National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 52 

1978 

BIOASSAY OF 

3-NITROPROPIONIC ACID 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 504-88-1 

NCI-CG-TR-52 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

3-NITROPROPIONIC ACID
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

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

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-1302
 

BIOASSAY OF
 

3-NITROPROPIONIC ACID
 

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-nitropropionic acid 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-nitropropionic acid was
 

conducted by The Dow Chemical Company, Indianapolis, Indiana,
 

initially under direct contract to NCI and currently under a
 

subcontract to Tracor Jitco, Inc., prime contractor for the NCI
 

Carcinogenesis Testing Program.
 

The experimental design and doses were determined by Dr. E. K.
 

Weisburger^. Dr. C. G. Gerbig^ supervised the preparation of the
 

gavage solutions and was responsible for animal care.
 

Histopathologic examinations were performed by Dr. J. L.
 

Emerson2»3, the principal investigator, and the diagnoses
 

included in this report represent his interpretations. Drs.
 

Emerson and Gerbig prepared the data for the methodology section
 

of this report.
 

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

Mason Research Institute . The statistical analyses were
 

performed by Dr. J. R. Joiner-3, using methods selected for the
 

bioassay program by Dr. J. J. Gart". Chemicals used in this
 

bioassay were analyzed under the direction of Dr. E. Murrill ,
 

and the analytical results were reviewed by Dr. S. S. Olin^.
 

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

NCI. Those responsible for the report at Tracor Jitco were
 

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

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

C. H. Williams, toxicologists; 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.
 

The statistical analysis was reviewed by members of the
 

Mathematical Statistics and Applied Mathematics Section of NCI":
 

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

Robert E. Tarone.
 

The following other scientists at NCI were responsible for
 

evaluating the bioassay experiment, interpreting the results, and
 

reporting the findings:
 

Dr. Kenneth C. Chu
 

Dr. Cipriano Cueto, Jr.
 

Dr. J. Fielding Douglas
 

Dr. Dawn G. Goodman
 

Dr. Richard A. Griesemer
 

Dr. Harry A. Milman
 

Dr. Thomas W. Orme
 

Dr. Robert A. Squire8
 

Dr. Jerrold M. Ward
 

^Carcinogenesis Testing Program, Division of. Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

iv
 
Dow Chemical Company, P.O. Box 68511, Indianapolis,
 

Indiana.
 

%ow with the Abbott Laboratories, D-469 AP 9, North Chicago,
 

Illinois.
 

Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

5

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

Maryland.
 

^Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division
 

of Cancer Cause and Prevention, National Cancer Institute,
 

National Institutes of Health, Bethesda, Maryland.
 

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

Missouri.
 

°Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of 3-nitropropionic acid (95% pure) 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-nitropropionic acid at one of the following doses by gavage 5
 

days per week. For male rats, the doses were 0.425 or 0.85
 

mg/animal/day; for females, they were 0.6 or 1.2 mg/animal/day.
 

For both sexes of mice, the doses were 0.375 or 0.75
 

mg/animal/day. The rats were administered the chemical for 110
 

weeks and the mice for 104 weeks. The controls consisted of 50
 

untreated rats and 50 untreated mice of each sex. All surviving
 

rats were killed at 111 weeks and all surviving mice at 104 or
 

105 weeks.
 

Mean body weights and mortality of the dosed animals were not
 

markedly affected by 3-nitropropionic acid under the conditions
 

of this bioassay, indicating that the maximum tolerated dose may
 

not have been reached. The various clinical signs observed were
 

common to both dosed and control groups.
 

In rats, the combination of neoplastic nodule of the liver and
 

hepatocellular carcinoma occurred in the males with a significant
 

dose-related trend (P = 0.010) and with a higher incidence (P =
 

0.012) in the high-dose group of animals than in the controls
 

(controls 0/49, low-dose 3/50, high-dose 6/49). All but one of
 

these tumors were neoplastic nodules. In the females, only two
 

neoplastic nodules occurred, one in each of the dosed groups.
 

Biliary hyperplasia occurred at a higher incidence in the dosed
 

males than in the corresponding controls (controls 19/50,
 

low-dose 32/50, high-dose 36/50), but the incidence of this
 

lesion in the dosed females was not increased as compared with
 

controls. There was also a dose-related trend (P = 0.033) in the
 

incidence of pancreatic islet-cell adenoma in the male rats
 

(controls 4/49, low-dose 6/50, high-dose 11/50); however, direct
 

comparisons of incidences in the dosed and control groups were
 

not statistically significant. The historical incidence of
 

vii
 
pancreatic islet-cell adenoma among 100 control Fischer 344 rats
 

at the laboratory was 7/100 (7%). In addition, focal myocardial
 

fibrosis was observed at a higher incidence in dosed rats than
 

among controls (males: controls 1/4, low-dose 17/49, high-dose
 

24/48; females: controls 2/48, low-dose 9/46, high-dose 9/50).
 

In mice, each type of neoplasm found in the dosed and control
 

mice has been encountered previously as a spontaneous lesion. No
 

specific tumor was found to occur at a statistically significantly higher incidence among dosed mice than among the respective
 

control groups.
 

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

was"an elevated incidence of hepatocellular neoplasms, primarily
 

benign, and of islet-cell adenomas of the pancreas in male
 

Fischer 344 rats receiving 3-nitropropionic acid as compared with
 

controls; however, there was no conclusive evidence that
 

3-nitropropionic acid was carcinogenic in these animals. The
 

chemical was not carcinogenic in female rats or in male or female
 

B6C3F1 mice.
 

viii
 
TABLE OF CONTENTS
 

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. Designs of Chronic Studies 8
 

G. Clinical and Pathologic Examinations 11
 

H. Data Recording and Statistical Analyses 12
 

III. Results -Rats 19
 

A. Body Weights and Clinical Signs (Rats) 19
 

B. Survival (Rats) 19
 

C. Pathology (Rats) 21
 

D. Statistical Analyses of Results (Rats) 24
 

IV. Results - Mice 27
 

A. Body Weights and Clinical Signs (Mice) 27
 

B. Survival (Mice) 27
 

C. Pathology (Mice) 30
 

D. Statistical Analyses of Results (Mice) 32
 

V. Discussion 35
 

VI. Bibliography 39
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms
 

in Rats Administered 3-Nitropropionic
 

Acid by Gavage 43
 

Table Al Summary of the Incidence of Neoplasms
 

in Male Rats Administered 3-Nitropropionic
 

Acid by Gavage 45
 

ix
 
Table A2 Summary of the Incidence of Neoplasms 

in Female Rats Administered 3-Nitropropionic 

Acid by Gavage 49 

Appendix B Summary of the Incidence of Neoplasms 

in Mice Administered 3-Nitropropionic 

Acid by Gavage 53 

Table Bl Summary of the Incidence of Neoplasms 

in Male Mice Administered 3-Nitropropionic 

Acid by Gavage 55 

Table B2 Summary of the Incidence of Neoplasms 

in Female Mice Administered 

3-Nitropropionic Acid by Gavage 59 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Administered 

3-Nitropropionic Acid by Gavage 63 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Administered 

3-Nitropropionic Acid by Gavage 65 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Administered 

3-Nitropropionic Acid by Gavage 74 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Administered 

3-Nitropropionic Acid by Gavage 83 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Administered 

3-Nitropropionic Acid by Gavage 85 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Administered 

3-Nitropropionic Acid by Gavage 91 

Appendix E Analyses of the Incidence of Primary 

Tumors in Rats Administered 

3-Nitropropionic Acid by Gavage.. 97 
Table El
 

Table E2
 

Appendix F
 

Table Fl
 

Table F2
 

Table 1
 

Table 2
 

Figure 1
 

Figure 2
 

Figure 3
 

Figure 4
 

Page 

Analyses of the Incidence of Primary 

Tumors in Male Rats Administered 

3-Nitropropionic Acid by Gavage 99 

Analyses of the Incidence of Primary 

Tumors in Female Rats Administered 

3-Nitropropionic Acid by Gavage 108 

Analyses of the Incidence of Primary 

Tumors in Mice Administered 

3-Nitropropionic Acid by Gavage 113 

Analyses of the Incidence of Primary 

Tumors in Male Mice Administered 

3-Nitropropionic Acid by Gavage 115 

Analyses of the Incidence of Primary 

Tumors in Female Mice Administered 

3-Nitropropionic Acid by Gavage 120 

TABLES 

Design of Chronic Studies of 

3-Nitropropionic Acid in Rats 9 

Design of Chronic Studies of 

3-Nitropropionic Acid in Mice 10 

FIGURES 

Growth Curves for Rats Administered 

3-Nitropropionic Acid by Gavage 20 

Survival Curves for Rats Administered 

3-Nitropropionic Acid by Gavage 22 

Growth Curves for Mice Administered 

3-Nitropropionic Acid by Gavage 28 

Survival Curves for Mice Administered 

3-Nitropropionic Acid by Gavage 29 

xi
 

I. INTRODUCTION
 

3-Nitropropionic acid (CAS 504-88-1; NCI C03076), also called
 

3-nitropropionic acid or hiptagenic acid, has been isolated from
 

plants, including a tropical forage plant (Cooke, 1955; Finnegan
 

and Mueller, 1965; Morris et al., 1954), and from nuts that are
 

eaten in the New Zealand area as a food staple (Bell, 1974;
 

Carter, 1951). It has been isolated from Streptomyces found in
 

soil (Anzai and Suzuki, 1960) and as a metabolite of certain
 

fungal species of Aspergillus (Bush et al., 1951; Iwasaki and
 

Kosikowski, 1973) and Penicillium (Hylin and Matsumoto, 1960;
 

Raistrick and Stossl, 1958). These species of fungi are commonly
 

present in several oriental fermented foodstuffs, both domestically and commercially produced, in which 3-nitropropionic acid has
 

been identified (Kinosita et al., 1968). Other fungal strains
 

that are frequent contaminants in many kinds of food have been
 

found to produce mycotoxins that have exhibited carcinogenic
 

activity in experimental animals (Butler et al., 1969; IARC,
 

1972; Wogan and Newberne, 1967).
 

3-Nitropropionic acid was selected for testing for carcinogenic
 

activity because it was known to demonstrate varying degrees of
 

toxicity in man and animals (Hutton et al. , 1958; Morris et al. ,
 

1954; Bell, 1974), and because its use in food preparations and
 
its identification as a contaminant in foods suggested there was
 

a possibility of long-term human exposure.
 
II. MATERIALS AND METHODS
 

A. Chemical
 

3-Nitropropionic acid, synthesized from B-propiolactone, was
 

obtained from Aldrich Chemical Co., Milwaukee, Wisconsin, in a
 

single batch (Lot No. 111627) for the chronic study.
 

Analysis at Midwest Research Institute confirmed the identity of
 

the chemical. Infrared and nuclear magnetic resonance (nmr)
 

spectra were as expected for 3-nitropropionic acid, with the
 

exception that the nmr spectra revealed a 5% impurity which was
 

identified as a dimeric ester of 3-hydroxypropionic acid.
 

Elemental analyses for carbon and hydrogen agreed with the
 

theoretical values for C3H5N04, the molecular formula for
 

3-nitropropionic acid, but the results for nitrogen were slightly
 

low. Titration with sodium hydroxide gave 100.7 + 0.3% of the
 

theoretical value. High-pressure liquid chromatography showed a
 

single peak (uv detector, 254 nm), whereas thin-layer
 

chromatography indicated two trace impurities. Water content by
 

Karl Fischer analysis was 0.35 +_ 0.01%. In summary, the analyses
 

indicated that the batch used for the chronic study was
 

approximately 95% pure, with a single major organic impurity,
 

apparently a dimeric ester of 3—hydroxypropionic acid, comprising
 

most of the remainder.
 
The chemical was stored at 4°C in the original glass container.
 

B. Dosage Preparation
 

3-Nitropropionic acid was administered in feed during the sub-


chronic study. Polarographic and chromatographic analyses of
 

extracts of samples of the test diets suggested partial
 

decomposition of the chemical. To maintain adequate doses during
 

the chronic study, the chemical was administered by gavage in an
 

aqueous solution. A 1-mg/ml solution of 3-nitropropionic acid in
 

distilled water was prepared once per day and used within 1-1/2
 

hours after preparation. This solution was stable for 3 hours at
 

ambient temperature, as verified by both high-pressure liquid
 

chromatographic and polarographic analyses.
 

C. Animals
 

Rats and mice of each sex, obtained through contracts of the
 

Division of Cancer Treatment, National Cancer Institute, were
 

used in these bioassays. The rats were of the Fischer 344 strain
 

obtained from A. R. Schmidt/Sprague-Dawley, Madison, Wisconsin,
 

and the mice were B6C3F1 hybrids obtained from Charles River
 

Breeding Laboratories, Inc., Wilmington, Massachusetts. The rats
 

and mice were approximately 28 days of age when received. On
 

arrival at the laboratory, all animals were quarantined (rats for
 

7 days, mice for 14 days) and then assigned to control or dosed
 
groups. All animals were individually identified: rats were
 

earmarked and mice were toe-clipped.
 

D. Animal Maintenance
 

All animals were housed in temperature- and humidity-controlled
 

rooms. The temperature was maintained at 22-25°C, and the
 

relative humidity was maintained at 45-55%. The room air was
 

changed 15 times per hour. Illumination was provided by
 

fluorescent lighting 14 hours per day. Wayne® Lab Blox animal
 

meal (Allied Mills, Inc., Chicago, 111.) and chlorinated well
 

water that was deionized were available ad libitum.
 

Initially, rats in the chronic study were housed individually in
 

suspended cages made of stainless-steel wire mesh (Ford Fence
 

Co., Indianapolis, Ind.). At week 45, all rats were housed three
 

per cage in suspended polycarbonate cages (Maryland Plastics,
 

Federalsburg, Md.) lined with autoclaved Absorb-Dri bedding (Lab
 

Products, Inc., Garfield, N. J.) and equipped with filters and an
 

automatic watering system. The cages were changed, washed, and
 

sanitized at 82°C twice per week. The feeders were changed,
 

washed, and sterilized once per week, and the filters were
 

changed every 2 weeks.
 

Mice were housed five per cage in filtered, prebedded cages made
 

of disposable polypropylene (Lab Products, Inc., Garfield, N.J.).
 
The cages were changed twice per week and the used cages were
 

incinerated. Feeders, water bottles, and cage lids were also
 

changed twice per week, and cage filters were changed once per
 

week. Feeders and sipper tubes were washed and sterilized prior
 

to use. Water bottles and cage lids were sanitized at 82°C.
 

Rats and mice were housed in separate rooms. The animal racks
 

were rotated once per week and the cages were kept in fixed
 

positions on the racks. The rats administered 3-nitropropionic
 

acid were housed in the same room as rats fed 2-amino5-nitrothiazole (CAS 121-66-4) and the positive control, N-9Hf luoren-2-ylacetamide (CAS 53-96-3), in the diet. The mice
 

administered 3-nitropropionic acid were housed in the same room
 

as mice fed 2-amino-5-nitrothiazole, N,N'-dicyclohexylthiourea
 

(CAS 1212-29-9), proflavine hydrochloride (CAS 952-23-8),
 

l,3-dichloro-5,5-dimethylhydantoin (CAS 118-52-5), and N-9Hfluoren-2-ylacetamide in the diet. The control animals were
 

housed in the same room with respective dosed animals.
 

E. Subchronic Studies
 

Subchronic feeding studies were conducted with rats and mice to
 

estimate the maximum tolerated doses of 3-nitropropionic acid, on
 

the basis of which low and high concentrations (hereinafter
 

referred to as "low doses" and "high doses") were determined for
 
administration in the chronic studies. In the subchronic
 

studies, 3-nitropropionic acid was added to the animal feed in
 

concentrations ranging from 100 to 900 ppm for rats and from 150
 

to 800 ppm for mice. Five males and five females of each species
 

were tested at the different doses, and equal numbers of males
 

and females were used as untreated controls. All animals were
 

fed the chemical for 6 weeks, then observed for 2 weeks. All
 

animals were necropsied and gross lesions were examined
 

histologically.
 

In male rats, mean body weight gain was 77% of that of the
 

controls at 100 ppm, 59% at 150 ppm, and 57% at 250 ppm. All
 

males at 500 and 900 ppm died. In females, mean body weight gain
 

was 97% of the controls at 100 ppm, 87% at 150 ppm, 71% at 250
 

ppm, and 62% at 500 ppm. Two females died at 250 ppm, four at
 

500 ppm, and five at 900 ppm. On histologic examination, testicular atrophy with spermatogenic arrest was found in male rats and
 

malacia in the midbrain in both sexes of rats given doses of 150
 

ppm and above. For male rats, the low and high doses for the
 

chronic studies were set at 25 and 50 ppm; for females, they were
 

set at 50 and 100 ppm.
 

In male mice, mean body weight gain of groups receiving 150 or
 

600 ppm was not affected. An early weight depression was
 

observed at 800 ppm; however, these animals recovered, and their
 
final weights were comparable to those of control mice. Mean
 

body weights in female mice were not markedly affected at any
 

dose tested. One male died at 600 ppm, and one male died at 800
 

ppm. Hydronephrosis was found in nine mice, but the incidence
 

was not dose related. For both male and female mice, the low and
 

high doses for the chronic studies were set at 75 and 150 ppm.
 

