BIOASSAY OF 3-NITROPROPIONIC ACID FOR POSSIBLE CARCINOGENICITY

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.
Dr.
Dr.
Dr.
Dr.
Dr.
Dr.
Dr.
Dr.
Kenneth C. Chu
Cipriano Cueto, Jr.
J. Fielding Douglas
Dawn G. Goodman
Richard A. Griesemer
Harry A. Milman
Thomas W. Orme
Robert A. Squire8
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 significant­
ly 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
B. Dosage Preparation.
C. Animals
D. Animal Maintenance
E. Subchronic Studies
F. Designs of Chronic Studies
G. Clinical and Pathologic Examinations
H. Data Recording and Statistical Analyses
III. Results -Rats
A.
B.
C.
D.
19
Body Weights and Clinical Signs (Rats)
Survival (Rats)
Pathology (Rats)
Statistical Analyses of Results (Rats)
IV. Results - Mice
A.
B.
C.
D.
3
4
4
5
6
8
11
12
19
19
21
24
27
Body Weights and Clinical Signs (Mice)
Survival (Mice)
Pathology (Mice)
Statistical Analyses of Results (Mice)
27
27
30
32
V.
Discussion
35
VI.
Bibliography
39
APPENDIXES
Appendix A
Table Al
Summary of the Incidence of Neoplasms
in Rats Administered 3-Nitropropionic
Acid by Gavage
43
Summary of the Incidence of Neoplasms
in Male Rats Administered 3-Nitropropionic
Acid by Gavage
45
ix
Table A2
Appendix B
Table Bl
Table B2
Appendix C
Table Cl
Table C2
Appendix D
Table Dl
Table D2
Appendix E
Summary of the Incidence of Neoplasms
in Female Rats Administered 3-Nitropropionic
Acid by Gavage
49
Summary of the Incidence of Neoplasms
in Mice Administered 3-Nitropropionic
Acid by Gavage
53
Summary of the Incidence of Neoplasms
in Male Mice Administered 3-Nitropropionic
Acid by Gavage
55
Summary of the Incidence of Neoplasms
in Female Mice Administered
3-Nitropropionic Acid by Gavage
59
Summary of the Incidence of Nonneoplastic
Lesions in Rats Administered
3-Nitropropionic Acid by Gavage
63
Summary of the Incidence of Nonneoplastic
Lesions in Male Rats Administered
3-Nitropropionic Acid by Gavage
65
Summary of the Incidence of Nonneoplastic
Lesions in Female Rats Administered
3-Nitropropionic Acid by Gavage
74
Summary of the Incidence of Nonneoplastic
Lesions in Mice Administered
3-Nitropropionic Acid by Gavage
83
Summary of the Incidence of Nonneoplastic
Lesions in Male Mice Administered
3-Nitropropionic Acid by Gavage
85
Summary of the Incidence of Nonneoplastic
Lesions in Female Mice Administered
3-Nitropropionic Acid by Gavage
91
Analyses of the Incidence of Primary
Tumors in Rats Administered
3-Nitropropionic Acid by Gavage..
97
Page
Table El
Table E2
Appendix F
Table Fl
Table F2
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
Table 1
Table 2
Design of Chronic Studies of
3-Nitropropionic Acid in Rats
9
Design of Chronic Studies of
3-Nitropropionic Acid in Mice
10
FIGURES
Figure 1
Figure 2
Figure 3
Figure 4
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.
3-Nitropropionic acid
INTRODUCTION
(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
Carter, 1951).
soil
Zealand
area as a food
1960)
1974;
and as a metabolite of certain
fungal species of Aspergillus
Raistrick
(Bell,
It has been isolated from Streptomyces found in
(Anzai and Suzuki,
Kosikowski,
staple
1973)
(Bush
and Penicillium
and Stossl, 1958).
et al., 1951; Iwasaki and
(Hylin
and Matsumoto,
1960;
These species of fungi are commonly
present in several oriental fermented foodstuffs, both domestical­
ly and commercially
been identified
produced, in which 3-nitropropionic acid has
(Kinosita et al. , 1968).
that are frequent contaminants
found
to produce
activity
in
mycotoxins
experimental
Other
fungal
strains
in many kinds of food have been
that
animals
have
exhibited
(Butler
et
carcinogenic
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.
A.
MATERIALS AND METHODS
Chemical
3-Nitropropionic
acid,
synthesized
from
B-propiolactone,
obtained from Aldrich Chemical Co., Milwaukee,
Wisconsin,
was
in a
single batch (Lot No. 111627) for the chronic study.
Analysis at Midwest Research Institute confirmed the identity of
the
chemical.
Infrared
spectra were as
and
expected
nuclear
for
magnetic
3-nitropropionic
resonance
acid,
(nmr)
with
the
exception that the nmr spectra revealed a 5% impurity which was
identified
Elemental
as
a
dimeric
ester
of
analyses
for
carbon
values
for
C3H5N04,
theoretical
and
3-hydroxypropionic
hydrogen
the
agreed
molecular
acid.
with
the
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
detector,
peak
(uv
chromatography
254
nm),
whereas
indicated two trace impurities.
thin-layer
Water content by
Karl Fischer analysis was 0.35 +_ 0.01%.
In summary, the analyses
indicated
the
that
approximately
the
batch
used
for
95% pure, with a single
major
chronic
organic
study
was
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
chronic
Polarographic
extracts
study.
of
samples
administered
of
the
decomposition of the chemical.
and
in feed during the sub-
chromatographic
test
diets
analyses
suggested
of
partial
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
verified
temperature,
as
by both high-pressure
liquid
chromatographic and polarographic analyses.
C.
Rats
Animals
and mice
Division
of each
of Cancer
sex,
obtained
Treatment,
used in these bioassays.
through contracts
National
Cancer
Institute,
the
mice
the
were
The rats were of the Fischer 344 strain
obtained from A. R. Schmidt/Sprague-Dawley, Madison,
and
of
were B6C3F1
hybrids
obtained
from Charles
Breeding Laboratories, Inc., Wilmington, Massachusetts.
and mice were approximately
Wisconsin,
River
The rats
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.
All
Animal Maintenance
animals were housed in temperature- and humidity-controlled
rooms.
The
relative
humidity was maintained
changed
15
fluorescent
meal
temperature was
times
per
lighting
(Allied
Mills,
maintained
hour.
at
at 45-55%.
Illumination
14 hours per day.
22-25°C,
and
the
The room air was
was
provided
by
Wayne® Lab Blox animal
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
Co., Indianapolis, Ind.).
per
cage
in
suspended
wire mesh
(Ford Fence
At week 45, all rats were housed three
polycarbonate cages
(Maryland
Federalsburg, Md.) lined with autoclaved Absorb-Dri
Plastics,
bedding (Lab
Products, Inc., Garfield, N. J.) and equipped with filters and an
automatic watering system.
sanitized
washed,
at 82°C
and
twice
sterilized
The cages were changed, washed, and
per week.
once
per
The feeders were
week,
and
the
changed,
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
incinerated.
changed twice per week and the used cages were
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.
were
rotated
once per week
positions on the racks.
acid
were
housed
5-nitrothiazole
in
(CAS
and
The animal
the cages were kept in fixed
The rats administered 3-nitropropionic
the
same
room
as
rats
fed
2-amino­
121-66-4) and the positive control,
f luoren-2-ylacetamide
racks
(CAS
53-96-3),
in
the
diet.
The
N-9H­
mice
administered 3-nitropropionic acid were housed in the same room
as
(CAS
mice
fed
2-amino-5-nitrothiazole,
1212-29-9),
proflavine
N,N'-dicyclohexylthiourea
hydrochloride
l,3-dichloro-5,5-dimethylhydantoin
fluoren-2-ylacetamide in the diet.
(CAS
(CAS
118-52-5),
952-23-8),
and
N-9H­
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.
fed
All animals were
the chemical for 6 weeks, then observed for 2 weeks.
animals
were
necropsied
and
gross
lesions
were
All
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.
males at 500 and 900 ppm died.
All
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, testicu­
lar 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.
body weights in female mice were not markedly
dose tested.
ppm.
Mean
affected at any
One male died at 600 ppm, and one male died at 800
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
decreased.
groups
increased,
the amounts per unit of body weight
Since water was used
administered
a vehicle
by
as
the
gavage
vehicle,
were
no
control
included.
The
control animals were those started with another chemical on test
at the same
only.
time in the diet; thus, they received control diet
Table 1.
S ex and
Test
Group
Design of Chronic Studies of 3-Nitropropionic Acid
in Rats
Initial
No. of
Animals3
3-Nitropro­
pionic Acid
Dose
(mg/animal/day)k
Time
on Study0
Observed
Dosed
(weeks)
(weeks)
Male
Control
50
Od
Low-Dose
50
0.425
110
1
High-Dose
50
0.85
110
1
Control
50
od
Low-Dose
50
0.6
110
1
High-Dose
50
1.2
110
1
111
Female
111
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.
Sex and
Test
Group
Design of Chronic Studies of 3-Nitropropionic Acid
in Mice
Initial
No. of
Animals3
3-Nitropro­
pionic Acid
Dose
(mg/animal/day)°
Time
on Study0
Observed
Dosed
(weeks)
(weeks)
Male
Control
50
Od
Low-Dose
50
0.375
104
1
High-Dose
50
0.75
104
1
Control
50
Od
Low-Dose
50
0.375
104
1
High-Dose
50
0.75
104
1
104
Female
104
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.
intervals.
Clinical observations
Animals
chat
examination were killed
were
were recorded at weekly
moribund
at
the
tiiae
and necropsied; however,
of
daily
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.
tissues were examined
The following
microscopically: skin, lungs and
bronchi,
trachea, bone marrow, spleen, lymph nodes, thymus, heart, sali­
vary
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.
Occasion­
ally, additional tissues were also examined microscopically.
The
different tissues were preserved in 10% buffered formalin, embed­
ded
in paraffin,
eosin.
sectioned,
and
stained
with
hematoxylin
and
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
cannibalized,
or
that died early.
judged
to
be
in
Also,
such
an
some animals were
advanced
autolysis as to preclude histopathologic evaluation.
number
of animals
state of
Thus, the
from which particular organs or tissues were
examined microscopically varies, and does not necessarily
sent
the number
of animals
that were
repre­
placed on study in each
group.
H.
Data Recording and Statistical Analyses
Pertinent data on this experiment have been recorded in an auto­
matic
data processing
system,
System (Linhart et al., 1974).
the Carcinogenesis
Bioassay Data
The data elements include descrip­
tive information on the chemicals, animals, experimental design,
clinical
observations,
pathologic
results,
as
survival,
body
recommended
Against Cancer (Berenblum, 1969).
by
weight,
and
individual
the International
Union
Data tables were generated for
verification of data transcription and for statistical review.
These
data
described
results
were
analyzed
in this section.
that
bear
on
the
using
the
statistical
techniques
Those analyses of the experimental
possibility
of
carcinogenicity
are
discussed in the statistical narrative sections.
Probabilities
of survival
were
estimated
12
by
the
product-limit
procedure of Kaplan and Meier
report
censored
in
the
form
of
as of the time
(1958) and are presented in this
graphs.
Animals
were
statistically
that they died of other than natural
causes . or were found to be missing; animals
causes were not statistically censored.
dying from natural
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
examin­
ation was required to detect lesions prior to histologic sampling
(e.g.,
skin
or
mammary
tumors),
appeared at multiple sites
or when
lesions
(e.g., lymphomas), the
consist of the numbers of animals
could
have
denominators
necropsied.
The purpose of the statistical analyses of tumor incidence is to
determine whether animals receiving the test chemical developed a
13
significantly
animals.
higher
proportion of tumors than did the control
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
dose level.
compared
that of a group of dosed animals at each
When results for a number of dosed groups (k) are
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.
cases
where
this correction was
narrative section.
used,
it is discussed
In
in the
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
tailed 0.05
curve is different
level of significance.
from zero
at the
one-
Unless otherwise noted, the
direction of the significant trend is a positive dose relation­
ship.
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.
such an early tumor was found, comparisons were based
When
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
incidence of tumors.
methods were
used
to analyze
the
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
methods
of
as
the
comparing
time
point
these
of
curves
tumor
were
observation.
used
for
Cox's
two groups;
Tarone's extension to testing for linear trend was used for three
groups.
used
The statistical tests for the incidence of tumors which
life-table
noted,
in
the
methods
were
direction
of
one-tailed
a
and,
positive
unless
dose
Significant departures from linearity (P < 0.05,
otherwise
relationship.
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
relative risk is defined as Pt/pc where pjprobability
of
the
group.
The
is the true binomial
incidence of a specific type of tumor in a
dosed group of animals
spontaneous
(Gart, 1971).
incidence
and
of
pc
is the
the same
true probability of
type
of
the
tumor in a control
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.
of
Values in excess
unity represent the condition of a larger proportion in the
dosed group than in the control.
The
lower
relative
and upper limits
risk have
analyses.
The
of
the confidence
been included
interval of
in the tables
the
of statistical
interpretation of the limits is that in approxi­
mately 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
experiment.
greater
When the lower limit of the confidence interval is
than
significant
from the
one,
result
it
can
be
inferred
that
(P < 0.025 one-tailed
a
statistically
test when the control
incidence is not zero, P < 0.050 when the control incidence
zero) has occurred.
is
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
16
while
the
upper
limit
indicates
that
there
is
a
theoretical
possibility
induction of tumors by the test chemical, which
detected under the conditions of this test.
17
of
the
could not be
III.
A.
RESULTS - RATS
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
glands.
The
clinical appearance was consistent
sialodacryoadenitis.
this
salivary
with that of
Both control and dosed animals developed
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.
The
Survival (Rats)
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
il
D D
sii
o
j. 250­
Q
O
CD
Z 200­
<
111
MALE RATS
5
[] CONTROL
O LOW DOSE
A HIGH DOSE
40
—1—
50
—I—
60
—I—
90
70
TIME ON STUDY (WEEKS)
n 11
h­
gi 8
a
>• 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.
male
rats,
low-dose
30/50
(60%)
of
the
controls,
group, and 30/50 (60%)
the last week of
the study.
controls, 26/50 (52%)
33/50
(66%)
of
the
of the high-dose group lived to
In females,
33/50
(66%)
of
the
of the low-dose group, and 32/50 (64%) of
the high-dose group survived to the last week of the study.
sufficient
number
In
of
rats
of
each
sex
was
at
risk
for
A
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
groups.
occurred in both the control and dosed
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
high-dose
in males:
5/49
(10%).
observed in 1/50
controls
0/49
(0%),
low-dose
3/50
(6%),
In female rats, neoplastic nodules were
(2%) of each dosed group, but in none of
controls.
