 3* ,.£ j'l , .-I, , 

:"-&- -.:^i 

BIOASSAY OF
 

3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

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

Public Health Service
 

National Institutes of Health
 

DREW Publication No. (NIH) 78-1345
 

BIOASSAY OF
 

3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

FOREWORD; This report presents the results of the bioassay of
 

3-(chloromethyl)pyridine hydrochloride conducted for the
 

Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute (NCI), National Institutes
 

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

experiments designed to determine whether selected environmental
 

chemicals have the capacity to produce cancer in animals.
 

Negative results, in which the test animals do not have a greater
 

incidence of cancer than control animals, do not necessarily mean
 

that the test chemical is not a carcinogen, inasmuch as the
 

experiments are conducted under a limited set of circumstances.
 

Positive results demonstrate that the test chemical is
 

carcinogenic for animals under the conditions of the test and
 

indicate that exposure to the chemical is a potential risk to
 

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

carcinogens requires a wider analysis.
 

CONTRIBUTORS; The bioassay of 3-(chloromethyl)pyridine
 

hydrochloride was conducted by Litton Bionetics, Inc.,
 

Kensington, Maryland, initially under direct contract to NCI and
 

currently under a subcontract to Tracor Jitco, Inc., the prime
 

contractor for the NCI Carcinogenesis Testing Program.
 

The experimental design and doses were chosen by Drs. E. K.
 

Weisburger1, J. H. Weisburger1'2, and N. P. Page1'^, NCI project
 

officers, and Dr. F. M. Garner'*, the principal investigator. The
 

administration of the test chemical and the observation of the
 

animals were supervised by Dr. Garner and Mr. S. Johnson, the
 

co-principal investigator, with the technical assistance of Mr.
 

R. Cypher^, Mr. H. D. Thornett^, and Mr. D. J. Howard^. Ms. J.
 

Blalock^ was responsible for assembly of data.
 

iii
 
Histopathologic examination of the rats was performed by Drs. R.
 

J. Montali4, H. Seibold4, N. J. Wosu4, and P. Hildebrandt4.
 

Mouse tissues were examined by Drs. Wosu and B. C. Zook4. All
 

diagnoses of tumors were reviewed by Dr. Montali, who prepared
 

the interpretive pathology narrative.
 

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

Mason Research Institute^. The statistical analyses were
 

performed by Dr. J. R. Joiner" and Ms. P. L. Yong^, using methods
 

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

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

Murrill°, dosage solutions were analyzed by Mr. H. Paulin^, and
 

the results of the analyses were reviewed by Dr. S. S. Olin".
 

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

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

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

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

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

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

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

P. J. Graboske.
 

The following other scientists at NCI were responsible for
 

evaluating the bioassay, interpreting the results, and reporting
 

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

Fielding Douglas, Dr. Dawn G. Goodman', Dr. Richard A. Griesemer,
 

Dr. Morton H. Levitt. Dr. Harry A. Milman, Dr. Thomas W. Orme,
 

Dr. Robert A. Squire^, Dr. Sherman F. Stinson, Dr. Jerrold M.
 

Ward, and Dr. Carrie E. Whitmire.
 

^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

with the Naylor Dana Institute for Disease Prevention,
 

American Health Fouundation, Hammond House Road, Valhalla,
 

New York.
 

iv
 
at the Environmental Protection Agency, 401 M Street, S.W. ,
 

Washington, D. C.
 

^Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington,
 

Maryland.
 

Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

6

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

Maryland.
 

'Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute, National
 

Institutes of Health, Bethesda, Maryland.
 

8

Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

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

Suite 660, Washington, D. C.
 

'Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of 3-(chloromethyl)pyridine hydrochloride for possible
 

carcinogenicity was conducted by administering the test chemical
 

by gavage to Fischer 344 rats and B6C3F1 mice.
 

Groups of 50 rats and 50 mice of each sex were administered
 

3-(chloromethyl)pyridine hydrochloride in a vehicle of distilled
 

water three times per week at one of the following doses, either
 

75 or 150 mg/kg body weight for the rats and either 100 or 200
 

mg/kg body weight for the mice. The low-dose rats were dosed for
 

103 weeks and the low-dose mice for 102 weeks. Because of early
 

deaths in the high-dose animals, the high-dose rats were dosed
 

for only 83 weeks and the high-dose mice for only 81 weeks.
 

Controls consisted of groups of 20 rats and 20 mice of each sex
 

which were administered the vehicle only for 104 weeks. All
 

surviving rats and mice were killed at 104 weeks.
 

Mean body weights of the male and female rats were lower in the
 

dosed groups than in the corresponding control groups, and the
 

depressions in weight were dose related. At the termination of
 

the administration of the test chemical to the high-dose groups
 

of rats, the mean body weights of these groups recovered rapidly.
 

The mean body weights of the male mice were unaffected by the
 

administration of the chemical; those of the females were only
 

slightly affected. Mortality was generally higher in the dosed
 

groups of rats and mice than in the corresponding control groups
 

and was dose related in all tests except those using the female
 

mice; however, sufficient numbers of animals of each species and
 

sex were at risk for the development of late-appearing tumors.
 

In rats, proliferative squamous-cell lesions of the forestomach
 

were observed in the dosed males (carcinomas: high-dose 1/50;
 

papillomas: low-dose 1/47, high-dose 2/50; hyperplasias: low-dose
 

1/47, high-dose 2/50) and the dosed females (carcinomas:
 

high-dose 1/48), but not in the male or female vehicle controls.
 

The results of the Fisher exact test were not significant for
 

squamous-cell papillomas or carcinomas. However, comparison of
 

the incidence of these tumors in the high-dose males with that in
 

99 historical vehicle controls shows that the probability that
 

three or more such tumors did not occur by chance, given that
 

vii
 
none have been observed in the controls in this laboratory, is
 

P = 0.014.
 

In mice, squamous-cell papillomas or carcinomas of the
 

forestomach occurred in the low- and high-dose groups of each
 

sex, but not in the corresponding control groups. The incidence
 

in the high-dose males was significantly higher (P = 0.025) than
 

that in the control males (males: vehicle controls 0/19, low-dose
 

2/43, high-dose 10/47 [21%]; females: vehicle controls 0/19,
 

low-dose 1/45, high-dose 5/48 [10%]). Comparison of the
 

incidences of these tumors in the high-dose males and females
 

with those observed in the corresponding groups of 100 historical
 

vehicle controls of each sex shows that the probability that
 

their occurrence was not due to chance is P < 0.001. Also, a
 

life- table analysis of the incidence in males indicated a
 

significant (P = 0.003) increase in tumors over the period of
 

observation (58 weeks to 104 weeks) in relation to an increase in
 

dose.
 

Although the incidence of squamous-cell papillomas and carcinomas
 

in male and female rats was significant only in males compared
 

with historical vehicle controls, these tumors are of the same
 

type as those appearing at the same site in male and female mice.
 

Because these tumors are rare and not found in controls, and
 

because they were found in dosed animals of both species, they
 

are considered to be related to administration of the test
 

chemical by gavage.
 

It is concluded that under the conditions of this bioassay,
 

3-(chloromethyl)pyridine hydrochloride was carcinogenic in male
 

Fischer 344 rats and in B6C3F1 mice of both sexes, producing
 

papillomas and carcinomas at the site of topical application, the
 

stomach.
 

viii
 
TABLE OF CONTENTS
 

Page
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dosage Preparation 4
 

C. Animals 4
 

D. Animal Maintenance 5
 

E. Subchronic Studies 6
 

F. Chronic Studies 8
 

G. Clinical and Pathologic Examinations 8
 

H. Data Recording and Statistical Analyses 11
 

III. Results - Rats 17
 

A. Body Weights and Clinical Signs (Rats) 17
 

B. Survival (Rats) 17
 

C. Pathology (Rats) 20
 

D. Statistical Analyses of Results (Rats) 21
 

IV. Results - Mice 25
 

A. Body Weights and Clinical Signs (Mice) 25
 

B. Survival (Mice) 25
 

C. Pathology (Mice) 28
 

D. Statistical Analyses of Results (Mice) 30
 

V. Discussion 33
 

VI. Bibliography 37
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms
 

in Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 39
 

Table Al Summary of the Incidence of Neoplasms
 

in Male Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 41
 

ix
 
Table A2
 

Appendix B
 

Table Bl
 

Table B2
 

Appendix C
 

Table Cl
 

Table C2
 

Appendix D
 

Table Dl
 

Table D2
 

Summary of the Incidence of Neoplasms
 

in Female Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 45
 

Summary of the Incidence of Neoplasms
 

in Mice Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 49
 

Summary of the Incidence of Neoplasms
 

in Male Mice Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 51
 

Summary of the Incidence of Neoplasms
 

in Female Mice Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 54
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 59
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Male Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 61
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Female Rats Administered 

3-(Chloromethyl)pyridine Hydrochloride 

by Gavage 66 

Summary of the Incidence of Nonneoplastic 

Lesions in Mice Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage... 71 

Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Administered 

3-(Ghloromethyl)pyridine Hydrochloride 

by Gavage 73 

Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Administered 

3-(Chloromethyl)pyridine Hydrochloride 

by Gavage 77 
Appendix E
 

Table El
 

Table E2
 

Appendix F
 

Table Fl
 

Table F2
 

Table 1
 

Table 2
 

Figure 1
 

Figure 2
 

Analyses of the Incidence of Primary
 

Tumors in Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage. 83
 

Analyses of the Incidence of Primary
 

Tumors in Male Rats Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 85
 

Analyses of the Incidence of Primary
 

Tumors in Female Rats Administered
 

3-(Chloromethyl)pyridine Hydrochloride
 

by Gavage 91
 

Analyses of the Incidence of Primary
 

Tumors in Mice Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage.... 95
 

Analyses of the Incidence of Primary
 

Tumors in Male Mice Administered
 

3-(Chloromethyl)pyridine Hydrochloride
 

by Gavage 97
 

Analyses of the Incidence of Primary
 

Tumors in Female Mice Administered
 

3-(Chloromethyl)pyridine Hydrochloride
 

by Gavage ....102
 

TABLES
 

Chronic Studies of 3-(Chloromethyl)pyridine Hydrochloride in Rats 9
 

Chronic Studies of 3-(Chloromethyl)pyridine Hydrochloride in Mice. 10
 

FIGURES
 

Growth Curves for Rats Administered
 

3-(Chloromethyl)pyridine Hydrochloride
 

by Gavage 18
 

Survival Curves for Rats Administered
 

3-(Chloromethyl)pyridine Hydrochloride
 

by Gavage 19
 

xi
 
Figure 3 Growth Curves for Mice Administered 

3-(Chloromethyl)pyridine Hydrochloride 

by Cavage 26 

Figure 4 Survival Curves for Mice Administered 

3-(Chloromethyl)pyridine Hydrochloride 

by Gavage 27 

xii
 
I. INTRODUCTION
 

3-(Chloromethyl)pyridine hydrochloride (CAS 6959-48-4; NCI
 

C03838) is an intermediate that
 

HCI
 

has been proposed for use in the
 

synthesi -u •s f of

 • agricultural i !
,
 

pharmaceutical, and veterinary
 

chemicals (Arnall and Clark, 3<ch'oromethyl)pyridine (hydrochloride)
 

1966). Its synthesis was achieved in the mid 1960's by the
 

Midland Tar Distillers, Ltd., in Great Britain, through a process
 

in which the alpha carbon of the alkyl side chain to the pyridine
 

ring is chlorinated preferentially (Arnall and Clark, 1966).
 

This discovery made it economically feasible for the first time
 

to synthesize large quantities of monochloroalkylpyridine
 

intermediates. This compound is neither manufactured in the
 

United States, nor imported, at the present time (Stanford
 

Research Institute, 1977; USITC, 1977a and 1977b), however, at
 

the time it was selected for bioassay, it was felt that it could
 

become a widely used industrial intermediate.
 

II. MATERIALS AND METHODS
 

A. Chemical
 

Three batches of the test chemical, hereinafter referred to as
 

3-(chloromethyl)pyridine hydrochloride, were obtained from
 

Columbia Organic Chemicals, Columbia, South Carolina, for these
 

studies. These batches were identified by the date of receipt at
 

Midwest Research Institute, Kansas City, Missouri, as Lot No.
 

C02-7-73, Lot No. C012-5-73, and Lot No. C02-25-75. All batches
 

were used during the chronic studies; Lot No. C02-7-73 was also
 

used during the Subchronic studies.
 

The identity of each batch was confirmed by infrared, nuclear
 

magnetic resonance, and ultraviolet spectral analyses. Elemental
 

analyses (C, H, N, Cl) for C^HyNC^ were slightly low for
 

chlorine in Lot Nos. C02-7-73 and C02-25-75. Trace impurities
 

were found in all three lots by thin-layer chromatography. High-


pressure liquid chromatography (ultraviolet detector, 254nm)
 

indicated the presence of a single impurity, accounting for 0.2%
 

of the total peak area in Lot No. C02-7-73, and of three minor
 

impurities (0.49%) in Lot No. C02-25-75. Lot No. C02-7-73
 

contained 0.69 + 0.04% water, Lot No. C012-5-73, < 0.17%, and Lot
 

No. C02-25-75, 0.82 + 0.11%, as determined by Karl Fischer
 
analysis. Throughout this report the term used to represent this
 

material is 3-(chloromethyl)pyridine hydrochloride.
 

These batches were stored at 4°C in the original containers.
 

B. Dosage Preparation
 

Solutions of 3-(chloromethyl)pyridine hydrochloride were prepared
 

in distilled water (Borden Polar Water Co., Beltsville, Md.) at
 

concentrations of 1 and 2% for mice and 0.75 and 1.50% for rats.
 

These were administered by gavage on the same day on which they
 

were prepared.
 

C. Animals
 

Fischer 344 rats and B6C3F1 mice of each sex were obtained from
 

Charles River Breeding Laboratories, Wilmington, Massachusetts,
 

under a contract with the Division of Cancer Treatment, NCI.
 