F. Designs of Chronic Studies
 

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

Because the test chemical was unstable in feed, the method of
 

administration used for the chronic study was gavage. Doses were
 

converted from parts per million to milligrams per animal per day
 

(mg/animal/day) based on an estimated food consumption of 17
 

g/day for male rats, 12 g/day for female rats, and 5 g/day for
 

mice (both sexes). The doses in mg/animal/day that are stated in
 

the tables were used throughout the study; thus, as the weights
 

of the animals increased, the amounts per unit of body weight
 

decreased. Since water was used as the vehicle, no control
 

groups administered a vehicle by gavage were included. The
 

control animals were those started with another chemical on test
 

at the same time in the diet; thus, they received control diet
 

only.
 
Table 1. Design of Chronic Studies of 3-Nitropropionic Acid
 

in Rats
 

S ex and
 

Test
 

Group
 

Male
 

Control
 

Low-Dose
 

High-Dose
 

Female
 

Control
 

Low-Dose
 

High-Dose
 

Initial
 

No. of
 

Animals3
 

50
 

50
 

50
 

50
 

50
 

50
 

3-Nitropropionic Acid
 

Dose
 

(mg/animal/day)k
 

Od
 

0.425
 

0.85
 

od
 

0.6
 

1.2
 

Time
 

Dosed
 

(weeks)
 

110
 

110
 

110
 

110
 

on Study0
 

Observed
 

(weeks)
 

111
 

1
 

1
 

111
 

1
 

1
 

a

Rats were approximately 50 days of age when placed on study.
 

"Animals were administered the chemical by gavage 5 days per week.
 

C

A11 animals were started on study on the same day.
 

"The control groups were not administered the chemical.
 
Table 2. Design of Chronic Studies of 3-Nitropropionic Acid
 

in Mice
 

Sex and
 

Test
 

Group
 

Male
 

Control
 

Low-Dose
 

High-Dose
 

Female
 

Control
 

Low-Dose
 

High-Dose
 

Initial
 

No. of
 

Animals3
 

50
 

50
 

50
 

50
 

50
 

50
 

3-Nitropropionic Acid
 

Dose
 

(mg/animal/day)°
 

Od
 

0.375
 

0.75
 

Od
 

0.375
 

0.75
 

Time
 

Dosed
 

(weeks)
 

104
 

104
 

104
 

104
 

on Study0
 

Observed
 

(weeks)
 

104
 

1
 

1
 

104
 

1
 

1
 

a

Mice were approximately 53 days of age when placed on study.
 

bAnimals were administered the chemical by gavage 5 days per week.
 

C

A11 animals were started on study on the same day.
 

d

The control groups were not administered the chemical.
 

10
 
G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity and
 

were weighed every 14 days during the first 3 months and every 28
 

days thereafter. Clinical observations were recorded at weekly
 

intervals. Animals chat were moribund at the tiiae of daily
 

examination were killed and necropsied; however, some, moribund
 

animals were isolated from their cage-mates for a few days prior
 

to being killed.
 

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, lungs and bronchi,
 

trachea, bone marrow, spleen, lymph nodes, thymus, heart, salivary gland, liver, gallbladder (mice), pancreas, esophagus,
 

stomach, small intestine, large intestine, colon, kidney, urinary
 

bladder, pituitary, adrenal, thyroid, parathyroid, mammary gland,
 

testis or ovary, prostate or uterus, brain, and eyes. 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.
 

11
 
A few tissues from some animals were not examined, particularly
 

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

cannibalized, or judged to be in such an advanced state of
 

autolysis as to preclude histopathologic evaluation. Thus, the
 

number of animals from which particular organs or tissues were
 

examined microscopically varies, and does not necessarily represent the number of animals that were placed on study in each
 

group.
 

H. Data Recording and Statistical Analyses
 

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

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

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

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

verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear on the possibility of carcinogenicity are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

12
 
procedure of Kaplan and Meier (1958) and are presented in this
 

report in the form of graphs. Animals were statistically
 

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

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

causes were not statistically censored. Statistical analyses for
 

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

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

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

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

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

the denominators included only those animals for which that site
 

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

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

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

consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

13
 
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. Under the
 

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

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


tailed 0.05 level of significance. Unless otherwise noted, the
 

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

14
 
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 >- f the proportions surviving without
 

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

The week during which an animal died naturally or was sacrificed
 

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

methods of comparing these curves were used for two groups;
 

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

groups. The statistical tests for the incidence of tumors which
 

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

noted, in the direction of a positive dose relationship.
 

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

were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared with its control was calculated
 

15
 
from the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as Pt/pc where pj- 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
 

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
 

16
 
indicates that there is a theoretical possibility of the
 

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

detected under the conditions of this test.
 

17
 

III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of both dosed groups of each sex were not
 

appreciably lower than those of the controls (figure 1).
 

Throughout the study, the dosed rats were generally comparable to
 

the controls in appearance and behavior. Early during the second
 

year of the study, approximately 75% of the rats, including the
 

controls, developed acute swelling of the submaxillary salivary
 

glands. The clinical appearance was consistent with that of
 

sialodacryoadenitis. Both control and dosed animals developed
 

this condition, which lasted for approximately 14 days. The
 

animals developed partial anorexia and rough coats, and in some
 

cases the animals lost weight. Unilateral and occasionally
 

bilateral cataracts appeared in both control and dosed rats at
 

the end of the first year and continued through the second year.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered 3-nitropropionic
 

acid by gavage at the doses of this bioassay, together with those
 

of the controls, are shown in figure 2.
 

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

19
 
D D il si i 

o 

j. 250­

Q 

O 

CD 

Z 200­

< 

111 

5 MALE RATS 

[] CONTROL 

O LOW DOSE 

A HIGH DOSE 

—1— —I— —I— 40 50 60 70 90 

TIME ON STUDY (WEEKS) 

n 1 1 ha gi 8 

>• 250­ Q 

O 

m § 200­

,H 

FEMALE RATS 

D CONTROL 

O LOW DOSE 

A HIGH DOSE 

40 50 60 70 

TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves for Rats Administered 3-Nitropropionic Acid by Gavage 

20 
mortality is not significant at the 0.05 level in either sex. In
 

male rats, 30/50 (60%) of the controls, 33/50 (66%) of the
 

low-dose group, and 30/50 (60%) of the high-dose group lived to
 

the last week of the study. In females, 33/50 (66%) of the
 

controls, 26/50 (52%) of the low-dose group, and 32/50 (64%) of
 

the high-dose group survived to the last week of the study. A
 

sufficient number of rats of each sex was 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 neoplasms occurred in both the control and dosed
 

groups. Each type of neoplasm represented in the tables has been
 

encountered previously as a spontaneous lesion in rats.
 

In male rats, only one hepatocellular carcinoma was observed;
 

this tumor was present in a high-dose animal. The incidence of
 

neoplastic nodules, as described by Squire and Levitt (1975), was
 

as follows in males: controls 0/49 (0%), low-dose 3/50 (6%),
 

high-dose 5/49 (10%). In female rats, neoplastic nodules were
 

observed in 1/50 (2%) of each dosed group, but in none of the
 

controls.
 

21
 
£• PROBABILITY OF SURVIVAL PROBABILIT

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The incidence of pancreatic islet-cell adenoma was dose related
 

in males (controls 4/49 [8%] , low-dose 6/50 [12%], high-dose
 

11/50 [22%]). This trend was not evident in females.
 

Nonneoplastic lesions consisted of degenerative, proliferative,
 

and inflammatory changes that are commonly observed in aging rats
 

(Davey and Moloney, 1970; Sass et al., 1975). These conditions
 

occurred in a random fashion and did not appear to be related to
 

administration of the chemical.
 

Focal myocardial fibrosis occurred in 1/48 (2%) control males,
 

17/49 (35%) low-dose males, 24/48 (50%) high-dose males; 2/48
 

(4%) control females, 9/46 (20%) low-dose females, and 9/50 (18%)
 

high-dose females.
 

Biliary hyperplasia occurred in 19/50 (38%) control males, 32/50
 

(64%) low-dose males, and 36/50 (72%) high-dose males; 15/50
 

(30%) control females, 17/50 (34%) low-dose females, and 18/50
 

(36%) high-dose females.
 

In the judgment of the pathologist, 3-nitropropionic acid was not
 

carcinogenic in Fischer 344 rats when administered under the
 

conditions of this study, although chemical administration may be
 

associated with a slightly increased incidence of benign tumors
 

of the pancreatic islets and of the liver in males.
 

23
 
D. Statistical Analyses of Results (Rats)
 

Tables El and E2 of Appendix E contain the statistical analyses
 

of the incidences of those primary tumors that are relevant to
 

adequate analysis as well as those primary tumors that occurred
 

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

least 5% in one or more than one group.
 

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

positive dose-related trend in combined tumor incidences of those
 

animals with either neoplastic nodules or hepatocellular
 

carcinoma of the liver are significant (P = 0.010) and the
 

results of the Fisher exact test show that the incidence in the
 

high-dose group is significantly higher (P = 0.012) than that in
 

the controls. At this laboratory, none out of a total of 100
 

control male rats receiving only the control diet used in this
 

study were observed to have neoplastic nodules or hepatocellular
 

carcinomas. The statistical analysis suggests that the incidence
 

of this combination of tumors in male rats is dose associated.
 

The results of statistical tests on the incidence of these tumors
 

in females are not significant.
 

In the analyses of the incidence of islet-cell adenoma of the
 

pancreatic islets in male rats, the result of the Cochran-


Armitage test is significant (P = 0.033). The Fisher exact test
 

24
 
shows a probability level of 0.049 when the incidence in the
 

high-dose group is compared with that in the controls, but this
 

level is above that of 0.025, which is required by the multiple
 

comparison criterion. The laboratory historical controls have an
 

incidence of 7/100 (7%) of islet-cell adenoma. No significant
 

incidence of islet-cell adenoma is obtained for the females, and
 

no islet-cell carcinoma was observed in either sex. No other
 

tumors appeared in significant incidences in the dosed groups
 

when compared with the control groups.
 

25
 

IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of both low- and high-dose males and females
 

were lower than those of the controls during the greater part of
 

the bioassay (figure 3). Fluctuations in the growth curve may be
 

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

body weight may be subject to wide variation. Throughout the
 

study, the dosed animals were generally comparable to the
 

controls in appearance and behavior. Focal alopecia, focal
 

dermatitis, and small palpable nodules in the perineal area were
 

observed in increasing numbers of male mice after 7 months on
 

study. These lesions were associated with fighting.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered 3-nitropropionic
 

acid by gavage at the doses of this bioassay, together with those
 

of the controls, are shown in figure 4.
 

In each sex, the result of the Tarone test for positive dose-


related trend in mortality is not significant at the 0.05 level.
 

In male mice, 38/50 (76%) of the controls, 36/50 (72%) of the
 

low-dose group, and 38/50 (76%) of the high-dose group lived to
 

27
 
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Dgt 

Q

s > 

MALE MICE 

D CONTROL 

O LOW DOSE 

A HIGH DOSE 

40 50 60 70 

TIME ON STUDY (WEEKS) 

D o o 0 D0 D D Q 

D 00°°°00 0 

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 35 

a

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30 

IU 20 

FEMALEMICE 

D CONTROl 

O LOW DOSE 

A HIGH DOSE 

40 50 60 70 

TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves for Mice Administered 3-Nitropropionic Acid by Gavage 

28 
10 20 30 40 50 60 70 80 90 100 110 

ft^^­

^ra1

 tO**** ~7^-

°1 &.-,...

^J 

L. ti ^0 

T \—h A 

TAA 

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3 

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0. 

FEMALE MICE 

D CONTROL 

0.20­ O LOW DOSE —' 

AHIG HOOSE 

n on­ 10 20 40 50 60 70 80 90 100 110 

TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves for Mice Administered 3-Nitropropionic Acid by Gavage 

29 
the end of the study. In females, 35/50 (70%) of the controls,
 

43/50 (86%) of the low-dose group, and 39/50 (78%) of the high-


dose group survived to termination of the study. A sufficient
 

number of mice of each sex was 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 neoplasms occurred in both the control and dosed
 

groups. Each of the types of neoplasms represented in the tables
 

has been encountered previously as a spontaneous lesion in the
 

mouse.
 

The incidences of hepatocellular carcinomas, hepatocellular
 

adenomas, and hyperplastic lesions (nodular hyperplasia and
 

hyperplastic nodule) of the liver in mice are summarized below:
 

30
 
MALES FEMALES 

Low High Low High 

Control Dose Dose Control Dose Dose 

Number of
 

animals with
 

tissues
 

examined
 

microscopically 49 50 49 49 50
 50
 

Hepatocellular
 

carcinoma 16 (33%) 8 (16%) 12 (24%) 1 (2%) 1 (2%) 2 (4%)
 

Hepatocellular
 

adenoma 4^(8%) 2 (4%) 4 (8%) 1 (2%) 0 (0%) 2 (4%)
 

Hyper-


plastic
 

lesions 1 (2%) 1 (2%) 0 (0%) 0 (0%) 0 (0%) 0 (0%)
 

The incidences of proliferative hepatocellular lesions were
 

greater in males than in females; however, there was no
 

indication that these lesions were related to administration of
 

the test chemical.
 

Other lesions that occurred among control and dosed groups were
 

considered to be spontaneous.
 

Several chronic inflammatory, degenerative, and proliferative
 

conditions were observed in all groups. These conditions were
 

considered to be of common occurrence, spontaneous, and not
 

related to administration of the test chemical.
 

31
 
In the judgment of the pathologist, 3-nitropropionic acid was not
 

carcinogenic in B6C3F1 mice when administered under the
 

conditions of this study.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 of Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

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

5% in one or more than one group.
 

There is no specific incidence of tumors in either sex of mice
 

for which the results of the Cochran-Armitage test or of the
 

Fisher exact test are significant at the 0.05 level in the
 

positive direction. In two instances the control groups had a
 

significantly higher incidence than dosed groups. The incidence
 

of hepatocellular adenoma or carcinoma in male mice is lower (P =
 

0.021) in the low-dose group than in the control group. In
 

female mice, the occurrence of the combination of tumors in the
 

hematopoietic system is lower (P = 0.015) in the high-dose group
 

than in the control group. These results in the negative
 

direction cannot be explained by differential mortality, since
 

survivals of these groups within each sex are comparable.
 

In each of the 95% confidence intervals, shown in the tables, the
 

value of one or less than one is included; this indicates the
 

32
 
absence of significant positive results. It should also be noted
 

that each of the intervals (except that for the incidence of
 

hepatocellular adenoma and carcinoma in the low-dose group of
 

male mice and that for the incidence of hematopoietic tumors in
 

the high-dose group of female mice) has an upper limit greater
 

than one, indicating the theoretical possiblity of the induction
 

of tumors by 3-nitropropionic acid, which could not be detected
 

under the conditions of this test.
 

33
 

V. DISCUSSION
 

Mean body weights and mortality of the dosed rats were not
 

markedly affected by 3-nitropropionic acid under the conditions
 

of the bioassay. Mean body weights of dosed mice were slightly
 

lower than those of controls throughout the greater part of the
 

bioassay. The various clinical signs observed were common to
 

both dosed and control groups.
 

In rats, the combination of neoplastic nodule of the liver and
 

hepatocellular carcinoma occurred in the males with a significant
 

dose-related trend (P = 0.010) and with a higher incidence (P =
 

0.012) in the high-dose group of animals than in the controls
 

(controls 0/49, low-dose 3/50, high-dose 6/49). All but one of
 

these tumors were neoplastic nodules. In the females, only two
 

neoplastic nodules occurred, one in each of the dosed groups.
 

Biliary hyperplasia occurred at a higher incidence in the dosed
 

males than in the corresponding controls (controls 19/50,
 

low-dose 32/50, high-dose 36/50), but the incidence of this
 

lesion in the dosed females was not increased as compared with
 

controls. There was also a dose-related trend (P = 0.033) in the
 

incidence of pancreatic islet-cell adenoma in the male rats
 

(controls 4/49, low-dose 6/50, high-dose 11/50); however, direct
 

comparisons of incidences in the dosed and control groups were
 

not statistically significant. The historical incidence of
 

35
 
pancreatic islet-cell adenomas among 100 control Fischer 344 rats
 

at the laboratory was 7/100 (7%). In addition, focal myocardial
 

fibrosis was observed at a higher incidence in dosed rats than
 

among controls (males: controls 1/4, low-dose 17/49, high-dose
 

24/48; females: controls 2/48, low-dose 9/46, high-dose 9/50).
 

In mice, each type of neoplasm found in the dosed and control
 

mice has been encountered previously as a spontaneous lesion. No
 

specific tumor was found to occur at a statistically significantly higher incidence among dosed mice than among the respective
 

control groups.
 

The minimum acute lethal dose of 3-nitropropionic acid has been
 

reported to be 100 mg/kg for rats (Bell, 1974). Rabbits treated
 

with a total of 5.5 g over a period of 34 days showed no toxic
 

effects (Hutton et al., 1958). There have been no previous
 

long-term toxicity studies of this chemical. The compound first
 

attracted attention when Morris et al. (1954) found that it was
 

present in a potential pasture legume (Indigofera endecaphylla)
 

grown in tropical countries. This legume was severely toxic to
 

grazing animals and the toxic principle was thought to be
 

3-nitropropionic acid. Hutton et al. (1958), however, fed the
 

leaves of the legume and also pure 3-nitropropionic acid to
 

rabbits and found the leaves caused severe liver damage, while
 

the pure acid had no effect on the liver. 3-Nitropropionic acid
 

36
 
is one of the metabolites of fungi such as Aspergillus flavus,
 

which is a widespread contaminant of foodstuffs.
 