21
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PROBABILITY OF SURVIVAL
The
incidence of pancreatic islet-cell
in males
(controls
4/49
adenoma was dose related
[8%] , low-dose
6/50
[12%],
11/50 [22%]).
This trend was not evident in females.
Nonneoplastic
lesions
consisted
of degenerative,
high-dose
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
17/49
(35%)
low-dose
males,
(4%) control females, 9/46
24/48
(20%)
(50%)
(2%)
control males,
high-dose
males;
2/48
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,
(30%)
control females,
and
36/50
17/50
(72%)
(34%)
high-dose
low-dose
males;
15/50
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
positive dose-related
animals
with
carcinoma
of
the
Cochran-Armitage
test
for
trend in combined tumor incidences of those
either
the
of
neoplastic
liver
results of the Fisher
are
nodules
significant
or
(P =
hepatocellular
0.010) and
the
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
pancreatic
islets
of
the
in
incidence of islet-cell adenoma of the
male
rats,
the
result
Armitage test is significant (P = 0.033).
24
of
the
Cochran-
The Fisher exact test
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.
incidence
of
7/100
The laboratory historical controls have an
(7%)
of islet-cell adenoma.
No
significant
incidence of islet-cell adenoma is obtained for the females, and
no
islet-cell
tumors
carcinoma was
observed
appeared in significant
in either sex.
incidences
when compared with the control groups.
25
No other
in the dosed groups
IV.
A.
RESULTS - MICE
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
body
weight
study,
controls
the
may
be
dosed
in
subject
animals
appearance
to wide
were
and
diminishes,
variation.
generally
behavior.
the
Throughout
comparable
Focal
mean
to
alopecia,
the
the
focal
dermatitis, and small palpable nodules in the perineal area were
observed
study.
B.
The
in increasing
numbers
of male mice after 7 months on
These lesions were associated with fighting.
Survival (Mice)
Kaplan
survival
and
Meier
curves
estimating
the
probabilities
for male and female mice administered
of
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,
low-dose group, and 38/50 (76%)
36/50 (72%)
of the high-dose
27
of the
group lived to
D D
AAA
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40­
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Dgt
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s>
MALE MICE
D CONTROL
O LOW DOSE
A HIGH DOSE
40
50
60
70
TIME ON STUDY (WEEKS)
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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
ft^^­
40
50
60
~7^-
70
80
^ra
1
90
tO****
°1
100
110
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m
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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.
43/50 (86%)
In females, 35/50 (70%) of the controls,
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
Appendix B, tables Bl and B2; findings
on nonneoplastic
in
lesions
are summarized in Appendix D, tables Dl and D2.
A variety of neoplasms
groups.
occurred in both
the control
and dosed
Each of the types of neoplasms represented in the tables
has been encountered
previously
as a spontaneous lesion
in the
mouse.
The
incidences
adenomas,
and
of
hepatocellular
hyperplastic
lesions
carcinomas,
(nodular
hepatocellular
hyperplasia
and
hyperplastic nodule) of the liver in mice are summarized below:
30
MALES
Low
Dose
Control
Number of
animals with
tissues
examined
microscop­
49
ically
50
Hepato­
cellular
carcinoma
16 (33%)
Hepato­
cellular
adenoma
Hyper-
plastic
lesions
The
Low
Dose
Control
49
49
50
High
Dose
50
8 (16%) 12 (24%)
1 (2%)
1 (2%)
2 (4%)
4^(8%)
2 (4%)
4 (8%)
1 (2%)
0 (0%)
2 (4%)
1 (2%)
1 (2%)
0 (0%)
0 (0%)
0 (0%)
0 (0%)
incidences
greater
FEMALES
High
Dose
in
of
males
proliferative
than
in
hepatocellular
females;
however,
lesions
there
was
were
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,
conditions were observed in all groups.
considered
to
be
of
common
occurrence,
These
proliferative
conditions
spontaneous,
related to administration of the test chemical.
31
and
and
were
not
In the judgment of the pathologist, 3-nitropropionic
carcinogenic
in
B6C3F1
mice
when
acid was not
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
Fisher
exact
test
positive direction.
are
significant
at
In two instances
the 0.05
level
the
in the
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
direction
the
control group.
cannot
These
results
be explained by differential
in
the
negative
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
32
the
absence of significant positive results.
that each of the intervals
hepatocellular
It should also be noted
(except that for
adenoma and carcinoma
the incidence of
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.
Mean
body
markedly
weights
and
DISCUSSION
mortality of
the
dosed
rats were
not
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
(controls 0/49,
low-dose
3/50,
high-dose 6/49).
these tumors were neoplastic nodules.
neoplastic
nodules
occurred,
than in the
one
All but one of
In the females, only two
in each
of
the dosed groups.
Biliary hyperplasia occurred at a higher incidence
males
than
low-dose
lesion
in
32/50,
the
corresponding
high-dose
controls
36/50),
controls
but
the
in the dosed
(controls
incidence
in the dosed females was not increased
19/50,
of
this
as compared with
controls.
There was also a dose-related trend (P = 0.033) in the
incidence
of
(controls 4/49,
comparisons
not
pancreatic
islet-cell
adenoma
low-dose 6/50, high-dose
in
the
male
rats
11/50); however, direct
of incidences in the dosed and control groups were
statistically
significant.
The
35
historical
incidence
of
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.
specific tumor was found to occur at a statistically
ly
higher
No
significant­
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).
long-term
There have
toxicity studies of this chemical.
been
no
previous
The compound first
attracted attention when Morris et al.
(1954) found that it was
present
(Indigofera
in a potential pasture legume
grown in tropical countries.
grazing
animals
3-nitropropionic
leaves
of
the
and
acid.
legume
the
This
legume was severely toxic to
toxic
principle
Hutton et al.
and
also
rabbits and found the leaves
endecaphylla)
pure
was
to
be
(1958), however,
fed the
3-nitropropionic
acid
caused severe liver
the pure acid had no effect on the liver.
36
thought
damage,
to
while
3-Nitropropionic acid
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,
3-nitropropionic
there
was
no
conclusive
acid was carcinogenic
evidence
that
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
identified
as
S-nitropropionic
JJH2): 133-136, 1960.
antibiotic bovinocidin,
acid.
J. Antibiotics
Armitage, P., Statistical Methods in Medical Research, John Wiley
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Bell,
M.
E., Toxicology
3-nitropropionic acid.
1974.
of karaka
kernel, karakin, and
New Zealand J. Sci. 17:327-334,
Berenblum, I., ed., Carcinogenicity Testing, A_ Report of the
Panel on Carcinogenicity of the Cancer Research Commission
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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.
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Carter, C. L. , The constitition of karakin.
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Cooke, A. R., The toxic constituent of Indigofera endecaphylla.
Arch. Biochem. Biophys. 5^:114-120, 1955.
Cox, D. R., Regression models and life tables.
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Cox, D. R., Analysis of Binary Data, Methuen & Co., Ltd., London,
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Investigations 23(3);327-334. 1970.
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toxic
extract
of
Indigofera
endecaphylla.
The
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a review of
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Toxicity for
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Hylin,
J.
W.
and
Matsumoto,
H. ,
The
biosynthesis
of
3-nitropropionic acid by Penicilluim
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IARC Monographs on
the Evaluation of the Carcinogenic Risk of Chemicals to Man,
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Statist.
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R.
G. ,
Jr., Simultaneous
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McGraw-Hill Book Co., New York, 1966, pp. 6-10.
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Metabolites of Pencillium atrovenetum G.
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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
HIGH DOSE
LOW DOSE
50
50
50
50
50
50
50
50
50
*SKIN
SQUAMOUS CELL PAPILLOMA
BASAL-CELL CAECINOMA
TRICHOEPITHELIOMA
(50)
1 (2%)
(50)
1 (2ft)
2 (4X)
(50)
*SOBCUT TISSU2
SQUAMOUS CELL CARCINOMA
BASAL-CELL CARCINOMA
FIBROMA
FIBROSARC0.1A
LIPOMA
MESENCHYBOHA, BENIGN
(50)
(50)
1 (2ft)
1 (2X)
1 (2%)
(50)
ANIMALS INITIALLY IN STUDY
ANIHALS NECROPSIED
ANIMALS EXAMINED HISTOPATHOLOGIC ALLY
I N T E G U M E N T A R Y SYSTEM
RESPIRATORY
1
(2%)
1 (2%)
1 (2%)
1 12%)
1 (2ft)
1 (2%)
1
(2%)
1 (2%)
SYSTEM
tLUNG
SQUAMOUS CELL CARCINOMA, METASTA
ALVEOLAR/BRONCHIOLAR A D E N O M A
ALVEOLAR/B20NCHIOLAR CARCINOMA
CORTICAL CARCINOHA, METASTATIC
C-CELL CARCINOMA, METASTATIC
(50)
3
(49)
(48)
1 (2ft)
1 (2ft)
1 (2X)
(6%)
1
1 (2%)
3
1
(6ft)
(2ft)
(2*)
HEMATOPOIETIC SYSTEM
*HULTIPLE O R G A N S
FER-TYPE
M A L I G . L Y M P H O M A , ON
M A L I G . L Y M P H O M A , LY
OCYTIC TYPE
U N D I F F E R E N 1 I A T E D L E D KEHIA
EHIA
LYMPHOCYIIC LEUKEMIA
GRANULOCYTIC LEUKEMIA
GRAfi.2kOCYTIC_SARCOMA_
(50)
1 (2%)
4 (8%)
2 (4X)
4 (8ft)
2 (4%)
(50)
1 (2%)
# N U M B E R O F A N I M A L S WITH T I S S U E E X A M I N E D M I C R O S C O P I C A L L Y
* N U M B E R OF A N I M A L S NECROPSIED
45
4
(8ft)
(50)
7
4
(14%)
(8X)
1 (2ft)
1 (2%)
2 (4»)
2 (4%)
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
(49)
HIGH DOSE
•S P L E E N
MALIG.LYMPHOMA. UNDIFFER-TYPE
H A L I G . L Y M P H O M A , HISTIOCYTIC TYPE
UNDIFFERENTIATED LEUKEMIA
(49)
1 (2K)
(49)
1 (2%)
#HEDIASTINAL L.NODE
A L V E O L A R / B R O N C H I O L A 8 CA,
(41)
(43)
1 (2%)
(43)
(48)
(49)
1 ( 2 % )
( 4 8 )
(50)
1 (2%)
1 (2%)
METASTA
CIRCULATORY SYSTEM
tHEART
HEMANGIOMA
ANITSCHKOW-CELL SARCOMA
1
( 2 X )
DIGESTIVE SYSTEM
*PALATE
SQUANOUS CELL CARCINOMA
(50)
1 (2%)
(50)
ILIVEH
(49)
(50)
NEOPLASTIC NODULE
HEPATOCELLULAR CARCINOMA
3 (6%)
(49)
5 (10%)
1 (2%)
URINARY SYSTEM
#URINARY BLADDER
TRANSITIONAL-CELL PAPILLOMA
(47)
(42)
1 (2X)
( 4 5 )
•PITUITARY
CARCINOMA,NOS
CHROHOPHOBE ADENOMA
(46)
(48)
(49)
•ADRENAL
CORTICAL CARCINOMA
PHEOCHROBOCYTOHA
PHEOCHROMOCYTOMA, M A L I G N A N T
(49)
1 (2%)
(50)
4 (8X)
1 (2%)
5
•THYROID
IOLLICULAR;CELL_CARCINOMA
(46)
ENDOCRINE SYSTEM
1
5
3 (7%)
t NUMBER OF ANIMALS WITH TISSUE EXAMINED
* NUMBER OF ANIMALS NECHOPSIED
(49)
MICROSCOPICALLY
46
(2%)
(10X)
4 (8%)
(10«)
(50)
1 (2%)
5 (10%)
(47)
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
3 (7%)
1 (2%)
C-CELL A D E N O M A
C-CELL C A R C I N O M A
8 (16%)
2 (4%)
HIGH DOSE
4 (9%)
(49)
4 (8%)
(50)
*MAMMARY GLAND
FIBBOADENOMA
(50)
1 (2%)
(50)
(50)
*PREPUTIAL GLAND
C A R C I N O M A , NOS
ADENOMA, NOS
(50)
1 (2%)
2 (4%)
(50)
(50)
HESTIS
INTERSTITIAL-CELL TUMOR
(50)
48 ( 9 6 % )
(49)
44 ( 9 0 % )
(«9)
(50)
(50)
1 ( 2 % )
(50)
(50)
(50)
(50)
1 (2%)
(50)
(50)
1 ( 2 % )
(50)
tMIDBRAIN
ASTROCYTOMA
(50)
1 (2%)
(50)
(50)
tCEHEBELLUM
ASTROCYTOMA
(50)
(50)
(50)
1 (2%)
(50)
(50)
1 (2%)
(50)
(50)
(50)
(50)
t P A N C R E A T I C ISLETS
ISLET-CELL ADENOMA
6 (12%)
(50)
11 ( 2 2 % )
R E P R O D U C T I V E SYSTEM
1 (2%)
4 (8%)
48 (98%)
NERVOUS SYSTEM
tBRAIN/IENINGES
S Q U A M O U S CELL C A R C I N O M A ,
METASTA
tCEREBRUM
ASTROCYTOMA
#BRAIN
S Q U A M O U S CELL C A R C I N O M A ,
METASTA
SPECIAL SENSE ORGANS
*EAR C A N A L
S Q U A M O U S CSLL
CARCINOMA
MOSCOLOSKELETAL SYSTEM
*SKELETAL MUSCLE
LIPOMA
* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
47
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)
CONTROL
BODY
LOW DOSE
HIGH DOSE
CAVITIES
*PEHITONEUM
MESOTHELIOMA, NOS
(50)
1 (2X>
(50)
3 (6»)
(50)
(50)
(50)
50
50
12
11
ALL OTHER SYSTEMS
*HULTIPLE O R G A N S
FIBROUS HISTIOCITOMA, M A L I G N A N T
MESOTHELIOMA, HALIGNANT
(50)
1 (2%)
1 (2%)
A N I M A L DISPOSITION S U M M A R Y
ANIMALS INITIALLY IN STUDY
N A T U R A L DEATHS
MORIBUND SACRIFICE
SCHEDULED SACRIFICE
ACCIDENTALLY KILLED
TERMINAL SACRIFICE
A N I M A L MISSING
50
15
5
30
32
27
TOTAL A N I M A L S WITH P8IMABY TUMORS*
TOTAL P R I M A R Y TUMORS
49
99
49
49
112
TOTAL ANIMALS WITH BENIGN TUMORS
TOTAL BENIGN TUMORS
48
72
48
76
48
TOTAL A N I M A L S WITH M A L I G N A N T TUMORS
TOTAL MALIGNANT TUMORS
23
26
14
17
24
1
3
1
d INCLUDES AUTOLYZED ANIMALS
TUMOR S U M M A R Y
TOTAL A N I M A L S WITH SECONDARY TUMORS*
TOTAL SECONDARY TUMORS
99
1
TOTAL A N I H A L o WITH TUMORS UNCERTAIN­
BENIGN
O
R MALIGNANT
TOTAL UNCERTAIN TUMORS
29
5
1
1
78
1
6
6
5
5
TOTAL A N I M A L S WITH TUMORS UNCERTAIN­
PRIMARY OR MSTASTATIC
TOTAL UNCERTAIN TUMORS
* P R I M A R Y TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
* SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT O R G A N
48
TABLE A2.