The animals were 28 days of age when received at the laboratory
 

and were quarantined for 2 weeks prior to the start of the
 

bioassay. Animals with clinical signs of disease and runts were
 

killed. The remaining animals were segregated into equal weight
 

groups and assigned to control or dosed groups in such a way that
 

the mean weights of animals in each cage within a particular
 

group were approximately the same.
 
D. Animal Maintenance
 

All animals were housed in temperature- and humidity-controlled
 

rooms. The temperature was maintained at 21-25°C and the
 

relative humidity at 45-55%. There were 10 changes of room air
 

per hour, and the incoming and exhaust air was filtered through
 

high efficiency particulate air (HEPA) filters (Flanders Filters,
 

McLean, Va.). The animal rooms were positively pressurized with
 

respect to the exit hall and negatively pressurized with respect
 

to the entrance hall. Rooms were illuminated by cool white
 

fluorescent lighting 8 hours per day.
 

Rats were housed four per cage and mice five per cage in solid
 

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

cage was covered with a wire mesh screen and a sheet of filter
 

paper and contained heat-treated hardwood chip bedding
 

(Absorb-Dri , Lab Products, Garfield, N.J.) in the bottom. Cages
 

and water bottles were sanitized two times per week, racks were
 

washed each month, and feed hoppers were sanitized once per week
 

at approximately 82°C; bedding was replaced two times per week
 

and filter paper was replaced each month.
 

The animals were fed Wayne® Lab Blox (Allied Mills, Inc.,
 

Chicago, 111.) and received fresh diets three times per week.
 
Water bottles contained tap water which had been acidified to pH
 

2.5.
 

Rats and mice were housed in separate rooms. Control and dosed
 

animals were housed in the same room. Animals administered
 

3-(chloromethyl)pyridine hydrochloride were housed in the same
 

room with animals administered the following chemicals:
 

Rats
 

Feed Studies
 

(CAS 105-55-5) N,N-diethylthiourea
 

(CAS 99-56-9) 4-nitro-o-phenylenediamine
 

(CAS 89-25-8) l-phenyl-3-methyl-5-pyrazolone
 

Mice
 

Feed Studies
 

(CAS 2735-04-8) 2,4-dimethoxyaniline
 

(CAS 140-49-8) 4-chloroacetylacetanilide
 

(CAS 139-94-6) nithiazide
 

(CAS 624-18-0) p-phenylenediamine dihydrochloride
 

(CAS 99-56-9) 4-nitro-o-phenylenediamine
 

(CAS 89-25-8) l-phenyl-3-methyl-5-pyrazolone
 

Gavage Studies
 

(CAS 512-56-1) trimethylphosphate
 

(CAS 4377-33-7) 2-(chloromethyl)pyridine hydrochloride
 

(CAS 1955-45-9) pivalolactone
 

E. Subchronic Studies
 

The LD5Q for 3-(chloromethyl)pyridine hydrochloride administered
 

orally to either Fischer 344 rats or B6C3F1 mice has been
 

reported as 316 rag/kg (Litton-Bionetics, Inc., 1973).
 

6
 
Subchronic studies were conducted with Fischer 344 rats and
 

B6C3F1 mice to estimate the maximum tolerated doses of
 

3-(chloromethyl)pyridine hydrochloride, on the basis of which two
 

doses (hereinafter called "low" and "high" doses) were determined
 

for the chronic studies. For the Subchronic studies, the test
 

chemical was administered by gavage, three times per week, at
 

doses of 68, 100, 147, 215, or 316 rag/kg to rats, and 100, 147,
 

215, 316, or 464 mg/kg to mice. Five males and five females were
 

tested at each dose, and groups of equal size served as vehicle
 

controls, receiving distilled water only. Following a 7-week
 

period of administration of the test chemical, the animals were
 

observed for 1 week and then killed and necropsied.
 

Within the first 3 weeks of the Subchronic test, two of the male
 

rats and all of the female rats died at the highest dose, 316
 

mg/kg. In those male rats that were alive at week 7, mean body
 

weights were depressed to the same extent in all groups, and in
 

no case did this exceed 15% of controls. In the surviving
 

females, there were no apparent effects on body weights.
 

In the mice, all males and females administered the highest dose,
 

464 mg/kg, died during the first week. At the end of week 7,
 

there were only small weight depressions in the surviving female
 

groups and no apparent weight depression in the males.
 
No signs of toxicity were found on gross pathologic examination
 

of the organs taken from rats and mice.
 

The low and high doses for the chronic studies using rats were
 

set at 75 and 150 mg/kg, and those for the chronic studies using
 

mice were set at 100 and 200 mg/kg, respectively.
 

F. Chronic Studies
 

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

chronic studies are shown in tables 1 and 2.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity and
 

were weighed and palpated for masses at regular intervals.
 

Animals that were moribund and those that survived to the
 

termination of the bioassay were killed using C02 and necropsied.
 

The pathologic evaluation consisted of gross and microscopic
 

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

from killed animals and from animals found dead. The following
 

tissues were examined microscopically: skin, lymph nodes, mammary
 

gland, salivary gland, bone marrow, trachea, lungs and bronchi,
 

heart, thyroid, parathyroid, esophagus, stomach, small intestine,
 

large intestine, liver, gallbladder (mice), pancreas, spleen,
 

kidney, adrenal, urinary bladder, prostate or uterus, testis or
 
Table 1. Chronic Studies of 3-(Chloromethyl)pyridine
 

Hydrochloride in Rats
 

3-(Chloromethyl )
 

pyridine
 

Sex and Initial Hydrochloride Time on Study
 

Test No. of Doseb Dosed Observed
 

Group Animals3 (mg/kg) (weeks) (weeks)
 

Male
 

Vehicle-Control0 20 0 104
 

Low -Dose 50 75 103 1
 

High-Dose 50 150 83d

 21
 

Female
 

Vehicle-Control0 20 0 104
 

Low-Dose 50 75 103 1
 

High-Dose 50 150 83d

 21
 

a

Rats were approximately 6 weeks of age when placed on study.
 

^3-(Chloromethyl)pyridine hydrochloride was administered three
 

times per week in distilled water at a volume of 1 ml per 100 gm
 

body weight. Doses were calculated using the mean body weight of
 

the group and were adjusted every month.
 

°Vehicle controls received distilled water three times per week
 

at a volume of 1 ml per 100 gm body weight.
 

d

Because of early deaths, the high-dose rats were dosed for only
 

83 weeks.
 
Table 2. Chronic Studies of 3-(Chloromethyl)pyridine
 

Hydrochloride in Mice
 

3-(Chloromethyl )
 

Sex and 

Test 

Group 

Initial

No. of

Animals3

pyridine 

 Hydrochloride 

Doseb 

 (mg/kg) 

Time o

Dosed

(weeks)

n Study 

 Observed 

 (weeks) 

Male 

Vehicle-Control0 20 0 104 

Low -Dose 50 100 102 2 

High-Dose 50 200 81d 23 

Female 

Vehicle-Control0 20 0 104 

Low-Dose 50 100 102 2 

High-Dose 50 200 81d

 23 

a

Mice were approximately 6 weeks of age when placed on study.
 

'33-(Chloromethyl)pyridine hydrochloride was administered three
 

times per week in distilled water at a volume of 1 ml per 100
 

gm body weight. Doses were calculated using the mean body weight
 

of the group and were adjusted every month.
 

°Vehicle controls received distilled water three times per week at
 

a volume of 1 ml per 100 gm body weight.
 

^Because of early deaths, the high-dose mice were dosed for only
 

81 weeks.
 

10
 
ovary, brain, and pituitary. Occasionally, additional tissues
 

were also examined microscopically. The different tissues were
 

preserved in 10% buffered formalin, embedded in paraffin,
 

sectioned, and stained with hematoxylin and eosin. Special
 

staining techniques were utilized when indicated for more
 

definitive diagnosis.
 

A few tissues from some animals were not examined, particularly
 

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

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

state of autolysis as to preclude histopathologic evaluation.
 

Thus, the number of animals from which particular organs or
 

tissues were examined microscopically varies, and does not
 

necessarily represent the number of animals that were placed on
 

study in each group.
 

H. Data Recording and Statistical Analyses
 

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

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

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

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

11
 
verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear on the possibility of carcinogenicity are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

report in the form of graphs. Animals were statistically
 

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

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

causes were not statistically censored. Statistical analyses for
 

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

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

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

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

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

the denominators included only those animals for which that site
 

12
 
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
 

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

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

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

where the Fisher exact P values are shown.
 

The Cochran-Armitage test for linear trend in proportions, with
 

continuity correction (Armitage, 1971), was also used when
 

appropriate. Under the assumption of a linear trend, this test
 

determines if the slope of the dose-response curve is different
 

13
 
from zero at the one-tailed 0.05 level of significance. Unless
 

otherwise noted, the direction of the significant trend is a
 

positive dose relationship. This method also provides a two-


tailed test of departure from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

first tumor was observed were excluded by basing the statistical
 

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

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

such an early tumor was found, comparisons were based exclusively
 

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

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

obtained, the standard procedures for analyses of the incidence
 

of tumors (Fisher exact tests, Cochran-Armitage tests, etc.) were
 

followed.
 

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

incidence of tumors. Curves of the proportions surviving without
 

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

The week during which an animal died naturally or was sacrificed
 

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

methods of comparing these curves were used for two groups;
 

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

14
 
groups. The statistical tests for the incidence of tumors which
 

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

noted, in the direction of a positive dose relationship.
 

Significant departures from linearity (P < 0.05, two-tailed test)
 

were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared to its control was calculated
 

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

relative risk is defined as P£/PC where pt is the true binomial
 

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

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

spontaneous incidence of the same type of tumor in a control
 

group. The hypothesis of equality between the true proportion of
 

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

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

of unity represent the condition of a larger proportion in the
 

dosed group than in the control.
 

The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical
 

analyses. The interpretation of the limits is that in
 

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

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

control group would be within the interval calculated from the
 

15
 
experiment. When the lower limit of the confidence interval is
 

greater than one, it can be inferred that a statistically
 

significant result (P < 0.025 one-tailed test when the control
 

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

zero) has occurred. When the lower limit is less than unity, but
 

the upper limit is greater than unity, the lower limit indicates
 

the absence of a significant result while the upper limit
 

indicates that there is a theoretical possibility of the
 

induction of tumors by the test chemical, which could not be
 

detected under the conditions of this test.
 

16
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

The mean body weights of the male and female rats were lower in
 

the dosed groups than in the corresponding control groups, and
 

the depressions in weight were dose related (figure 1). At the
 

termination of the administration of the test chemical to the
 

high-dose groups of rats (83 weeks), the mean body weights of
 

these groups increased. Fluctuation in the growth curve may be
 

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

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

related to administration of the test chemical were reported.
 

B

' Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered 3-(chloromethyl)pyridine hydrochloride by gavage at the doses of this
 

bioassay, together with those of the vehicle controls, are shown
 

in figure 2. The result of the Tarone test for positive dose-


related trend in mortality is significant (P < 0.001) in each
 

sex.
 

In male rats, 21/50 (42%) of the high-dose group, 38/50 (76%) of
 

the low-dose group, and 19/20 (95%) of the control group survived
 

17
 
(J 300­

1-

X 

o 

UJ 250­

Q

A 

A

D

D O

 A A 

D u 

o °

 A

 ° 

A A A A A 

Q 

O 200-| 

MALE RATS 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

10 20 40 50 60 70 

TIME ON STUDY (WEEKS) 

90 100 110 

a

IX 

o 

UJ

5 

o 

O

 300­

 250­

 200-^ 

0

A 

D D

D O °

g ,Bssiii: ^ 

g A A A A 

o

A § A 

g 150­

FEMALE RATS 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

20 30 40 50 60 70 

TIME ON STUDY (WEEKS) 

80 90 100 110 

Figure 1. Growth Curves for Rats Administered 3-(Chloromethyl)Pyridine Hydrochloride by Gavage 

18 
•g

3

ro PROBABILITY OF SURVIVAL PROBABILITY OF SUR

v» > o a o o -• O O O O O

 O

O 

p

 £ S 2 p i 3 8 3 fc !S 8 8 S g fe S S S 

i

S_

n

gO LO

W DOSE 

D > O CD

^ E 4? ^ 

m 

....>--:

c I 1 i 

' i f £ 

i

|

a 

 E 9- m 

tf 3 i i; 

? 5 

 r8­

) 6) 71

 

 S 

; 53 i5

8. 

w_ 

^f 

-t g. ^

I

S

a. m ^ S

m CO O >•' O Z a. Z tn 

JU.

STUDY (WEEKS]

 

n

5°

3

CD

3­

1

3 >•

i !

!..»! 

0 7

 

D

•* 1^1 > (WEEKS 

f~ 

I 9 r' 1 

CD g. >vt 

^ § , 

i

V 

f

O 9-' S- _ 

o^ 

n i S 

2. c^>r* >• n. 

CD

>-: 

ty 1

<• e> ^6 i e> 

CT

o>

S 
to termination of the study. In females, 26/50 (52%) of the
 

high-dose group, 40/50 (80%) of the low-dose group, and all 20 of
 

the control group lived to termination of the study.
 

Sufficient numbers of rats of each sex were at risk for the
 

development of late-appearing tumors.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

are summarized in Appendix C, tables Cl and C2.
 

A variety of neoplastic lesions were evident in the control and
 

dosed rats. Except for those of the forestomach, the tumors
 

occurred in a random fashion in all groups. In the stomachs of
 

dosed rats, there were both neoplastic and hyperplastic lesions
 

as noted in the following table:
 

Male Rats Female Rats
 

Vehicle Low High Vehicle Low High
 

Control Dose Dose Control Dose Dose
 

Number of Animals
 

with Stomach Examined
 

Microscopically 19 47 50 20 45 48
 

Squamous-cell
 

carcinoma 1(2%) 1(2%)
 

Squamous-cell
 

papilloma 1(2%) 2(4%)
 

Squamous-cell
 

hyperplasia 1(2%) 2(4%)
 

20
 
Grossly, the gastric nodules were described as single, white,
 

pinpoint to 1 mm or 2 mm in diameter, warty growths, limited to
 

the squamous portion of the stomach. Microscopically, the
 

squamous-cell carcinomas were well differentiated and consisted
 

of nests of squamous cells, some with keratin formation, that
 

obliterated or invaded the muscularis mucosa but did not extend
 

below the submucosa. The papillomas consisted of raised areas of
 

spikes of hyperplastic squamous epithelium with hyperkeratotic
 

caps. Chronic inflammation occurred in the submucosa beneath
 

some of these lesions.
 