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

was an elevated incidence of hepatocellular neoplasms, primarily
 

benign, and of islet-cell adenomas of the pancreas in male
 

Fischer 344 rats receiving 3-nitropropionic acid as compared with
 

controls; however, there was no conclusive evidence that
 

3-nitropropionic acid was carcinogenic in these animals. The
 

chemical was not carcinogenic in female rats or in male or female
 

B6C3F1 mice.
 

37
 

VI. BIBLIOGRAPHY
 

Anzai, K. and Suzuki, S., A new antibiotic bovinocidin, 

identified as S-nitropropionic acid. J. Antibiotics 

JJH2): 133-136, 1960. 

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

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

Bell, M. E., Toxicology of karaka kernel, karakin, and
 

3-nitropropionic acid. New Zealand J. Sci. 17:327-334,
 

1974.
 

Berenblum, I., ed., Carcinogenicity Testing, A_ Report of the
 

Panel on Carcinogenicity of the Cancer Research Commission
 

of the UICC, Vol. _2, International Union Against Cancer,
 

Geneva, 1969.
 

Bush, M. T. , Touster, 0., and Brockman, J. E., The production of
 

3-nitropropionic acid by a strain of Aspergillus flavis. J.
 

Biol. Chem. 188(2);685-693, 1951.
 

Butler, W. H., Greenblatt, M., and Lijinsky, W., Carcinogenesis
 

in rats by aflatoxins Bj, Gj and B£. Cancer Res.
 

.29:2206-2211, 1969.
 

Carter, C. L. , The constitition of karakin. J. Sci. Food Agric.
 

.2:54-55, 1951.
 

Cooke, A. R., The toxic constituent of Indigofera endecaphylla.
 

Arch. Biochem. Biophys. 5^:114-120, 1955.
 

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.
 

Davey, F. R. and Moloney, W. C., Postmortem observations on
 

Fischer rats with leukemia and other disorders. Lab.
 

Investigations 23(3);327-334. 1970.
 

Finnegan, R. A. and Mueller, W. H., Chemical examination of a
 

toxic extract of Indigofera endecaphylla. The
 

endecaphyllins. J. Pharmaceut. Sci. 54:1136-1144, 1965.
 

39
 
Gart, J. J., The comparison of proportions: a review of
 

significance tests, confidence limits and adjustments for
 

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

Mutton, E. M., Windrum, G. M. and Kratzing, C. C., Studies on the
 

toxicity of Indigofera endecaphylla, I. Toxicity for
 

rabbits. J_._ Nutrition 64:321-333, 1958.
 

Hylin, J. W. and Matsumoto, H. , The biosynthesis of
 

3-nitropropionic acid by Penicilluim atrovenetum. Arch.
 

Biophys. 93:542-545. 1960.
 

International Agency for Research on Cancer, IARC Monographs on
 

the Evaluation of the Carcinogenic Risk of Chemicals to Man,
 

Vol. J^, World Health Organization, Geneva, 1972, pp.
 

145-156, 1972.
 

Iwasaki, T. and Kosikowski, F. V., Production of 3-nitropropionic
 

acid in foods. J. Food Science 38:1162-1165, 1973.
 

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

incomplete observations. J. Amer. Statist. Assoc.
 

53_: 457-481, 1958.
 

Kinosita, R., Ishiko, T. , Sugiyama, S. , Seto, T. , Igarasi, S.,
 

and Goetz, I. E., Mycotoxins in fermented food, Cancer Res.
 

.28:2296-2307, 1968.
 

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

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

Res. 7:230-248, 1974.
 

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

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

Morris, M. P., Pagan, C., and Warmke, H. E. , Hiptagenic acid, a
 

toxic component of Indigofera endecaphylla. Science
 

U9_: 322-323, 1954.
 

Raistrick, H. and Stossl, A., Studies in the biochemistry of
 

micro-organisms. Metabolites of Pencillium atrovenetum G.
 

Smith: 6-nitropropionic acid, a major metabolite. Biochem.
 

Journal 68:647-653. 1958.
 

40
 
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 ResL 32;1073-1081, 1972.
 

Sass, B., Rabstein, L. S., Madison, R., Nims, R. M., Peters, R.
 

L., and Kelloff, G. J., Incidence of spontaneous neoplasms
 

in F344 rats throughout the natural life-span. J. NCI
 

_54 (6): 1449-1453, 1975.
 

Squire, R. A. and Levitt, M. H., Report of a workshop on
 

classification of specific hepatocellular lesions in rats.
 

Cancer Res. 35:3214-3223, 1975.
 »
 

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

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

Wogan, G. N. and Newberne, P. M., Dose-response characteristics
 

of aflatoxin B^ carcinogenesis in the rat. Cancer Res.
 

27:2370-2376, 1967.
 

41
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS ADMINISTERED 3-NITROPROPIONIC ACID
 

BY GAVAGE
 

43
 

TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY 50 50 50
 

ANIHALS NECROPSIED 50 50 50
 

ANIMALS EXAMINED HISTOPATHOLOGIC ALLY 50 50 50
 

INTEGUMENTAR Y SYSTEM 

*SKIN 

SQUAMOUS CELL PAPILLOMA 

BASAL-CELL CAECINOMA 

TRICHOEPITHELIOMA 

(50) 

1

1

 (2%) 

(2%) 

(50) 

1 (2ft) 

2 (4X) 

(50) 

1 (2ft) 

1 (2%) 

*SOBCUT TISSU2 

SQUAMOUS CELL CARCINOMA 

BASAL-CELL CARCINOMA 

FIBROMA 

FIBROSARC0.1A 

LIPOMA 

MESENCHYBOHA, BENIGN 

(50) 

1

1

1

 (2%) 

(2%) 

12%) 

(50) 

1 (2ft) 

1 (2X) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

RESPIRATORY SYSTEM
 

tLUNG (50) (49) (48) 

SQUAMOUS CELL CARCINOMA, METASTA 1 (2ft) 

ALVEOLAR/BRONCHIOLAR ADENOMA 3 (6%) 1 (2ft) 

ALVEOLAR/B20NCHIOLAR CARCINOMA 1 (2X) 3 (6ft) 

CORTICAL CARCINOHA, METASTATIC 1 (2ft) 

C-CELL CARCINOMA, METASTATIC 1 (2%) 1 (2*) 

HEMATOPOIETIC SYSTEM 

*HULTIPLE ORGANS (50) (50) (50) 

MALIG.LYMPHOMA , ON FER-TYPE 1 (2%) 1 (2%) 7 (14%) 

MALIG.LYMPHOMA , LY OCYTIC TYPE 4 (8%) 4 (8ft) 4 (8X) 

UNDIFFEREN1IATED LEDKEHI EHIA A 2 (4X) 1 (2ft)

LYMPHOCYIIC LEUKEMI A 4 (8ft) 2 (4») 

GRANULOCYTIC LEUKEMIA 2 (4%) 1 (2%) 2 (4%) 

GRAfi.2kOCYTIC_SARCOMA_ 

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

45 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

• SPLEE N (49) (49) (49) 

MALIG.LYMPHOMA . UNDIFFER-TYP E 1 (2K) 1 (2%) 

HALIG.LYMPHOMA , HISTIOCYTIC TYPE 1 (2%) 

UNDIFFERENTIATE D LEUKEMI A 1 (2%) 

#HEDIASTINA L L.NOD E (41 ) (43 ) (43) 

ALVEOLAR/BRONCHIOLA 8 CA, METAST A 1 (2%) 

CIRCULATORY SYSTEM
 

tHEART (48) (49) (48) 

HEMANGIOM A 1 (2%) 

ANITSCHKOW-CEL L SARCOMA 1 (2X) 

DIGESTIVE SYSTEM
 

*PALATE (50) (50) (50)
 

SQUANOUS CELL CARCINOMA 1 (2%)
 

ILIVE H (49) (50) (49) 

NEOPLASTIC NODUL E 3 (6%) 5 (10%) 

HEPATOCELLULA R CARCINOM A 1 (2%) 

URINAR Y SYSTE M 

#URINAR Y BLADDE R (47) (42) (45) 

TRANSITIONAL-CELL PAPILLOMA 1 (2X) 

ENDOCRINE SYSTEM
 

•PITUITARY	 (46) (48) (49) 

CARCINOMA,NOS 1 (2%) 

CHROHOPHOBE ADENOMA 3 (7%) 5 (10X) 4 (8%) 

•ADRENAL	 (49) (50) (50) 

CORTICAL CARCINOMA 1 (2%) 1 (2%) 

PHEOCHROBOCYTOHA 4 (8X) 5 (10«) 5 (10%) 

PHEOCHROMOCYTOMA, MALIGNANT 1 (2%) 

•THYROID	 (46) (49) (47)
 

IOLLICULAR;CELL_CARCINOMA
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECHOPSIED
 

46
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

C-CELL ADENOM A 3 (7%) 8 (16%) 4 (9%) 

C-CELL CARCINOM A 1 (2%) 2 (4%) 

tPANCREATIC ISLETS (49) (50) (50) 

ISLET-CELL ADENOM A 4 (8%) 6 (12%) 11 (22%) 

REPRODUCTIVE SYSTEM 

*MAMMAR Y GLAN D (50) (50) (50)

 

FIBBOADENOM A 1 (2%)

 

*PREPUTIA L GLAN D (50) (50) (50) 

CARCINOMA , NOS 1 (2%) 

ADENOMA , NO S 2 (4%) 1 (2%) 4 (8%) 

HESTIS (50) (49) («9) 

INTERSTITIAL-CELL TUMO R 48 (96%) 44 (90%) 48 (98%) 

NERVOU S SYSTEM 

tBRAIN/IENINGE S (50) (50) (50) 

SQUAMOU S CELL CARCINOMA , METASTA 1 (2%) 

tCEREBRU M (50) (50) (50) 

ASTROCYTOM A 1 (2%) 

#BRAIN (50) (50) (50)

 

SQUAMOU S CELL CARCINOMA , METASTA 1 (2%) 

tMIDBRAI N (50) (50) (50) 

ASTROCYTOMA 1 (2%) 

tCEHEBELLU M (50) (50) (50) 

ASTROCYTOM A 1 (2%) 

SPECIAL SENSE ORGAN S 

*EAR CANA L (50) (50) (50)

 

SQUAMOU S CSLL CARCINOM A 1 (2%) 

MOSCOLOSKELETAL SYSTEM
 

*SKELETAL MUSCLE (50) (50) (50)
 

LIPOMA
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

47
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

BODY CAVITIES
 

*PEHITONEUM (50) (50) (50)
 

MESOTHELIOMA, NOS 1 (2X> 3 (6»)
 

ALL OTHER SYSTEMS 

*HULTIPLE ORGANS

FIBROUS HISTIOCITOMA, MALIGNANT

MESOTHELIOMA, HALIGNANT

 (50) 

1

 1

 (2%) 

 (2%) 

(50) (50) 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 NATURAL DEATHS 15 12
 

MORIBUND SACRIFICE 5 11
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 30 32 27
 

ANIMAL MISSING
 

d INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMAR Y
 

TOTAL ANIMALS WITH P8IMABY TUMORS* 49 49 49
 TOTAL PRIMARY TUMORS 99 99 112
 

TOTAL ANIMALS WITH BENIGN TUMORS 48 48 48
 TOTAL BENIGN TUMORS 72 76 78
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 23 14 24
 

TOTAL MALIGNANT TUMORS 26 17 29
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 1 3 1
 

TOTAL SECONDARY TUMORS 1 5 1
 

TOTAL ANIHALo WITH TUMORS UNCERTAINBENIG O N R MALIGNANT 1 6 5
 

TOTAL UNCERTAIN TUMORS 1 6 5
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR MSTASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

48
 
TABLE A2. 

* 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

ADMINISTERED 3-NITROPROPIOWC ACID BY GAVAGE 

CONTROL LOW DOSE HIGH DOSE 

iNIHALS INITIALLY IN STUDY 50 50 50
 

ANIHALS NECROP3IED 50 50 50
 

iNIHALS EXAMINED HISTOPATHOLOGIC ALLY 50 50 50
 

INTEGUMENTAR Y SYSTEM 

*SKIN (50) (50) (50) 

PAPILLOMA , NOS 1 (2%) 

SEBACEOUS ADENOM A 1 (2%) 

*SUBCUT TISSUS (50) (50) (50) 

SQUAMOI1S CSLL CARCINOM A 1 (2X) 

RESPIHATOR Y SYSTEM 

tLUNG (50) (49) (1*9) 

SQUAMOUS CSLL CARCINOMA , METAST A 1 (2*) 

C-CELL CARCINOHA, METASTATIC 1 (2%) 

LIPOSARCOMA, HETASTATIC 1 (2%) 

HEMATOPOI5TIC SYSTEM
 

*HULTIPLE ORGANS (50) (50) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

MALIG. LYMPHOMA, UN DIFPES-TYP E 4 (8%) 1 (2S) 4 (3%)
 

MALIG. LYMPHOSA, LYMPHOCYTIC TYPE 1 (2%) 2 (U%) 2 (4%)
 LEUKEMIA, NOS 1 (2%)
 

LYMPHOCYTIC LEUKEMIA 1 (2X)
 

GRANULOCYTIC LEUKEMIA 2 (HI) 1 (2%)
 

MONOCYTIC LEUKEMIA 1 (2%)
 

*SPLEEN (50) (50) (49)
 

MALIG. LYMPHOMA, UNDIFFER-TY PE 1 (2%) 1 (2%)
 

GRANULOCYTIC LEUKEMIA 1 (2%)
 

#LYMPH NODE (44) (41) (45)
 

* NUMBER OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

49
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

#MANDIBULAR L. NODE

SQUANOUS C3LL CARCINOMA, METASTA

 (44) (41) (45) 

1 (2%) 

CIRCULATORY SYSTEM 

HONE 

DIGESTIVE SYSTEM 

*TONGUE

SQUAMOUS CBLL CARCINOMA, METASTA

 (50) (50)

 1 (2*) 

 (50) 

•LIVER

NEOPLASTIC NODULE

 (49) (50)

 1 (2%)

 (50) 

1 (2%) 

URINARY SYSTEM 

IKIDNEY

MIXED TUNOH, MALIGNANT

 (49) (50)

 1 (2%) 

 (50) 

ENDOCRINE SYSTEM 

#PITUITARY 

CHROMOPHOBE ADENOMA 

(45) 

19 (4295) 

(46) 

15 (33%) 

(47) 

20 (43%) 

•ADRENAL 

PHEOCHROMOCYTOMA 

(49) 

3 (6%) 

(50) 

1 (2%) 

(49) 

1 (2%) 

tTHYROID 

FOLLICULAR-CELL ADENOMA 

FOLLICULAR-CELL CARCINOMA 

C-CELL ADENOMA 

C-CELL CARCINOMA 

(50) 

1

3

2

 (2%) 

 (6X) 

 (4%) 

(44) 

3 (7*) 

1 (2%) 

(44) 

1 (2%) 

3 (7%) 

2 (5%) 

#PANCREATIC ISLETS 

ISLET-CELL ADENOMA 

(49) 

1 (2X) 

(49) (47) 

1 (2%) 

EEPHODUCTIVE SYSTEM 

*MAMMARY GLAND 

ADE NOM A ^NOS 

(50) (50) (50) 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

50
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ADENOCARCINOMA, NOS 1 (2%) 3 (6X) 

FIBROADENOHA 12 (2<*X) 14 (2d%) 13 (26%) 

*PREPUTIAL GLAND (50) (50) (50)

 

CARCINOMA,NOS 1 (2%)

 

ADENOMA , NO S 2 2 (>4%]

 

#U-TEBUS (50) (48) (49) 

LEIOBYOMA 1 (2%) 

ENDOMETRIA L STROMAL POLYP 2 (14%) 4 (8X) 5 (10%) 

ICERVIX UTEBI (50) (48) (49) 

FIBROSARCOMA 1 (2%) 

tOVAR Y (50) (47) (48) 

SERTOLI-CELL TUHOR 1 (2X) 

HERVOUS SYSTEM 

tCEREBRUM (49) (50) (50) 

OLIGODENDROGLIOMA 1 (2%) 

SPECIAL SENSE ORGANS 

*ZYMBAL' S GLAN D (50) (50) (50) 

SQUAMOO S CELL CARCINOM A 1 (2%} 

BUSCULOSKELETAL SYSTEM 

*HftNDIBLE (50) (50) (50) 

SQUAHOUS CELL CARCINOMA 1 (2%) 

BODY CAVITIES
 

*BESENTERY (50) (50) (50)
 

FIBROSARCOMA 1 (2%)
 

fcLL OTHER SYSTEMS 

LOBBOSAC6AL REGION 

L£POSARCOMA 1 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* HUHBER OF ANIMALS NECROPSIED 

51 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL HISSING
 

INCLUDES AOTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 

TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMT3RS
 

TOTAL MALIGNANT TUMOES
 

TOTAL ANIMALS WITH SECONDARY TUMORSt


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMOBS UNCERTAIN­ BENIGN OP. MALIGNANT


TOTAL UNCERTAIN TUMORS


TOTAL ANIHALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

50
 

4
 

13
 

33
 

40
 

59
 

35
 

45
 

11
 

14
 

3


 3


50 50 

15 9 

10 11 

25 30 

30 39 

50 66 

27 29 

39 46 

10 19 

10 19 

1 2 

1 3 

1 1 

1 1 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 • SECONDARY TUTORS: METASTATIC TUMORS CR TUMORS INVASIVE INTO AN ADJACENT OEGAN
 