*
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS
ADMINISTERED 3-NITROPROPIOWC ACID BY GAVAGE
CONTROL
LOW DOSE
HIGH DOSE
50
50
50
50
50
50
50
50
50
*SKIN
P A P I L L O M A , NOS
SEBACEOUS A D E N O M A
(50)
(50)
(50)
*SUBCUT TISSUS
SQUAMOI1S CSLL C A R C I N O M A
(50)
(50)
(50)
1 (2X)
(50)
(49)
(1*9)
1 (2*)
1 (2%)
(50)
1 (2%)
1 (2S)
2 (U%)
(50)
iNIHALS INITIALLY IN STUDY
ANIHALS NECROP3IED
iNIHALS EXAMINED HISTOPATHOLOGIC ALLY
I N T E G U M E N T A R Y SYSTEM
1 (2%)
1 (2%)
RESPIHATORY SYSTEM
tLUNG
S Q U A M O U S CSLL C A R C I N O M A , M E T A S T A
C-CELL C A R C I N O H A , M E T A S T A T I C
LIPOSARCOMA, HETASTATIC
HEMATOPOI5TIC
1
(2%)
SYSTEM
*HULTIPLE ORGANS
M A L I G N A N T LYMPHOMA, NOS
MALIG. LYMPHOMA, UN DIFPES-TYP E
MALIG. LYMPHOSA, LYMPHOCYTIC TYPE
LEUKEMIA, NOS
LYMPHOCYTIC LEUKEMIA
GRANULOCYTIC LEUKEMIA
MONOCYTIC LEUKEMIA
(50)
*SPLEEN
MALIG. LYMPHOMA, UNDIFFER-TY PE
GRANULOCYTIC LEUKEMIA
(50)
(50)
1 (2%)
1 (2%)
(49)
1 (2%)
#LYMPH NODE
(44)
(41)
(45)
4 (8%)
1 (2%)
1 (2X)
2 (HI)
1 (2%)
1 (2%)
* NUMBER OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
* N U M B E R OF ANIMALS NECROPSIED
49
4 (3%)
2 (4%)
1 (2%)
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED)
CONTROL
#MANDIBULAR L. NODE
SQUANOUS C3LL CARCINOMA, METASTA
LOW DOSE
HIGH DOSE
(44)
(41)
(45)
1 (2%)
*TONGUE
SQUAMOUS CBLL CARCINOMA, METASTA
(50)
(50)
1 (2*)
(50)
•LIVER
NEOPLASTIC NODULE
(49)
(50)
1 (2%)
(50)
1 (2%)
(49)
(50)
1 (2%)
(50)
#PITUITARY
CHROMOPHOBE ADENOMA
(45)
19 (4295)
(46)
15
(47)
20 (43%)
•ADRENAL
PHEOCHROMOCYTOMA
(49)
3
(50)
1 (2%)
(49)
1 (2%)
tTHYROID
FOLLICULAR-CELL ADENOMA
FOLLICULAR-CELL CARCINOMA
C-CELL A D E N O M A
C-CELL CARCINOMA
(50)
(44)
(44)
1 (2%)
#PANCREATIC ISLETS
ISLET-CELL ADENOMA
(49)
1 (2X)
(49)
(47)
1 (2%)
(50)
(50)
(50)
CIRCULATORY SYSTEM
HONE
DIGESTIVE SYSTEM
U R I N A R Y SYSTEM
IKIDNEY
MIXED TUNOH, MALIGNANT
ENDOCRINE SYSTEM
(6%)
1 (2%)
3 (6X)
2 (4%)
(33%)
3 (7*)
1 (2%)
3 (7%)
2 (5%)
EEPHODUCTIVE SYSTEM
* M A M M A R Y GLAND
AD E NOM A ^NO S
I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
50
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED)
CONTROL
ADENOCARCINOMA,
FIBROADENOHA
NOS
1
12
LOW DOSE
(2%)
(2<*X)
14 (2d%)
(50)
HIGH DOSE
3 (6X)
13 (26%)
*PREPUTIAL GLAND
CARCINOMA,NOS
ADENOMA, NOS
(50)
1 (2%)
#U-TEBUS
LEIOBYOMA
E N D O M E T R I A L STROMAL P O L Y P
(50)
ICERVIX UTEBI
FIBROSARCOMA
(50)
(48)
(49)
1 (2%)
tOVARY
SERTOLI-CELL TUHOR
(50)
1 (2X)
(47)
(48)
(49)
(50)
1 (2%)
(50)
(50)
(50)
(50)
1 (2%}
(50)
(50)
1 (2%)
(50)
(50)
(50)
(50)
1 (2%)
2
2
(50)
(>4%]
(49)
(48)
1 (2%)
2 (14%)
4
(8X)
5 (10%)
HERVOUS SYSTEM
tCEREBRUM
OLIGODENDROGLIOMA
SPECIAL SENSE O R G A N S
*ZYMBAL'S GLAND
S Q U A M O O S CELL C A R C I N O M A
BUSCULOSKELETAL SYSTEM
*HftNDIBLE
SQUAHOUS CELL C A R C I N O M A
BODY CAVITIES
*BESENTERY
FIBROSARCOMA
fcLL OTHER SYSTEMS
LOBBOSAC6AL REGION
L£POSARCOMA
1
• NUMBER OF ANIMALS WITH TISSUE E X A M I N E D MICROSCOPICALLY
* H U H B E R OF ANIMALS NECROPSIED
51
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
ANIMAL DISPOSITION SUMMARY
13
50
15
10
33
25
30
TOTAL ANIMALS WITH PRIMARY TUMORS*
TOTAL PRIMARY TUMORS
40
59
30
50
39
66
TOTAL ANIMALS WITH BENIGN TUMORS
TOTAL BENIGN TUMORS
35
45
27
39
29
46
TOTAL ANIMALS WITH MALIGNANT TUMT3RS
TOTAL MALIGNANT TUMOES
11
10
10
19
19
1
2
ANIMALS INITIALLY IN STUDY
NATURAL DEATHS
MORIBUND SACRIFICE
SCHEDULED SACRIFICE
ACCIDENTALLY KILLED
TERMINAL SACRIFICE
ANIMAL HISSING
50
4
50
9
11
INCLUDES AOTOLYZED ANIMALS
TUMOR S U M M A R Y
TOTAL ANIMALS WITH SECONDARY TUMORSt
TOTAL SECONDARY TUMORS
14
3
3
TOTAL ANIMALS WITH TUMOBS UNCERTAIN­
BENIGN OP. MALIGNANT
TOTAL UNCERTAIN TUMORS
1
1
3
1
1
1
TOTAL ANIHALS WITH TUMORS UNCERTAIN­
PRIMARY OR METASTATIC
TOTAL UNCERTAIN TUMORS
* 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
ANIHALS INITIALLY IN STUDY
A N I H A L S NECROPSIED
ANIMALS EXAMINED HI STOPATH OLOGIC ALLY
LOW DOSE
HIGH DOSE
50
50
50
50
49
49
50
50
50
(49)
(50)
I N T E G U M E N T A R Y SYSIEM
*SUBCUT TISSUE
FIBROMA
FIBROSARC05U
HEMANGIOMA
HEMANGIOSARCOMA
(50)
1 (2*)
2 ((*»)
1 (2*)
2 (4%)
1 (2%)
RESPIRATORY SYSTEM
#LUNG
HEPATOCELLULAR CARCINOMA. METAST
ALVEOLAR/BRONCHIOLAR A D E N O M A
ALVEOLAR/BRONCHIOLAR CARCINOMA
CORTICAL CARCINOMA, BETASTATIC
FIBROSARC051A, METASTATIC
(49)
3
10
4
1
(43)
3 (6%)
5 (10%)
3 (6*)
(6S)
(20%)
(8%)
(2%)
(50)
8 (16X)
3 (6%)
1 .12*)
HEMATOP01ETIC SYSTEM
(50)
*MULTIPLE ORGANS
MALIGNANT LYMPHOMA, NOS
MAL1G.LYMPHOMA, LYMPHOCYTIC TYPE
M A L I G . L Y M P H O M A , HISTI3CYTIC T Y P E
LYMPHOCYTIC LEUKEMIA
GRANOLOCYTIC LEUKEMIA
MONOCYTIC L E U K E M I A
GEANULOCYTIC SARCOMA
(49)
*SUBCUT TISSUE
MAST-CELL TUMOR
(49)
(50)
(50)
1 (2%)
#SPLEEN
(46)
(50)
(46)
(50)
1 (2%)
5 (10%)
4 (8%)
2 (4%)
1 (2«)
2 (4%)
1 (2X)
4 (8%)
1 (2S)
1(2%)
1 (2%)
t N U M B E R OF A N I M A L S WITH TISSUE EXAMINED MICROSCOPICALLY
* N U M B E R OF ANIMALS NECROPSIED
55
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED)
CONTROL
4
HEMANGIOSAHCOMA
MALIG.LYMPHOMA, HISTIOCYTIC TYPE
LOW DOSE
HIGHD>OSE
2
(«*)
1
(2%)
(9X)
1 (2%)
(30)
#MESENTERIC L. NODE
MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
(40)
1 (3S)
(31)
tLIVER
GRANOLOCYTIC LEUKEMIA
(49)
(50)
(49)
tPEYERS PATCH
MALIGNANT LYMPHOMA, NOS
MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
(47)
(49)
(49)
1
(2%)
3
(6*)
#THYMUS
M A L I G N A N T LYMPHOMA, NOS
(35)
(38)
1 (3X)
(41)
JLIVER
HEPATOCELLULAR ADENOMA
HEPATOCELLULAR CARCINOMA
CORTICAL CARCINOMA, METASTATIC
HEMANGIOMA
HEMANGIOSARCOMA
ANGIOSARCOMA
(49)
4 (8%)
16 (33X)
(50)
2 (4£)
8 (16%)
(49)
4
(8%;
12
(24*)
tSTOMACH
ADENOMATOUS POLYP, NOS
(48)
(50)
(46)
1
(2%)
(46)
(49)
(50)
1
(3X)
1 (2%)
1
(2X)
CIRCULATORY SYSTEM
NONE
DIGESTIVE SYST2M
1 (2%)
1 (2%)
2 (H%)
1
(2*)
4
(8?)