In organs other than the stomach, there were some degenerative
 

and inflammatory lesions of the type usually encountered in aged
 

rats, but none of the lesions were attributed to the test
 

chemical. Chemically related lesions were not found in rats
 

dying early in the study.
 

Based on the histopathologic examination, it was concluded that
 

the gastric squamous-cell neoplastic lesions may be associated
 

with the administration of 3-(chloromethyl)pyridine hydrochloride
 

in rats under the conditions of this bioassay.
 

D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

21
 
least two animals of one group and at an incidence of at least 5%
 

in one or more than one group. The early deaths in the high-dose
 

group resulted in a shortened dosage period which obscures the
 

meaning of trend analyses. Thus, these analyses have been
 

omitted in the tables.
 

The results of the Fisher exact tests comparing the incidences of
 

tumors in each of the dosed groups with that in the control group
 

are not significant in the positive direction in either sex. The
 

combined incidence of squamous-cell papillomas or carcinomas of
 

the stomach in male rats was 0/19 in the controls, 1/47 (2%) in
 

the low-dose group, and 3/50 (6%) in the high-dose group. The
 

records of control animals at this laboratory indicate no such
 

tumors occurred in 99 historical gavage vehicle-control male
 

rats. The results of the Fisher exact test of these incidences
 

are not significant, but under the estimate of 1% incidence in
 

male control rats, the binomial probability (Fears, 1977) of
 

three or more such tumors in 50 male rats is significant (P =
 

0.014). This analysis suggests an association between the
 

occurrence of squamous-cell tumors in the high-dose group and the
 

administration of the chemical. There was one squamous-cell
 

carcinoma in the high-dose group of female rats compared with
 

none in the 100 gavage vehicle-control female rats seen at this
 

laboratory.
 

22
 
The results of the Fisher exact test on incidences of tumors of
 

the pancreas and tumors of the testis in dosed male rats were
 

significant in the negative direction. The higher incidences of
 

tumors in the control group than in the dosed groups may have
 

occurred because the dosed animals did not live as long as the
 

control animals.
 

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

in the tables, the value of one or less than one is included;
 

this indicates the absence of significant positive results. It
 

should also be noted that each of the intervals has an upper
 

limit greater than one (except those of the incidences of islet-


cell tumors and of tumors of the testis in high-dose male rats),
 

indicating the theoretical possibility of the induction of tumors
 

by 3-(chloromethyl)pyridine hydrochloride, which could not be
 

detected under the conditions of this test.
 

23
 

IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

The mean body weights of the male mice were unaffected by
 

administration of the 3-(chloromethyl)pyridine hydrochloride, and
 

those of the females were only slightly affected (figure 3).
 

Fluctuation in the growth curve may be due to mortality; as the
 

size of a group diminishes, the mean body weight may be subject
 

to variation. No other clinical signs related to administration
 

of the test chemical were reported.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered 3-(chloromethyl)pyridine hydrochloride by gavage at the doses of this
 

bioassay, together with those of the vehicle controls, are shown
 

in figure 4. In male mice, the result of the Tarone test for
 

positive dose-related trend in mortality is significant (P =
 

0.006). In females, the result of the Tarone test is not
 

significant.
 

There were 23/50 (46%) of the male high-dose group, 30/50 (60%)
 

of the low-dose group, and 15/20 (75%) of the control group
 

surviving to termination of the study. There were 30/50 (60%) of
 

25
 
A D 

g g o 

g 

8 g g 8 n 8n o a n Sa O D A a Q 

o

D 

(9 

uj 26­

a

o 

Q 

g 

4 

UJ 

5 15-1 

MALE MICE 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

10 20 40 50 60 70 90 100 

TIME ON STUDY (WEEKS) 

a a

D D 

ogg £ D A o 

D no 

S A OAA 

O OD 

(9 

Ul

S 

 25­

8 

D a a 

8 

Q

§ »-

Z H 

4 

ui 

5 15-1 

FEMALE MICE 

D VEHICLE CONTROL 

O LOW DOSE 

A HIGH DOSE 

10 20 40 50 60 70 80 90 100 110 

TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves for Mice Administered 3-(Chloromethyl)Pyridine Hydrochforide by Gavage 

26 
a 0­ A-A-- ,

- A A, ^% A n A , °^' L i" ' : O"L_ 1 *> L 

1 

> A o T 

• o 

CO 

u. 

o ngn_ 

j V 

to 040. 

0) 

o 

DC 

F flALE MCE 

/EHICLE CO 

O OW DOSE

 

A rUGH DOSE

 

n nn10 20 40 50 60 10 80 90 100 110 

TIME ON STUDY (WEEKS) 

6— i. A- t_>" ^ 0.90­

/ S" 0­

"&b= 

0.80­

0—o A

A­ [b—D _l 0.70­ A A > ""\. •••••A DC 0.60­

U9 

FEMALE VIICE 

O vEHICLE CO

 

O L OW DOSE

 

A H IGH DOSE 

20 30 40 50 60 70 90 100 

TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves for Mice Administered 3-(Chloromethyl)Pyridine Hydrochloride by Gavage PROBABILIT

Y OF 

O O O O O O s s s s § s 1 1 1 1 1 t 

27 
the female high-dose group, 39/50 (78%) of the low-dose group,
 

and 15/20 (75%) of the control group surviving to termination of
 

the study.
 

Sufficient numbers of mice of each sex were at risk for the
 

development of late-appearing tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

are summarized in Appendix D, tables Dl and D2.
 

A variety of neoplastic lesions were evident in control and dosed
 

mice. Except for those in the stomach, they were spontaneous
 

tumors occurring in a random fashion.
 

In the stomach of dosed mice, there were hyperplastic and
 

neoplastic lesions that did not occur in the controls. The
 

lesions were limited to the squamous portion of the stomach
 

(forestomach) and occurred at the following incidence:
 

28
 
Male Mice Female Mice
 

Vehicle Low High Vehicle Low High
 

Control Dose Dose Control Dose Dose
 

Number of Animals
 

with Stomach Examined
 

Microscopically 19 43 47 19 45
 48
 

Squamous-cell
 

carcinoma 2(4%) 2(4%)
 

Squamous-cell
 

papilloma 2(5%) 8(17%) 1(2%) 3(6%)
 

Squamous-cell
 

hyperplasia 1(2%) 2(4%) 3(6%)
 

One of the squamous-cell carcinomas in a high-dose male mouse was
 

described grossly as a large, hard mass in the stomach.
 

Microscopically, this tumor, which arose from the forestomach,
 

had invaded the liver and pancreas and metastasized to the
 

mesenteric lymph node. The other squamous-cell carcinomas were
 

well differentiated and consisted of epithelial nests that
 

invaded through the muscularis mucosa or into the stalk but did
 

not extend beyond the gastric submucosa.
 

Grossly, the papillomas were described as single, pinpoint to
 

small white nodules arising from the squamous gastric mucosa.
 

Histologically, in favorable sections, they consisted of tall
 

hyperplastic spikes of squamous epithelium with hyperkeratotic
 

caps that lined connective tissue stalks derived from the stomach
 

wall.
 

The hyperplastic areas consisted of increased layers of basophi29
 
lie squamous cells and hyperkeratosis. In some of the affected
 

mice there was mild to moderate chronic inflammation in the
 

submucosa underlying the papillomas and hyperplastic areas.
 

Other tumors observed were those commonly recorded as spontaneous
 

neoplasms of mice. These included a few hepatocellular and
 

pulmonary tumors and some hematopoietic neoplasms, all with a
 

slightly elevated incidence in the dosed mice. The lowered
 

survival rates of the dosed mice made it difficult to interpret
 

the incidences of these tumors, particularly when the differences
 

in incidences between the dosed and control groups were marginal.
 

Other nonneoplastic lesions were of the type usually found in
 

aged mice, and none was attributed to the test chemical.
 

Chemical-related lesions did not occur in mice dying early in the
 

study.
 

Based on the histopathologic examination, it was concluded that
 

the squamous-cell tumors of the forestomach were associated with
 

the administration of 3-(chloromethyl)pyridine hydrochloride in
 

B6C3F1 mice under the conditions of this bioassay.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

30
 
least two animals of one group and at an incidence of at least 5%
 

in one or more than one group. The early deaths in the high-dose
 

group resulted in a shortened dosage period which obscures the
 

meaning of trend analyses. Thus, these analyses have been omitted
 

in the tables.
 

The probability level of the Fisher exact test comparing the
 

combined incidence of squamous-cell papillomas or carcinomas of
 

the stomach in the high-dose group of male mice with that in the
 

control group of male mice is at the upper limit (P = 0.025) of
 

that required for significance by the Bonferroni inequality
 

criterion for multiple comparisons. In females, the results of
 

the Fisher exact test are not significant; however, these tumors
 

occurred only in the dosed groups. No such tumors have been
 

observed at this laboratory in any of 100 historical gavage
 

vehicle-control male or female mice. Under the estimate of the
 

binomial parameter (Fears, 1977) of an incidence of 1% in control
 

mice, the binomial probability of 10/47 (21%) or higher incidence
 

in the high-dose male mice and of 5/48 (10%) incidence or higher
 

in the high-dose female mice is significant at a level less than
 

P = 0.001.
 

In addition to the analysis described above, a life-table
 

analysis was performed, utilizing the time at which a tumor was
 

observed (see section H, p. 13) and based on the time-weighted
 

31
 
dose calculated over the 104 weeks of the bioassay. This
 

analysis of the incidence in males (controls 0/19, low-dose 2/43
 

[5%], high-dose 10/47 [21%]) ranging from the first tumor
 

observed in the high-dose group (58 weeks) to the end of the
 

bioassay (104 weeks) indicated a significant (P = 0.003) increase
 

in the observation of tumors over this time period in relation to
 

increase in dose. These analyses indicate an association of
 

squamous-cell papillomas or carcinomas of the stomach with the
 

administration of the test chemical.
 

32
 
V. DISCUSSION
 

3-(Chloromethyl)pyridine hydrochloride was toxic for Fischer 344
 

rats and B6C3F1 mice inasmuch as mean body weights were depressed
 

in the dosed rats and mortality was generally higher in both
 

dosed rats and dosed mice than in corresponding control groups.
 

The nature of the toxic effect could not be established
 

histopathologically. The depression in mean body weight was dose
 

related in both sexes of rats, and the mortality was dose related
 

in rats and male mice. In female mice only the survival of the
 

high-dose group was affected. Because of early deaths in the
 

high-dose groups of both the rats and the mice, administration of
 

the test chemical was terminated about 20 weeks earlier for these
 

groups than for the low-dose groups of both species. Sufficient
 

numbers of animals of each species and sex were at risk, however,
 

for the development of late-appearing tumors.
 

In rats, proliferative squamous-cell lesions of the stomach were
 

observed in the dosed males (carcinomas: high-dose 1/50;
 

papillomas: low-dose 1/47, high-dose 2/50; hyperplasias: low-dose
 

1/47, high-dose 2/50) and a dosed female (carcinomas: high- dose
 

1/48), but not in the male or female vehicle controls. The
 

results of the Fisher exact test were not significant for
 

squamous-cell papillomas or carcinomas. However, comparison of
 

the incidence of these tumors in the high-dose males with that in
 

33
 
99 historical gavage vehicle controls shows that the probability
 

that three or more such tumors did not occur by chance, given
 

that none have been observed in the controls in this laboratory,
 

is 0.014.
 

In mice, squamous-cell papillomas or carcinomas of the stomach
 

occurred in the low- and high-dose groups of each sex, but not in
 

the corresponding control groups of the 100 historical controls
 

of each sex. The incidence in the high-dose males was
 

significantly higher (P = 0.025) than that in the control males
 

(males: vehicle controls 0/19, low-dose 2/43, high-dose 10/47
 

[21%]; females: vehicle controls 0/19, low-dose 1/45, high-dose
 

5/48 [10%]). Comparison of the incidences of these tumors in the
 

high-dose males and females with those observed in the
 

corresponding groups of 100 historical vehicle controls of each
 

sex shows that the probability that their occurrence was not due
 

to chance is P < 0.001. Also, a life-table analysis of the
 

incidence in males indicated a significant (P = 0.003) increase
 

in tumors over the period of observation (58 weeks to 104 weeks)
 

in relation to an increase in dose.
 

Although the incidence of squamous-cell papillomas and carcinomas
 

in male rats was significant only in comparison with historical
 

vehicle controls, these tumors are of the same types as those
 

that appeared at the same site in male and female mice. Because
 

34
 
these tumors are rare and not found in controls, and because they
 

were found in dosed animals of both species, they are considered
 

to be related to administration of the test chemical by gavage in
 

both species.
 

It is concluded that under the conditions of this bioassay,
 

3-(chloromethyl)pyridine hydrochloride was carcinogenic in male
 

Fischer 344 rats and B6C3F1 mice of both sexes, producing
 

papillomas and carcinomas of the forestomach. Neoplastic lesions
 

related to chemical administration were restricted to the site of
 

topical application, the stomach.
 

35
 

VI. BIBLIOGRAPHY
 

Arnall, P. and Clark, N. R., Monochloroalkylpyridines.
 

Manufacturing Chemist and Aerosol News 37(7);39-41, 1966.
 

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

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

Berenblum, I., ed., Carcinogenic!ty Testing: A Report of the
 

Panel of Carcinogenic!ty oj the Cancer Research Commission
 

of UIGC, Vol. _2, International Union Against Cancer, Geneva,
 

1969.
 

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

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

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

1970, pp. 48-52.
 

Fears, T. R., Tarone, R. E., and Chu, K. C., False-positive and
 

false-negative rates for carcinogenicity screens. Cancer
 

Res. 37;1941-1945, 1977.
 