52
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE ADMINISTERED 3-NITROPROPIONIC ACID
 

BY GAVAGE
 

53
 

TABLE B1. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE 

CONTROL LOW DOSE 

ANIHALS INITIALLY IN STUDY 50 50
 

ANIHALS NECROPSIED 49 50
 ANIMALS EXAMINED HI STOPATH OLOGIC ALLY 49 50
 

INTEGUMENTAR Y SYSIEM 

*SUBCU T TISSU E (49) (50) 

FIBROMA 1 (2*) 

FIBROSARC05U 2 (4%) 2 ((*») 

HEMANGIOM A 1 (2*) 

HEMANGIOSARCOM A 

RESPIRATORY SYSTEM 

#LUNG (49) (43) 

HEPATOCELLULAR CARCINOMA. METAST 

ALVEOLAR/BRONCHIOLAR ADENOMA 

3 

10 

(6S) 

(20%) 

3 

5 

(6%) 

(10%) 

ALVEOLAR/BRONCHIOLAR CARCINOMA 4 (8%) 3 (6*) 

CORTICAL CARCINOMA, BETASTATIC 

FIBROSARC051A, METASTATIC 

1 (2%) 

1 .12*) 

HEMATOP01ETIC SYSTE M 

*MULTIPLE ORGAN S (49) (50) 

MALIGNAN T LYMPHOMA , NO S 1 (2%) 

MAL1G.LYMPHOMA, LYMPHOCYTIC TYPE 4 (8%) 5 (10%) 

MALIG.LYMPHOMA , HISTI3CYTIC TYPE 2 (4%) 

LYMPHOCYTIC LEUKEMI A 1 (2«) 

GRANOLOCYTIC LEUKEMI A 2 (4%) 

MONOCYTIC LEUKEMI A 1 (2X) 

GEANULOCYTI C SARCOM A 1 (2%) 

*SUBCUT TISSU E (49) (50) 

MAST-CELL TUMOR 

#SPLEEN (46) (50)
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

55 

HIGH DOSE 

50
 

50
 

50
 

(50) 

1 (2%) 

(50)
 

8 (16X)
 

3 (6%)
 

(50) 

4 (8%) 

1 (2S) 

1 (2%) 

(50)
 

1 (2%)
 

(46)
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE
 HIGHD >OSE
 

HEMANGIOSAHCOMA 4 (9X)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2%)
 

#MESENTERIC L. NODE (40) (31)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (3S)
 

tLIVER (49) (50)
 

GRANOLOCYTIC LEUKEMIA 1 (2%)
 

tPEYERS PATCH (47) (49)
 

MALIGNANT LYMPHOMA, NOS
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (2X)
 

#THYMUS (35) (38)
 

MALIGNANT LYMPHOMA, NOS 1 (3X)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYST2M
 

JLIVER (49) (50)
 

HEPATOCELLULAR ADENOMA 4 (8%) 2 (4£)
 

HEPATOCELLULAR CARCINOMA 16 (33X) 8 (16%)
 

CORTICAL CARCINOMA, METASTATIC 1(2%)
 

HEMANGIOMA 1(2%)
 

HEMANGIOSARCOMA 2 (H%) 1 (2*)
 

ANGIOSARCOMA
 

tSTOMACH (48) (50)
 

ADENOMATOUS POLYP, NOS
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

fADRENAL (46) (49)
 

CORTICAL_CARCINOMA_
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

2
 

1
 

(30)
 

1
 

(49)
 

(49)
 

1
 

3
 

(41)
 

(49)
 

4
 

12
 

4
 

1
 

(46)
 

1
 

(50)
 

(«*)
 

(2%)
 

(3X)
 

(2%)
 

(6*)
 

(8%;
 

(24*)
 

(8?)
 

(2%)
 

(2%)
 

56
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ttHYBOID

FOLLICULAR-CELL ADENOMA

 (US) (44) (<*8) 

1 (2%) 

REPRODUCTIVE SYS1EM 

#TESTIS

INTEBSTITIAL-CELL TDBOR

 (<»7)

 1 (2*) 

 (49) (50) 

NERVOUS SYSTEM 

HONE 

SPECIAL SENSE ORGANS 

*HABDERIAN GLAND

PAPILLARY ADENOMA

PAPILLARY CYSTADENOMA, NOS

 (49)

 1 (2*) 

 (50)

 1 (2S)

 (50) 

2 («*) 

HUSCULOSKELETAL SYSTEM 

*SKULL

OSTEOMA

 (U9) (50) (50) 

1 (2*) 

BODY CAVITIES 

*ABDOHINAL CAVITY

CORTICAL CARCINOMA, METASTATIC

 (49)

 1 (2*) 

 (50) (50) 

ALL OTHER SYSTEMS 

THORAX 

FIBROSARCOMA, HETASTATIC

DIAPHRAG M 

IIBRQSAJBCQMAJ._METASTAIIC

t NUMBER OF ANIBALS WITH TISSUE * NUMBER Of ANIMALS NECBOPSIED  EXAMINED MICROSCOPICALLY 

1 

1 

57 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

ANIMAL DISPOSITION SUflMABY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

NATURAL DEATHS 10 11 12
 

MORIBUND SACRIFICE 2 3
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 38 36 38
 

ANIMAL MISSING
 

» INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 39 28 38
 

TOTAL PRIMARY TUMORS 54 39 55
 

TOTAL ANIMALS WITH BENIGN TUMORS 15 9 19
 

TOTAL BENIGN TUMORS 17 11 19
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 31 24 27
 

TOTAL MALIGNANT TUMORS 37 28 35
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 4 4
 

TOTAL SECONDARY TUMORS 6 6
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN 08 MALIGNANT 1
 

TOTAL UNCERTAIN TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

* SECONDARY TUMORS: METASTA'IIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

58
 
TABLE B2. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE 

CONTROL LOW DOSE HIGH DOSE 

ANIMAL S INITIALL Y IN STUD Y 5 0 50 50 

ANIMAL S NECROPSIE D 50 50 50 

ANIMAL S EXAMIN2 D HISTOPATHOLOGICALLY 50 50 50 

INTEGUMENTARY SYSTEM
 

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

FIBROSARCOMA 1 (21) 2 (US) 

8ESPIRATORY SYSTE M 

tLUNG (47) l'»3) (49)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 2 (4%) 4 (ax/ 1 (2X)
 

ALVEOLAR/BHONCHIOLAR CARCINOMA 2 (4%) 2 (4X)
 

FIBROSARCOMA, METASTATIC 1 (2%)
 

OSTEOSARCOMA 1 (2S)
 

HBHATOPOIETIC SYSTEM 

*MOLTIPLE ORGAN S (50) (50) (50) 

8ALIG.LYMPHOMA , LYMPHOCYTIC TYP E 11 (22%) 10 (20%) 7 (14%) 

MALIG.LYMPHOMA , HISTIOCYTIC TYP E 6 (12% ) 2 (4X) 

UNDIFFEBENTIATED LEDKEMI A 

LYHPHOCYTIC LEUKEMI A 2 (4%) 

GBANULOCYTI C LEUKEMI A 1 (2%) 2 (4*) 

GRANULOCYTIC SARCOHA 1 (2«) 

tBONE MARRO W (46) (4b ) (50) 

GRANULOCYTIC SARCOM A 1 (2%) 

•SPLEEN (47) (50) (50) 

HEMANGIOSAHCOM A 1 (2%) 3 (6%) 

MALIG.LYMPHOMA , LYMPHOCYTIC TYPE 1 (2%) 

HALIG. LYMPHOHA, HISTIOCYTIC TYPH 

•LYMP H NOD E (38) (36) (33)

 

GRANULOCYTIC_SARCOMA 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

59
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

tMESENTER1C L. NODE (38) (36) (33) 

MALIG.LYMPHOMA , HISTIOCYTIC TYPE 1 (3%) 1 (3%) 

*PEYERS PATC H (48) (48) (49) 

MALIG.LYMPHOHA , LYMPHOCYTIC TYPE 3 (6*) 1 (2%) 

CIRCULATORY SYSIEfl 

NON E 

DIGESTIVE SYSTE M 

tLIVER (49) (50) (50)
 

HEPATOCELLULAR ADENOMA 1 (2%) 2 (4%)
 

HEPATOCELLULAR CARCINOMA 1 (2%) \ (2%) 2 (4%)
 

HEMANGIOSARCOMA 1 (2*)
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

tPITOITARY (43) (48) (42)
 

CH80MOPHOBE ADENOMA 2 (5X) 4 (8%) 1 (2%)
 

#ADRENAL (48) (50) (49)
 

CORTICAL CARCINOMA 1 (2«)
 

PHEOCHROMOCYTOMA 1 (2%)
 

tTHYfiOID (40) (47) (45) 

FOLLICULAR-CELL ADENOM A 1 (2%) 

REPRODUCTIV E SYSTEM 

*MAMBAR Y GLAN D (50) (50) (50) 

ADENOCARCINOMA , NOS 1 (2)5) 1 (2%} 

FIB80ADENOMA 1 (2X) 

*VAGIN A (50) (50) (50) 

SOJ&MOUS_CfiLL_CARCINOflA 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

60
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

•UTERUS

LEIOMYOSARCOB A

 (47)

 2 (4)5)

 (49)

 1 (2%) 

 (48) 

IOVARY

PAPILLARY CYSTADENOMA, NOS

GRANULOSA-CELL TUMOR

 (39)

 1 (3X)

 (47) (47) 

1 (2%) 

1 (2%) 

NERVOUS SYSTEM 

tBRAIN/HENINGSS

OSTEOSARCOHA, METASTATIC

 (47) (50) (50) 

1 (2%) 

SPECIAL SENSE ORGANS 

*HARDERIAN GLAND

PAPILLARY CYSTADENOMA, NOS

 (50) (50) (50) 

1 (28) 

HUSCULOSKELETAL SYSTEM 

*SKELETAL MUSCLE

FIBROSARCOBA

 (50) (50)

 1 (2%) 

 (50) 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

DIAPHRAGM 

OSTEOSARCOMA, METASTATIC 1 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY

NATURAL DEATH2

MORIBUND SACRIFICE

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE

ANIMAL MISSING 

 50

 14

 1 

 35

 50

 7

 43

 50 

 11 

 39 

d_IlCLUDES_AUTj}LYZEp_ANIMAI.S 

* NUMBER,OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

61
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMOBS* 26 32 26
 

TOTAL PRIMARY TUMORS 31 39 33
 

TOTAL ANIMALS WITH BENIGN TUMORS 3 9 8
 

TOTAL BENIGN TUMORS 5 9 8
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 23 25 19
 

TOTAL MALIGNANT TUMORS 25 30 24
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2
 

TOTAL SECONDARY TUMORS 3
 

TOTAL ANIHALS WITH TUMORS UNCEKTAINBENIGN OR MALIGNANT 1 1
 

TOTAL UNCERTAIN TUMORS 1 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR MSTASTATIC
 

TOTAL UNCERTAIN TUMORS
 

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

62
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS ADMINISTERED 3-NITROPROPIONIC ACID
 

BY GAVAGE
 

63
 

TABLE C1. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

ADMINISTERED 3 NITROPROPIONIC ACID BY GAVAGE 

CONTROL LOW DOSE
 

ANIMALS INITIALLY IN STUDY 50 50
 

ANIMALS NECROPSIED 50 50
 

AHIHALS EXAMINED HISTOPATHOLOGICALLY 50 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (50) (50)
 

CYST, NOS 1 (2%)
 

EPIDERMAL INCLUSION CYST
 

HYPERKERATOSIS 2 (4%)
 

ACANIHOSIS
 

*SUBCUT TISSUE (50) (50)
 EPIDERMAL INCLUSION CYST
 

RESPIRATORY SYSTEM
 

*NASAL CAVITY (50) (50)

 

INFLAMMATION, CHRONIC 1 (2%) 

#TRACHEA (1*9) (50) 

INFLAMMATION, NOS 17 (35%) 22 (44%) 

INFLAMMATION, CHRONIC 1 (2%) 3 (6%) 

HYPERPLASIA, LYMPHOID 3 (6%) 2 (4% ) 

tLUNG/BRONCHUS (50) (49 ) 

BRONCHIECTASIS 4 (8%) 2 

INFLAMMATION, SUPPURATIVE 

HYPERPLASIA, FOCAL 2 (4«) 

HYPERPLASIA, LYMPHOID 6 (16%) 19 (39%) 

tLUNG (50) (49) 

ATELECTASIS 1 (2%) 

CONGESTION , NOS 2 (4%) 1 (2%) 

INFLAMMATION , SUPPURATIV E 1 (2*) 

INFLAMMATION , ACUT E SUPPURATIV E 

BRONCHOPNEUHONIA ACUTE SUPPURATI 1 (2%) 2 (4%) 

PNEUMONIA t _CHRgNIC_MURIN E 

* NUMBEfi OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIED 

65
 

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 

(47)
 

14	 (30%)
 

2 (4%)
 

1 (2%)
 

(48)
 

2 (4%)
 

1 (2%)
 

24 (507.)
 

(48)
 

1 (2%)
 

1 (2%)
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

INFLAMMATION. GRANULOMATOUS 1 (2%)
 

GRANULOHA, NOS 1 (2%)
 

INFLAMMATION, FOCAL GRAN0LOMATOU 1 (2%)
 

FIBROSIS 1 (2*)
 

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

PIGMENTATION, NOS 1 (2%)
 

HEMOSIDEROSIS 1 (2»)
 

ALVEOLAR MACROPHAGES 5 (10%) 2 (4%)
 

tLUNG/ALVEOLI (50) (49) (48)
 

CONGESTION, NOS 1 (2%) 1 (2%)
 

EDEMA, NOS 1 (2%) 1 (2%)
 

HEMORRHAGE 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

#BONE MARROW (49) (48) (50)
 

HYPERPLASIA, NOS 4 (8%) 1 (2%)
 

HYPERPLASIA, HEMATOPOIETIC 4 (8%) 6 (17%) 16 (32%)
 

HYPERPLASIA, ERYTHROID 1 (2%) 1 (2%) 1 (2%)
 

HYPERPLASIA, GRANULOCYTIC 1 (2%)
 

HYPOPLASIA, ERYTHHOID 1 (2%)
 

tSPLEEN (49) (49) (49)
 

CONGESTION, NOS 1 (2%) 2 (4%)
 

FIBROSIS 1 (2%)
 

HEMOSIDEROSIS 23 (47*) 36 (733S) 31 (63%)
 

ATROPHY, NOS 1 (2%)
 

LEUKEMOID REACTION 1 (2X) 1 (2%)
 

HYPERPLASIA, RETICDLUM CELL 1 (2%)
 

HEMATOPOIESIS 25 (51%) 39 (80%) 34 (69%)
 

ERYTHBOPOIESIS 1 (2%) 1 (2%)
 

GRANULOPOIESIS 1 (2%)
 

#LYMPH NODE (41) (43) (43)
 

HEMOSIDEROSIS 1 (2%)
 

#SUBMANDIBOLA R L.NODE (41) (43) (43) 

LYMPHANGIECTASI S 1 (2%) 

IMANDIBULAR L. NODE (41) (43) (43)
 

LYMPHANGIECTASIS 2 (5%) 1 (2%)
 

tBHONCHIAL LYMPH NODE (41) (43) (43)
 

LIMPHANGIECTASIS
 

* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

66
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

(43)
 

(29)
 

1 (3«)
 

1 (3«)
 

(48)
 

1 (2»)
 

(48)
 

5 (10%)
 

24 (50%)
 

1 (2%)
 

1 (2%)
 

(48)
 

(50)
 

2 (4«)
 

(49)
 

4 (8%)
 

(50)
 

1 (2«)
 

(46)
 

1 (2%)
 

(49)
 

•MESENTERIC L. NODE
 

HEMOSIDEHOSIS
 

tTHYMUS
 

LYMPH4NGIECTASIS
 

HEHOSIDEBOSIS
 

CIBCOLATORY SYSTEM
 

•HEART
 

FIBBOSIS, DIFFUSE
 

•MYOCARDIUM
 

INFLAMMATION, FOCAL
 

INFLAMMATION, INTEBSTITIAL
 

ABSCESS, NOS
 

FIB80SIS
 

FIBBOSIS, FOCAL
 

SCAR
 

DEGENERATION, NOS
 

CALCIFICATION, DYSTBOPHIC
 

•BNDOCABDIUM
 

INFLAMMATION, FOCAL
 

*POLMONARY ARTEHY
 

MEDIAL CALCIFICATION
 

CALCIFICATION, FOCAL
 

•HEPATIC SINUSOID
 

CONGESTION, NOS
 

DIGESTIVE SYSTEM
 

*HOUTH
 

ABSCESS, NOS
 

•SALIVARY	 GLAND
 

EDEMA, NOS
 

•LIVER
 

CONGESTION, NOS
 

CONTROL
 

(<t1)
 

(37)
 

(48)
 

(48)
 

2 (4*)
 

1 (2X)
 

4 (8%)
 

1 (2X)
 

6	 (13»)
 

(48)
 

2 (U%)
 

(50)
 

(49)
 

(50)
 

(U9)
 

(49)
 

1 ii»l _
 

LOW DOSE
 

(43)
 

1 (2X)
 

(28)
 

(49)
 

(49)
 

6 (12%)
 

17	 (35«)
 

1 (2*)
 

(49)
 

(50)
 

1 (2%)
 

1 (2X)
 

(50)
 

1 (2«)
 

(50)
 

(49)
 

(50)
 

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

67
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

HEMORHHAGE 1 (2%)
 