1
(2%)
U R I N A R Y SYSTEM
N O N E
ENDOCRINE SYSTEM
fADRENAL
CORTICAL_CARCINOMA_
# NUMBER OF ANIMALS WITH TISSUE EXAMINED
* N U M B E R OF ANIMALS NECROPSIED
MICROSCOPICALLY
56
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED)
CONTROL
ttHYBOID
FOLLICULAR-CELL ADENOMA
LOW DOSE
HIGH DOSE
(US)
(44)
(<*8)
1 (2%)
(<»7)
(49)
(50)
(50)
(50)
REPRODUCTIVE SYS1EM
#TESTIS
INTEBSTITIAL-CELL TDBOR
1 (2*)
NERVOUS SYSTEM
HONE
SPECIAL
SENSE ORGANS
*HABDERIAN GLAND
PAPILLARY ADENOMA
PAPILLARY CYSTADENOMA, NOS
(49)
1 (2*)
1 (2S)
2 («*)
HUSCULOSKELETAL SYSTEM
*SKULL
(U9)
(50)
OSTEOMA
(50)
1 (2*)
BODY CAVITIES
*ABDOHINAL CAVITY
CORTICAL CARCINOMA, METASTATIC
(49)
1 (2*)
(50)
ALL OTHER SYSTEMS
THORAX
FIBROSARCOMA, HETASTATIC
1
DIAPHRAGM
IIBRQSAJBCQMA J ._METASTAIIC
1
t NUMBER OF ANIBALS WITH TISSUE EXAMINED MICROSCOPICALLY
* N U M B E R Of ANIMALS NECBOPSIED
57
(50)
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
ANIMAL DISPOSITION SUflMABY
50
10
2
50
11
3
50
12
38
36
38
TOTAL ANIMALS WITH PRIMARY TUMORS*
TOTAL PRIMARY TUMORS
39
54
28
39
38
55
TOTAL ANIMALS WITH BENIGN TUMORS
TOTAL BENIGN TUMORS
15
17
9
11
19
19
TOTAL ANIMALS WITH MALIGNANT TUMORS
TOTAL MALIGNANT TUMORS
31
37
24
28
27
35
4
6
4
ANIMALS INITIALLY IN STUDY
NATURAL DEATHS
MORIBUND SACRIFICE
SCHEDULED SACRIFICE
ACCIDENTALLY KILLED
TERMINAL SACRIFICE
ANIMAL MISSING
» INCLUDES AUTOLYZED ANIMALS
TUMOR S U M M A R Y
TOTAL ANIMALS WITH SECONDARY TUMORS*
TOTAL SECONDARY TUMORS
TOTAL ANIMALS WITH TUMORS UNCERTAIN­
BENIGN 08 MALIGNANT
TOTAL UNCERTAIN TUMORS
6
1
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
A N I M A L S INITIALLY IN STUDY
ANIMALS NECROPSIED
ANIMALS EXAMIN2D HISTOPATHOLOGICALLY
LOW DOSE
HIGH DOSE
50
50
50
50
50
50
50
50
50
INTEGUMENTARY SYSTEM
*SUBCUT TISSUE
FIBROSARCOMA
(50)
(50)
1
(21)
(50)
2
(US)
(49)
1
2
1
1
(2X)
(4X)
(2%)
(2S)
(50)
7
(14%)
(50)
1
(2%)
8ESPIRATORY SYSTEM
tLUNG
ALVEOLAR/BRONCHIOLAR A D E N O M A
ALVEOLAR/BHONCHIOLAR CARCINOMA
FIBROSARCOMA, METASTATIC
OSTEOSARCOMA
HBHATOPOIETIC
(47)
2 (4%)
l'»3)
4
(ax/
2
(4%)
SYSTEM
(50)
*MOLTIPLE O R G A N S
8ALIG.LYMPHOMA, LYMPHOCYTIC TYPE
M A L I G . L Y M P H O M A , HISTIOCYTIC T Y P E
UNDIFFEBENTIATED LEDKEMIA
LYHPHOCYTIC L E U K E M I A
GBANULOCYTIC LEUKEMIA
G R A N U L O C Y T I C SARCOHA
(50)
1 1 (22%)
6 (12%)
tBONE MARROW
GRANULOCYTIC SARCOMA
(46)
•SPLEEN
HEMANGIOSAHCOMA
M A L I G . L Y M P H O M A , LYMPHOCYTIC T Y P E
HALIG. L Y M P H O H A , HISTIOCYTIC T Y P H
(47)
(50)
1 (2%)
(50)
3 (6%)
1 (2%)
• L Y M P H N O D E
GRANULOCYTIC_SARCOMA
(38)
(36)
(33)
2
1
10
2
(4%)
(2%)
(4X)
2 (4*)
1 (2«)
(4b)
* NUMBER OF A N I M A L S WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
59
(20%)
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
t M E S E N T E R 1 C L. NODE
M A L I G . L Y M P H O M A , HISTIOCYTIC T Y P E
(38)
1 (3%)
(36)
(33)
1 (3%)
*PEYERS PATCH
MALIG.LYMPHOHA, LYMPHOCYTIC TYPE
(48)
(48)
3 (6*)
(49)
1 (2%)
(49)
1 (2%)
1 (2%)
1 (2*)
(50)
\ (2%)
(50)
2 (4%)
2 (4%)
tPITOITARY
CH80MOPHOBE ADENOMA
(43)
2 (5X)
(48)
4 (8%)
(42)
1 (2%)
#ADRENAL
CORTICAL CARCINOMA
PHEOCHROMOCYTOMA
(48)
(50)
(49)
1 (2«)
1 (2%)
tTHYfiOID
FOLLICULAR-CELL ADENOMA
(40)
(47)
(45)
1 (2%)
*MAMBARY GLAND
A D E N O C A R C I N O M A , NOS
FIB80ADENOMA
(50)
(50)
1 (2)5)
1 (2X)
(50)
1 (2%}
*VAGINA
SOJ&MOUS_CfiLL_CARCINOflA
(50)
(50)
(50)
CIRCULATORY SYSIEfl
NONE
DIGESTIVE SYSTEM
tLIVER
HEPATOCELLULAR ADENOMA
HEPATOCELLULAR CARCINOMA
HEMANGIOSARCOMA
U R I N A R Y SYSTEM
NONE
ENDOCRINE
SYSTEM
REPRODUCTIVE SYSTEM
* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
60
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
•UTERUS
LEIOMYOSARCOBA
(47)
2 (4)5)
(49)
1 (2%)
IOVARY
PAPILLARY CYSTADENOMA, NOS
GRANULOSA-CELL TUMOR
(39)
(47)
(47)
1 (2%)
1 (2%)
(47)
(50)
(50)
1 (2%)
(50)
(50)
(50)
1 (28)
(50)
(50)
1 (2%)
(50)
1 (3X)
(48)
NERVOUS SYSTEM
tBRAIN/HENINGSS
OSTEOSARCOHA, METASTATIC
SPECIAL SENSE ORGANS
*HARDERIAN G L A N D
PAPILLARY CYSTADENOMA, NOS
HUSCULOSKELETAL SYSTEM
*SKELETAL MUSCLE
FIBROSARCOBA
BODY CAVITIES
NONE
ALL OTHER SYSTEMS
DIAPHRAGM
OSTEOSARCOMA, METASTATIC
1
ANIMAL DISPOSITION SUMMARY
ANIMALS INITIALLY IN STUDY
N A T U R A L DEATH2
MORIBUND SACRIFICE
SCHEDULED SACRIFICE
ACCIDENTALLY KILLED
TERMINAL SACRIFICE
ANIMAL MISSING
50
14
1
50
35
d_IlCLUDES_AUTj}LYZEp_ANIMAI.S
* NUMBER,OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
61
7
50
11
43
39
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
TUMOR SUMMARY
TOTAL ANIMALS WITH PRIMARY TUMOBS*
TOTAL PRIMARY TUMORS
TOTAL ANIMALS WITH BENIGN TUMORS
TOTAL BENIGN TUMORS
TOTAL ANIMALS WITH MALIGNANT TUMORS
TOTAL MALIGNANT TUMORS
31
32
39
26
33
3
5
9
9
8
8
23
25
25
30
19
24
26
TOTAL ANIMALS WITH SECONDARY TUMORS*
TOTAL SECONDARY TUMORS
TOTAL ANIHALS WITH TUMORS UNCEKTAIN­
BENIGN OR MALIGNANT
TOTAL UNCERTAIN TUMORS
2
3
1
1
1
1
TOTAL ANIMALS WITH TUMORS UNCERTAIN­
PRIMARY 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
ANIMALS INITIALLY IN STUDY
ANIMALS NECROPSIED
AHIHALS EXAMINED HISTOPATHOLOGICALLY
50
50
50
LOW DOSE
HIGH DOSE
50
50
50
50
50
50
(50)
(50)
INTEGUMENTARY SYSTEM
*SKIN
CYST, NOS
EPIDERMAL INCLUSION CYST
HYPERKERATOSIS
ACANIHOSIS
(50)
1
(2%)
2
(4%)
*SUBCUT TISSUE
EPIDERMAL INCLUSION CYST
(50)
(50)
(50)
1 (2%)
*NASAL CAVITY
INFLAMMATION, CHRONIC
(50)
(50)
(50)
#TRACHEA
INFLAMMATION, NOS
INFLAMMATION, CHRONIC
HYPERPLASIA, LYMPHOID
(1*9)
17 (35%)
1 (2%)
3 (6%)
(50)
22 ( 4 4 % )
3 (6%)
2 (4%)
(47)
14
2
tLUNG/BRONCHUS
BRONCHIECTASIS
INFLAMMATION, SUPPURATIVE
HYPERPLASIA, FOCAL
HYPERPLASIA, LYMPHOID
(50)
4 (8%)
(49)
2
(48)
2 (4%)
1 (2%)
tLUNG
ATELECTASIS
CONGESTION, NOS
INFLAMMATION, SUPPURATIVE
INFLAMMATION, ACUTE SUPPURATIVE
B R O N C H O P N E U H O N I A ACUTE S U P P U R A T I
PNEUMONIAt_CHRgNIC_MURINE
(50)
1 (2%)
2 (4%)
RESPIRATORY
1
1
1
(2%)
(2%)
(2%)
SYSTEM
1 ( 2 % )
6
2
19
(16%)
* N U M B E f i O F A N I M A L S WITH T I S S U E E X A M I N E D M I C R O S C O P I C A L L Y
* N U M B E R OF ANIMALS NECROPSIED
24
(2%)
(507.)
(48)
(49)
1 (2%)
65
(4«)
(39%)
1
(30%)
(4%)
1 (2%)
1
(2*)
2
(4%)
1
1
(2%)
(2%)
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
INFLAMMATION. GRANULOMATOUS
GRANULOHA, NOS
INFLAMMATION, FOCAL GRAN0LOMATOU
FIBROSIS
NECROSIS, FOCAL
PIGMENTATION, NOS
HEMOSIDEROSIS
ALVEOLAR MACROPHAGES
LOW DOSE
HIGH DOSE
1 (2%)
1 (2%)
1 (2%)
1 (2*)
1 (2%)
1 (2%)
1 (2»)
5 (10%)
1 (2%)
2
(4%)
(50)
1 (2%)
1 (2%)
1 (2%)
(49)
(48)
1 (2%)
1 (2%)
#BONE MARROW
HYPERPLASIA, NOS
HYPERPLASIA, HEMATOPOIETIC
HYPERPLASIA, ERYTHROID
HYPERPLASIA, GRANULOCYTIC
HYPOPLASIA, ERYTHHOID
(49)
4
(48)
1 (2%)
6 (17%)
1 (2%)
(50)
tSPLEEN
CONGESTION, NOS
FIBROSIS
HEMOSIDEROSIS
ATROPHY, NOS
LEUKEMOID REACTION
HYPERPLASIA, RETICDLUM CELL
HEMATOPOIESIS
ERYTHBOPOIESIS
GRANULOPOIESIS
(49)
1 (2%)
1 (2%)
23 (47*)
1 (2%)
1 (2X)
#LYMPH NODE
HEMOSIDEROSIS
(41)
#SUBMANDIBOLAR L.NODE
LYMPHANGIECTASIS
tLUNG/ALVEOLI
CONGESTION,
EDEMA, NOS
HEMORRHAGE
NOS
HEMATOPOIETIC SYSTEM
(8%)
4 (8%)
1 (2%)
16 (32%)
1 (2%)
1 (2%)
1 (2%)
(49)
25 (51%)
(49)
2 (4%)
(63%)
36 (733S)
31
1
1
39
1
34 (69%)
1 (2%)
(2%)
(2%)
(80%)
(2%)
1 (2%)
(43)
(43)
(41)
(43)
(43)
1 (2%)
IMANDIBULAR L. NODE
LYMPHANGIECTASIS
(41)
(43)
2 (5%)
(43)
1 (2%)
tBHONCHIAL LYMPH NODE
LIMPHANGIECTASIS
(41)
(43)
(43)
* NUMBER OF ANIMALS HITH TISSUE
* NUMBER OF ANIMALS NECROPSIED
1 (2%)
EXAMINED MICROSCOPICALLY
66
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
LOW DOSE
CONTROL
HIGH DOSE
•MESENTERIC L. NODE
HEMOSIDEHOSIS
(<t1)
(43)
1 (2X)
(43)
tTHYMUS
LYMPH4NGIECTASIS
HEHOSIDEBOSIS
(37)
(28)
(29)
1 (3«)
1 (3«)
•HEART
FIBBOSIS, DIFFUSE
(48)
(49)
(48)
1 (2»)
•MYOCARDIUM
INFLAMMATION, FOCAL
INFLAMMATION, INTEBSTITIAL
ABSCESS, NOS
FIB80SIS
FIBBOSIS, FOCAL
SCAR
DEGENERATION, NOS
CALCIFICATION, DYSTBOPHIC
(48)
2 (4*)
(49)
(48)
•BNDOCABDIUM
INFLAMMATION, FOCAL
(48)
2 (U%)
(49)
(48)
*POLMONARY ARTEHY
MEDIAL CALCIFICATION
CALCIFICATION, FOCAL
(50)
(50)
1 (2%)
1 (2X)
(50)
•HEPATIC SINUSOID
CONGESTION, NOS
(49)
(50)
1 (2«)
(49)
4 (8%)
*HOUTH
ABSCESS, NOS
(50)
(50)
(50)
1 (2«)
•SALIVARY GLAND
EDEMA, NOS
(U9)
(49)
(46)
1 (2%)
•LIVER
CONGESTION, NOS
(49)
1 ii»l
(50)
(49)
CIBCOLATORY SYSTEM
1 (2X)
4 (8%)
1 (2X)
6 (12%)
5 (10%)
17 (35«)
24 (50%)
1 (2%)
1 (2%)
6 (13»)
1 (2*)
2 (4«)
DIGESTIVE SYSTEM
_
• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
67
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
LOW DOSE
CONTROL
HIGH DOSE
1 (2%)
HEMORHHAGE
NECROSIS, NOS
NECEOSIS, COAGULATIVE
1
METAMORPHOSIS FATTY
1 12%}
FOCAL CELLULAR CHANGE
PHAGOCYTIC CELL
ANGIECTASIS
HYPEHPLASIA, RETICULUM CELL
HEMATOPOIE5I3
1
(2%)
2 (4«)
1 (2%)
1 (2%)
5 (10X)
3 (6*)
3 (6S)
1 (2%)
4 (8%)
(2%)
4 (8»)
*LIVER/CENTHILOBULAR
NECROSIS, NOS
NECROSIS, FOCAL
NECROSIS, COAGULATIVE
METAMORPHOSIS FATTY
PIGMENTATION. NOS
(49)
*BILE DUCT
INFLAMMATION, FOCAL
HYPERPLASIA, NOS
(50)
(50)
1 (2X)
1 (2%)
3 (6%)
2 (4*)
1 (2*)
(50)
1 (2%)
18 (36%)
HYPERPLASIA, FOCAL
2 (4X)
30 (60%)
HYPERPLASIA, DIFFUSE
#PANCREAS
EDEMA, NOS
PE3IAHTERITIS
HEMOSIDEHOSIS
(49)
1 (2%}
1 (2%)
(50)
IPANCREATIC DUCT
HYPERPLASIA, NOS
HYPERPLASIA, EPITHELIAL
(49)
(50)
1
(2S)
1
(2%)
(50)
1 (2%)
3U (68%)
2 (4«)
(50)
1 (2X)
1 (2%)
1 (2%)
8 (16%)
2 («)
HYPERPLASIA, FOCAL
(49)
(50)
1 (2%)
1 (2%)
6 (12%)
(46)
(44)
(45)
1 (2%)
ISTOHACH
ULCER, NOS
ULCER, FOCAL
EROSION
(49)
1 (2%)
1 (2S)
1 (2%)
(50)
(49)
•PEYERS PATCH
HYPEHPLASIft, LYMPHOID
(49)
E> (10»)
(50)
(45)
6 (13%)
SILEUH
MUCOCELE
(49)
1
.I2*L_
#ESOPHAGUS
PERFORATION, INFLAMMATORY
f NUHBER OF ANIMALS HITH TISSUE EXAMINED
* NUHBER OF ANIMALS NECROPSIED
3 (6X)
(50)
_.