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

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Statist. Inst. 39(2);148-169,
 

1971.
 

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

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

1958.
 

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

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

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

Litton-Bioiietics, Inc., Progress Report for Contract No.
 

NIH-NCI-E-C-72-3252 submitted to the National Cancer
 

Institute, August 15, 1973 (revised October 10, 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. 12:1073-1081, 1972.
 

Stanford Research Institute, Directory of Chemical Producers,
 

Menlo Park, Calif., 1977.
 

37
 
Tarone, R. E., Tests for trend in life table analysis. Biometrika
 

jx2 (3): 679-682, 1975. 

United States International Trade Commission, Imports of 

Benzenoid Chemicals and Products, 1976. USITC Publication 

828, United States International Trade Commission, 

Washington, D. C., 1977a.
 

United States International Trade Commission, Synthetic Organic
 

Chemicals, United States Production and Sales, 1976. USITC
 

Publication 833, United States International Trade
 

Commission, Washington D. C., 1977b.
 

38
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN RATS
 

ADMINISTERED 3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

BY GAVAGE
 

39
 

TABLE A1. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

ANIMAL S INITIALLY IN STUDY 20 50 50 

ANIMAL S NECBOPSIED 20 50 50 

ANIMAL S EXAMINE D HI STOPATHOLOGICALLY 20 49 50 

INTEGUMENTAR Y SYSTEM 

*SUBCUT TISSUE (20) (50) (50) 

FIBEOHA 1 (2%) 

FIEROSRRCOMA 1 (2») 2 (4K) 

NEUROFIBBOBA 1 (2%) 

RESPIRATORY SYSTEM 

tLUN G (20) (49) (49) 

ALVECLAE/flEONCHIOLA R CARCINOMA 2 (10X) 3 (6») 

C-CELL CARCINOMA, HETASTATIC 1 (5%) 

FIEROSARCOMA, METASTATIC 1 (2X) 

HEMATOPOIETIC SYSTEB 

*BULTIPLE ORGAN S (20) (50) (50) 

LEUKEMIA,NOS 1 (5%) 2 (4S) 

UNDIFFERENTIATE D LEUKEBI A 1 (5%) 1 (2X) 

CIRCULATOR Y SYSTEM 

*ELOOD VESSEL (20) (50) (50) 

C-CJLL CARCINOMA, BETASTATIC 1 (2X) 

DIGESTIVE SYSTaM 

tLIVER (19) (48) (48)

 

NEOELASTIC NODULE 1 (5X)

 

HEPATOCELLULAR CARCINOM A 1 (2%)

 

tSTOBACH (19) (H7) (50)

 

SQUAHOyS CELL PAPILLOHA 1 (2X1

 

* NUHBEB OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* SOMBEfi. OF ANIHAL S NECROPSIED 

41 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

SQUAHOU S CELL CARCINOM A 1 <2X) 

URINAfi Y SYSTEM 

NONE 

ENDOCRIN E SYSTEM 

#PITUITAR Y 

CHRCMOPHOBE

CHROHOPHOBE

 ADENOM A 

 CARCINCB A 

(19) 

1 (5%) 

(42) 

4 (10X) 

3 (1%) 

(41) 

1 (2X) 

1 (2%) 

*ADRENA L (19) (49) (49) 

CORTICAL ADENOM A 

CO&TICAL CARCINOM A 

PHEOCHROMOCYTOM A 

1 (5%) 

1 15%) 

2 (11X) 

1 (2*) 

2 (4%) 

1

1

 (2%) 

(2%) 

#THYRCIE 

FOLIICULAR-CELL ADENOM A 

(") (39) 

1 (3%) 

(41) 

FOLLICULAR-CELL CARCINOM A 1 (6%) 

C-CELL

C-CELL

 ADENOM A 

 CARCINOMA 1 (6*) 

1

1

 PS) 

 OX) 

1 (2%) 

# PANCREATIC ISLETS 

IS L IT-CELL ADENOM A 

(20) 

3 (15X) 

(48) 

2 (H%) 

(49) 

ISLET-CELL CARCINOM A 1 (2*) 

REPRODUCTIVE SYSTEM 

*PREPUTIAL GLAN D (20) (50) (50) 

ADEKCHA , NOS 1 (5%) 

tTlSTIS (19) (46) (47) 

INTERSTITIAL-CELL TUMOR 17 (89X) 38 (83%) 20 (43X) 

NERVOU S SYSTEM 

IBEAIN (20) (46) (47) 

6LICBLASTOMA MULTIFORM E 1 (2%) 

SPECIAL SENSE ORGANS 

__NOJ! 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMALS NECEOPSIEC 

42 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

MUSCULCSKEIETA L SYSTE M 

*«ANDIBL E (20) (50) (50) 

FIEKOSARCOM A 1 (2%) — — — — — — — — —••— — — •———• — — — — — — •— — — — — — — — — — — — — — ..———__.— -.— — _-—.._._,— — ___—____-._»_._.____,— ».___,— __„_, 

BOD Y CAVITIES 

*MESENTER Y (20) (50) (50) 

LIECM A 1 (2%) 

*TONIC A VAGINALI S (20) (50) (50) 

MESOTHELIOMA , NOS 1 (2%) 

ALL OTHER SYSTEMS 

NON E 

ANIMA L EISPOSITION SUMMAR Y 

ANIMA1 S INITIALLY IN STUDY 20 50 50 

NATUFA I DEATHS 1 10 27 

MORIEUN D SACRIFICE 1 2 

SCHELULE 0 SACRIFICE 

ACCIDENTALL Y KILLE D 1 

TERMINA L SACRIFICE 19 38 21 

ANIKA L MISSING 

£_!NCLUDES_AUTOLYZED_AJ!_MALJ 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIHALS NECROPSIED
 

43
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

TOMOfi SUMMAR Y 

TOTAL ANIMAL S WITH PRIMAR Y TUMORS* 

TOTA L PRIMAR Y TUMOE S 

20 

33 

<»5 

65 

24 

34 

TOTAL ANIMALS WITH BENIGN

TOTAL BFNIG N TUMOR S 

 TOMCRS 19 

25 

41 

52 

21 

26 

TOTAL ANIMAL S WIT H MALIGNAN T

TOTAL MALIGNAN T TUMOR S 

 TOMOBS 7 

7 

11 

12 

8 

8 

TOTAL ANIMAL S KITH SECONDAR Y TUHORS# 

TOTAL SECONDAR Y TUMOR S 

1 

1 

2 

2 

TOTAL ANIMALS KITH TUMORS

BENIG K OR MALIGNAN T 

TOTAL UNCERTAI N TUMORS 

 UNCERTAIN1 

1 

1 

1 

TOTAL ANIMALS WITH TUMORS

EEIMABY CR METASTATIC 

TOTAL UNCERTAIN TUMORS 

 UNCERTAIN­

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

44
 
TABLE A2. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

VEHICLE 

CONTROL LOW DOSE 

ANIHALS INITIALLY IN STUDY

ANIMALS NECFOPSIED

ANIHALS EXAMINED HISTOPATHOLOGICALLY

 20 

 20 

 20 

250 

49 

49 

INTEGUMENTAR Y SYSISH 

*SKIN (20) (19) 

TRICHOSPITHELIOMA 1 (2%)

SEBACEOU S «DENGCARCINOM A 1 (2%) 

KEfiATOACANTHOMA 1 (2%)

FIEPOM A 1 (2X) 

*SUECUT TISSUE (20) (49)


LIPOMA 1 (2%)
 

MIXED KESENCHYHAL TUMOR, MALIGNA


RESPIRATORY SYSTEM
 

#LUNG (20) (48)

ALVECLAR/BRONCHIOLAf i CARCINOM A 1 (5X)

CORTICA L CARCINOMA , USTASTATIC 1 (5%) 

HEMATOPOIETIC SYSTE M 

#EEAI N (19) («9)

MALIGNAN T RETICULOSIS

*MULTIFL E OfiGAN S (20) (49)

UNDIFFEBENTIATED LEUKEMIA 1 (5%) 2 (tX)

LYMPHOCYT1C LEUKEMI A 1 (2X) 

CIRCULATOR Y SYSTE M 

NON E 

DIGESTIVE SYSTEM 

#STOMACE (20) (45)

SO.UA«OUS_CELL_CARCINC.MA

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUflBEF. OF ANIMALS NECBOFSIED 

oi 50 ANIMAL S WiiRE INITIALLY IN THE STUDY, BUT ONE ANIHA L HAS FOUND

A MALE IN A FEMALE GROUP. 

HIGH DOSE
 

50 

50 

50 

(50)
 

 (50)
 

1 (2X)
 

 (50) 

1 (2X) 

 (50) 

1 (2X) 

 (50) 

2 (4X) 

 (48) 

J 

 TO BE 

45 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

URINAR Y SYSTEM 

((URINAR Y BLADDER 

FAFILLCMA , NOS 

ENDOCRIN E SYSTEH 

tPITUITAF Y 

CHRCMOFHOB E A0ENOH A 

CHEOMOPHOBE C4RCINCM A 

#ADRENA L 

CORTICAL CABCINOM A 

PHEOCHROMOCYTOMA 

#THYPOI E 

FOLIICULAR-CELL ADENOMA 

C-CELL ADENOM A 

C-CELL CARCINOMA 

REPRODUCTIV E SYSTE M 

*MAMMAP Y GLAND 

SARCCMA , NOS 

FIEROADENOMA 

*EREPUTIA L SLAN D 

ADEtiCMA , NOS 

*VAGINA 

LEICMYCM A 

#UTERO S 

PAPILLAR Y ADENOM A 

SARCOMA , NOS 

ENDOMETPiA L STROMA L

HEKANGIOPEEICYIOMA,

#OVARY 

PAPILLAR Y ADENOM A

 

NERVOU S SYSTEM 

*BRAI N 

 POLYP 

 NOS 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

(16) (40) (35) 

1 (3S) 

(19) (43) (40) 

8 (42%) 

1 (5%) 

11 (26%) 

1 (2%) 

13

1

 (33%) 

 (3%) 

(19) (49) (50) 

1 (5%) 

1 (2%) 2 (»«) 

(17) (45) 

1 (2%) 

(38) 

1 (2%) 

1 (3%) 

(20) (49) (50) 

2 (10%) 

1 (2X) 

4 (8%) 1 (2%) 

(20) (49) (50) 

2 (10%) 1 (2%) 

(20) (49) (50) 

1 (2%) 

(19) (47) (49) 

1 (2%) 

2 (11%) 

1 (2%) 

8 (17%) 2 (4%) 

1 (2%) 

(18) («5) («8) 

1 (2%) 

(19) (49) (50) 

____ C H R CB0 EH OB E_CA RCI N CJ A_,__I Nl A SI VE 

# NUMBE E OF ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMALS NECROPSIED 

46 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

1 (2X) 

SPECIAL SENSE ORGAN S 

NON E 

MUSCULQSKEIETA L SYSTEM 

NONE 

BODY CAVITIES 

*HEBIASTINUM

HESCTHILIOHA, NOS

*MESEN1EBY

LIFCMA

 (20)

 1 (5X) 

 (20)

 (49)

 (49)

 (50) 

 (50) 

1 (2X) 

ALL OTHER SYSTEMS 

SITE UNKNOH N 

LIECMA 1 

ANIMA L IISPOSITION SUMMAR Y 

ANIHAL S INITIALLY IN STUD Y 20 50 50 

NATURA L DEATH3 21 

MORIBUN D SACRIFICE 3 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINA L SACKIFICE 20 HO 26 

ANIMA L MISSING 

ANIMA L DELETED(WRON G SEX) 1 

g_HLgkU.P-gS_AUTOLYZED AKIMAL S 

# NUMBER OF ANIMALS WITH TISSUE EXAMINE D HICBOSCOPICALLJT 

* NUMBER OF ANIMAL S NECROPSIED 

47 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

TUMO R SUMMAR Y 

TOTAL ANIMAL S WIT H PBIKAR Y TUMORS* 14 31 23

 

TOTAL PRIMAR Y TUMOE S 19 H3 29

 

TOTAL ANIMALS WIT H BENIGN TUHCRS 12 26 18 

TOTAL BENIGN TUMOP S lit 3H 21 

TOTA L ANIMAL S WIT H BAIIGNAST TUMORS 4 8 5 

TOTAL MALIGNAN T TUMORS U 8 8 

TOTAL ANIMAL S WITH SECONDARY TUMORS* 2 

TOTA L SECONDARY TUMORS 2

 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIG N O E MALIGNAN T 1 1

 

TOTAL UNCERTAI N TUMORS 1 1 

TOTAL ANIMALS WITH TUMORS UNCERTAINFEIMABY OP METiSTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUM08S EXCEPT SECONDARY TOHOBS
 

* SECONCARY TUMORS: METASTATIC TUMORS OB TUMOKS IHVASIYE INTO AN ADJACENT ORGAN
 

48
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MICE
 

ADMINISTERED 3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

BY GAVAGE
 

49
 

TABLE 81.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE

 

VEHICLE 

CONTROL LOW DOSE 

ANIHAI S INITIALLY IN STUD Y 20 50 

ANIMALS HISSING 2 

ANIMALS NECBOPSIED 20 48 

ANIHAI S EXAMINE D HIS TOEATHOLOGICALL Y 20 48 

INTIGUMENTAK i SISTER 

NONE 

EESEIHA1CSY SYSTEM 

*LUNG (20) (45) 

HEFATOCELLULAR CARCINOMA , METAST 1 (5%) 

ALVECLAR/oEONCHIOLAR ADENOM A 1 (5%) 

ALVEOLAE/BRONCHIOLAR CASCINGMA 

OSTEOSAECOMA. HETASTATIC 

1 (5X) 

1 (5*) 

4 (98) 

HEMATOFOIETIC SYSTEM 

*MOLTIPLE ORGANS (20) (48)
 

MALIGNANT LYMPHOMA, NOS 1 (5S) 2 (4%)
 

MALIGNANT i-YHPHOMA, MIXED TYPE 1 (5%) 1 (2X)
 

LEUKEMIA, NOS
 

ONCIFFEFENTIATED LEUKEMIA 1 <5X) 3 (6%)
 

LYMPHOCYTIC LEUKEMIA 1 (2X)
 

#EBONCFIAL Li'MPH NODE (16) (33)
 

ALVICLAR/BROSCHIOLAP. CA, METASTA
 

tMESENTEEIC L. NODE (16) (33)
 

SQUAMOUS CELL CARCINCMA, METASTA
 

HALIGNANT LYHPHOMA , NOS
 

.1ALIGNANT LYKPHOMA, MIXED TYPE 2 (6X)
 

CIRCULATOPY SYSTEM 

* BLOOD VESSEL (20) (48)

, ALVECLAEZBaO£CHIOLAR_CA_t_MJTASlA 1 (2 SI 

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE E OF ANIMALS NECROPSIED 

HIGH DOSE 

50 

50 

50 

(48) 

5 (10?!) 