NECROSIS, NOS 1 (2%)
 

NECEOSIS, COAGULATIVE 1 (2%)
 

METAMORPHOSIS FATTY 1 12%} 2 (4«) 5 (10X)
 

FOCAL CELLULAR CHANGE 1 (2%) 3 (6*)
 

PHAGOCYTIC CELL 1 (2%)
 

ANGIECTASIS 1 (2%)
 

HYPEHPLASIA, RETICULUM CELL 3 (6S) 4 (8%)
 

HEMATOPOIE5I3 4 (8»)
 

*LIVER/CENTHILOBULAR (49) (50) (49)
 

NECROSIS, NOS 1 (2X)
 

NECROSIS, FOCAL 1 (2S)
 

NECROSIS, COAGULATIVE 1 (2%)
 

METAMORPHOSIS FATTY 2 (4*) 3 (6%) 1 (2%)
 

PIGMENTATION. NOS 1 (2*)
 

*BILE DUCT (50) (50) (50)


INFLAMMATION, FOCAL 1 (2%)
 

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

HYPERPLASIA, FOCAL 18 (36%) 30 (60%) 3U (68%)
 

HYPERPLASIA, DIFFUSE 2 (4«)
 

#PANCREAS (49) (50) (50)
 EDEMA, NOS 1 (2%} 1 (2X)
 

PE3IAHTERITIS 1 (2%) 1 (2%)
 

HEMOSIDEHOSIS 1 (2%)
 

IPANCREATIC DUCT (49) (50) (50)
 

HYPERPLASIA, NOS 1 (2%)
 

HYPERPLASIA, EPITHELIAL 1 (2%)
 

HYPERPLASIA, FOCAL 2 («) 8 (16%) 6 (12%)
 

#ESOPHAGUS (46) (44) (45)
 

PERFORATION, INFLAMMATORY 1 (2%)
 

ISTOHACH (49) (50) (49)
 

ULCER, NOS 1 (2%)
 

ULCER, FOCAL 1 (2S)
 

EROSION 1 (2%)
 

•PEYERS PATCH (49) (50) (45)
 

HYPEHPLASIft, LYMPHOID E> (10») 3 (6X) 6 (13%)
 

SILEUH (49) (50) (45)
 

MUCOCELE 1
 .I2*L_ _.
 

f NUHBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUHBER OF ANIMALS NECROPSIED
 

68
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

tCOLON (32) (30) (39)
 

NEMATODIASIS 3 (9X) 2 (7 it; 6 (15%)
 

ORINARY SYSTEM
 

•KIDNEY	 (50) (49) (49)
 

CAST, NOS 1 (2X) 1 (2S)
 

CONGESTION, NOS 1 (2X) 1 (2X)
 

INFLAMMATION,. INTERSTITIAL 1 (2X) 1(2%)
 

ABSCESS, NOS 1 (2X)
 

INFLAMMATION, CHRONIC 8 (16%) 9 (18X) 10 (20%)
 

INFLAMMATION, CHRONIC FOCAL 26 (52X) 29 (59Z) 25 (51X)
 

INFLAMMATION, CHRONIC DIFFUSE 1 (2X) 1 (2X)
 

SCLEROSIS 1 (2%)
 

NEPHRO5IS, NOS 1 (2%)
 

GLOMERULOSCLEROSIS, NOS 1 (2X)
 

CALCIFICATION, NOS 1 (2X)
 

•KIDNEY/CAPSULE	 (50) (49) (49)
 

CYST, NOS 1 (2%)
 

#KIDNEY/CORTEX (50) (49) (49)
 

CAST, NOS 1 (2X)
 

CYST, NOS 1 (2%)
 

PIGMENTATION, NOS 6 (12%)
 

•KIDNEY/TUBULE	 (50) (49) (49)
 

CAST, NOS 1 (2X) 1 (2%)
 

PIGMENTATION, NOS 3 (6X)
 

fCONVOLUTED TUBULES (50) (49) (49)
 

DEGENERATION, HYALINE 1 (2%)
 

PIGMENTATION, NOS 3 (6X) 1 (2X)
 

IKIDNEY/PELVIS (50) (49) (U9)
 

INFLAHHATION, SOPPORATIVE 1 (2X)
 

•URINARY BLADDER (47) (42) (45)
 

HEMORRHAGE 1 (2X)
 

INFLAMMATION, HEHORRHAGIC 1 (2X)
 

tO. BLADDER/SUBMOCOSA (47) (42) (45)
 

HEMORRHAGE . 1 (2X) J_i2£l_ _
 

* NUMBER OF ANIMALS WITH TISSUE EXAHIHED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

69
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

ENDOCRIN E SYSTE M 

tPITUITASY (46) (48) (49) 

CYST, NOS 1 (2%) 

HEMORRHAG E 1 (2») 1 (2%) 

HEHOSIDEROSIS 1 (2%) 

HYPERPLASIA, NOS 1 (2%) 

HYPERPLASIA, FOCAL 1 (2%) 

ANGIECTASIS 2 (4S) 4 (8%) 3 (6%) 

tADRENA L (49) (50) (50) 

ANGIECTASIS 1 (2%) 1 (2«) 4 (8«) 

tADHENA L CORTEX (49) (50) (50)

 

HYPERPLASIA, NODULAR 1 (2*) 1 (2*) 

tADRENA L MEDULLA (49) (50) (50) 

HYPERPLASIA, NODULA R 2 (4%) 

HYPERPLASIA, NOS 1 (2«) 

HYPERPLASIA, FOCAL 1 (2*) 5 (10*) 9 (18S) 

ITHYROID (46) (49) (47) 

CYSTIC FOLLICLES u (8«) 2 (4S) 

PIGMENTATION, NOS 1 (2«) 

HYPEHPLASIA, C-CELL 23 (50%) 29 (59X) 26 (55%) 

HYPERPLASIA, FOLLICULAR-CELL 1 (2%) 2 (4%) 

tTHYROID FOLLICLE (46) (49) (47) 

PIGMENTATION , NOS 3 (6*) 2 (4K) 

REPRODUCTIVE SYSTEM 

*MAMMAR Y GLAN D (50) (50) (50) 

GALACTOCELE 1 (2%) 

*PREPDTIAL GLAN D (50) (50) (50) 

INFLA8HATION, SUPPURATIVE 2 (4») 1 (2%) 5 (10%) 

INFLAMMATION, CHHOHIC 2 (4%) 

NECROSIS, NOS 1 (2*) 

tPROSTATE (44) (44) (U7) 

INFLAMMATION, FOCAL 4 (9%) 

INFLAMMATION^ SOPPUBATIVE 2_^5%1 ,4_19%) i_12*J 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

70
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

NECROSIS, SOS 1 (2X) 

tTESTIS 

CALCIFICATION, FOCAL 

ATROPHY, NOS 

ATROPHY, FOCAL 

ASPERMATOGENESIS 

HYPERTROPHY, NOS 

HYPEHPLASIA, INTERSIITIAL CELL 

(50) 

32 (6UX) 

7 (1l*X) 

4 (8X) 

1 (2X) 

(1*9) 

1 (2X) 

39 (80X) 

1 (2X) 

1 (2X) 

1 (2X) 

(1*9) 

38 (78X) 

2 (4X) 

3 (6X) 

1 (2X) 

6 (12X) 

tTESTIS/TOBULE 

CALCIFICATION, NOS 

CALCIFICATION, FOCAL 

(50) 

1 (2X) 

(i»9) 

1 (2X) 

C*9) 

NERVOUS SYSTEM 

*BRAIN/HENISGES 

THROMBOSIS, NOS 

(50) 

1 (2X) 

(50) (50) 

IBRAIN STEM 

HEMORRHAGE 

NECROSIS, NOS 

MALACIA 

(50) 

1 (2X) 

(50) (50) 

1

1

 (2X) 

 (2») 

IHIDBRAIN 

NECROSIS, NOS 

HALACIA 

(50) 

1

1

 (2X) 

 (2X) 

(50) (50) 

*SPINAL CORD 

HEMORRHAGE 

DEGENERATION, NOS 

BALACIA 

(50) (50) 

1 (2%) 

(50) 

1 (2X) 

1 (2X) 

1 (2X) 

SPECIAL SENSE ORGANS 

*EYE 

HEMORRHAGE 

INFLAMBATION, NOS 

PERIVASCULITIS 

DEGENERATION. NOS 

CATARACT 

HEHOSIDEROSIS _ ANEMIA^ NOS 

(50) 

1

13

 (2X) 

 (26X) 

(50) 

1 (2X) 

1 (2X) 

1 (2X) 

16 (32X) 

_

(50) 

12 (24X) 

1 (2X) 

1 I2X) 

f NO«BER

* NOHBER

 OF

 OF

 AHIBALS

 ANIMALS

 WITH TISSUE

 NECHOPSIED 

 EXAMINED MICROSCOPICALLY 

71 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

*EYE/CORNEA (50) (50) (50)
 

OLCER, NOS 1 (2%)
 

INFLAMMATION, INTERSTITIAL 1 (2%) 2 (4%)
 

*LENS CAPSULE (50) (50) (50)
 

CALCIFICATION, SOS 1 (2»)
 

*BIDDLE EAR (50) (50) (50)
 

INFLAHMATIQN, NOS 1 (2*)
 

HOSCULOSKELETAL SYSTEM
 

*SKELETAL MUSCLE (50) (50) (50)
 

ATROPHY, NOS 1 (2«)
 

*HUSCLE HIP/THIGH (50) (50) (50)
 

ATROPHY, NOS 1 (2%)
 

BODY CAVITIES
 

*MEDIASTINUM (50) (50) (50)
 

THROMBOSIS, NOS 1 (2%)
 

*ABDOMINAL CAVITY (50) (50) (50)
 

NECEOSIS, FAT 1 (2X)
 

*PLEURA (50) (50) (50)
 

HYDROTHORAX 1 (2%)
 

*HESENTERY (50) (50) (50)
 

INFLAMMATION, NOS 1 (2%)
 

FIBROSIS 1 (2%) 1 (2%)
 

PERIABTERITIS 1 (2%)
 

NECROSIS, FAT 2 (4«)
 

ALL OTHER SYSTEMS
 

*MULTIPLE ORGANS (50) (50) (50)
 

CONGESTION, NOS 2 (US)
 

JAUNDICE, NOS 1 (2%)
 

DIAPHRAGM
 

HERNIA*. NOS 1
 

t KUMBEfi OF ANIMAL S HITH TISSUE EXAMINE D MICROSCOPICALLY * NUMBE R OF ANIMAL S NECROPSIED 

72 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

ADIPOSE TISSUE
 

INFLAHH&TION, NOS
 

INFLAMMATION, FOCAL
 

SPECIAL MORPHOLOGY SUMMARY 

NONE
 

t NUMBER OF ANIMALS SITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECfiOPSIED
 

73
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE 

CONTROL LOW DOSE 

ANIMALS INITIALLY IN STUDY 50 50
 

ANIMALS NECHOPSIED 50 50
 

ANIMALS EXANIN3D HISTOPATHOLOGIC ALLY 50 50
 

INTEGUMENTAR Y SYSTEM 

*SKIN (50) (50) 

LYMPHOCYTIC INFLABMATOR Y INFILTR 1 (2%) 

NECROSIS, FOCAL 1 (2%) 

*SUBCUT TISSUS (50) (50) 

ABSCESS, NOS 1 (2%) 

RESPIRATORY. SYSTEM 

#TRACHEA 

INFLAMMATION, NOS 

INFLAMMATION, SUPPURATIVE 

INFLAMMATION, CHRONIC 

HYPERPLASIA, LYMPHOID 

(49) 

17

1

 (35S) 

 (2%) 

(48) 

10

1

 (2U) 

 (2%) 

*LUNG/BRONCHU3 

BRONCHIECTASIS 

INFLAMMATION, NOS 

INFLAMMATION, SUPPURATIVE 

HYPERPLASIA, NOS 

HYPERPLASIA, FOCAL 

HYPERPLASIA, LYBPHOID 

(50) 

2 (4X) 

1 (2%) 

27 (5U») 

(49) 

2

1

27

 (4%) 

 (2%) 

 (55X) 

tLUNG 

CONGESTION, NOS 

EDEMA, NOS 

BRONCHOPNEUHONIA, NOS 

INFLAMMATION, FOCAL 

INFLAMMATION, INTERSTITIAL 

PNEUMONIA, ASPIRATION 

BRONCHOPNEUBONIA SUPPURATIVE 

BRONCHOPNEOMONIAj. ACUTE 

(50) 

1 <2X) 

(49) 

1

1

1

1

1

1

 (2«) 

 (2X) 

 (2%) 

 {2*> 

 (2*) 

 J2«l 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

74
 

HIGH DOSE 

50
 

50
 

50
 

(50)
 

(50)
 

(49)
 

11	 (22%}
 

1 (2%)
 

3 (6«)
 

2 (4%)
 

(49)
 

1 (2%)
 

1 (2%)
 

31 (63%)
 

(49)
 

1 (2%)
 

1 (2%)
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE
 

INFLAMMATION, ACUTE SUPPURATIVE 1 (2X)
 

BRONCHOPNEUMOHIA ACUTE SUPPURATI 1 (2%)
 

PNEUMONIA, CHRONIC SORINE 5 (10%) 4 (8%)
 

INFLAMMATION, FOCAL GRANULOMATOU 1 (2%)
 

PEBIVASCULAR CUFFING 2 (451)
 

HEHOSIDEROSIS 1 (2%)
 

ALVEOLAR MACROPHAGES 2 (4%J 5 (10*)
 

HYPERPLASIA, LYMPHOID 1 (2%)
 

tLUNG/ALVEOLI (50) (49)
 

CONGESTION, NOS 1 (2%) 3 (6%)
 

EDEMA, NOS 3 (6%)
 

HEMATOPOIETIC SYSTEM
 

IBONE MABROW	 (50) (47)
 

HEHOSIDEROSIS
 

HYPOPLASIA, NOS	 1 (2%)
 

HYPEHPLASIA, NOS	 1 (2%) 1 (2%)
 

HYPERPLASIA, HEMATOPOIETIC 3 (6%) 2 (4*)
 

HYPERPLASIA, GRANULOCYTIC 2 (4%)
 

ERYTHROPOIESIS 2 (4%)
 

GRANULOPOISSIS 1 (2%)
 

•SPLEEN	 (50) (50)
 

ECTOPIA 1 (2X)
 

CONGESTION, NOS 1 (2%) 1 (2%)
 

HEMOSIDEHOSIS 34 (68%) 44 (88%)
 

LEUKEMOID REACTION 1 (2%)
 

HYPERPLASIA, RETICULUM CELL 1 (2%)
 

HEMATOPOIESIS 40 (80%) 37 (74X)
 

EHYTHROPOISSIS 3 (6%)
 

GRANULOPOISSIS 2 (4%)
 

tSPLENIC CAPSULE (50) (50) 

INFLAMMATION, FOCAL 1 (2%) 

tLYMPH NODE (44) (41)
 

LYMPHANGIECTASIS
 

HEHOSIDEROSIS 1 (2%)

 

*HANDIBULAR L. NODE (44) (41) 

PIGMENTATION, NOS 1 (2%) 

tCERVICAL LYMPH NODE (44) (41)
 

CONGEST IONj._NOS
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUHBEB OF ANIMALS NECROPSIED 

75 

HIGH DOSE
 

1 (2*)
 

1 (2%)
 

4 (8%)
 

1 (2%)
 

1 (2%)
 

(49)
 

3 (6%)
 

(50)
 

1 (2%)
 

6 (12%)
 

1 (2%)
 

(49)
 

40	 (82%)
 

36 (73%)
 

2 (4%)
 

2 (4%)
 

(49)
 

(45)
 

1 (2%)
 

(45)
 

(45)
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

HEMOSIDEROSIS 1 (2X)
 

ITHYMUS (39) (30) (31)
 

LYMPHANGIECTASIS 1 (3%)
 

CONGESTION, NOS 1 (3%)
 

HEMOSIDEROSIS 1 (3»)
 

CIRCULATORY SYSTEM
 

IHEABT/ATRIUM (48) (46) (50)
 

THROMBOSIS, NOS 1 (2X)
 

tMYOCARDIUM (48) (46) (50)
 

INFLAMMATION, FOCAL 2 (4X)
 

INFLAMMATION, INTERSTITIAL 7 (1536) 2 (4%)
 

FIBROSIS 1 (2X)
 

FIBROSIS, FOCAL 2 (4X) 9 (20X) 9 (1835)
 

SCAR 1 (2X)
 

IENDOCARDIUM (48) (46) (50)
 

INFLAMMATION, FOCAL 1 (2%)
 

*PULMONARY ARTERY (50) (50) (50)
 

CALCIFICATION, NOS 1 (2*)
 

CALCIFICATION, FOCAL 1 (2%)
 

tHEPATIC SINUSOID (49) (50) (50)
 

CONGESTION, NOS 4 (835) 1 (235)
 

HYPEBPLASIA, GRANULOCYTIC 1 (2X)
 

DIGESTIVE SYSTEM
 

*TONGOE (50) (50) (50)
 

HYPERKERATOSIS 1 (2X)
 

ACANTHOSIS 1 (235)
 

tLIVER (49) (50) (50)
 

HERNIA, NOS 1 (2X) 1 (2%)
 

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

METAMORPHOSIS FATTY 9 (18X) 11 (22X) 3 (6%)
 

PIGMENTATION, NOS 1 (2%)
 

FOCAL CELLULAR CHANGE 1 (2%)
 