MICROSCOPICALLY
68
(45)
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
(32)
3 (9X)
(30)
•KIDNEY
CAST, NOS
CONGESTION, NOS
INFLAMMATION,. INTERSTITIAL
ABSCESS, NOS
INFLAMMATION, CHRONIC
INFLAMMATION, CHRONIC FOCAL
INFLAMMATION, CHRONIC DIFFUSE
SCLEROSIS
NEPHRO5IS, NOS
GLOMERULOSCLEROSIS, NOS
CALCIFICATION, NOS
(50)
1
1
1
1
8
26
1
(49)
•KIDNEY/CAPSULE
CYST, NOS
(50)
(49)
1 (2%)
(49)
#KIDNEY/CORTEX
CAST, NOS
CYST, NOS
PIGMENTATION, NOS
(50)
(49)
1 (2X)
(49)
•KIDNEY/TUBULE
CAST, NOS
PIGMENTATION, NOS
(50)
1 (2X)
3 (6X)
(49)
(49)
1 (2%)
fCONVOLUTED TUBULES
DEGENERATION, HYALINE
PIGMENTATION, NOS
(50)
(49)
(49)
1 (2%)
1 (2X)
IKIDNEY/PELVIS
INFLAHHATION, SOPPORATIVE
(50)
(49)
(U9)
1 (2X)
•URINARY BLADDER
HEMORRHAGE
INFLAMMATION, HEHORRHAGIC
(47)
(42)
(45)
1 (2X)
1 (2X)
tO. BLADDER/SUBMOCOSA
HEMORRHAGE
(47)
. 1 (2X)
(42)
(45)
tCOLON
NEMATODIASIS
2 (7 it;
HIGH DOSE
(39)
6 (15%)
ORINARY SYSTEM
(2X)
(2X)
(2X)
(2X)
(16%)
(52X)
(2X)
(49)
1 (2S)
1 (2X)
1 (2%)
9 (18X)
29 (59Z)
1 (2X)
1 (2%)
1 (2%)
10 (20%)
25 (51X)
1 (2X)
1 (2X)
1 (2%)
6 (12%)
3 (6X)
* NUMBER OF ANIMALS WITH TISSUE EXAHIHED
* NUMBER OF ANIMALS NECROPSIED
MICROSCOPICALLY
69
J_i2£l_ _
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
(48)
(49)
ENDOCRINE SYSTEM
tPITUITASY
CYST, NOS
HEMORRHAGE
HEHOSIDEROSIS
H Y P E R P L A S I A , NOS
H Y P E R P L A S I A , FOCAL
ANGIECTASIS
(46)
1
1
tADRENAL
ANGIECTASIS
(49)
1 (2%)
(50)
t A D H E N A L CORTEX
H Y P E R P L A S I A , NODULAR
(49)
1 (2*)
(50)
1 (2*)
(50)
t A D R E N A L MEDULLA
HYPERPLASIA, NODULAR
HYPERPLASIA, NOS
H Y P E R P L A S I A , FOCAL
(49)
2 (4%)
(50)
(50)
ITHYROID
CYSTIC FOLLICLES
P I G M E N T A T I O N , NOS
H Y P E H P L A S I A , C-CELL
HYPERPLASIA, FOLLICULAR-CELL
(46)
tTHYROID FOLLICLE
P I G M E N T A T I O N , NOS
(46)
(49)
3 (6*)
(47)
2 (4K)
*MAMMARY GLAND
GALACTOCELE
(50)
(50)
(50)
*PREPDTIAL GLAND
INFLA8HATION, SUPPURATIVE
I N F L A M M A T I O N , CHHOHIC
NECROSIS, NOS
(50)
2 (4»)
tPROSTATE
INFLAMMATION, FOCAL
INFLAMMATION^ SOPPUBATIVE
(44)
(2%)
(2»)
1 (2%)
1 (2%)
1
(2%)
2
(4S)
(8%)
1 (2%)
3 (6%)
1 (2«)
(50)
4 (8«)
4
1 (2«)
5 (10*)
1 (2*)
(49)
u (8«)
9 (18S)
(47)
2 (4S)
1 (2«)
29 (59X)
1 (2%)
23 (50%)
26 (55%)
2 (4%)
REPRODUCTIVE SYSTEM
1 (2%)
2
(50)
1 (2%)
(50)
5
(10%)
(4%)
1 (2*)
(44)
2_^5%1
t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
70
,4_ 19%)
(U7)
4 (9%)
i_12*J
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
1
(2X)
(1*9)
1
39
1
1
(2X)
(80X)
(2X)
(2X)
1
(2X)
NECROSIS, SOS
tTESTIS
CALCIFICATION, FOCAL
ATROPHY, NOS
ATROPHY, FOCAL
ASPERMATOGENESIS
HYPERTROPHY, NOS
HYPEHPLASIA, INTERSIITIAL CELL
(50)
tTESTIS/TOBULE
CALCIFICATION, NOS
CALCIFICATION, FOCAL
(50)
1 (2X)
32 (6UX)
7 (1l*X)
4 (8X)
1
HIGH DOSE
LOW DOSE
(2X)
(1*9)
38 (78X)
2 (4X)
3 (6X)
1 (2X)
6 (12X)
(i»9)
1
C*9)
(2X)
NERVOUS SYSTEM
*BRAIN/HENISGES
THROMBOSIS, NOS
(50)
1 (2X)
(50)
(50)
IBRAIN STEM
HEMORRHAGE
NECROSIS, NOS
MALACIA
(50)
(50)
(50)
1 (2X)
IHIDBRAIN
NECROSIS, NOS
HALACIA
(50)
1 (2X)
1 (2X)
(50)
(50)
*SPINAL CORD
HEMORRHAGE
DEGENERATION, NOS
BALACIA
(50)
(50)
(50)
1 (2X)
1 (2X)
1 (2X)
1
(2X)
1 (2»)
1 (2%)
SPECIAL SENSE ORGANS
*EYE
HEMORRHAGE
INFLAMBATION, NOS
PERIVASCULITIS
DEGENERATION. NOS
CATARACT
HEHOSIDEROSIS
_ ANEMIA^ NOS
(50)
(50)
1 (2X)
1 (2X)
1 (2X)
1 (2X)
13 (26X)
(50)
16 (32X)
_
f NO«BER OF A H I B A L S WITH TISSUE E X A M I N E D MICROSCOPICALLY
* N O H B E R OF A N I M A L S NECHOPSIED
71
12 (24X)
1 (2X)
1 I2X)
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
(50)
1 (2%)
2 (4%)
(50)
(50)
1 (2»)
(50)
(50)
(50)
(50)
1 (2*)
(50)
*SKELETAL MUSCLE
ATROPHY, NOS
(50)
(50)
(50)
1 (2«)
*HUSCLE HIP/THIGH
ATROPHY, NOS
(50)
(50)
(50)
1 (2%)
*MEDIASTINUM
THROMBOSIS, NOS
(50)
(50)
(50)
1 (2%)
*ABDOMINAL CAVITY
NECEOSIS, FAT
(50)
1 (2X)
(50)
(50)
*PLEURA
HYDROTHORAX
(50)
1 (2%)
(50)
(50)
*HESENTERY
INFLAMMATION, NOS
FIBROSIS
PERIABTERITIS
NECROSIS, FAT
(50)
(50)
(50)
1 (2%)
1 (2%)
*EYE/CORNEA
OLCER, NOS
INFLAMMATION, INTERSTITIAL
(50)
*LENS CAPSULE
CALCIFICATION, SOS
*BIDDLE EAR
INFLAHMATIQN, NOS
1 (2%)
HOSCULOSKELETAL SYSTEM
BODY CAVITIES
1 (2%)
1 (2%)
2 (4«)
ALL OTHER SYSTEMS
*MULTIPLE ORGANS
CONGESTION, NOS
JAUNDICE, NOS
(50)
(50)
DIAPHRAGM
HERNIA*. NOS
(50)
2 (US)
1 (2%)
1
t K U M B E f i OF A N I M A L S HITH TISSUE E X A M I N E D MICROSCOPICALLY
* N U M B E R OF A N I M A L S NECROPSIED
72
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
ADIPOSE TISSUE
INFLAHH&TION, NOS
INFLAMMATION, FOCAL
SPECIAL MORPHOLOGY SUMMARY
N O N E
t NUMBER OF ANIMALS SITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECfiOPSIED
73
HIGH DOSE
TABLE C2.
SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS
ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE
CONTROL
ANIMALS INITIALLY IN STUDY
ANIMALS NECHOPSIED
ANIMALS EXANIN3D HISTOPATHOLOGIC ALLY
50
50
50
LOW DOSE
HIGH DOSE
50
50
50
50
50
50
(50)
1 (2%)
(50)
(50)
1
(50)
I N T E G U M E N T A R Y SYSTEM
*SKIN
LYMPHOCYTIC INFLABMATORY INFILTR
N E C R O S I S , FOCAL
(50)
* S U B C U T TISSUS
A B S C E S S , NOS
(50)
1 (2%)
(2%)
RESPIRATORY. SYSTEM
#TRACHEA
INFLAMMATION, NOS
INFLAMMATION, SUPPURATIVE
INFLAMMATION, CHRONIC
HYPERPLASIA, LYMPHOID
(49)
17
*LUNG/BRONCHU3
BRONCHIECTASIS
INFLAMMATION, NOS
INFLAMMATION, SUPPURATIVE
HYPERPLASIA, NOS
HYPERPLASIA, FOCAL
HYPERPLASIA, LYBPHOID
(50)
2 (4X)
1
(2%)
tLUNG
CONGESTION, NOS
EDEMA, NOS
BRONCHOPNEUHONIA, NOS
INFLAMMATION, FOCAL
INFLAMMATION, INTERSTITIAL
PNEUMONIA, ASPIRATION
BRONCHOPNEUBONIA SUPPURATIVE
BRONCHOPNEOMONIAj. ACUTE
(50)
1
(48)
10 (2U)
(35S)
1
(2%)
(49)
2
1
27
27 (5U»)
1
(2%)
(49)
11 (22%}
1 (2%)
3 (6«)
2 (4%)
(49)
1
(2%)
1
(2%)
(4%)
(2%)
(55X)
(49)
1 (2«)
1 (2X)
31
(49)
1 (2%)
<2X)
1
1
(2%)
{2*>
1
1 (2*)
1 J2«l
t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
74
(63%)
(2%)
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
INFLAMMATION, ACUTE SUPPURATIVE
BRONCHOPNEUMOHIA ACUTE SUPPURATI
PNEUMONIA, CHRONIC SORINE
INFLAMMATION, FOCAL GRANULOMATOU
PEBIVASCULAR CUFFING
HEHOSIDEROSIS
ALVEOLAR MACROPHAGES
HYPERPLASIA, LYMPHOID
LOW DOSE
1 (2X)
1 (2%)
4 (8%)
1 (2%)
5 (10%)
HIGH DOSE
1 (2*)
1 (2%)
4
(8%)
1 (2%)
2 (451)
1 (2%)
5 (10*)
2 (4%J
1 (2%)
1 (2%)
(50)
1 (2%)
(49)
3 (6%)
3 (6%)
(49)
3 (6%)
IBONE M A B R O W
HEHOSIDEROSIS
HYPOPLASIA, NOS
HYPEHPLASIA, NOS
HYPERPLASIA, HEMATOPOIETIC
HYPERPLASIA, GRANULOCYTIC
ERYTHROPOIESIS
GRANULOPOISSIS
(50)
(47)
(50)
1 (2%)
•SPLEEN
ECTOPIA
CONGESTION, NOS
HEMOSIDEHOSIS
LEUKEMOID REACTION
HYPERPLASIA, RETICULUM CELL
HEMATOPOIESIS
EHYTHROPOISSIS
GRANULOPOISSIS
(50)
tSPLENIC CAPSULE
INFLAMMATION, FOCAL
(50)
(50)
1 (2%)
(49)
tLYMPH NODE
LYMPHANGIECTASIS
HEHOSIDEROSIS
(44)
(41)
(45)
1 (2%)
*HANDIBULAR L. NODE
PIGMENTATION, NOS
(44)
(41)
1 (2%)
(45)
tCERVICAL LYMPH NODE
CONGEST IONj._NOS
(44)
(41)
(45)
tLUNG/ALVEOLI
CONGESTION, NOS
EDEMA, NOS
HEMATOPOIETIC SYSTEM
1
(2%)
1 (2%)
2 (4*)
1 (2%)
3 (6%)
2 (4%)
6 (12%)
1 (2%)
2 (4%)
1 (2%)
1 (2%)
34 (68%)
(50)
1 (2X)
1 (2%)
44 (88%)
(49)
40 (82%)
1 (2%)
1 (2%)
40 (80%)
37 (74X)
3 (6%)
2 (4%)
36 (73%)
2 (4%)
2 (4%)
1 (2%)
* NUMBER OF ANIMALS WITH TISSUE EXAMINED
* NUHBEB OF ANIMALS NECROPSIED
75
MICROSCOPICALLY
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
HEMOSIDEROSIS
ITHYMUS
LYMPHANGIECTASIS
CONGESTION, NOS
HEMOSIDEROSIS
LOW DOSE
HIGH DOSE
1 (2X)
(39)
(30)
1 (3%)
(31)
1 (3%)
1
(3»)
CIRCULATORY SYSTEM
IHEABT/ATRIUM
THROMBOSIS, NOS
(48)
(46)
1 (2X)
(50)
tMYOCARDIUM
INFLAMMATION, FOCAL
INFLAMMATION, INTERSTITIAL
FIBROSIS
FIBROSIS, FOCAL
SCAR
(48)
(46)
(50)
2 (4X)
2 (4%)
IENDOCARDIUM
INFLAMMATION, FOCAL
(48)
(46)
1 (2%)
(50)
*PULMONARY ARTERY
CALCIFICATION, NOS
CALCIFICATION, FOCAL
(50)
(50)
(50)
1 (2*)
1 (2%)
tHEPATIC SINUSOID
CONGESTION, NOS
HYPEBPLASIA, GRANULOCYTIC
(49)
(50)
4 (835)
1 (2X)
(50)
1 (235)
*TONGOE
HYPERKERATOSIS
ACANTHOSIS
(50)
(50)
1 (2X)
1 (235)
(50)
tLIVER
HERNIA, NOS
NECROSIS, FOCAL
METAMORPHOSIS FATTY
PIGMENTATION, NOS
FOCAL CELLULAR CHANGE
ANGIECTASIS
(49)
(50)
1 (2X)
1 (2X)
11 (22X)
(50)
1 (2%)
1 (2%)
3 (6%)
1 (2%)
7 (1536)
1
(2X)
2 (4X)
9 (20X)
1 (2X)
9
(1835)
DIGESTIVE SYSTEM
9 (18X)
3 (6X)
t NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
76
1 (2%)
1 (2X)
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
LOW DOSE
CONTROL
HYPERPLASIA, RETICULUM CELL
HEMATOPOIE3IS
ERYTHHOPOISSIS
4 (8«)
2 (4X)
1 (2%)
\ (2%)
HIGH DOSE
1 (2%)
ILIVEH/CENTRILOBULAR
NECROSIS, FOCAL
HETAHORPHOSIS FATTY
(49)
1 (2%)
2 (4%)
(50)
tLIVER/PERIPORTAL
METAMORPHOSIS FATTY
(49)
1 (2%)
(50)
(50)
1 (2%)
(50)
(50)
*BILE DOCT
INFLAMMATION, FOCAL
HYPERPLASIA, NOS
HYPERPLASIA, FOCAL
2 (4%)
*
(50)
(50)
2 (4%)
2 (4%)
17 (34X)
15 (30%)
(49)
2 (4%)
16 (32S)
(47)
tPAHCREAS
LYBPHOCYTIC INFLAMHATORY INFILTR
PERI ART EBI IIS
(49)
1 (2%)
tPANCREATIC DUCT
HYPERPLASIA, FOCAL
(49)
5 (10*)
(49)
6 (12%)
(47)
9 (19%)
tPANCREATIC ACINUS
NODULE
(49)
(49)
1 (2%)
(47)
tESOPHAGUS
ABSCESS, NOS
(49)
(48)
1 (2%)
(42)
tSTOHACH
ULCER, NOS
ULCER, FOCAL
(50)
1 (2%)
(50)
(49)
tCARDIAC STOMACH
ULCER, NOS
EROSION
(50)
1 (2*)
tPEYEKS PATCH
HYPERPLASIA, LYMPHOID
(49)
4 (8%)
(45)
2 (4%)
(50)
tCOLON
NEMATODIASIS
HYPERPLASIA, RETICULDS CELL
__HY £E 8 £ LA SI A ^LY HPHO ID
(35)
5 (14%)
(39)
4 (10%)
1 (3%)
1_J3%1 _ .