(50)
 

2 (4%)
 

1 (2X)
 

1 (2X)
 

3 (6X)
 

1 (2X)
 

(35)
 

1 (3X)
 

(35)
 

1 (3X)
 

1 (3S)
 

1 (3X)
 

 (50) 

51 
TABLE 81. MALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

DIGESTIVE SYSTEM 

#IIVER (20) (16) (49) 

SQUABOUS CELL CARCINOMA , INVASI V 1 (2%) 

HEPATOCELLULA B ADENOH A 1 (5X) 

HEPATOCELLULAR CARCINOM A 2 (10%) 5 (11*) 9 (18%) 

* PA DC BIAS (18) (33) (36) 

SQOABOUS CELL CAECINCBA, INVASIV 1 (3*) 

*S1CBACH (19) (l»3) (47) 

SQUAMOU S CEL L PAPILLOHA 2 (5%) 6 (17%) 

SQUAMOU S CELL CARCINOM A 2 (4%) 

URINAR Y SYSTEM 

NON E 

ENDOCRIN E SYSTEM 

#THYROIC (10) (31) (35) 

FOUICULAR-CELL CABCINOH A 1 (3%) 

REPRODUCTIVE SYSTEM
 

ftTESTIS (19) (45) (44) 

INTEFSTITIAL-CELL TUMO R 1 (2X) 

NERVOU S SYSTEM 

NON E 

SPICIAL SENSE OEGAN S 

NONE 

MUSCULOSKELETA L SYSTEB 

NOJE 

* NUBBEE OF ANIBALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBES OB ANIGALS NECRCPSIED
 

52
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

BODY CAVITIES 

NOME 

ALL CTHEfi SYSTEMS 

SITE UNKNOW N 

SARCCHA, NOS 

ANIMAL tISFCSITION SUMMAR Y 

ANIMAL S INITIALLY IB STUDY 20 50 50 

NATURA L DEATHS l» 17 25 

MORIBUND SACEIFICE 1 1 2 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 15 30 23 

ANIKAL HISSING 2 

H INCLUDES AUTOLYZED ANIMALS 

TUMO R SUMMAR Y 

TOTAL ANIMALS WITH PRIMARY TUMCRS *

TOTAL PRIMAR Y TUMORS

 7 

8 

20 

22 

28 

35 

TOTAL ANIMALS HITH BENIGN TUMORS

TOTAL BENIGN TUMORS

 2 

2 

3 

3 

8 

8 

TOTAL ANIMALS HITH MALIGNANT TUMORS

TOTAL MALIGNAN T TUMORS

 5 

6 

19 

19 

24 

27 

TOTAL ANIMALS WITH SECONDARY

TOTAL SECONDARY TUMORS

 2 TUMORS*

2

1 

1

2 

» 

TOTAL ANIMALS HITH TUMOBS UNCERTAINBENIGN OP MALIGNANT 

TOTAL UNCEfiTAIN TUMOBS 

TOTAL ANIMALS HITH TDMOHS UNCERTAINEBIMABY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

* SECONDARY TUMORS: MEIASTATIC TUMORS OB TUMORS INVASIVE INTO AN ADJACENT OBGAN 

53 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

'—*-—*—-• TM* —TM --T-. 

— ——— j —— —— 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

ANIHALS INITIALLY I N STUDY 20 50 50 

ANIMAL S MISSING 1 

ANIMALS NECROPSIED 20 50 49 

ANIHAL S EXAMINE D HI STO PATHOLOGIC ALLY 20 50 U9 

INTEGUMENTAR Y SYSTEM 

NONE 

RESPIRATOR Y SYSTEM 

tLUN G (18) (50) (49) 

HEfATOCELLULA R CARCINOMA, HETAST 1 (2%) 

ALVECLAR/BKONCHJOLAE ADENOMA 1 (2%) 

ALVEOLAR/BRONCHIOLAR CARCINOMA 1 (6%) 3 (6%) 

OSTEOSARCOMA, METASTATIC 1 (2%) 

HEMATOPOIETIC SYSTEM 

*MULTIPLE ORGANS (20) (50) (U9) 

MALIGNANT LYMPHOMA, NOS 1 (5X) 2 (<»X) 1 (2%) 

MALIG. iYHPHOHA, HISTIOCYTIC TYPE 1 (2») 

MALIGNAN T LYMPHOMA , MIXED TYPE 2 (<t%) 

LEDKEMIA,NOS 1 (2S) 

UN DIFFERENTIATED LEOKEBIA 2 (UX) 2 (US) 

LYMPHOCYTIC LEUKEMIA 1 (5X) 1 (2X) 

# LYMPH NODE (16) (i»2) (*»0) 

ALVECLAR/BRONCHIOLAB CA, METASTA 1 OX) 

tHANDIBOLA R L. NODE (16) (t2) C»0) 

MALIGNANT LYMPHOBA, HIXED TYPE 1 OX) 

#MESEN1FRIC L. NODE (16) («) (40) 

MALIGNANT LYHPHOMA, HIXED TYPE 4 (10%) 2 (5X) 

CIRCULATORY SYSTEM 

*BLOOD VESSEL (20) (50) (49) 

FOL'LICOLAH-CELL CAECIHOHA. IHVIS 1 (2X) 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

54 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

DIGESTIV E SYSTEM 

*IIVEB


HEFATOCFLLULAR CARCINOMA


#STCMACH


SQUAMOUS CELL PAPILLCMA


SQUAMOUS CELL CARCINOMA


URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

*PITUITAF Y

CHECHOPHOBE ADENOM A

tADRENA L

COETICAL ADENOM A

#THYRCI C

FCLLICULAR-CEL L CAECINOM A

HEPRCDUCTIV E SYSTEM 

#UTERUS


ENECBETEIAL STROMAL EOLYP


#CVAR Y

FAFILLAP.Y ADENOM A

NERVOU S SYSTE M 

NON E 

SPECIAL SENS E ORGAN S 

*EYE/LACRIMA L GLAN D

AD£NOCAPCINOMA J L _NO S 

# NUMBE R OF ANIMAL S WIT H TISSUE

* NUMBE R OF ANIMAL S NECECPSIED 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

 (20) (50) («9) 

1 (555) 2 (U%) 

 (19) (45) (<t8) 

1 (2%) 3 (6X) 

. 2 {<»») 

 (9) (25) (29) 

2 (7%) 

 (16 ) (tl) («3) 

1 (2«) 

 (8) (15) (36) 

1 (7X) 

 (18) («7) (U7) 

1 (2%) 

 (6) (13) (33) 

1 (8X) 

 (20) (50) (U9 ) 

 EXAMINE D MICROSCOPICALLY 

55 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

MUSCULCSKELETA L SYSTEM 

NON E 

BOC Y CAVITIES 

*ABDOMINA L CAVITY

SAECCHA , NOS

 (20)

 1 (5%) 

 (50) (49) 

A LL OTHEE SYSTEMS 

*MUlTIfL£ ORGAN S

SAFCCHA, NOS

 (20) (50)

 1 (2%) 

 (<»9) 

ANIMA L IISEOSITION SUMMAP Y 

ANIMAL S INITIALLY IN STUDY

NATURA L DEATHS

MORIBUN D SACRIFICE

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLEE 

TERMINA L SACRIFICE

ANIMA L MISSING

 20

 U

 1

 15

 50

 11

 39

 50 

 17 

2 

30 

1 

al INCLUDES AUTOLYZED ANIMALS 

# NUMBE R

* NUMBE R

 OF ANIMAL S

 OF iNIHALS

 WIT H TISSUE

 NECROPSIED 

 EXAMINE D MICROSCOPICALLY 

56 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

TUMO R SUMMAR Y 

TOTAL ANIMALS WITH PRIMAR Y

TOTAL FRIMAEY TOMOES

 TUSORS* 5 

5 

14 

15 

20 

2H 

TOTAL ANIHAL S WIT H BENIG N TUMOR S 

TOTAL BENIG N TUMOR S 

U 

a 

6 

6 

TOTAL ANIMALS WITH MALIGNAN T TUHOBS

TOTAL MALIGNANT TUMORS

 5 

5 

11 

11 

17 

18 

TOTAL ANIMAL S WITH SECONDAR Y TUMORS t 

TOTAL SECONDAR Y TUMORS 

2 

2 

2 

2 

TOTAL ANIMALS WITH TDHORS

BENIG N CR MALIGNANT 

TOTAL UNCERTAI N TUMORS 

 UNCERTAINTOTAL ANIMAL S WIT H TUMORS UNCERTAIN ­

EBIHA5Y CP METASTATIC 

TOTAL UNCERTAIN TUMORS 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TDMORS
 

# SECONCARY TUMORS: METASTATIC TUHORS OR TDMORS INVASIVE INTO AN ADJACENT ORGAN
 

57
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN
 

RATS ADMINISTERED 3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

BY GAVAGE
 

59
 

TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

-—-———-— "— •' -"-*—— VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

ANIMAL S INITIALLY IN STUDY 20 50 50 

ANIMAL S NfCBOPSIED 20 50 50 

ANIMALS EXAMINE D HIS: rOFATHOLOGICALLY 20 49 50 

INTEGUMENTAR Y SYSTEM 

NONE 

RESEIRA10EY SYSTEM 

tLUSG/EBONCHUS (20) (49) (49)

 

POLYEOID HYPERPLASI A 1 (2*)

 

tLUN G (20) (49) (49)

CONGESTION, NOS 7 (14*) 

EDEMA, NOS 1 (2X) HEMORRHAGE 1 (2X) 

PNEUMONIA , CHRONIC MURIN E 1 (2X) GRANULOMA, FOREIGN BODY 1 (2%)

HYPERPLASIA, ADENOMATOOS 2 (4*) 1 (2X) 

HEBATOPOIETIC SYSTEM 

• SPLEEN (20) (48) (48) 

INFARCT, NOS 1 (2*) 

HEMOSIDEROSIS 1 (2%) 

HEM ATO POXES IS 1 (5*) 

tMESENTIRIC L. NODE (19) (47) (43)

 

HISTIOCYTOSIS 1 (2X)

 

CIKCULATORY SYSTEM 

* HE ART (19) (48) (46) 

CALCIFICATION, NOS 1 (2%) 

IHYOCAEDIOH (19) (48) (46) 

* NUMBER OF ANIMALS WITH TISSDE EXAMINED MICROSCOPICALLY 

* NUMBEB OF A HI HALS NECROPSIED 

61 
TABLE C1. MALE RATS: NONNEOPLASTIC

INFLAMMATION , CHEONIC FOCAL 

FIEPOSIS 

FIEROSIS, FOCA L 

*BLOOE VESSEL 

CALCIFICATION, NOS 

*ARTER Y 

INFLAMMATION , CHRONIC 

*AORTA 

INFLAMMATION , CHRONIC 

DIGESTIV E SYSTEM 

#SALIVAR Y GLAN D 

INFLAMMATION , NO S 

FIBBOSIS, DIFFUSE 

#LIVE R 

CONGESTION , PASSIV E 

INFLAMMATION , ACUT E FOCAL 

INFLAMMATION , GR ANULOMATOU S 

CIRRHOSIS, NOS 

METAMORPHOSI S FATT Y 

EOSINOPH1LIC CYTO CHANG E 

*HEFATIC CAPSUL E 

HYFIPFLASIA . FOCAL 

*BILE EUCT 

HYPERPLASIA , NO S

 

tPANCRIA S 

ATRCEHY , NO S 

fPANCKEATI C ACINU S 

ATECFHY , NO S 

ATROPHY , FOCAL 

f ESOPKAGU S 

INFLAMMATION , ACUTE 

#S1CMACH 

DILATATION, . NOS 

 LESIONS (CONTINUED) 

VEHICLE 

CONTROL 

1 (5%) 

2 (11%) 

(20) 

(20) 

(20) 

(19) 

(19) 

1 (5%) 

2 (11X) 

1 (5X) 

(19) 

(20) 

17 (85K) 

(20) 

(20) 

(20) 

(19) 

HIGH DOSE 

2 (UX) 

1 (2%) 

3 (736) 

(50) 

1 (2X) 

(50) 

1 (2X) 

(50) 

(44) 

1 (2X) 

1 (2X) 

(48) 

2 (4X) 

(48) 

(50) 

22 (44*) 

(49) 

1 (2X) 

(49) 

(44) 

(50) 

_JLJ2iL__. 