ANGIECTASIS 3 (6X) 1 (2X)
 

t NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

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

CONTROL LOW DOSE
 

HYPERPLASIA, RETICULUM CELL 4 (8«) 

HEMATOPOIE3IS 1 (2%) 2 (4X) 

ERYTHHOPOISSIS \ (2%) 

ILIVEH/CENTRILOBULAR (49) (50) 

NECROSIS, FOCAL 

HETAHORPHOSIS FATTY 

1 (2%) 

2 (4%) 2 (4%) 

tLIVER/PERIPORTAL (49) (50) 

METAMORPHOSIS FATTY 1 (2%) 

*BILE DOCT *

(50) (50) 

INFLAMMATION, FOCAL 2 (4%) 

HYPERPLASIA, NOS 

HYPERPLASIA, FOCAL 15 (30%) 17 (34X) 

tPAHCREAS (49) (49) 

LYBPHOCYTIC INFLAMHATORY INFILTR 1 (2%) 

PERI ART EBI IIS 

tPANCREATIC DUCT (49) (49) 

HYPERPLASIA, FOCAL 5 (10*) 6 (12%) 

tPANCREATIC ACINUS (49) (49) 

NODULE 1 (2%) 

tESOPHAGUS (49) (48) 

ABSCESS, NOS 1 (2%) 

tSTOHACH (50) (50) 

ULCER, NOS 1 (2%) 

ULCER, FOCAL 1 (2X) 

tCARDIAC STOMACH (50) (50) 

ULCER, NOS 

EROSION 

1 (2*) 

1 (2%) 

tPEYEKS PATCH 

HYPERPLASIA, LYMPHOID 

(49) 

4 (8%) 

(45) 

2 (4%) 

tCOLON (35) (39) 

NEMATODIASIS 5 (14%) 4 (10%) 

HYPERPLASIA, RETICULDS CELL 1 (3%) 

__HY£E 8 £LA SIA ^LY HPHO ID . 1_J3%1 _. 

t NUHBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

77
 

HIGH DOSE
 

1 (2%)
 

(50)
 

2 (4%)
 

(50)
 

1 (2%)
 

(50)
 

2 (4%)
 

16 (32S)
 

(47)
 

1 (2%)
 

(47)
 

9 (19%)
 

(47)
 

(42)
 

(49)
 

(49)
 

(50)
 

4 (8%)
 

(39)
 

5 (13%)
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

URINARY SYSTEM 

#KIDNEY (U9) (50) (50)
 

CAST, NOS 1 (2%)
 

INFLAMMATION, FOCAL 1 (2%)
 

INFLAMHATION, INTERSTITIAL 1 (2X) 1 (2%)
 

INFLAMMATION, CHRONIC 2 (4X) 2 (4S) 2 (US)
 

INFLABMATION. CHRONIC FOCAL 12 (24X) 7 (14X) 13 (26%)
 

INFLAMMATION, CHRONIC DIFFUSE 1 (2%)
 

SCAR 1 (2X)
 

NEPHROSIS, NOS 1 (2%)
 

INFARCT, ACUTE 1 (2X)
 

PIGHENTATION, NOS 2 (4«) 2 (4«) 1 (2%)
 

HYPERPLASIA, TUBULAR CELL 1 (2X)
 

*KIDNEY/CORTEX (49) (50) (50)
 

SCAR 1 (2XJ
 

INFARCT, NOS 1 (2X)
 

PIGMENTATION, NOS 17 (35X) 26 (52X) 20 (40X)
 

HYPERPLASIA, LYHPHOID 1 (2X)
 

#KIDNEY/TUBULE (49) (50) (50)
 

CAST, NOS 1 (2*)
 

PIGHENTATION, NOS 2 (4X) 2 (OX) 1 (2XJ
 

ATROPHY, NOS 1 (2X)
 

tCONVOLUTED TUBULES (49) (50) (50)
 

CAST, NOS 1 (2X)
 

HYALINE MEMBRANE 1 (2X)
 

PIGHENTATION, NOS 1 (2%)
 

tKIDNEY/PELVIS (49) (50) (50)
 

CALCIFICATION, FOCAL 1 (2%) 1 (2%)
 

#URINARY BLADDER (35) (42) (41)
 

CALCULUS, NOS 1 (3X)
 

INFLABMATION, CHRONIC 1 (3*)
 

HYPERPLASIA, EPITHELIAL 1 (3X)
 

ENDOCRINE SYSTEM 

tPITUITARY (45) (46) (47)
 

CYST A _NOS 

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

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

HIGH DOSE 

2 (4%) 

3 (6*) 

20 (43%) 

(49) 

1 (2%) 

7 (14%) 

(49) 

(49) 

2 (4%) 

(44) 

1 (2%) 

21 (43%) 

(44) 

(50) 

10 (20%) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 

1 (2%) 

2 (4%) 

(50) 

HEMORRHAGE 

HEMORRHAGIC CYST 

HEHOSIDEROSIS 

HYPEHPLASIA, NOS 

HYPERPLASIA,

ANGIECTASIS 

•ADRENAL 

CYST, NOS 

DEGENERATION,

HEMOSIDEROSIS 

ANGIECTASIS 

•ADRENAL CORTEX 

HEMORRHAGE 

IADRENAL MEDULL A 

HYPERPLASIA,

tTHYROID 

 FOCAL 

 NOS 

 FOCAL 

CYSTIC FOLLICLES 

LYBPHOCYTIC INFLAHMATORY INFILTR 

HYPERPLASIA, C-CELL 

tTHYROID FOLLICLE 

PIGMENTATION, NOS 

REPRODUCTIVE SYSTEM 

*MAMMARY GLAND 

GALACTOCELE 

NECROSIS, FOCAL 

METAPLASIA, SQUAMOUS

 

ADENOSIS 

*PREPUTIAL GLAND 

INFLAHHATION, SUPPURATIVE 

INFLAMMATION, ACUTE SUPPUHATIVE 

NECROSIS, NOS 

HYPERPLASIA, NOS 

HYPERPLASIA, CYSTIC 

*V AGIN A 

HEHATOBAX_NOS_ 

CONTROL 

2 (4%) 

2 («} 

1 (2%) 

3 (7%) 

1 (2%) 

3 (7%) 

(49) 

1 (2%) 

3 (6%) 

(49) 

1 (2%) 

(49) 

(50) 

1 (2%) 

39 (78%) 

(50) 

(50) 

5 (10%) 

1 (2%) 

(50) 

7 (14%) 

1 (2%) 

(50) 

LOW DOSE 

1 (2%) 

3 (7%) 

3 (7%) 

1 (2%) 

1 (2%) 

14 (30%) 

(50) 

1 (2%) 

6 (12%) 

(50) 

(50) 

1 (2%) 

(44) 

1 (2%) 

1 (2« 

24 (55%) 

(44) 

1 (2%) 

(50) 

4 (8%) 

1 (2ft) 

(50) 

2 (4%) 

1 (2%) 

(50) 

1 _[2X]_ 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED 8ICROSCOPICALLY
 

* NUHBEE OF ANIMALS NECROPSIED
 

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

CONTROL LOW DOSE
 

tUTERUS (50) (48) 

HEHORRHAGE 1 (2«) 

INFLAMMATION, NOS 

INFLAMMATION, SUPPORATIVE 

1

1

 (2X) 

 (2*) 

PYOHETRA 1 (2%) 

NECROSIS, NOS 1 (2%) 

ICEBVIJC UTERI (50) (48) 

HYPERPLASIA, NOS 1 (2%) 

*UTERUS/ENDOMETRIUM (50) C*8) 

CYST, NOS 1 (2%) 

HEMORRHAGE 1 (2%) 

INFLAMMATION, FOCAL 1 (2%) 

ULCER, FOCAL 1 (2%) 

LYMPHOCYTIC INFLAMMATORY INFILT8 1 (2%) 

INFLAMMATION, SOPPURATIVE 3 (16%) 8 (17X) 

INFLAMMATION, VESICULAR 1 (2*) 

INFLAMMATION, CHRONIC SUPPURATIV 1 (2%) 

NECROSIS, NOS 

HYPERPLASIA, CYSTIC 2 (4*) 1 (2*) 

#OVARY/OVIDOCT (50) (148) 

INFLAMMATION, NOS 1 (2%) 

INFLAMMATION, SUPPURATIVE 5 (10X) 13 (21%) 

tOVARY (50) (47) 

CYST, NOS 9 (18%) 8 (17K) 

FOLLICOLAR CYST, NOS 

INFLAMMATION, NOS 

NERVOUS SYSTEM
 

*BRAIN/MENINGES (49) (50)
 

INFLAMMATION, SUPPURATIVE 1 (2%)
 

ICEREBRUM (49) (50)
 

INFLAMMATION, SUPPURATIVE 1 (2%)
 

ABSCESS, NOS
 

tBRAIN (49) (50)
 

COMPRESSION 2 (4«)
 

INFLAMMATION^ NOS i_i2«J. _
 

* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

80
 

HIGH DOSE
 

(49)
 

(49)
 

(49)
 

14 (29«)
 

2 (4«)
 

2 (4»)
 

(49)
 

12 (24%)
 

(48)
 

10	 (2H)
 

1 (2%)
 

1 (2%)
 

(50)
 

(50)
 

1 (2%)
 

(50)
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE
 

MALACIA 1 (2%)
 

*MIDBRAIN (U9) (50)
 

COMPRESSION
 

GLIOSIS 1 (2%)
 

*SPINAL CORD (50) (50)
 

HEMORRHAGE 1 (2%)
 

SPECIAL SENSE ORGANS
 

*EYE (50) (50)
 

HEMORRHAGE
 

INFLAMMATION, NOS
 

INFLAMMATION, INTERSTITIAL
 

PUS
 

INFLAMMATION, SUPPURATIVE 1 (2%)
 

CATARACT 11 (22%) 15 (30*)
 

*EYE/CORNEA (50) (50)
 

ULCER, NOS 1 (2%)
 

INFLAMMATION, SUPPURATIVE 1 (2%)
 

*EAR (50) (50)
 

INFLAMMATION, NOS
 

*EAR CANAL (50) (50)
 

INFLAMMATION, SUPPORATIVE 1 (2%)
 

MUSCOLOSKELETAL SYSTEM
 

*SKELETAL MUSCLE (50) (50)
 

ATROPHY, NOS 1 (2%)
 

BODY CAVITIES
 

*MESENTERY (50) (50)
 

FIBROSIS 2 (4%)
 

ALL OTHER SYSTEMS
 

*MULTIPLE ORGANS (50) (50)
 

____ CONGEST ION j._NOS
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

81
 

HIGH DOSE
 

(50)
 

2 (H%)
 

(50)
 

(50)
 

1 (2%)
 

1 (2%)
 

1 (2S)
 

1 (2%)
 

16 (32%)
 

(50)
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

(50)
 

(50)
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

JAUNDICE, NOS 

THORAX 

HEMORRHAGE 

1 (2*) 

1 

DIAPHRAGM 

HERNIA, NOS 1 1 2 

ADIPOSE TISSUE 

INFLAMMATION, NOS 2 

SPECIAL MORPHOLOGY SUMMARY 

NONE 

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

82
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE ADMINISTERED 3-NITROPROPIONIC ACID
 

BY GAVAGE
 

83
 

TABLE 01. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE 

CONTROL LOW DOSE 

ANIMALS INITIALLY IN STDDY 50 50 

ANIMALS NECROPSIED 49 50 

ANIMALS EXAMINED HISTOPATHOLOGIC ALLY 49 50 

INTEGUMENTARY SYSTEH
 

*SKIN (49) (50)
 

CYST, NOS 1 (2%)
 

ULCER, NOS 1 (2%)
 

HYPEHPLASIA, NOS 1 (2%)
 

*SUBCUT TISSUE (49) (50)
 

ABSCESS, NOS 1 (2X)
 

INFLAMMATION, CHRONIC 1 (2X)
 

RESPIRATORY SYSTEM
 

tLUNG/BRONCHUS (49) (48)
 

METAPLASIA, SQUAMOOS 1 (2«)
 

HYPERPLASIA, LYMPHOID 11 (22%) 1 (2%)
 

ILUNG (49) (48)
 

CONGESTION, NOS 1 (2%)
 

HEMORRHAGE 1 (2%)
 

PNEUMONIA, ASPIRATION
 

PERIVASCULITIS 1 (2*)
 

ALVEOLAR MACROPHAGES
 

HYPERPLASIA, ADENOMATOUS 1 (2%)
 

HYPERPLASIA, ALVEOLAR EPITHELIUM
 

tLUNG/ALVEOLI (49) (48)
 

CONGESTION, NOS
 

HEHATOPOIETIC SYSTEM
 

*BONE HARROW (46) (49)
 

HYPERPLASIAX_HOS
 

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

85
 

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

(50)
 

(50)
 

2 (4%)
 

(50)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

HYPERPLASIA, HEHATOPOIETIC 2 (4X) 2 (4*)
 

HYPERPLASIA, GR ANOLOCYTIC 2 (4%) 1 (2X)
 

*SPLEEN (46) (50) (46)
 

HEMORRHAGE 1 (2%)
 

ANGIECTASI3 1 (2%)
 

HYPERPLASIA. HEMATOPOIETIC 1 (2%)
 

HYPERPLASIA, RETICULOS CELL 2 (4*) 1 (2%)
 

HYPERPLASIA, LYMPHOID 2 (4X) 5 (10%)
 

HEKATOPOIESIS 24 (52%) 23 (46X) 17 (37 X)
 

ERYTHBOPOIESIS 2 (4») 1 (2X) 2 (4%)
 

GRANOLOPOIESIS 1 (2%)
 

tLYHPH NODE (40) (31) (30)
 

HEMATOPOIESIS 1 (3%)
 

#ME"5ENTERIC L. NODE (40) (31) (30)
 

THROHBOSIS, NOS 1 (3%)
 

CONGESTION, NOS 3 (8%)
 

HEMOSIDEROSIS 1 (3%)
 

ERYTHROPHAGOCYTOSIS 2 (7*)
 

HYPEBPLASIA, HEHATOPOIETIC 1 (3%)
 

HYPERPLASIA, LYHPHOID 1 (3X)
 

#THYMOS (35) (38) (41)
 

HYPERPLASIA., RETICOLUH CELL 1 (3%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEH
 

*TONGUE (49) (50) (50)
 

HYPERPLASIA, EPITHELIAL 1 (2%)
 

HYPERKERATOSIS 1 (2%)
 

ACANTHOSIS 1 12*)
 

tLIVER (49) (50) (49)
 

HEMORRHAGE 1 (2%) 1 (21)
 

FIBROSIS, FOCAL 1 (2%)
 

DEGENERATION, NOS 1 (2%)
 

NECROSISA_FOCAL
 

* NOMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

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

CONTROL LOW DOSE HIGH DOSE 

HETAHORPHOSIS FATTY 

CLEAR-CELL CHANGE 

HYPERPLASIA, MODULAR 

HYPERPLASTIC HODBLE 

ANGIECTASIS 

HYPERPLASIA, HEH ATOPOIETIC 

HYPERPLASIA, RETICULOB CELL 

HYPERPLASIA, LYHPHOID 

HEBATOPOIESIS 

ERYTHROPOIESIS 

4 (8%) 

1 (2%) 

1 (2%) 

1 <2X) 

1 (2%) 

1 (2%) 

1

1

1

3

1

1

 (2*) 

 (2%) 

 (2%) 

 (6*) 

 (2%) 

 (2*) 

3 (6S) 

1 (2%) 

2 (4%) 

1 (2») 

#HEPATIC CAPSULE 

HEBATOHA, NOS 

FIBROSIS, FOCAL 

(49) 

1 (2%) 

(50) 

1 (2X) 

(49) 

tLIVER/CENTRILOBULAR 

NECROSIS, NOS 

BETAHORPHOSIS FATTY 

(U9) 

1 (2%) 

(50) (49) 

1 (2%) 

*LIVER/PEBIPORTAL 

HYPERPLASIA, LYBPHOID 

(49) 

1 (2%) 

(50) (49) 

tLIVER/HEPATOCYTES 

NECROSIS, NOS 

NECROSIS, FOCAL 

(49) (50) 

2 (4S) 

(49) 

1 (2%) 

*BILE DUCT 

CYST, NOS 

INFLABBATION, NOS 

INFLABBATION, FOCAL 

INFLABBATION, SUPPURATIVE 

FOCAL CELLULAR CHANGE 

HYPERPLASIA, NOS 

HYPERPLASIA, FOCAL 

(49) 

1 (2%) 

4 (8%) 

(50) 

1

1

1

1

 (2%) 

 (2%) 

 (2«) 

 (236) 

(50) 

1 (2%) 

1 (2%) 

2 (4«) 

tPANCREAS 

CYSTIC DUCTS 

FIBROSIS 

NECEOSIS, NOS 

(48) 

1 (2%) 

1 (2%) 

1 (2%) 

(50) (47) 

tP&NCREATIC DUCT 

CYST, NOS 

HYPERPLASIA, FOCAL 

(48) 

1 (2%) 

(50) 

3 (6*) 

(47) 

#PEYERS PATCH 

INFLABBATION^ SUPPURATIVE 

(47) (49) (49) 

1 i2%L 

» NUBBER OF ANIBALS WITH TISSUE EXABINED BICROSCOPICALLY
 * NUBBER OF ANIBALS NECROPSIED
 

87
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE
 

HYPERPLASIA, NOS	 1 (2%)
 

HYPERPLASIA, LYMPHOID	 2 (4%) 10 (20%)
 

#COLON (22) (44)
 

NEMATODIASIS 4 (18%) 6 (14%)
 

URINARY SYSTEM
 

•KIDNEY	 (48) (49)
 

HYDRONEPHBOSIS 1 (2%)
 

LYNPHOCYTIC INFLAMHATOHY INFILTR 2 (4%)
 

INFLAMMATION, INTERSTITIAL 1 (2%)
 

INFLAMMATION, CHRONIC 1 (2%)
 

FIBROSIS 1 (2%)
 

HYPERPLASIA, LYMPHOID 1 (2%) 1 (2%)
 

tKIDNEY/CORTEX (48) (49)
 

LYMPHOCYTIC INFLAHMATORY INFILTR 1 (2%)
 

INFARCT, NOS 1 (2%)
 

tU. BLADDER/SUBMUCOSA (47) (46)
 

HYPERPLASIA, LYMPHOID
 

ENDOCRINE SYSTEM
 

•ADRENAL/CAPSULE	 (46) (49)
 

HYPERPLASIA, FOCAL 28 (61%) 41 (84%)
 

•ADRENAL CORTEX (46) (49)
 

HYPERPLASIA, NOS 2 (4%)
 

•THYROID	 (43) (44)
 

CYSTIC FOLLICLES 1 (2%)
 

HYPERPLASIA, FOLLICULAR-CELL 2 (5%) 2 (5%)
 

•PANCREATIC ISLETS (48) (50)
 

HYPESPLASIA, NOS 2 (4%)
 

REPRODUCTIVE SYSTEM
 

*PREPUTIAL GLAND (49) (50)
 

CYST, NOS 3 I6%}.__ .
 

• NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 * NURBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 

5 (10%)
 

(35)
 

1 (3%)
 

(50)
 

1 (2%)
 

(50)
 

(49)
 

1 (2%)
 

(50)
 

38 (76%)
 

(50)
 

(48)
 

1 (2%)
 

2 (4%)
 

(47)
 

(50)
 

88
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

INFLAMMATION, SUPPURATIVE 2 (<»X) 2 (4X) 1 (2*)
 

INFLAMMATION, CHRONIC 1 (2*)
 

INFLAMMATION, CHBONIC SOPPUBATIV 1 (2S)
 

HYPERKERATOSIS 1 (2»)
 

tTESTIS (47) (49) (50)
 

GRANULOMA, SPERMATIC 1 (2*)
 

ATROPHY, NOS 2 (4X)
 

ATROPHY, FOCAL 4 (8*) 1 (2%)
 

ASPERMAIOGENESIS 1 (2%) 

*EPIDIDYMIS (U9) (50)

INFLAMMATION, SUPPURATIVE 1 (2*) 

FIBHOSIS 1 (2») 

FIBROSIS, FOCAL

NECROSIS, FAT 1 (2*) ------------------------------------------------------------------------------­

 (50) 

1 (2X) 

J 

HERVOUS SYSTEM 

IBRAIN/HENINGES

INFLAMMATION, NOS

 C*8) (49)

 1 (2*) 

 (49) 

SPECIAL SENSE ORGANS 

*EYE

CATARACT

 (49) (50)

 1 (2*) 

 (50) 

*EYE/CORNEA

INFLAMMATION, NOS

 (49) (50)

 1 (2X) 

 (50) 

*LENS CAPSULE

DEGENERATION, NOS

 (49) (50)

 1 (2*) 

 (50) 

HOSCDLOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

*PERITONEOM

____ HEMORRHAGE 

 (49) (50) (50) 

t NDHBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

89 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

INFLAHBATION, NOS 1 (2%)
 

NECROSIS, FOCAL 1 (2%)
 

*ABDOBINAL VISCERA (1*9) (50) (50)
 

ADHESION, NOS 1 (2%)
 

*PLEURA (49) (50) (50)
 

HYDROTHORAX 1 (2%)
 

HEBOTHORAX 2 (UX)
 

INFLAMMATION, FOCAL 1 (2%)
 

*MESENTERY (49) (50) (50)
 

F1BROSIS 4 (8*) 1 (2%)
 

fIBROSIS, FOCAL 1 (2%)
 

NECROSIS, FAT 2 (4S) 3 (6S) 3 (6%)
 

ALL OTHER SYSTEMS
 

ADIPOSE TISSUE
 

INFLAHMATION, NOS 2 1
 

FIBROSIS 1
 

SPECIAL MOBPHOLOGY SUMMARY
 

NO LESION REPORTED 1 1
 

AUTO/NECROPSY/HISTO PERF 1
 

AUTOLYSIS/NO NECROPSY 1
 

t HUMBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

90
 
TABLE D2. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE

 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY 50	 50 50
 

UNIMALS NECROPS1ED	 50 50 50
 ANIBALS EXAMINED HISTOPATHOLOGICALLY 50	 50 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (50) (50) (50)
 

ALOPECIA 1 (2S)
 

HYPERKERATOSIS 1 (2%)
 

RESPIRATORY SYSTEM
 

•LUNG/BRONCHUS	 (47) (49) (49)
 

HYPERPLASIA, LYMPHOID 18 (38%) 3 (6X) 3 (6%)
 

#LUNG (47) (49) (<*9)
 

CONGESTION, NOS 1 (21) 2 (4%)
 

INFLAMMATION, FOCAL 1 (2%)
 

INFLAMMATION, SUPPURATIVE 1 (25)
 

ALVEOLAB MACROPHAGES 1 (2%)
 

HYPEBPLASIA, LYHPHOID 1 (2%) 1 (2S)
 

HEMATOPOIETIC SYSTEM
 

tBONE MARROW (46) (43) (50)

 

HYPERPLASIA, HEMATOPOIETIC 3 (1%) (2%) 

HYPERPLASIA, GEANULOCYTIC 1 (2%}	 1 (2%)
 

GRANULOPOIESIS	 1 (2%)
 

ISPLEEN (47) (50) (50)
 

RUPTURE 1 (2%)
 

THROMBOSIS, NOS 1 (2%)
 

LEUKEMOID REACTION 1 (2%) 2 (<U)
 

HYPERPLASIA, LYMPHOID 6 (13%) 8 (16*) 11 (22%)
 

HEMATOPOIESIS 19 (40%) 22 (44X) 22 (44%)
 

EHYTHROPOIESIS 1 (2%)
 

MYELOPOIESIS 1 12%l _ _
 

# NOHBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

91
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

(33)
 

1 (3X)
 

(33)
 

1 (3%)
 

(33)
 

1 (3%)
 

(35)
 

(49)
 

1 (2%)
 

(49)
 

1 (2%)
 

(50)
 

(50)
 

1 (2X)
 

1 (2%)
 

2 (4»)
 

1 (2%)
 

1 (2X)
 

(50)
 

_ 3 (6%)
 

»LY«PH NODE
 

EDEMA, NOS
 

HYPERPLASIA, LYMPHOID
 

#KAHDIBULAR L. NODE
 

HEMORRHAGE
 

#NESENTERIC L. NODE
 

INFLAMMATION, GRANULOMATOUS
 

HYPERPLASIA, LYMPHOID
 

#THYMUS
 

HYPEHPLASIA, LYMPHOID
 

CIRCULATORY SYSTEM
 

•HEART/ATRIUM
 

THROMBOSIS, NOS
 

#CAEDIAC VALVE
 

PIGMENTATION, NOS
 

*UTEBINE ARTERY
 

THROMBOSIS, NOS
 

DIGESTIVE SYSTEM
 

tLIVER
 

HEMOBRHAGIC CYST
 

NECROSIS, FOCAL
 

NECROSIS, ISCHEMIC
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

LEUKEMOID REACTION
 

HYPE8PLASIA, RETICULOM CELL
 

HYPERPLASIA, LYMPHOID
 

HEMATOPOIE3IS
 

tLIVER/HEPATOCYTES
 

DEGENERATION, NOS
 

NECROSIS, NOS
 

NECROSIS^. FOCAL
 

CONTROL
 

(38)
 

1 (3X)
 

(38)
 

(38)
 

1 (3X)
 

(38)
 

1 (3X)
 

(49)
 

(49)
 

(50)
 

1 (2%)
 

(49)
 

1 (2X)
 

2 («X)
 

1 (2X)
 

2 (4X)
 

3 (6X)
 

(49)
 

1 J2X}.
 

LOW DOSE
 

(36)
 

(36)
 

(36)
 

(42)
 

1 (2*)
 

(50)
 

(50)
 

1 (2X)
 

(50)
 

(50)
 

1 (2X)
 

2 (4X)
 

1 (2%)
 

(50)
 

1 (2«)
 

1 (2«)
 

1 (2X1
 

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

92
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

•PANCREATIC DUCT (UU) (t9) (50)
 

CYST, NOS 1 (2*)
 

tPEYERS PATCH (48) (48) (49)
 

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

IDUODENUM (48) (48) (49)
 

HYPERPLASIIV, LYNPHOID 1 (2%)
 

•COLON	 (36) (38) (35)
 

NEMATODIASIS 1 (3J)
 

URINARY SYSTEM 

tKIDNEY (49) (50) (50) 

CONGESTION, NOS 1 (2X) 

GLOMERULONEPHRITIS, NOS 1 (2%) 

PYELONEPHRITIS, NOS 

PYELONEPHRITIS DIFFUSE 

1

1

 (2%) 

 (2%) 

INFLAMMATION, CHRONIC FOCAL 

GLOMERULOSCLEROSIS, NOS 

1 (2*) 

1 (2%) 

HYPERPLASIA, LYBPHOID 10 (20%) 4 (8«) 

tKIDNEY /CORTEX 

SCAR 

(49) 

1 (2%) 

(50) (50) 

DEGENERATION, HYALINE 1 (2%) 

fKIDNEY/TUBULS 

DEGENERATION, HYALINE 

(49) (50) 

1 (2S) 

(50) 

1 (2K) 

ENDOCRINE SYSTEM
 

IPITUITARY (43) (48) (42)
 

HEMORRHAGIC CYST 1 (2%)
 

HYPERPLASIA, NOS 1 (2X) 2 (5ft)
 

ANGIECTASIS 2 (4%) 2 (5»)
 

IADRENAL/CAPSOLE (48) (50) (49)
 

HYPERPLASIA, FOCAL 43 (90%) 47 (94S) 42 (86%)
 

•ADRENAL	 CORTEX (48) (50) (49)
 

HYPEBPLASIAX FOCAL ._ _ _i_im
 

* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

93
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE
 

tTHYROID (40) (47)
 

CYSTIC FOLLICLES 1 (3%)
 

INFLAMMATION, FOCAL
 

HYPERPLASIA, FOLLICUL AR-CELL 6 (15%) 13 (23%)
 

8EP80DUOTIVE SYSTEM
 

fMJTERUS (47) (49)
 

HYDROMETRA 1 (2%)
 

HEMORRHAGE 1 (2%)
 

METAPLASIA, SQOAMOUS 2 (4*)
 

tUTERUS/ENDOMETRIUM (47) (49)
 

HEMATOMA, NOS 1 (2%)
 

INFLAMMATION, SUPPORATIVE 3 (6%) 1 (2%)
 

HYPEBPLASIA, NOS 1 (2%)


HYPERPLASIA, CYSTIC 19 (40%) 37 (76%)
 

•OVARY/OVIDUCT	 (47) (49)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 1 (2%) 1 (2/S)
 

INFLAMMATION, SUPPURATIVE 3 (6%)
 

NECROSIS, NOS 1 (2%)
 

•OVARY	 (39) (47)
 

CYST, NOS 4 (10*) 10 (2H)
 

FOLLICULAR CYST, NOS 4 (9%)
 

HEM08RHAGIC CYST 1 (3%)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 1 (2%)
 

INFLAMMATION, SUPPORATIVE 1 (3%)
 

INFLAMMATION, CHRONIC 1 <3«)
 

CALCIFICATION, FOCAL
 

HYPERPLASIA, LYMFHOID
 

NERVOUS SYSTEM
 

IBRAIN/MENING3S (47) (50)
 

PERIVASCULAR CUFFING 1 (2K)
 

*CEBEBROM (47) (50)
 

EPIDERMAL INCLUSION CYST
 

SPECIAL SENSE ORGANS
 

• EIE (50) (50)
 

PHTHISIS BULBI 1 _I2%]_
 

t NUMBER. ,OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE R OF ANIMAL S NECBOPSIED
 

HIGH DOSE
 

(45)
 

1
 

1
 

5
 

(48)
 

1
 

(48)
 

1
 

30
 

(43)
 

(47)
 

12
 

4
 

1
 

1
 

1
 

1
 

(50)
 

(50)
 

1
 

(50)
 

(2%)
 

(2%)
 

(11*)
 

(2%)
 

(2%)
 

£63%)
 

(26%)
 

(9%)
 

(2%)
 

(2%)
 

(2*)


(2%)
 

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

CONTROL LOW DOSE HIGH DOSE 

_ .. _•_•_ _ — __ _«________—_»_ w —_—_—_- . — — —_ _____— — — — — _ __ — _ _ _ -, —»—_ — _—._-.— «i_ _ — — — — — — •— —— — — 

*HARDERIAN GLAND (50) (50) (50) 

ABSCESS, NOS 1 (2X) 

INFLAMMATION, CHRONIC 1 (2%) 

BUSCULOSKELEIAL SYSTEM
 

NONE
 

BODY CAVITIES
 

•PERITONEUM	 (50) (50) (50)
 

CYST, NOS 1 (2X)
 

HEMORRHAGE 1 (2ft)
 

*PLEORA	 (50) (50) (50)
 

HYDROTHORAX 1 (2%)
 

HEHOTHORAX 1 (2%)
 

&LL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

AUTO/NECROPSY/HISTO PERF 1
 

* SOMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

95
 

APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN
 

RATS ADMINISTERED 3-NITROPROPIONIC ACID
 

BY GAVAGE
 

97
 

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

Administered 3-Nitropropionic Acid by Gavagea
 

Low
 

Topography: Morphology Control Dose
 

All Sites: Skin Tumorsb 2/50 (4) 6/50 (12)
 

P Valuesc>d N.S. N.S.
 

Relative Risk (Control)f 3.000
 

Lower Limit 0.569
 

Upper Limit 29.254
 

Weeks to First Observed Tumor 111 91
 

Hematopoietic System: Malignant Lymphoma,
 

Undiff erentiated Leukemia, or
 

Lymphocytic Leukemiab 11/50 (22) 6/50 (12)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Control)f 0.545
 

Lower Limit 0.179
 

Upper Limit 1.477
 

Weeks to First Observed Tumor 96 96
 

High
 

Dose
 

2/50 (4)
 

N.S.
 

1.000
 

0.075
 

13.326
 

111
 

14/50 (28)
 

N.S.
 

1.273
 

0.597
 

2.785
 

83 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

Low High
 

Topography; Morphology Control Dose Dose
 

Hematopoietic System:
 

All Neoplasms'3 13/50 (26) 7/50 (14) 16/50 (32)
 

P Values0 »d	 N.S. N.S. N.S.
 

Departure from Linear Trend6	 P = 0.043
 

Relative Risk (Control)f 0.539 1.231
 

Lower Limit 0.199 0.624
 

Upper Limit 1.323 2.474
 

o 

o Weeks to First Observed Tumor 90 96 83 

Liver: Neoplastic Nodule^ 0/49 (0) 3/50 (6) 5/49 (10) 

P Valuesc»d P = 0.022 N.S. P = 0.028 

Relative Risk (Control)f Infinite Infinite 

Lower Limit 0.590 1.262
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 111 109
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

Low High 

Topography^ Morphology Control Dose Dose 

Liver: Neoplastic Nodule or 

Hepatocellular Carcinomab 0/49 (0) 3/50 (6) 6/49 (12) 

P Valuesc » d P = 0.010 N.S. P = 0.012 

Relative Risk (Control)f Infinite Infinite 

Lower Limit 0.590 1.601 

Upper Limit Infinite Infinite 

Weeks to First Observed Tumor — 111 109 

Pituitary: Chromophobe Adenoma*5 3/46 (7) 5/48 (10) 4/49 (8) 

P Valuesc » d N.S. N.S. N.S. 

Relative Risk (Control)f 1.597 1.252 

Lower Limit 0.331 0.224 

Upper Limit 9.773 8.137 

Weeks to First Observed Tumor 111 101 95 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats 

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

Low High
 

Topographyj Morphology Control Dose Dose
 

Adrenal: Pheochromocytoma^ 4/49 (8) 5/50 (10) 5/50 (10)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Control)f 1.225 1.225
 

Lower Limit 0.280 0.280
 

Upper Limit 5.833 5.833
 

Weeks to First Observed Tumor 88 79 106
 

g Thyroid: Follicular-cell Carcinomab 1/46 (2) 0/49 (0) 4/47 (9)
 

N>
 

P Valuesc>d N.S. N.S. N.S.
 

Relative Risk (Control)f 0.000 3.915
 

Lower Limit 0.000 0.407
 

Upper Limit 17.510 188.454
 

Weeks to First Observed Tumor 111 99
 —
 
__ 

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

Administered 3-Nitropropionic

(continued) 

Topography: Morphology Control
 

Thyroid: C-cell Carcinomab 1/46 (2)
 

P Valuesc»d N.S.
 

Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 111
 

Thyroid: C-cell Adenoma or Carcinomab 4/46 (9)
 

o 

CO 

P Valuesc>d N.S.
 

Departure from Linear Trend6 P = 0.044
 

Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 111
 

 Acid by Gavage3 

Low
 

Dose
 

2/49 (4)
 

N.S.
 