(39)
1 (2%)
1 (2X)
(50)
(49)
1 (2%)
.
t NUHBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
77
4
(8%)
5 (13%)
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
U R I N A R Y SYSTEM
#KIDNEY
CAST, NOS
INFLAMMATION, FOCAL
INFLAMHATION, INTERSTITIAL
INFLAMMATION, CHRONIC
INFLABMATION. CHRONIC FOCAL
INFLAMMATION, CHRONIC DIFFUSE
SCAR
NEPHROSIS, NOS
INFARCT, ACUTE
PIGHENTATION, NOS
HYPERPLASIA, TUBULAR CELL
(U9)
*KIDNEY/CORTEX
SCAR
INFARCT, NOS
PIGMENTATION, NOS
HYPERPLASIA, LYHPHOID
(49)
#KIDNEY/TUBULE
CAST, NOS
PIGHENTATION, NOS
ATROPHY, NOS
(49)
tCONVOLUTED TUBULES
CAST, NOS
HYALINE M E M B R A N E
PIGHENTATION, NOS
(49)
1 (2X)
1 (2X)
tKIDNEY/PELVIS
CALCIFICATION, FOCAL
(49)
#URINARY BLADDER
CALCULUS, NOS
INFLABMATION, CHRONIC
HYPERPLASIA, EPITHELIAL
(50)
1 (2%)
(50)
1 (2%)
1
2
7
1
1
1 (2X)
2 (4X)
12 (24X)
(2%)
(4S)
(14X)
(2%)
(2X)
2 (US)
13 (26%)
1 (2%)
1 (2X)
2 (4«)
1 (2X)
2 (4«)
(50)
1 (2XJ
17 (35X)
1 (2X)
2 (4X)
26 (52X)
1 (2%)
(50)
1 (2X)
20 (40X)
(50)
1 (2*)
2 (OX)
1 (2X)
(50)
(50)
(50)
1 (2XJ
1 (2%)
(50)
1 (2%)
(50)
(35)
1 (3X)
1 (3*)
1 (3X)
(42)
(41)
(45)
(46)
(47)
1 (2%)
ENDOCRINE SYSTEM
tPITUITARY
CYST A _NOS
f N U M B E R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
78
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
HEMORRHAGE
HEMORRHAGIC CYST
HEHOSIDEROSIS
HYPEHPLASIA, NOS
HYPERPLASIA,
FOCAL
ANGIECTASIS
2
LOW DOSE
(4%)
1
3
3
1
1
14
2 («}
1 (2%)
3 (7%)
1 (2%)
3 (7%)
(2%)
(7%)
(7%)
(2%)
(2%)
(30%)
HIGH DOSE
2 (4%)
3 (6*)
20 (43%)
•ADRENAL
CYST, NOS
DEGENERATION, NOS
HEMOSIDEROSIS
ANGIECTASIS
(49)
(49)
1 (2%)
•ADRENAL CORTEX
HEMORRHAGE
(49)
1 (2%)
(50)
(49)
IADRENAL MEDULLA
HYPERPLASIA, FOCAL
(49)
(50)
1 (2%)
(49)
2 (4%)
tTHYROID
CYSTIC FOLLICLES
LYBPHOCYTIC INFLAHMATORY INFILTR
H Y P E R P L A S I A , C-CELL
(50)
1 (2%)
(44)
1 (2%)
(44)
tTHYROID FOLLICLE
PIGMENTATION, NOS
(50)
(50)
1 (2%)
1 (2%)
3 (6%)
6 (12%)
1 (2«
24 (55%)
39 (78%)
7 (14%)
1
(2%)
21 (43%)
(44)
(44)
1 (2%)
REPRODUCTIVE SYSTEM
* M A M M A R Y G L A N D
GALACTOCELE
NECROSIS, FOCAL
METAPLASIA, SQUAMOUS
ADENOSIS
(50)
5 (10%)
*PREPUTIAL GLAND
INFLAHHATION, SUPPURATIVE
I N F L A M M A T I O N , ACUTE SUPPUHATIVE
NECROSIS, NOS
HYPERPLASIA, NOS
H Y P E R P L A S I A , CYSTIC
(50)
7 (14%)
*V AGIN A
HEHATOBA X _NOS_
(50)
(50)
4 (8%)
(50)
10 (20%)
1 (2%)
1 (2%)
1 (2ft)
1 (2%)
(50)
2
(4%)
(50)
1 (2%)
1 (2%)
1 (2%)
2 (4%)
1 (2%)
1 (2%)
(50)
1 _[2X]_
* NUMBER OF ANIMALS WITH TISSUE EXAMINED 8ICROSCOPICALLY
* NUHBEE OF ANIMALS NECROPSIED
79
(50)
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
tUTERUS
HEHORRHAGE
INFLAMMATION, NOS
INFLAMMATION, SUPPORATIVE
PYOHETRA
NECROSIS, NOS
(50)
ICEBVIJC UTERI
HYPERPLASIA, NOS
(50)
*UTERUS/ENDOMETRIUM
CYST, NOS
HEMORRHAGE
INFLAMMATION, FOCAL
ULCER, FOCAL
LYMPHOCYTIC INFLAMMATORY INFILT8
INFLAMMATION, SOPPURATIVE
INFLAMMATION, VESICULAR
INFLAMMATION, CHRONIC SUPPURATIV
NECROSIS, NOS
HYPERPLASIA, CYSTIC
(50)
#OVARY/OVIDOCT
INFLAMMATION, NOS
INFLAMMATION, SUPPURATIVE
(50)
tOVARY
CYST, NOS
FOLLICOLAR CYST, NOS
INFLAMMATION, NOS
(50)
9
LOW DOSE
HIGH DOSE
(48)
(49)
1 (2«)
1 (2X)
1 (2*)
1 (2%)
1 (2%)
(48)
(49)
1 (2%)
C*8)
(49)
1 (2%)
1 (2%)
1 (2%)
1 (2%)
1 (2%)
8 (17X)
1 (2*)
3 (16%)
14 (29«)
1 (2%)
2 (4*)
1 (2*)
(148)
(49)
1 (2%)
13 (21%)
12 (24%)
(47)
8 (17K)
(48)
10 (2H)
5 (10X)
(18%)
2 (4«)
2 (4»)
1 (2%)
1
(2%)
NERVOUS SYSTEM
*BRAIN/MENINGES
INFLAMMATION, SUPPURATIVE
(49)
ICEREBRUM
INFLAMMATION, SUPPURATIVE
ABSCESS, NOS
(49)
tBRAIN
COMPRESSION
INFLAMMATION^ NOS
(49)
(50)
(50)
1 (2%)
(50)
(50)
1 (2%)
1 (2%)
(50)
2 (4«)
i_i2«J. _
* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
80
(50)
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
MALACIA
1
LOW DOSE
HIGH DOSE
(2%)
(50)
(50)
2 (H%)
*MIDBRAIN
COMPRESSION
GLIOSIS
(U9)
*SPINAL CORD
HEMORRHAGE
(50)
1 (2%)
(50)
(50)
(50)
(50)
(50)
1
1
1
1
1 (2%)
SPECIAL SENSE ORGANS
*EYE
HEMORRHAGE
INFLAMMATION, NOS
INFLAMMATION, INTERSTITIAL
PUS
INFLAMMATION, SUPPURATIVE
CATARACT
11
1 (2%)
15 (30*)
(22%)
(2%)
(2%)
(2S)
(2%)
16 (32%)
*EYE/CORNEA
ULCER, NOS
INFLAMMATION, SUPPURATIVE
(50)
(50)
1 (2%)
1 (2%)
(50)
*EAR
INFLAMMATION, NOS
(50)
(50)
(50)
1 (2%)
*EAR CANAL
INFLAMMATION, SUPPORATIVE
(50)
(50)
1 (2%)
(50)
(50)
1 (2%)
(50)
(50)
(50)
(50)
2 (4%)
(50)
(50)
(50)
(50)
MUSCOLOSKELETAL SYSTEM
*SKELETAL MUSCLE
ATROPHY, NOS
BODY CAVITIES
*MESENTERY
FIBROSIS
ALL OTHER SYSTEMS
*MULTIPLE ORGANS
____ CONGEST ION j._NOS
t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
81
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
JAUNDICE, NOS
1
THORAX
HEMORRHAGE
DIAPHRAGM
HERNIA, NOS
HIGH DOSE
(2*)
1
1
ADIPOSE TISSUE
INFLAMMATION, NOS
1
2
SPECIAL MORPHOLOGY SUMMARY
NONE
t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
82
2
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
ANIMALS INITIALLY IN STDDY
ANIMALS NECROPSIED
ANIMALS EXAMINED HISTOPATHOLOGIC ALLY
50
49
49
LOW DOSE
HIGH DOSE
50
50
50
50
50
50
(50)
(50)
INTEGUMENTARY SYSTEH
*SKIN
CYST, NOS
ULCER, NOS
HYPEHPLASIA, NOS
(49)
*SUBCUT TISSUE
ABSCESS, NOS
INFLAMMATION, CHRONIC
(49)
(50)
1 (2X)
1 (2X)
(50)
tLUNG/BRONCHUS
METAPLASIA, SQUAMOOS
HYPERPLASIA, LYMPHOID
(49)
(48)
(50)
ILUNG
CONGESTION, NOS
HEMORRHAGE
PNEUMONIA, ASPIRATION
PERIVASCULITIS
ALVEOLAR MACROPHAGES
HYPERPLASIA, ADENOMATOUS
HYPERPLASIA, ALVEOLAR EPITHELIUM
(49)
1 (2%)
1 (2%)
tLUNG/ALVEOLI
CONGESTION, NOS
(49)
(48)
(50)
1 (2%)
(46)
(49)
(50)
1
(2%)
1 (2%)
1 (2%)
RESPIRATORY SYSTEM
1 (2«)
11 (22%)
1
(2%)
(48)
2 (4%)
(50)
1
(2%)
1
(2%)
1
(2%)
1 (2*)
1
(2%)
HEHATOPOIETIC SYSTEM
*BONE HARROW
HYPERPLASIAX_HOS
* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* BOMBER OF ANIMALS NECROPSIED
85
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
HYPERPLASIA, HEHATOPOIETIC
HYPERPLASIA, GR ANOLOCYTIC
LOW DOSE
HIGH DOSE
2 (4*)
2 (4X)
2 (4%)
1 (2X)
*SPLEEN
HEMORRHAGE
ANGIECTASI3
HYPERPLASIA. HEMATOPOIETIC
HYPERPLASIA, RETICULOS CELL
HYPERPLASIA, LYMPHOID
HEKATOPOIESIS
ERYTHBOPOIESIS
GRANOLOPOIESIS
(46)
1 (2%)
1 (2%)
tLYHPH NODE
(40)
1 (3%)
(31)
(30)
#ME"5ENTERIC L. NODE
THROHBOSIS, NOS
CONGESTION, NOS
HEMOSIDEROSIS
ERYTHROPHAGOCYTOSIS
HYPEBPLASIA, HEHATOPOIETIC
HYPERPLASIA, LYHPHOID
(40)
1 (3%)