LOW DOSE 

6 (13X) 

3 (6S) 

4 (8X) 

(50) 

(50) 

(50) 

1 (2%) 

(48) 

(48) 

1 (2X) 

1 (2%) 

(48) 

1 (2X) 

(50) 

34 (68X) 

(48) 

(48) 

1 <2X) 

2 (4X) 

(47) 

1 (2X) 

(47) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

62
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HYEEFPLASIA , EPITHELIAL 

#GASTEIC MUCOSA 

DILATATION, NOS 

#COLON 

PAEASITISM 

URINAE Y SYSTEM 

• KIDNE Y 

CONGESTION, NOS 

GLOMEROLONEPHBITIS , NOS 

INFLAMMATION , CHBONIC 

NEPHEOSIS , HEMOGLOEINURIC 

CALCIFICATION, NOS 

tKIDNEY/TUBULE 

NECPCSIS, NOS 

NECEOSIS, FOCAL 

ENDOCRIN E SYSTEM 

#PITUITAR Y 

CYST, NOS 

HYPEEPLASIA , CHEOMCPHOBE-CELL 

tADEENA I 

CYTCFLASMIC VACOOLIZ ATI CN 

HYPEKPLASIA , NODULA R 

ANGIECTASIS 

tTKYROI E 

CYSTIC FOLLICLES 

INFLAMMATION , CHRONIC FCCAL 

HYPERPLASIA , C-CELL 

#PARATHYROI D 

HYPERPLASIA , ADENOMATOU S 

REPRODUCTIV E SYSTEM 

* MAMMAR Y GLAN D 

CYSTIC DUCTS 

VEHICLE 

CONTROL 

(19) 

12 (63%) 

(20) 

14 (70%) 

(20) 

12 (60%) 

(20) 

1 (5%) 

(19) 

(19) 

1 (5%) 

(17) 

2 (12%) 

(8)

1 (13%) 

(20) 

LOW DOSE 

1 (2%) 

(47) 

23 (49%) 

(47) 

18 (38%) 

(49) 

28 (57%) 

(49) 

(42) 

1 (2%) 

2 (5%) 

(49) 

1 (2%) 

6 (12%) 

(39) 

1 (3%) 

(19) 

(50) 

1 (2%) 

HIGH DOSE 

2 (4%) 

(50) 

15 (30%) 

(48) 

10 (21%) 

(49) 

6 (12%) 

13 (27%) 

1 (2%) 

1 (2%) 

1 (2%) 

(49) 

1 (2%) 

(41) 

1 (2%) 

(49) 

2 (4% ) 

1 (2%) 

2 (4%) 

(41) 

1 (2%) 

1 (2%) 

1 (2%) 

(17) 

(50) 

# NUMBER OF ANIMALS WITH TISSDE EXAMINED MICROSCOPICALLY
 

* NOMBEE OF ANIMALS NECROPSIED
 

63
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

tFKOSTATE (17) (43) (41) 

INFLAMMATION . SOPPUHATIVE 1 (236) 

INFLAMMATION , ACUTE FOCAL 1 (6%) 

HYPERPLASIA , NOS 1 (2X) 

tfTESTIS (19) (t6) (47) 

TORSIO N 1 (28) 

ATROPHY, NOS 2 (11X) 4 (9X) 1 (2*) 

HYPERPLASIA , INTERSTITIA L CELL 1 (2%) 

#TESTIS/TUBULE (19) (46) (47) 

CALCIFICATION, NOS 1 (2X) 

NERVOU S SYSTEM 

NONE 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

NCNE
 

BODY CAVITIES 

*HESENTERY (20) (50) (50) 

PERIAR1ERITIS 1 (5X) 

NECROSIS, FAT 1 (2«) 

ALL OTHER SYSTEMS 

NON E 

SPECIAL BOEPHOLOGY SUMMAR Y 

NO-US ION_BEPgRTJD 1 JO 

# NUMBE R OF iHIMALS BITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMALS NECBOPSIID 

64 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

JUTO/NECROPSY/HISTO PEEF 2 

AUTC/NECROPSY/NO HISTO 1 

* NUMBE R OF ANIMALS WITH TISSUE EXABINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

65 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

VEHICLE ———— • 

CONTROL LOWE >OSE HIGH DOSE 

ANIMAL S INITIALL Y IN STUD Y 20 350 50 

ANIMAL S KECROPSIED 20 49 50 

ANIMAL S EXAMINE D HI STOEATHOLOGIC ALLY 2 0 49 50 

INTEGUMENTAR Y SYSTEM 

*SUBCU T TISSUE (20) (49) (50) 

EPIDERMA L INCLUSION CYST 1 (2X) 

RESPIRATOR Y SYSTEM 

* TRACHEA (17) (44) (41) 

INFLAMMATION , CHRONIC 1 (2X ) 

HYPERPLASIA , EPITHELIA L 1 (2%) 

METAPLASIA , SQUAMOO S 1 (2X) 

*LUNG (20) (48) (50) 

CONGESTION , NO S 8 (16X) 

ABSCESS, NOS 1 (2X) 

PNEUMONIA , CHRONIC MURIN E 1 (5%) 1 (2%) 2 (4X) 

INFLAMMATION , GR ANULOMATOU S 1 <2X) 

GRANULOMA , NOS 1 (5%) 

GRANULCMA , FOREIG N BODY 1 (5%) 1 (2X) 

HYPEEPLASIA , ADENOBATOU S 2 (4X) 

HYPERPLASIA , ALVEOLA R EPITHELIU M 1 (2%) 1 (2X) 

HISTIOCYTOSIS 2 (4X) 1 (2X ) 

LEUKEMOID REACTION 1 (2X) 

HEMATOPOIETIC SYSTEM 

*SPLEEN (20) (48) (49) 

HEMATOMA , ORGANIZE D 1 (5%) 

FIEROSIS, FOCA L 1 (2X) 

INFARCT, NOS 1 (2X) 

HEMOSIDEROSI S 1 (2%) 

HEMATOPOIESIS 2 [4X1 

f NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECEOPSIED 

a) 50 ANIMALS WERE INITIALLY IN THE STUEY, BUT ONE ANIMA L HAS FOUND TO BE A MA1E 

IN A FEHAIE GROUP. 

66 
CIKCOLATOR Y SYSTEM 

tMYOCARDIUM 

INFLAHBATION , ACUTJ FOCAL 

INFLAMMATION , CHRONIC FOCAL 

FIEROSIS, FOCAL 

DIGESTIVE SYSTEM 

ILIVEE 

CONGESTION, NOS 

CONGESTION, CHRONIC PASSIVE 

INFLAMMATION , NECROTIZING 

GRANULOBA , NOS 

NECROSIS, FOCAL 

METAMORPHOSI S FATTY 

BASOPHILIC CYTO CHANGE 

*BILE DUCT 

INFLAMMATION , CHRONIC 

HYPERPLASIA, NOS 

f PA NCR IAS 

INFLAMMATION , INTERSTITIAL 

tEANCREATI C ACINU S 

ATBCEHY , NO S 

ATROPHY , FOCAL 

#GASTRIC MOCOSA 

DILATATION, NOS 

ICOLON 

PARASITISM 

URINAR Y SYSTEM 

KKIDNE Y 

CONGFSTION, NOS 

GLOMERULONEPHRITIS , NOS 

#KIENIY/CORTE X 

CYST* NOS _ _ _ 

TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE 

(20) (47) 

1* (20%) 6 (13%) 

1 (5%) 1 (2%) 

(20) (48) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

(20) (49) 

1 (2%) 

5 (25*) 21 (43%) 

(20) (46) 

1 (2%) 

(20) (46) 

1 (2%) 

1 (5%) 1 (2%) 

(20) (45) 

12 (60%) 27 (60%) 

(20) (46) 

7 (35%) 17 (37%) 

(19) (49) 

1* (21%) 4 (8%) 

(19) (49) 

1 (2%) 

t NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECBCPSIED 

HIGH DOSE 

(49) 

1

8

1

(49) 

1

2

1

(50) 

11

(49) 

(49) 

1

2

(48) 

23

(49) 

1C

(50) 

7

3

(50) 

 (2%) 

 (16%) 

 (2%) 

 (2%) 

 (4%) 

 (2%) 

 (22%) 

 (2%) 

 (4%) 

 (48%) 

 (20%) 

 (14%) 

 (6%) 

67 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

#KIDNEY/TUBUL E (19) (50) 

NECRCSIS, NOS 1 (5%)

 

NECROSIS, FOCAL 1 (2X)

 

ENDOCRIN E SYSTEM 

#PITUITAR Y (19) (43) C»0) 

CYST, NOS 1 (5X) 4 (9X) 1 (3X) 

NECROSIS, FOCAL 1 (2X) 

HYPERPLASIA , CHROMOP KOBE-CELL 1 (2X) 

ANGIECTASIS 1 (2X) 

tADRENA I (19) (49) (50) 

CYTCFLASMIC VACUOLI Z ATI CN 1 (5X) 4 (8X) 3 (6X) 

ANGIECTASIS 3 (6X) 

HEMATOPOIESIS 1 (2X) 

fTHYBCID (45) (38) 

INFLAMMATION , CHRONIC FCCAL 1 (6X) 

HYPERPLASIA, C-CELL 3 (8X) 

HYPERPLASIA, FO LLICOLAR-CELL 1 (3X) 

REPBODUCTIVE SYSTEM 

*BA«MAR Y GLAN D (20) (49) (50) 

CYST, NOS 1 (5X)

 

CYSTIC DUCTS 1 (5X)

 

INFLAMMATION, CHRONIC 

HYPERPLASIA, NOS 1 (2X) 

*HAMBABY DUCT (20) (49) (50) 

HYPERPLASIA, NOS 1 (2X) 

#UTERU S (19) (47) (49) 

HYEROMETRA 1 (2X) 2 (4X) 

#CEFVIX UTEHI (19) (47) (49)

 

CYST, NOS 1 (5X)

 

tOTERUS/ENDOBETRIUH (19) (47) (49) 

ABSCESS, NOS 1 (2X) 

HY PIEPi*SI4A-Q.ISIIC. J_i5Xi_ 1 (2X1 1 (2X1 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NDHBEfi OF ANIMAL S NECKOPSIED 

68 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

tOVAR Y (18) (45) (48) 

CYST, NOS 1 (6%) 3 (7X) 2 (4») 

NERVOU S SYSTEM 

•BRAIN (19) (49) (50) 

HEMORRHAGE 1 (2X) 

SPECIAL SENSE ORGAN S 

*EYE (20) (49) (50) 

HEMORRHAG E 1 (2%) 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

*ABEOHINAL CAVITY (20) (49) (50) 

ABSCESS, NOS 1 (2%) 

NECROSIS, NOS 1 (2%) 

*EPICAEDIUM (20) (49) (50) 

INFLAMMATION, FIBRINOUS 1 (2%) 

ALL OTHER SYSTEMS 

SITE DNKNOM N 

NEC SCSIS, FAT 1 

SPECIAL MORPHOLOGY SUHHARY 

NO LESION REPORTED 1 2 8 

* NUMBE R OF ANIMALS WITH TISSOE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

69 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN
 

MICE ADMINISTERED 3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

BY GAVAGE
 

71
 

TABLE D1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

ANIMAL S 

ANIMALS 

ANIMAL S 

ANIMAL S 

INITIALL Y I 

KISSING 

NECROPSIE D 

EXAMINE D H I 

N S1UEY 

STOPATHOLOGICALL Y 

INTEGUMENTAR Y SYSTE M 

NON E 

RESPIRATOR Y SYSTEM 

#LUN G 

CONGESTION, NOS 

ERCNCHCPNEUMONIA , ACUT E 

PNEUMONIA , CHRONI C MURIN E 

PERIVASCULA R CUFFIN G 

HYPERPLASIA , ADENOHATOU S 

HISTICCYTOSIS 

LEOKEMOI D REACTIO N 

HYPERPLASIA , BASOPHILIC 

tLUNG/ALVEOL I 

CRYSTALS, NOS 

HISTIOCYTOSIS 

HEMATOPOIETIC SYSTEM 

#SPLEE N 

AMYlCICOoI S 

ANGIECTASI S 

HEMA'T'OPOIjiSIS 

#MESENTERI C L. NODE 

CONGESTION . NOS 

HEMOERHAG E 

HEMOSIDEfiOSIS 

HISTIOCYTOSIS 

MASTOCYTOSI S 

# NUMBE R OF ANIMAL S WITH TISSUE

* NUMBE R OF AtilflALS NECROPSIE D 

— VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

20 50 50 

2 

20 48 50 

20 <*8 50 

(20) (1*5) (48) 

2 7 (15S) 

1 (5%) 

3 (15X) 2 (4XJ 1 (2% ) 

1 (5*) 1 (2%) 

2 

3 

1 (2«) 

1 (2X) 

(20) (1*5) (48) 

1 (2X) 

1 (5%) 

(17) (41 ) (41) 

1 (2X) 

1 (2%) 

3 <7X ) 1 (2«) 

(16) (33) (35) 

4 (12%) 1 (355) 

1 (3X) 1 (3X) 

1 (3%) 

 EXAMINE D MICROSCOPICALL Y 

73 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

CIRCULATORY SYSTEM 

*POHTAL VEIN 

THROMBOSIS, NOS 

(20) (48) 

1 (2%) 

(50) 

DIGESTIVE SYSTEM 

tSALIVARY GLAND 

PEBIVASCULA6 CUFFIN G 

(19) 

1 (5%) 

(39) 

2 (5X) 

(44) 

2 (5%) 

#LIVER 

CYST, NOS 

CONGESTION, NOS 

HEMATOMA , NOS 

INFLAMMATION , FOCAL 

INFLAMMATION , CHRONIC 

INFLAMMATION , CHRONIC FOCAL 

GRANULATION , TISSUE 

THROMBOPHLEBITIS 

FIEROSIS 

PERIVASCULA R CUFFIN G 

NECROSIS, FOCAL 

INFARCT, NOS 

CALCIUM DEPOSIT 

CALCIFICATION, NOS 

EASOPHILIC CYTO CHANG E 

ATROPHY, NOS 

(20) 

1 (5%) 

1 (5«) 

1 (5%) 

1 (556) 

1 (5*) 

1 (5%) 

1 (5%) 

(46) 

1 (2X) 

1 (2%) 

1 (2X) 

4 (9%) 

1 (2%) 

1 (2X) 

1 (2X) 

(4%) 

(2») 

(2%) 

(49) 

1 (2X) 

1 (2% ) 

#LIVER/HEPATOCYTES 

CYTCPLASMIC CHANGE , NOS 

(20) (46) 

1 (2X) 

(49) 

#PANCFEA S 

INFLAMMATION , INTERSTITIA L 

(18) (33) (36) 

1 (3X) 

#EANCKEATIC DUCT 

NECROSIS, NOS 

(18) 