1.878
 

0.101
 

108.485
 

111
 

10/49 (20)
 

N.S.
 

2.347
 

0.735
 

9.593
 

107
 

High
 

Dose
 

0/47 (0)
 

N.S.
 

0.000
 

0.000
 

18.240
 

4/47 (9)
 

N.S.
 

0.979
 

0.193
 

4.955
 

103 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued)
 

Low High
 

Topography: Morphology Control Dose Dose
 

Pancreatic Islets: Islet-cell Adenoma^ 4/49 (8) 6/50 (12) 11/50 (22)
 

P Valuesc'd P = 0.033 N.S. P = 0.049
 

Relative Risk (Control)f 1.470 2.695
 

Lower Limit 0.372 0.865
 

Upper Limit 6.681 10.868
 

Weeks to First Observed Tumor 88 87 83
 

o
 Preputial Gland: Adenoma, NOS
 

-P­ (not otherwise specified)" 2/50 (4) 1/50 (2) 4/50 (8)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Control)f 0.500 2.000
 

Lower Limit 0.009 0.301
 

Upper Limit 9.290 21.316
 

Weeks to First Observed Tumor 111 111 106
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued)
 

Low
 

Topography: Morphology Control Dose
 

Testis: Interstitial-cell Tumorb 48/50 (96) 44/49 (90)
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Control) f 0.935
 

Lower Limit 0.871
 

Upper Limit 1.058
 

Weeks to First Observed Tumor 84 65
 

Peritoneum: Mesothelioma, NOSb 1/50 (2) 3/50 (6)
 

o
01
 

P Values0 »d N.S. N.S.
 

Relative Risk (Control)f 3.000
 

Lower Limit 0.287
 

Upper Limit 74.701
 

Weeks to First Observed Tumor 111 79
 

High
 

Dose
 

48/49 (98)
 

N.S.
 

1.020
 

0.954
 

1.061
 

0/50 (0)
 

N.S.
 

0.000
 

0.000
 

18.658
 

—
 

82 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-Nitropropionic Acid by Gavage3
 

(continued)
 

Topography: Morphology


Lung: Alveolar/Bronchiolar Carcinoma*3


P Valuesc»d


Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar Adenoma
 

or Carcinomab


P Valuesc>d


Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Low High 

 Control Dose Dose 

 0/50 (0) 1/49 (2) 3/48 (6) 

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

Infinite Infinite 

0.055 0.627 

Infinite Infinite 

— 111 86 

 3/50 (6) 2/49 (4) 3/48 (6) 

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

0.680 1.042 

0.059 0.146 

5.680 7.419 

 102 111 86 

a

Treated groups received doses of 0.425 or 0.85 mg/animal/day.
 

^Number of tumor-bearing animals/number of animals examined at site (percent),
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Administered 3-Nitropropionic Acid by Gavage3
 

(continued)
 

c

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


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the incidence of tumors in each treated group is the probability level for the Fisher exact test for the
 

comparison of that treated 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 treated group than in the control group.
 

e

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

95% confidence interval of the relative risk between each treated group and the control group.
 
Table E2. Analyses of the Incidence of Primary

Administered 3-Nitropropionic Acid

Topography: Morphology Control
 

Hematopoietic System: Malignant Lymphoma,
 

Lymphocytic Leukemia, or
 

Leukemia, NOSb 5/50 (10)
 

P Valuesc»d N.S.
 

Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 98
 o 

00 

Hematopoietic System:
 

All Neoplasms'3 7/50 (14)
 

P Values0 »d N.S.
 

Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 98
 

 Tumors in

 by Gavagea 

Low
 

Dose
 

5/50 (10)
 

N.S.
 

1.000
 

0.245
 

4.082
 

106
 

6/50 (12)
 

N.S.
 

0.857
 

0.268
 

2.684
 

105
 

 Female Rats 

High
 

Dose
 

8/50 (16)
 

N.S.
 

1.600
 

0.497
 

5.808
 

83
 

10/50 (20)
 

N.S.
 

1.429
 

0.535
 

4.072
 

83
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued)
 

Low
 

Topography; Morphology Control Dose
 

Pituitary: Chromophobe Adenomab 19/45 (42) 15/46 (33)
 

P Values0 »d N.S. N.S.
 

Relative Risk (Control)f 0.772
 

Lower Limit 0.423
 

Upper Limit 1.393
 

Weeks to First Observed Tumor 90 96
 

Adrenal: Pheochromocytoma^ 3/49 (6) 1/50 (2)
 o
 

vO
 

P Valuesc»d N.S. N.S.
 

Relative Risk (Control)f 0.327
 

Lower Limit 0.013
 

Upper Limit 3.417
 

Weeks to First Observed Tumor 107 103
 

High
 

Dose
 

20/47 (43)
 

N.S.
 

1.008
 

0.596
 

1.711
 

95
 

1/49 (2)
 

N.S.
 

0.333
 

0.013
 

3.486
 

107
 
Table E2. Analyses of the Incidence of Primary Tumors in

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

Low
 

Topography; Morphology Control Dose
 

Thyroid: C-cell Carcinoma^ 2/50 (4) 1/44 (2)
 

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

Relative Risk (Control)f 0.568
 

Lower Limit 0.010
 

Upper Limit 10.516
 

Weeks to First Observed Tumor 111 111
 

Thyroid: C-cell
 

Adenoma or Carcinoma^ 5/50 (10) 4/44 (9)
 

d
 P Values0 » N.S. N.S.
 

Relative Risk (Control)f 0.909
 

Lower Limit 0.191
 

Upper Limit 3.952
 

Weeks to First Observed Tumor 111 111
 

 Female Rats 

High
 

Dose
 

2/44 (5)
 

N.S.
 

1.136
 

0.085
 

15.083
 

59
 

5/44 (11)
 

N.S.
 

1.136
 

0.279
 

4.608
 

59
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued)
 

Topography: Morphology


Mammary Gland: Fibroadenomab


P Valuesc»d


Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observe4 Tumor


Uterus: Endometrial Stromal Polypb


P Valuesc»d


Relative Risk (Control) ^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Low High 

 Control Dose Dose 

 12/50 (24) 14/50 (28) 13/50 (26) 

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

1.167 1.083 

0.560 0.509 

2.475 2.334 

 94 67 88 

 2/50 (4) 4/48 (8) 5/49 (10) 

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

2.083 2.551 

0.314 0.441 

22.174 25.786 

 111 96 111 

a

Treated groups received doses of 0.6 or 1.2 mg/animal/day.
 

^Number of tumor-bearing animals/number of animals examined at site (percent).
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued)
 

G

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

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

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

the comparison of that treated 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 treated group than in the control group.
 

e

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

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

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN
 

MICE ADMINISTERED 3-^ITROPROPIONIC ACID
 

BY GAVAGE
 

113
 

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

 

Administered 3-Nitropropionic Acid by Gavagea

 

Topography; Morphology
 

Lung: Alveolar/Bronchiolar
 

Carcinoma"
 

P Valuesc»d
 

Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma^3
 

P Values0 »d
 

Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low 

Control Dose 

4/49 (8) 3/48 (6) 

N.S. N.S. 

0.766 

0.118 

4.285 

104 96 

14/49 (29) 8/48 (17) 

N.S. N.S. 

0.583 

0.234 

1.345 

85 96 

High
 

Dose
 

3/50 (6)
 

N.S.
 

0.735
 

0.113
 

4.120
 

105
 

10/50 (20)
 

N.S.
 

0.700
 

0.309
 

1.525
 

105
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice 

Administered 3-Nitropropionic Acid by Gavage3 

(continued) 

Low High
 

Topography: Morphology Control Dose Dose
 

Hematopoietic System: Malignant Lymphoma
 

or Lymphocytic Leukemia'1 6/49 (12) 10/50 (20) 11/50 (22)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Control)f 1.633 1.797
 

Lower Limit 0.585 0.664
 

Upper Limit 5.059 5.463
 

Weeks to First Observed Tumor 91 73 91
 

Hematopoietic System: All Neoplasmsb 8/49 (16) 12/50 (24) 12/50 (24)
 

P Valuesc»d N.S.. N.S. N.S.
 

Relative Risk (Control)f 1.470 1.470
 

Lower Limit 0.608 0.608
 

Upper Limit 3.783 3.783
 

Weeks to First Observed Tumor 91 73 91
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued) 

Topography: Morphology Control 

Low 

Dose 

High 

Dos e 

Liver: Hepatocellular Carcinoma^ 16/49 (33) 8/50 (16) 12/49 (24) 

P Valuesc » d N.S. P = 0.044 (N) N.S. 

Relative Risk (Control)f 

Lower Limit 

Upper Limit 

0.490 

0.201 

1.094 

0.750 

0.364 

1.504 

Weeks to First Observed Tumor 97 92 87 

Liver: Hepatocellular Adenoma 

or Carcinomab 20/49 (41) 10/50 (20) 16/49 (33) 

P Valuesc » d N.S. P = 0.021 (N) N.S. 

Departure from Linear Trend6 P = 0.037 

Relative Risk (Control)f 

Lower Limit 

Upper Limit 

0.490 

0.231 

0.975 

0.800 

0.446 

1.419 

Weeks to First Observed Tumor 97 92 87 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Administered 3-Nitropropionic Acid by Gavage3
 

(continued)
 

Low High
 

Topography: Morphology Control Dose Dose
 

All Sites: Hemangiosarcomab 5/49 (10) 1/50 (2) 6/50 (12)
 

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

Relative Risk (Control)f 0.196 1.176
 

Lower Limit 0.004 0.320
 

Upper Limit 1.665 4.565
 

Weeks to First Observed Tumor 85 96 99
 

All Sites: Hemangioma or
 

OC Hemangiosarcoma" 6/49 (12) 2/50 (4) 8/50 (16)
 

d
 P Values0 » N.S. N.S. N.S.
 

Relative Risk (Control)f 0.327 1.307
 

Lower Limit 0.033 0.430
 

Upper Limit 1.723 4.243
 

Weeks to first Observed Tumor 85 96 80
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice 

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

a

Treated groups received doses of 0.375 or 0.75 mg/animal/day.
 

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

c

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

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

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

the comparison of that treated 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 treated group than in the control group.
 

P eThe probability level for departure from linear trend is given when P < 0.05 for any comparison,
 vo
 

95% confidence interval of the relative risk between each treated group and the control group.
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered 3-Nitropropionic Acid by Gavagea
 

Low High
 

Topography; Morphology Control Dose Dose
 

Lung: Alveolar/Bronchiolar
 

Carcinoma** 0/47 (0) 2/49 (4) 2/49 (4)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Control)f Infinite Infinite
 

Lower Limit 0.284 0.284
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 105 105
 

NO Lung: Alveolar/Bronchiolar
 O Adenoma or Carcinoma'5 2/47 (4) 6/49 (12) 3/49 (6)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Control)f 2.878 1.439
 

Lower Limit 0.547 0.173
 

Upper Limit 28.023 16.603
 

Weeks to First Observed tumor 104 105 105
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice 

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

Low
 

Topography: Morphology^ Control Dose
 

Hematopoietic System: Malignant Lymphoma,
 

Undiff erentiated Leukemia, or
 

Lymphocytic Leukemia^ 20/50 (40) 18/50 (36)
 

P Valuesc»d P = 0.021 (N) N.S.
 

Relative Risk (Control)f 0.900
 

Lower Limit 0.516
 

Upper Limit 1.561
 

KJ Weeks to First Observed Tumor 79 84 

Hematopoietic System: All Neoplasmsb 21/50 (42) 21/50 (42) 

P Valuesc»d P = 0.014 (N) N.S. 

Relative Risk (Control)f 1.000 

Lower Limit 0.603
 

Upper Limit 1.659
 

Weeks to First Observed Tumor 79	 84
 

High
 

Dps e
 

10/50 (20)
 

P = 0.024 (N)
 

0.500
 

0.235
 

0.996
 

100
 

10/50 (20)
 

P = 0.015 (N)
 

0.476
 

0.226
 

0.939
 

100
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered 3-Nitropropionic Acid by Gavagea
 

(continued)
 

Low High
 

Tqp_ograj>hy: Morphology Control Dose Dose
 

Liver: Hepatocellular Carcinomab 1/49 (2) 1/50 (2) 2/50 (4)
 

P Valuesc>d N.S. N.S. N.S.
 

Relative Risk (Control)f 0.980 1.960
 

Lower Limit 0.013 0.106
 

Upper Limit 75.404 113.310
 

Weeks to First Observed Tumor 104 105
 —
 

N>
 All Sites: Hemangiosarcoma^ 1/50 (2) 1/50 (2) 3/50 (6)
 S3
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Control)f 1.000 3.000
 

Lower Limit 0.013 0.251
 

Upper Limit 76.970 154.270
 

Weeks to First Observed Tumor 104 105 101
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Administered 3-Nitropropionic Acid by Gavage3
 

(continued)
 

Topography; Morphology


All Sites: Sarcoma of All Kindsb


P Valuesc»d


Relative Risk (Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Pituitary: Chromophobe Adenoma


P Valuesc'd


Relative Risk (Control) ^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Low High 

 Control Dose Dose 

 2/50 (4) 4/50 (8) 6/50 (12) 

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

2.000 3.000 

0.301 0.569 

21.316 29.254 

 104 96 98 

 2/43 (5) 4/48 (8) 1/42 (2) 

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

1.792 0.512 

0.272 0.009 

19.046 9.452 

 104 105 105 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice 

Administered 3-Nitropropionic Acid by Gavagea 

(continued) 

a

Treated groups received doses of 0.375 or 0.75 mg/animal/day.
 

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

c

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

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

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

the comparison of that treated 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 treated group than in the control group.
 

,_• eThe probability level for departure from linear trend is given when P < 0.05 for any comparison.
 NJ
 

*­ f

r

The 95% confidence interval of the relative risk between each treated group and the control group.
 
Review of the Bioassay of 3-Nitroproprionic Acid* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

January 18, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976 under the authority of the National
 

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

bioassay program to identify and 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 organic chemistry, biochemistry, biostatistics, toxicology, pathology, and epide­ miology. Representatives of various Governmental agencies
 

participate as ad hoc members. The Data Evaluation/Risk
 

Assessment Subgroup of the Clearinghouse is charged with the
 

responsibility of providing a peer review of NCI bioassay
 

reports on chemicals studied for carcinogenicity. In this
 

context, below is the edited excerpt from the minutes of the
 

Subgroup's meeting at which 3-Nitroproprionic Acid was
 

reviewed.
 

The primary reviewer briefly described the experimental
 

design under which 3-Nitroproprionic Acid was tested. He
 

noted that there was no marked effect on weight gain or
 

mortality in the treated animals. He agreed with the con­ clusion in the report that 3-Nitroproprionic Acid was not
 

carcinogenic in either sex of mice or female rats, however,
 

he pointed out a dose-related trend in the incidence of
 

hepatic neoplasms and pancreatic islet-cell adenomas. Based
 

on the neoplasms in the treated male rats, the primary
 

reviewer questioned the conclusion that the evidence was
 

insufficient to state that 3-Nitroproprionic Acid was not
 

carcinogenic.
 

. A Program staff member pointed out that there was also
 

a significant increase in the incidence of hepatocellular
 

carcinomas in previous studies where a chemical induced
 

neoplastic nodules and was classified as a carcinogen. In
 

this study only a single hepatocellular carcinoma was found
 

in the treated male rats. Despite the lack of evidence for
 

125
 
the carclnogenicity of 3-Nitroproprionic Acid, he continued,
 

that the benign liver tumors were clearly treatment-related.
 

He pointed out, however, that the biological effect was
 

restricted to one species, one sex, and one organ site.
 

A Subgroup member argued that hyperplastic nodules and
 

carcinomas should be combined for the purposes of analysis,
 

since the former may represent a premalignant lesion. He
 

added that the ratio of hyperplastic nodules to hepatocellular
 

carcinomas is a function of the strength of the carcinogen
 

and the time to tumor detection. Since the ratio of hyper-


plastic nodules to liver carcinomas is higher in the case
 

of 3-Nitroproprionic Acid than for the other organochlorine
 

carcinogens, he concluded that it was not as powerful a
 

carcinogen as the others. Further discussion ensued as to
 

the appropriateness of combining benign and malignant tumors
 

for the purposes of statistical analysis.
 

The secondary reviewer opined that the evidence was
 

inconclusive as to the carcinogenicity of 3-Nitroproprionic
 

Acid in the treated male rats. He pointed out that the
 

chemical was tested at the same time and in the same room
 

with a number of other compounds (some of which were carcinogenic) and, as a result, cross-contamination may have occurred,
 

It was moved that the conclusion in the report be
 

accepted with an addition noting that the hyperplastic
 

nodules, which occurred in a statistically significant
 

Incidence, are generally thought to be premalignant. The
 

motion was seconded and, in further discussion, a Subgroup
 

member objected to combining neoplastic nodules and hepatoceullular carcinomas for the purposes of obtaining a statistically significant result. He opined that this could
 

set a bad precedent for combining benign and malignant tumors.
 

Voting in favor of the motion were Dr. Wolfe, Dr. Highland,
 

Dr. Strong, Dr. Brown, and Mr. Samuels. Those opposed
 

to the motion were Mr. Garfinkel, Dr. Kensler, and Dr. Rowe.
 

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.
 

«US. GOVERNMENT PRINTING OFFICE: 197S 260-839/3043 1-3 126 

DHEW Publication No. (NIH) 78-1302