3 (8%)
(31)
(30)
#THYMOS
HYPERPLASIA., RETICOLUH CELL
(35)
(38)
1 (3%)
(41)
*TONGUE
HYPERPLASIA, EPITHELIAL
HYPERKERATOSIS
ACANTHOSIS
(49)
(50)
(50)
1 (2%)
1 (2%)
1 12*)
tLIVER
HEMORRHAGE
FIBROSIS, FOCAL
DEGENERATION, NOS
NECROSISA_FOCAL
(49)
1 (2%)
1 (2%)
(50)
(49)
1 (21)
HEMATOPOIESIS
2
(46)
(50)
1
2
5
23
1
(4X)
24 (52%)
2 (4»)
1 (2%)
(2%)
(4*)
(10%)
(46X)
(2X)
1 (2%)
17 (37 X)
2
(4%)
1 (3%)
2 (7*)
1
1
(3%)
(3X)
CIRCULATORY SYSTEM
NONE
DIGESTIVE SYSTEH
1 (2%)
* NOMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
86
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
HETAHORPHOSIS FATTY
CLEAR-CELL CHANGE
HYPERPLASIA, MODULAR
HYPERPLASTIC HODBLE
ANGIECTASIS
HYPERPLASIA, HEH ATOPOIETIC
HYPERPLASIA, RETICULOB CELL
HYPERPLASIA, LYHPHOID
HEBATOPOIESIS
ERYTHROPOIESIS
LOW DOSE
4 (8%)
1 (2*)
1 (2%)
1 (2%)
1 (2%)
1 (2%)
1 <2X)
1
1
(49)
1 (2%)
tLIVER/CENTRILOBULAR
NECROSIS, NOS
BETAHORPHOSIS FATTY
(U9)
*LIVER/PEBIPORTAL
HYPERPLASIA, LYBPHOID
3 (6S)
1 (2%)
(2%)
(2%)
#HEPATIC CAPSULE
HEBATOHA, NOS
FIBROSIS, FOCAL
HIGH DOSE
3 (6*)
2 (4%)
1 (2%)
1 (2*)
1 (2»)
(50)
(49)
1 (2X)
(50)
(49)
1 (2%)
(49)
1 (2%)
(50)
(49)
tLIVER/HEPATOCYTES
NECROSIS, NOS
NECROSIS, FOCAL
(49)
(50)
(49)
1 (2%)
*BILE DUCT
CYST, NOS
INFLABBATION, NOS
INFLABBATION, FOCAL
INFLABBATION, SUPPURATIVE
FOCAL CELLULAR CHANGE
HYPERPLASIA, NOS
HYPERPLASIA, FOCAL
(49)
tPANCREAS
CYSTIC DUCTS
FIBROSIS
NECEOSIS, NOS
(48)
1 (2%)
1 (2%)
1 (2%)
(50)
(47)
tP&NCREATIC DUCT
CYST, NOS
HYPERPLASIA, FOCAL
(48)
(50)
3 (6*)
(47)
#PEYERS PATCH
INFLABBATION^ SUPPURATIVE
(47)
(49)
(49)
1 i2%L
1 (2%)
2 (4S)
1
(50)
1 (2%)
1 (2%)
1 (2«)
(50)
1 (2%)
(2%)
1 (2%)
4 (8%)
1 (236)
1
(2%)
» NUBBER OF ANIBALS WITH TISSUE EXABINED BICROSCOPICALLY
* NUBBER OF ANIBALS NECROPSIED
87
2 (4«)
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
1 (2%)
2 (4%)
10 (20%)
(22)
4 (18%)
(44)
6 (14%)
(35)
1 (3%)
•KIDNEY
HYDRONEPHBOSIS
LYNPHOCYTIC INFLAMHATOHY INFILTR
INFLAMMATION, INTERSTITIAL
INFLAMMATION, CHRONIC
FIBROSIS
HYPERPLASIA, LYMPHOID
(48)
(49)
1 (2%)
(50)
tKIDNEY/CORTEX
LYMPHOCYTIC INFLAHMATORY INFILTR
INFARCT, NOS
(48)
tU. BLADDER/SUBMUCOSA
HYPERPLASIA, LYMPHOID
HYPERPLASIA, NOS
HYPERPLASIA, LYMPHOID
#COLON
NEMATODIASIS
5 (10%)
URINARY SYSTEM
2
1
1
1
1
(4%)
(2%)
(2%)
(2%)
(2%)
1 (2%)
1 (2%)
(49)
1 (2%)
(50)
(47)
(46)
(49)
1 (2%)
(46)
28 (61%)
(49)
41 (84%)
(50)
38 (76%)
•ADRENAL CORTEX
HYPERPLASIA, NOS
(46)
2 (4%)
(49)
(50)
•THYROID
CYSTIC FOLLICLES
HYPERPLASIA, FOLLICULAR-CELL
(43)
1 (2%)
2 (5%)
(44)
(48)
•PANCREATIC ISLETS
HYPESPLASIA, NOS
(48)
2 (4%)
(50)
(47)
(49)
(50)
3 I6%}.__ .
(50)
1 (2%)
ENDOCRINE SYSTEM
•ADRENAL/CAPSULE
HYPERPLASIA, FOCAL
1 (2%)
2 (5%)
2 (4%)
REPRODUCTIVE SYSTEM
*PREPUTIAL GLAND
CYST, NOS
• NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
* NURBER OF ANIMALS NECROPSIED
88
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
INFLAMMATION, SUPPURATIVE
INFLAMMATION, CHRONIC
INFLAMMATION, CHBONIC SOPPUBATIV
HYPERKERATOSIS
LOW DOSE
2 (4X)
2 (<»X)
tTESTIS
GRANULOMA, SPERMATIC
ATROPHY, NOS
ATROPHY, FOCAL
ASPERMAIOGENESIS
(47)
*EPIDIDYMIS
(U9)
(49)
4
1
(8*)
HIGH DOSE
1
1
1
1
(50)
1 (2*)
2 (4X)
1 (2%)
(2%)
(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)
(49)
(50)
(50)
SPECIAL SENSE ORGANS
*EYE
CATARACT
*EYE/CORNEA
1 (2*)
(49)
(50)
INFLAMMATION, NOS
*LENS CAPSULE
DEGENERATION, NOS
(50)
1 (2X)
(49)
(50)
1 (2*)
(50)
(49)
(50)
(50)
HOSCDLOSKELETAL SYSTEM
NONE
BODY CAVITIES
*PERITONEOM
____ HEMORRHAGE
t NDHBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
89
(2*)
(2*)
(2S)
(2»)
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
INFLAHBATION, NOS
NECROSIS, FOCAL
LOW DOSE
HIGH DOSE
1 (2%)
1 (2%)
*ABDOBINAL VISCERA
ADHESION, NOS
(1*9)
(50)
(50)
1 (2%)
*PLEURA
HYDROTHORAX
HEBOTHORAX
INFLAMMATION, FOCAL
(49)
(50)
1 (2%)
2 (UX)
(50)
*MESENTERY
F1BROSIS
fIBROSIS, FOCAL
NECROSIS, FAT
(49)
1
(50)
4 (8*)
(50)
1 (2%)
1 (2%)
2 (4S)
3 (6S)
2
1
1
3
ALL OTHER SYSTEMS
ADIPOSE TISSUE
INFLAHMATION, NOS
FIBROSIS
SPECIAL MOBPHOLOGY SUMMARY
NO LESION REPORTED
AUTO/NECROPSY/HISTO PERF
AUTOLYSIS/NO NECROPSY
(2%)
1
1
t HUMBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
90
1
1
(6%)
TABLE D2.
SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE
ADMINISTERED 3-NITROPROPIONIC ACID BY GAVAGE
CONTROL
ANIMALS INITIALLY IN STUDY
UNIMALS NECROPS1ED
ANIBALS EXAMINED HISTOPATHOLOGICALLY
LOW DOSE
HIGH DOSE
50
50
50
50
50
50
50
50
50
(50)
(50)
1 (2S)
(50)
(49)
3 (6%)
INTEGUMENTARY SYSTEM
*SKIN
ALOPECIA
HYPERKERATOSIS
1 (2%)
RESPIRATORY SYSTEM
•LUNG/BRONCHUS
HYPERPLASIA, LYMPHOID
(47)
18 (38%)
(49)
#LUNG
CONGESTION, NOS
INFLAMMATION, FOCAL
INFLAMMATION, SUPPURATIVE
ALVEOLAB MACROPHAGES
HYPEBPLASIA, LYHPHOID
(47)
(49)
3 (6X)
(<*9)
1 (21)
2 (4%)
1 (2%)
1 (25)
1 (2%)
1
1 (2S)
(2%)
HEMATOPOIETIC SYSTEM
tBONE MARROW
H Y P E R P L A S I A , HEMATOPOIETIC
HYPERPLASIA, GEANULOCYTIC
GRANULOPOIESIS
(46)
ISPLEEN
RUPTURE
THROMBOSIS, NOS
LEUKEMOID REACTION
HYPERPLASIA, LYMPHOID
HEMATOPOIESIS
EHYTHROPOIESIS
MYELOPOIESIS
(47)
1
(50)
(43)
3 (1%)
(2%)
1 (2%)
(2%}
1 (2%)
1
6
(50)
(2%)
(13%)
8 (16*)
19 (40%)
22 (44X)
(50)
1
1
2
11
22
(2%)
(2%)
(<U)
(22%)
(44%)
1 (2%)
1 12%l
_ _
# NOHBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
* NUMBER OF ANIMALS NECROPSIED
91
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
»LY«PH NODE
EDEMA, NOS
HYPERPLASIA, LYMPHOID
(38)
#KAHDIBULAR L. NODE
HEMORRHAGE
LOW DOSE
HIGH DOSE
(36)
(33)
1 (3X)
(38)
(36)
(33)
1 (3%)
#NESENTERIC L. NODE
INFLAMMATION, GRANULOMATOUS
HYPERPLASIA, LYMPHOID
(38)
1 (3X)
(36)
(33)
#THYMUS
HYPEHPLASIA, LYMPHOID
(38)
1 (3X)
(42)
1 (2*)
(35)
•HEART/ATRIUM
THROMBOSIS, NOS
(49)
(50)
(49)
1 (2%)
#CAEDIAC V A L V E
PIGMENTATION, NOS
(49)
(50)
1 (2X)
(49)
1 (2%)
*UTEBINE ARTERY
THROMBOSIS, NOS
(50)
1 (2%)
(50)
(50)
tLIVER
HEMOBRHAGIC CYST
NECROSIS, FOCAL
NECROSIS, ISCHEMIC
METAMORPHOSIS FATTY
ANGIECTASIS
LEUKEMOID REACTION
HYPE8PLASIA, RETICULOM CELL
HYPERPLASIA, LYMPHOID
HEMATOPOIE3IS
(49)
(50)
1 (2X)
(50)
tLIVER/HEPATOCYTES
DEGENERATION, NOS
NECROSIS, NOS
NECROSIS^. FOCAL
(49)
1 (3X)
1 (3%)
CIRCULATORY SYSTEM
DIGESTIVE SYSTEM
1 (2X)
1 (2X)
2 («X)
2 (4X)
1 (2%)
1 (2%)
1 (2X)
2 (4X)
3 (6X)