1 (6%) 

(33) (36) 

tFANCREATI C ACINU S 

NECRCSIS, NOS 

ATROPHY , NOS 

(18) 

1 (6% ) 

1 (6%) 

(33) (36) 

#STCMACH (19) (43) 

-

(47) 

____ !_J2J1__ 

# NUMBF. E

* NUMBE R

 OF ANIMAL S

 OF ANIMAL S

 WIT H TISSUE

 NECROPSIED 

 EXAMINE D MICROSCOPICALLY 

74 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH 

INFLAMMATION , NOS 1 

HYPESPLASIA , EPITHELIA L 1 (2*) 2 

•PEYEES PATCH (17) (42) 

HYEEBPLASIA . NOS 1 (6%) 2 <5S) 

*COLON (18) (44) (47) 

PARASITISM 3 (17*) 12 (27X) 13 

UBINAR Y SYSTEM 

IKIDNE Y (20) (46) (47) 

CONGESTION, NOS 2 (4X) 7 

PERIVASCULA E CUFFIN G 3 (15*) 7 (15X) 5 

tKIDNEY/MEDULL A (20) (46) (47) 

CYST, NOS 1 

IPERIRENA L TISSUE (20) (46) (47) 

NECEOSIS, FOCAL 1 (2X) 

#KIENEY/TUBUL E (20) (46) (47) 

HYPIRPLASIA , FOCAL 1 (5*) 

#KIENEY/PFLVI S (20) (46) (47) 

INFLAMMATION , CHRONIC 1 

PEEIVASCULAH CUFFING 1 (2X) 

tURINAR Y BLADDER (13) (33) (37) 

MUCOCELE 1 (3X) 

INFLAMMATION , CHRONIC 1 

PERIVASCULAR CUFFING 2 (6X) 

EMDOCHINE SYSTEM 

NONE 

REPRODUCTIVE SYSTEM 

tPBOSTATE (19) (40) (46) 

INFLAHMATION^CHRONIC SUPPORATIV 

t NUMBER OF ANIHALS BITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIHALS NECBCPSIED 

DOSE 

(2X) 

(4X) 

(28X) 

(15X) 

(11X) 

(2X) 

(2X) 

(3X) 

75 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

NERVOU S SYSTEM 

#BEAIN

INFARCT, NOS

CALCIFICATION, FOCAL

 (20)

 6 (30%)

 (46)

 7 (15%)

 (50) 

1 (2X) 

4 (8«) 

SPECIAL

NON E 

 SENSfi ORGAN S 

MUSCOLOSKELETAL

NONE 

 SYSTEM 

BOEY CAVITIES 

NONE 

ALL OTHER

NOM E 

 SYSTEMS 

SPECIAL MORPHOLOGY SUMMAH Y 

NO LESION REPORTED

ANieiL MISSING/NO NECROPSY

ACITC/NECBOPSY/HISTO PERF

* NUMBE F Of ANIMALS WITH TISSU E

* NUMBEE Of ANIMALS NECROPSIED 

 EXAMINE D

 3 11

 2 

1 

 MICROSCOPICALLY 

 12 

76 
TABLE 02. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

ADMINISTERED 3(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE BY GAVAGE 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY

ANIMALS MISSING 

ANIMALS NECROPSIED

ANIMALS EXAMINED HISTOFATHOLOGICALLY

 20 

 20 

 20 

50 

50 

50 

50 

1 

49 

49 

INTEGUMENTAR Y SYSTE M 

NON E 

RESPIRATOR Y SYSTEM 

*LUNG/BRCNCHU S 

CRYSTALS , NOS 

(18) (50) 

1 (2%) 

(49) 

tfLUN G 

ATF.IECTASIS 

CONGESTION , NOS 

PNEUMONIA , CHRONIC MURIN E 

INFLAMMATION , GR ANULOMATOU S 

PERIVASCULA R CUFFING 

HIS1IOCYTOSIS 

(18) 

1

3

 (6%) 

 (17%) 

(50) 

1

11

 (2%) 

 (22S) 

(49) 

8 

3 

1 

1 

1 

(16X) 

(6X) 

(2S) 

(2%) 

(255) 

fcLUNG/ALVEOLI 

CRYSTALS , NOS 

PHAGOCYTIC CELL 

(18) (50)

1

1

 (2X) 

(2%) 

(49) 

HEMATOPOIETIC SYSTE M 

#BONE MARRO W 

HYEF.EPIASIA , HEMATCFOIETIC 

(9) (46)

3 (7%) 

(46) 

1 (25!) 

#SPLEE N 

HYFIRPLASIA , RETICULUB

HEMATOPOiESIS 

 CELL 

(17) 

4 (24%) 

(46) 

6 (13S) 

(45) 

2 

2 

(4%) 

(<**) 

#«ANCIBULA R i, . NODE 

INFIAMMATION , GEANU L OMATCU S 

(16) (42) 

1 (2X) 

(40) 

*B8CNCHIAL LYHP H NODE (16) (42) (40) 

# NUBBE E

* NUMBE R

 OF ANIMAL S WIT H TISSUE

 OF ANIMAL S NEC8CPSIED 

 EXAMINE D MICROSCOPICALLY 

77 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

1 . — ~——————— ___—. 

VEHICLE 

CONTROL LOW DOSE 

iHESENTERIC L. NODE (16) (42) 

HEMCFRHAG E 

INFLAMMATION , GRA NULCMATOU S 1 (2X) 

CYTOMEGALY 1 (2X) 

HYPEEPLASIA, RETICOLO M CEIL 1 (2X) 

HEHATOPOIESIS 1 (2X) 

CIRCULA10RY SYSTEM 

#HEART (18) (t8) 

CALCIFICATION, FOCAL 1 (6X) 

*BLCOC VESSEL (20) (50) 

INFLAMMATION , CHEONIC 1 (2X) 

THEOMEOPHLEBITIS 

*CCEONAE Y ARTER Y (20) (50) 

INFLAMMATION , CHRONIC 

*UTERIN E VEIN (20) (50) 

THEOMBOSIS, NOS 1 (5%) 

#hEEATIC SINUSOID (20) (50) 

THECMBOSIS, NOS 

DIGJiSTIVF SYSTEM 

#SALIVAE Y GLAN D (19) (U8 ) 

INFLAMMATION , CHRONI C FOCAL 1 (2%) 

PERIVASCULA R CUFFIN G 2 (11X) 1 (2X) 

#LIVEE (20) (50) 

INFLAMMATION . NO S 1 (2X) 

INFLAMMATION , CHRONIC FOCAL 2 (10S) 1 (2%) 

INFLAMMATION , GR ANULOMATOO S 

PERIVASCULA R CUFFIN G 1 (5«) 3 (6X) 

DEGENEEATION , NOS 1 <5X) 

NECROSIS, FOCAL 1 (5%) 1 (2X) 

INFARCT, NOS 1 (2X) 

METAMORPHOSIS FATT Y 1 (5X) 

HYPEFPLASIA, NOS 1 (5X) 

ANGIECTASIS 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECEOPSIED
 

HIGH DOSE 

(«0) 

1 OX) 

(48) 

(49) 

1 (2X) 

(49) 

1 (2X) 

(49) 

(49) 

1 (2X) 

(44) 

2 (5% ) 

(49) 

1 (2X) 

1 (2%) 

1 (2X) 

1 (2X) 

1_J2?1 

78
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

HISTIOCYTOSIS 

#LIVER/CENTHILOBULA R 

NECECSIS, NOS 

"*EILE DUCT 

INFLAMMATION, CHRONIC
 

INFLAMMATION, CHRONIC FOCAL
 

tPANCEEAS
 

CYSTIC DUCTS 

tPANCEPATIC ACINU S 

INFLAMMATION , CHEONIC 

ATEOPHY, NOS 

ATROPHY, FOCAL 

#STOHACH 

CYST, NOS 

ABSCESS, NOS 

CALCIFICATION, FOCAL 

HYPERPLASIA , EPITHELIAL 

MASTOCYTOSIS 

#GASTEIC SUBMUCOSA 

INFLAMMATION , ACUTE/CHRONIC 

#COLON 

PARASITISM 

URINARY SYSTEM
 

#KIDNEY
 

CONGESTION, NOS 

INFI»MHATION, CHEONIC 

PERIVASCULA8 CUFFING 

NEPHBOSIS, NOS 

AMYLOIDOSIS 

HETAPLASIA, OSSEOUS 

#KIDNEY/TUBULE
 

CALCIFICATION, NOS
 

ATROPHY, NOS 

BEGEljERATIOIl. NOS 

# HUMBEE OF ANIMAL S WITH TISSUE

* BOHBEE OF ANIMALS NECEOPSIED 

VEHICLE 

CONTROL LOW DOSE 

1 (5%) 

(20) (50) 

2 (4X) 

(20) (50) 

1 (2X) 

1 (2%) 

(13) (34) 

2 (6%) 

(13) (34) 

1 (3X) 

1 (3X) 

1 (3X) 

(19) (45) 

(19) (45) 

(19) (45) 

3 (16X) 7 (16S) 

(19) (49) 

2 (4X) 

1 <2X) 

5 (10X) 

1 (2X) 

1 (2X) 

(19) (49) 

1 (2X) 

1 (2X) 

 EXAMINE D MICROSCOPICALLY 

HIGH DOSE 

(49) 

(49) 

1 (2%) 

(38) 

(38) 

1 (3X) 

(48) 

1 (2X) 

1 (2X) 

1 (2X) 

3 (6X) 

1 (2X) 

(48) 

1 (2X) 

(47) 

4 <9X) 

(49) 

9 (18X) 

1 (2X) 

4 (8X) 

1 (2X) 

(49) 

79 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE 

•URINAR Y BLADDER 

INFLAMMATION , CHRONIC 

(15) 

1 (7%) 

(29) 

PERIVASCULA R CUFFIN G 2 (7X) 

LYBPHOCYTOSIS 

ENDOCRIN E SYSTEM 

• THYROI D (8) (15) 

HYFERCHROMATIS M 1 (7X) 

REFRODUCTIVF SYSTEM 

•UTERUS (18) (47) 

CYST, NOS 4 (22X) 1 (2X) 

PYOMETR A 

#UTEROS/ENDOMETRIU M (18) (47) 

CYST, NOS 4 (9X) 

INFLAMMATION , SUPPURATIV E 1 (2X) 

ABSCESS, NOS 1 (6X) 

INFLAMMATION, CHRONIC 1 (2X) 

HYPERPLASIA, CYSTIC 4 (22X) 9 (198) 

#OVARY/OVIDUCT (18) (47) 

INFLAMMATION , NOS 1 (2%) 

HYPEEPLASIA, NOS 1 (2X) 

tOVAR Y (6) (13) 

CYST, NOS 1 (17X) 5 (38X) 

HEMORRHAGIC CYST 

NERVOU S SYSTEM 

#BRAIN (18) (50) 

PERIVASCULITIS 

CORPORA AMYLACE A 1 (6X) 

CALCIFICATION, NOS 1 (6*) 

CALCIFICATION, FOCAL 2 (11%) 9 (18S) 

tMIDBBAIN (18) (50) 

CALCIFICATION, FOCAL 

SPECIAL SENSE ORGANS 

NONE ,._„_. ­

# NUMBER OF ANIMALS WITH TISSUE EIAHINED MICROSCOPICALLY 

* NUMBE R OF ANIMALS NECROPSIEC 

80 

HIGH DOSE 

(36) 

1

1

(36) 

(47) 

3

(47) 

1

1

(47) 

(33) 

4

2

(49) 

1

1

7

(49) 

1

 (3X) 

 (3X) 

 (6X) 

 <2X) 

 (2X) 

 (12X) 

 (6X) 

 (2X) 

 (2X) 

 (14X) 

 (2X) 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

VEHICLE 

CONTROL LOW DOSE HIGH DOSE 

HUSCULCSKELETAL SYSTEM 

*BONE 

FIEBOUS OS1EODYSTRCPHY 

(20) 

7 (35%) 

(50)

22 (44X)

 (49) 

 18 (37S) 

OSTEOSCLEROSIS 1 (2») 

BODY CAVITIES 

*MEDIASTINUM 

INFLAMMATION , CHRONIC 

(20) 

1 (5%) 

(50) (49) 

*PERITCNEAL CAVIT Y (20) (50) (49) 

GRANULOMA, NOS 1 (2S) 

*ELEURA 

INFLAMMATION , CHRONIC 

(20)

1 (5%) 

(50) (49) 

INFLAMMATION, CHRONIC FOCAL 1 (2X) 

ALL OTHER SYSTEMS 

NON E 

SPECIAL MORPHOLOGY SUMMARY 

NO 1ESION EXPORTED 

ANIKA L MISSING/NO NECROPSY 

AOTC/NECBOPSY/HISTO PERF 

* NOMBE P CF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBEE CF ANIMALS NECROPSIED 

81 

APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN RATS
 

ADMINISTERED 3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

BY GAVAGE
 

83
 

__ 

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3
 

Vehicle Low
 

Topography: Morphology Control Dose
 

Lung: Alveolar/Bronchiolar
 

Carcinomab 2/20 (10) 3/49 (6)
 

P Values0 »d N.S.
 

Relative Risk6 0.612
 

Lower Limit 0.078
 

Upper Limit 6.996
 

Weeks to First Observed Tumor 104 104
 

oo
 

Hematopoietic System: Leukemia13 2/20 (10) 1/50 (2)
 

P Values0 »d N.S.
 

Relative Risk6 0.200
 

Lower Limit 0.004
 

Upper Limit 3.681
 

Weeks to First Observed Tumor 103 104
 

High
 

Dose
 

0/49 (0)
 

N.S.
 

0.000
 

0.000
 

1.372
 

2/50 (4)
 

N.S.
 

0.400
 

0.032
 

5.277
 

101 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

(continued)
 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Stomach: Squamous-cell Papilloma
 

or Carcinomab 0/19 (0) 1/47 (2) 3/50 (6)
 

P Values0 »d N.S. N.S.
 

Relative Risk6 Infinite Infinite
 

Lower Limit 0.022 0.238
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — 67 76
 

oo
 

Pituitary: Chromophobe Carcinomab 0/19 (0) 3/42 (7) 1/41 (2)
 

P Values0 >d N.S. N.S.
 

Relative Risk6 Infinite Infinite
 

Lower Limit 0.284 0.026
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — 89 104
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3
 

(continued)
 

Topography; Morphology
 

Pituitary: Chromophobe Adenoma
 

or Carcinoma"
 

P Values0 >d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CD
 

Adrenal: Pheochromocytoma°
 

P Values0 »d
 

Relative Risk6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Vehicle Low 

Control Dose 

1/19 (5) 7/42 (17) 

N.S. 

3.167 

0.460 

138.815 

104 86 

2/19 (11) 2/49 (4) 

N.S. 

0.388 

0.031 

5.108 

104 104 

High
 

Dose
 

2/41 (5)
 

N.S.
 

0.927
 

0.052
 

53.355
 

104
 

1/49 (2)
 

N.S.
 

0.194
 

0.003
 

3.563
 

101
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

(continued)
 

Vehicle Low High
 

Topography; Morphology Control Dose Dose
 

Adrenal: Cortical Adenoma or
 

Carcinoma" 2/19 (11) 1/49 (2) 1/49 (2)
 

P Valuesc»d N.S. N.S.
 

Relative Risk6 0.194 0.194
 

Lower Limit 0.003 0.003
 

Upper Limit 3.563 3.563
 

Weeks to First Observed Tumor 104 104 104
 

00
 

00 Thyroid: C-cell Adenoma or
 

Carcinoma'3 1/17 (6) 2/39 (5) 1/41 (2)
 

P Valuesc»d N.S. N.S.
 

Relative Risk6 0.872 0.415
 

Lower Limit 0.050 0.006
 

Upper Limit 50.118 31.786
 

Weeks to First Observed Tumor 104 104 104
 
__ 

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3

 

(continued) 

Topography: Morphology

Vehicle 

 Control 

Pancreatic Islets: 

Islet-cell Adenomab 3/20 (15) 

P Values0 » d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 104 

00 

VO Pancreatic Islets: Islet-cell 

Adenoma or Carcinoma"3 3/20 (15) 

P Values0>d 

Relative Risk6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 104 

Low
 

Dose
 

2/48 (4)
 

N.S.
 

0.278
 

0.025
 

2.278
 

104
 

3/48 (6)
 

N.S.
 

0.417
 

0.062
 

2.915
 

103
 

High
 

Dose
 

0/49 (0)
 

P = 0.022(N)
 

0.000
 

0.000
 

0.673
 

0/49 (0)
 

P = 0.022 (N)
 

0.000
 

0.000
 

0.673
 

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

(continued) 

Vehicle Low High 

Topography; Morphology Control Dose Dose 

Testis: Interstitial-cell Tumor13 17/19 (89) 38/46 (83) 20/47 (43)
 

P Values0 >d N.S. P < O.OOl(N)
 

Relative Risk6 0.923 0.476
 

Lower Limit 0.800 0.394
 

Upper Limit 1.235 0.728
 

Weeks to First Observed Tumor 104 67 76
 

a

Dosed groups received 75 or 150 mg/kg by gavage.
 

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

G

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher exact
 

test for the comparison of that dosed group with the control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

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

e

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

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

Administered 3-(Chloromethyl)pyridine


Vehicle 

Topography: Morphology Control 

Hematopoietic System: Leukemia" 1/20 (5) 

P Values0 

Relative Risk"* 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 104
 

Pituitary: Chromophobe Carcinomab 1/19 (5)
 

P Values0
 

Relative Risk^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 104
 

 Hydrochloride by Gavagea
 

Low
 

Dose
 

3/49 (6)
 

N.S.
 

1.224
 

0.108
 

62.958
 

76
 

1/43 (2)
 

N.S.
 

0.442
 

0.006
 

33.913
 

85
 

High
 

Dose
 

2/50 (4)
 

N.S.
 

0.800
 

0.045
 

46.273
 

98
 

1/40 (3)
 

N.S.
 

0.475
 

0.006
 

36.387
 

95
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

(continued)
 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Pituitary: Chromophobe Adenoma
 

or Carcinomab 9/19 (47) 12/43 (28) 14/40 (35)
 

P Values0	 N.S. N.S.
 

Relative Riskd 0.589 0.739
 

Lower Limit 0.293 0.385
 

Upper Limit 1.342 1.619
 

Weeks to First Observed Tumor 104	 85 72
 

VO
 

NJ	 Mammary Gland: Fibroadenomab

 2/20 (10) 4/49 (8) 1/50 (2)
 

P Values0 N.S. N.S.
 

Relative Riskd 0.816 0.200
 

Lower Limit 0.131 0.004
 

Upper Limit 8.603 3.681
 

Weeks to First Observed Tumor 104 104 104
 
__ 

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

(continued)
 

Vehicle Low High
 

Topography: Morphology Control Dose Dose
 

Preputial Gland:
 

Adenoma, NOSb

 2/20 (10) 1/49 (2) 0/50 (0)
 

P Values0 N.S. N.S.
 

Relative Riskd 0.204 0.000
 

Lower Limit 0.004 0.000
 

Upper Limit 3.754 1.345
 

Weeks to First Observed Tumor 104 104
 

Uterus: Endometrial Stromal Polypb 2/19 (11) 8/47 (17) 2/49 (4)
 

OJ
 

P Values0 N.S. N.S.
 

Relative Riskd 1.617 0.388
 

Lower Limit 0.370 0.031
 

Upper Limit 14.802 5.108
 

Weeks to First Observed Tumor 104 76 104
 

a

Dosed groups received 75 or 150 mg/kg by gavage.
 

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

c

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher exact
 

test for the comparison of that dosed group with the control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

d

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

group.
 

APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN
 

MICE ADMINISTERED 3-(CHLOROMETHYL)PYRIDINE HYDROCHLORIDE
 

BY GAVAGE
 

95
 

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3
 

Topography; Morphology 

Vehicle

Control

 Low 

 Dose 

High 

Dose 

Lung: Alveolar/Bronchiolar 

Carcinoma" 1/20 (5) 4/45 (9) 5/48 (10) 

P Values0 N.S. N.S. 

Relative Riskd 

Lower Limit 

Upper Limit 

1.778 

0.195 

85.520 

2.083 

0.259 

96.358 

Weeks to First Observed Tumor 104 84 102 

Lung: Alveolar/Bronchiolar 

Adenoma or Carcinoma'-' 2/20 (10) 4/45 (9) 5/48 (10) 

P Values0 N.S. N.S. 

Relative Riskd 

Lower Limit 

Upper Limit 

0.889 

0.143 

9.340 

1.042 

0.192 

10.410 

Weeks to First Observed Tumor 104 84 102 
00 

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3
 

(continued)
 

Topography: Morphology


Hematopoietic System:
 

Undifferentiated Leukeraiab


P Values0
 

Relative Riskd
 

Lower Limit
 

Upper Limit
 

Weeks to Fir,st Observed Tumor


Hematopoietic System: Malignant
 

Lymphoma, Lymphocytic Leukemia
 

or Undifferentiated Leukemiab


P Values0
 

Relative Risk**
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Vehicle Low 

 Control Dose 

 1/20 (5) 3/48 (6) 

N.S. 

1.250 

0.110 

64.251 

 73 94 

 3/20 (15) 9/48 (19) 

N.S. 

1.250 

0.361 

6.662 

 73 61 

High
 

Dose
 

3/50 (6)
 

N.S.
 

1.200
 

0.106
 

61.724
 

63
 

9/50 (18)
 

N.S.
 

1.200
 

0.346
 

6.408
 

63
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3 

(continued) 

Vehicle Low High 

Topography; Morphology Control Dose Dose 

Hematopoietic System: 

All Leukemias or Lymphomas" 3/20 (15) 9/48 (19) 10/50 (20) 

P Values0 N.S. N.S. 

Relative Riskd 1.250 1.333 

Lower Limit 0.361 0.398 

Upper Limit 6.662 7.002 

Weeks to First Observed Tumor 73 61 63 

Liver: Hepatocellular Carcinomab 2/20 (10) 5/46 (11) 9/49 (18) 

P Values0 N.S. N.S. 

Relative Riskd 1.087 1.837 

Lower Limit 0.200 0.434 

Upper Limit 10.845 16.572 

Weeks to First Observed Tumor 102 58 99 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered 3-(Chloromet.hyl)pyridine Hydrochloride by Gavagea
 

(continued)
 

Topography: Morphology


Liver: Hepatocellular Adenoma
 

or Carcinomab


P Values0
 

Relative Riskd
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


o
 

o	 Stomach: Squamous-cell Papillomab


P Values0
 

Relative Riskd
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Vehicle Low 

 Control Dose 

 3/20 (15) 5/46 (11) 

N.S. 

0.725 

0.160 

4.348 

 94 58 

 0/19 (0) 2/43 (5) 

N.S. 

Infinite 

0.136 

Infinite 

— 61 

High
 

Dose
 

9/49 (18)
 

N.S.
 

1.224
 

0.354
 

6.533
 

99
 

8/47 (17)
 

N.S.
 

Infinite
 

0.966
 

Infinite
 

58
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered 3-(Chlororaethyl)pyridine Hydrochloride by Gavagea
 

(continued)
 

Topography: Morphology

Vehicle 

 Control 

Low 

Dose 

High 

Dose 

Stomach: Squamous-cell Papilloma 

or Carcinomab 0/19 (0) 2/43 (5) 10/47 (21) 

P Values0 N.S. P = 0.025 

Relative Riskd 

Lower Limit 

Upper Limit 

Infinite 

0.136 

Infinite 

Infinite 

1.259 

Infinite 

Weeks to First Observed Tumor — 61 58 

a

Dosed groups received 100 or 200 mg/kg by gavage.
 

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

G

Beneath the incidence of tumors in a dosed group is the probability level for the Fisher exact
 

test for the comparison of that dosed group with the control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

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

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

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

Topography; Morphology
 

Lung: Alveolar/Bronchiolar
 

Carcinoma^3
 

P Values0
 

Relative Riskd
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma"
 

P Values0
 

Relative Risk^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Vehicle Low 

Control Dose 

1/18 (6) 0/50 (0) 

N.S. 

0.000 

0.000 

6.729 

65 

1/18 (6) 1/50 (2) 

N.S. 

0.360 

0.005 

27.724 

65 104 

High
 

Dose
 

3/49 (6)
 

N.S.
 

1.102
 

0.098
 

56.666
 

50
 

3/49 (6)
 

N.S.
 

1.102
 

0.098
 

56.666
 

50
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavagea
 

(continued) 

Topography: Morphology

Vehicle 

 Control 

Low 

Dose 

High 

Dose 

Hematopoietic System: Malignant 

Lymphoma, Lymphocytic Leukemia 

or Undif ferentiated Leukemiab 2/20 (10) 9/50 (18) 9/49 (18) 

P Values0 N.S. N.S. 

Relative Riskd 

Lower Limit 

Upper Limit 

1.800 

0.426 

16.255 

1.837 

0.434 

16.572 

Weeks to First Observed Tumor 93 76 98 

Hematopoietic System: 

All Lymphomas or Leukemiasb 2/20 (10) 9/50 (18) 10/49 (20) 

P Values0 N.S. N.S. 

Relative Riskd 

Lower Limit 

Upper Limit 

1.800 

0.426 

16.255 

2.041 

0.498 

18.154 

Weeks to First Observed Tumor 93 76 96 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3
 

(continued)
 

Topography: Morphology

Vehicle 

 Control 

Low 

Dose 

High 

Dose 

Pituitary:

Adenomab

 Chromophobe 

 0/9 (0) 0/25 (0) 2/29 (7) 

P Values0 N.S. N.S. 

Relative Riskd 

Lower Limit 

Upper Limit 

— Infinite 

0.103 

Infinite 

Weeks to First Observed Tumor 104
 —
 

Stomach: Squamous-cell Papillomab 0/19 (0) 1/45 (2) 3/48 (6)
 

P Values0 N.S. N.S.
 

Relative Riskd Infinite Infinite
 

Lower Limit 0.023 0.248
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — 104 104
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered 3-(Chloromethyl)pyridine Hydrochloride by Gavage3
 

(continued)
 

Topography: Morphology

Vehicle 

 Control 

Low 

Dose 

High 

Dose 

Stomach: Squamous-cell Papilloma 

or Carcinomab 0/19 (0) 1/45 (2) 5/48 (10) 

P Values0 N.S. N.S. 

Relative Riskd 

Lower Limit 

Upper Limit 

Infinite 

0.023 

Infinite 

Infinite 

0.522 

Infinite 

Weeks to First Observed Tumor — 104 104 

a

Dosed groups received 100 or 200 mg/kg by gavage.
 

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

°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 control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

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

group.
 

Review of the Bioassay of 3-(Chloromethyl)pyridine Hydrochloride* 

for carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be exposed.
 The members of the Clearinghouse have been drawn from
 

academia, industry, organized labor, public interest groups,
 

State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 

biostatistics, toxicology, pathology, and epidemiology.
 

Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

Subgroup of the Clearinghouse is charged with the responsibility of providing a peer review of reports prepared on
 

NCI-sponsored bioassays of chemicals studied for carcinogenic­ ity. It is in this context that the below critique is given
 

on the bioassay of 3-(ChloromethyDpyridine Hydrochloride
 

for carcinogenicity.
 

The reviewer agreed with the conclusion In the report
 

that 3-(Chloromethyl)pyridine Hydrochloride was carcinogenic
 

under the conditions of test. After a brief review of the
 

experimental design, the reviewer said that the study was
 

adequate to support the conclusion on the compound's carcino­ genicity. The review of the bioassay of 3-(Chloromethyl)
 

pyridine Hydrochloride was accepted without objection.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical U.S.A.
 Michael B. Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

107
 

*U.S. GOVERNMEN T PRINTIN G OFF!CE : 1978-260-899/3,86
 

DHEW Publication No. (NIH) 78-1345