1 J2X}.
* NUMBER OF ANIMALS WITH TISSUE EXAMINED
* NUMBER OF ANIMALS HECROPSIED
92
2
(4»)
1 (2%)
1 (2X)
(50)
1 (2«)
1 (2«)
1 (2X1
MICROSCOPICALLY
(50)
_ 3 (6%)
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
LOW DOSE
HIGH DOSE
•PANCREATIC DUCT
CYST, NOS
(UU)
(t9)
1 (2*)
(50)
tPEYERS PATCH
HYPERPLASIA, LYHPHOID
(48)
(48)
(49)
3 (6%)
IDUODENUM
HYPERPLASIIV, LYNPHOID
(48)
(48)
(49)
1 (2%)
•COLON
NEMATODIASIS
(36)
(38)
(35)
1 (2*)
1 (3J)
URINARY SYSTEM
(50)
1 (2X)
(50)
tKIDNEY
CONGESTION, NOS
GLOMERULONEPHRITIS, NOS
PYELONEPHRITIS, NOS
PYELONEPHRITIS DIFFUSE
INFLAMMATION, CHRONIC FOCAL
GLOMERULOSCLEROSIS, NOS
HYPERPLASIA, LYBPHOID
(49)
tKIDNEY /CORTEX
SCAR
DEGENERATION, HYALINE
(49)
1 (2%)
1 (2%)
(50)
(50)
fKIDNEY/TUBULS
DEGENERATION, HYALINE
(49)
(50)
1 (2S)
(50)
1 (2K)
IPITUITARY
HEMORRHAGIC CYST
HYPERPLASIA, NOS
ANGIECTASIS
(43)
(48)
(42)
1 (2%)
2 (5ft)
2 (5»)
IADRENAL/CAPSOLE
HYPERPLASIA, FOCAL
(48)
43
•ADRENAL CORTEX
HYPEBPLASIAX FOCAL
(48)
1
(2%)
1
1
(2%)
(2%)
1
(2%)
1 (2*)
10
(20%)
4 (8«)
ENDOCRINE SYSTEM
1 (2X)
2 (4%)
* NUMBER OF ANIMALS HITH TISSUE EXAMINED
* NUMBER OF ANIMALS NECROPSIED
(50)
47 (94S)
(90%)
(50)
MICROSCOPICALLY
93
(49)
42
(49)
(86%)
._ _ _i_im
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
tTHYROID
CYSTIC FOLLICLES
INFLAMMATION, FOCAL
HYPERPLASIA, FOLLICUL AR-CELL
CONTROL
LOW DOSE
(40)
1 (3%)
(47)
6 (15%)
13
(23%)
HIGH DOSE
(45)
1
(2%)
1
(2%)
5
(11*)
8EP80DUOTIVE SYSTEM
fMJTERUS
HYDROMETRA
HEMORRHAGE
METAPLASIA, SQOAMOUS
(47)
tUTERUS/ENDOMETRIUM
HEMATOMA, NOS
INFLAMMATION, SUPPORATIVE
HYPEBPLASIA, NOS
HYPERPLASIA, CYSTIC
(47)
•OVARY/OVIDUCT
LYMPHOCYTIC INFLAMMATORY INFILTR
INFLAMMATION, SUPPURATIVE
NECROSIS, NOS
(47)
1 (2%)
•OVARY
(39)
4 (10*)
CYST, NOS
FOLLICULAR CYST, NOS
HEM08RHAGIC CYST
LYMPHOCYTIC INFLAMMATORY INFILTR
INFLAMMATION, SUPPORATIVE
INFLAMMATION, CHRONIC
CALCIFICATION, FOCAL
HYPERPLASIA, LYMFHOID
(49)
1 (2%)
1 (2%)
2 (4*)
3 (6%)
1 (2%)
(49)
1 (2%)
1 (2%)
37 (76%)
19 (40%)
(48)
1
(2%)
(48)
1
(2%)
30
£63%)
(49)
1 (2/S)
(43)
(47)
(47)
12
(26%)
4
(9%)
1
(2%)
3 (6%)
1 (2%)
10 (2H)
4 (9%)
1 (3%)
1
(2%)
1 (3%)
1 <3«)
1
(2%)
1
(2*)
1
(2%)
NERVOUS SYSTEM
IBRAIN/MENING3S
PERIVASCULAR CUFFING
(47)
(50)
1 (2K)
(50)
*CEBEBROM
(47)
(50)
(50)
1
(2%)
(50)
(50)
1 _I2%]_
(50)
EPIDERMAL INCLUSION CYST
SPECIAL SENSE ORGANS
• EIE
PHTHISIS BULBI
t NUMBER. ,OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
* N U M B E R OF A N I M A L S NECBOPSIED
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)
CONTROL
_ .. _ • _ • _ _ — _ _ _ « _ _ _ _ _ _ _ _ — _ » _
w
LOW DOSE
HIGH DOSE
— _ — _ — _ - . — — — _ _ _ _ _ _ — — — — — _ _ _ — _ _ _ -, — » — _ — _ — . _ - . — «i_ _ — — — — — — •— — — — —
*HARDERIAN GLAND
ABSCESS, NOS
INFLAMMATION, CHRONIC
(50)
(50)
1 (2X)
1 (2%)
(50)
•PERITONEUM
CYST, NOS
HEMORRHAGE
(50)
1 (2X)
(50)
(50)
*PLEORA
HYDROTHORAX
HEHOTHORAX
(50)
1 (2%)
BUSCULOSKELEIAL SYSTEM
NONE
BODY CAVITIES
1 (2ft)
(50)
1 (2%)
&LL OTHER SYSTEMS
N O N E
SPECIAL MORPHOLOGY SUMMARY
AUTO/NECROPSY/HISTO PERF
(50)
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.
Topography:
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavagea
Morphology
All Sites: Skin Tumorsb
P Valuesc>d
Control
2/50 (4)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Hematopoietic System: Malignant Lymphoma,
Undif f erentiated Leukemia, or
Lymphocytic Leukemiab
P Valuesc»d
6/50 (12)
N.S.
3.000
0.569
29.254
111
11/50 (22)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Low
Dose
91
6/50 (12)
N.S.
0.545
0.179
1.477
96
96
High
Dose
2/50 (4)
N.S.
1.000
0.075
13.326
111
14/50
N.S.
1.273
0.597
2.785
83
(28)
Table El.
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavage a
(continued)
Low
Topography;
Morphology
Hematopoietic System:
All Neoplasms'3
P Values0 » d
Departure from Linear Trend6
Control
13/50 (26)
N.S.
Neoplastic Nodule^
P Valuesc»d
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
N.S.
0.539
0.199
1.323
90
Weeks to First Observed Tumor
Liver:
7/50 (14)
High
Dose
16/50 (32)
N.S.
P = 0.043
Relative Risk (Control)f
Lower Limit
Upper Limit
o
o
Dose
0/49
(0)
P = 0.022
1.231
0.624
2.474
83
96
3/50 (6)
N.S.
Infinite
0.590
Infinite
111
5/49
(10)
P = 0.028
Infinite
1.262
Infinite
109
Table El.
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavage a
(continued)
Topography^ Morphology
Control
Liver: Neoplastic Nodule or
Hepatocellular Carcinomab
0/49 (0)
P Valuesc»d
P = 0.010
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Pituitary:
Chromophobe Adenoma*5
P Valuesc»d
—
3/46 (7)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
111
Low
Dose
3/50 (6)
High
Dose
6/49
(12)
N.S.
P = 0.012
Infinite
0.590
Infinite
Infinite
1.601
Infinite
111
5/48
(10)
109
4/49 (8)
N.S.
N.S.
1.597
0.331
9.773
1.252
0.224
8.137
101
95
Table El.
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavagea
(continued)
Low
Topographyj
Adrenal:
Morphology
Pheochromocytoma^
P Valuesc»d
Control
4/49 (8)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
g
Thyroid:
Follicular-cell Carcinomab
Dose
5/50 (10)
N.S.
1.225
0.280
5.833
88
1/46 (2)
79
High
Dose
5/50 (10)
N.S.
1.225
0.280
5.833
106
0/49 (0)
4/47 (9)
N.S.
N.S.
N>
P Valuesc>d
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
0.000
0.000
17.510
111
—
3.915
0.407
188.454
99
Table El.
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavage3
(continued)
Low
Topography:
Thyroid:
Morphology
C-cell Carcinomab
P Valuesc»d
Dose
High
Dose
1/46 (2)
2/49 (4)
0/47 (0)
N.S.
N.S.
N.S.
Control
Relative Risk (Control)f
Lower Limit
Upper Limit
1.878
0.101
108.485
0.000
0.000
18.240
__
Weeks to First Observed Tumor
Thyroid:
C-cell Adenoma or Carcinomab
111
111
4/46 (9)
10/49 (20)
N.S.
N.S.
4/47 (9)
o
CO
P Valuesc>d
Departure from Linear Trend6
P = 0.044
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
N.S.
2.347
0.735
9.593
111
107
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
Topography:
Morphology
Pancreatic Islets:
Islet-cell Adenoma^
P Valuesc'd
Control
4/49 (8)
P = 0.033
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
o
-P­
Preputial Gland: Adenoma, NOS
(not otherwise specified)"
P Valuesc»d
6/50 (12)
N.S.
1.470
0.372
6.681
88
87
High
Dose
11/50 (22)
P = 0.049
2.695
0.865
10.868
83
2/50 (4)
1/50 (2)
4/50 (8)
N.S.
N.S.
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Dose
0.500
0.009
9.290
111
111
2.000
0.301
21.316
106
Table El.
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavagea
(continued)
Topography:
Testis:
Morphology
Interstitial-cell Tumorb
P Valuesc»d
Control
48/50 (96)
N.S.
Relative Risk (Control) f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Peritoneum:
o
01
Mesothelioma, NOSb
P Values0 » d
84
1/50 (2)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Low
Dose
44/49
(90)
48/49
N.S.
N.S.
0.935
0.871
1.058
1.020
0.954
1.061
65
3/50 (6)
N.S.
3.000
0.287
74.701
111
High
Dose
79
(98)
82
0/50 (0)
N.S.
0.000
0.000
18.658
—
Table El.
Analyses of the Incidence of Primary Tumors in Male Rats
Administered 3-Nitropropionic Acid by Gavage3
(continued)
Low
Topography:
Lung:
Morphology
Alveolar/Bronchiolar Carcinoma*3
P Valuesc»d
Dose
High
Dose
0/50 (0)
1/49 (2)
3/48 (6)
N.S.
N.S.
N.S.
Infinite
0.055
Infinite
Infinite
0.627
Infinite
Control
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Lung: Alveolar/Bronchiolar Adenoma
or Carcinomab
P Valuesc>d
—
86
3/50 (6)
2/49 (4)
3/48 (6)
N.S.
N.S.
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
111
0.680
0.059
5.680
102
111
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),
1.042
0.146
7.419
86
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 inci­
dence 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.
Topography:
Analyses of the Incidence of Primary Tumors in Female Rats
Administered 3-Nitropropionic Acid by Gavage a
Morphology
Hematopoietic System: Malignant Lymphoma,
Lymphocytic Leukemia, or
Leukemia, NOSb
P Valuesc»d
Control
5/50 (10)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
o
Weeks to First Observed Tumor
Low
Dose
5/50 (10)
N.S.
1.000
0.245
4.082
98
106
7/50 (14)
6/50 (12)
High
Dose
8/50 (16)
N.S.
1.600
0.497
5.808
83
00
Hematopoietic System:
All Neoplasms'3
P Values0 » d
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
N.S.
0.857
0.268
2.684
98
105
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
Chromophobe Adenomab
Pituitary:
P Values0 » d
Control
19/45
(42)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
o
vO
Adrenal:
Pheochromocytoma^
P Valuesc»d
High
Dose
15/46 (33)
20/47 (43)
N.S.
N.S.
0.772
0.423
1.393
90
96
1.008
0.596
1.711
95
3/49 (6)
1/50 (2)
1/49 (2)
N.S.
N.S.
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Dose
0.327
0.013
3.417
107
103
0.333
0.013
3.486
107
Table E2.
Analyses of the Incidence of Primary Tumors in Female Rats
Administered 3-Nitropropionic Acid by Gavagea
(continued)
Low
Dose
High
Dose
2/50 (4)
1/44 (2)
2/44 (5)
N.S.
N.S.
N.S.
Topography; Morphology
Control
Thyroid:
C-cell Carcinoma^
P Values0 > d
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Thyroid: C-cell
Adenoma or Carcinoma^
P Values0 » d
0.568
0.010
10.516
111
111
59
5/50 (10)
4/44 (9)
5/44 (11)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
1.136
0.085
15.083
N.S.
0.909
0.191
3.952
111
111
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)
Low
Topography:
Morphology
Mammary Gland:
Fibroadenomab
P Valuesc»d
Dose
High
Dose
12/50 (24)
14/50 (28)
13/50 (26)
N.S.
N.S.
N.S.
Control
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observe4 Tumor
Uterus:
Endometrial Stromal Polypb
P Valuesc»d
1.167
0.560
2.475
94
Weeks to First Observed Tumor
67
2/50 (4)
4/48 (8)
N.S.
N.S.
2.083
0.314
22.174
Relative Risk (Control) ^
Lower Limit
Upper Limit
111
1.083
0.509
2.334
96
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).
88
5/49
(10)
N.S.
2.551
0.441
25.786
111
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
Low
Dose
High
Dose
Topography; Morphology
Control
Lung: Alveolar/Bronchiolar
Carcinoma"
4/49 (8)
3/48 (6)
3/50 (6)
N.S.
N.S.
N.S.
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
0.766
0.118
4.285
104
14/49 (29)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
96
8/48 (17)
N.S.
0.583
0.234
1.345
85
96
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
Topography:
Morphology
Hematopoietic System: Malignant Lymphoma
or Lymphocytic Leukemia'1
P Valuesc»d
Control
6/49 (12)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Hematopoietic System: All Neoplasmsb
P Valuesc»d
10/50
(20)
N.S.
91
8/49 (16)
N.S..
N.S.
1.470
0.608
3.783
91
1.797
0.664
5.463
73
12/50
73
11/50 (22)
N.S.
1.633
0.585
5.059
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
High
Dose
Dose
91
(24)
12/50
N.S.
1.470
0.608
3.783
91
(24)
Table Fl.
Analyses of the Incidence of Primary Tumors in Male Mice
Administered 3-Nitropropionic Acid by Gavagea
(continued)
Topography:
Morphology
Liver: Hepatocellular Carcinoma^
P Valuesc»d
Control
16/49
(33)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
P Valuesc»d
Departure from Linear Trend6
97
20/49
(41)
N.S.
(16)
P = 0.044 (N)
92
10/50 (20)
P = 0.021 (N)
High
Dos e
12/49
(24)
N.S.
0.750
0.364
1.504
87
16/49 (33)
N.S.
P = 0.037
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
8/50
0.490
0.201
1.094
Weeks to First Observed Tumor
Liver: Hepatocellular Adenoma
or Carcinomab
Low
Dose
0.490
0.231
0.975
97
92
0.800
0.446
1.419
87
Table Fl.
Analyses of the Incidence of Primary Tumors in Male Mice
Administered 3-Nitropropionic Acid by Gavage3
(continued)
Topography:
Morphology
All Sites: Hemangiosarcomab
P Values0 > d
Control
5/49 (10)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
OC
All Sites: Hemangioma or
Hemangiosarcoma"
P Values0 » d
1/50 (2)
N.S.
0.196
0.004
1.665
85
6/49 (12)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to first Observed Tumor
Low
Dose
96
2/50 (4)
N.S.
0.327
0.033
1.723
85
96
High
Dose
6/50 (12)
N.S.
1.176
0.320
4.565
99
8/50 (16)
N.S.
1.307
0.430
4.243
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
vo
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 F2.
Analyses of the Incidence of Primary Tumors in Female Mice
Administered 3-Nitropropionic Acid by Gavagea
Low
Topography;
Morphology
Lung: Alveolar/Bronchiolar
Carcinoma**
P Valuesc » d
Dose
High
Dose
0/47 (0)
2/49 (4)
2/49 (4)
N.S.
N.S.
N.S.
Infinite
0.284
Infinite
Infinite
0.284
Infinite
Control
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
NO
O
Lung: Alveolar/Bronchiolar
Adenoma or Carcinoma'5
P Valuesc»d
2/47 (4)
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed tumor
105
105
6/49 (12)
3/49 (6)
N.S.
2.878
0.547
28.023
104
105
N.S.
1.439
0.173
16.603
105
Table F2.
Analyses of the Incidence of Primary Tumors in Female Mice
Administered 3-Nitropropionic Acid by Gavagea
(continued)
Low
Topography:
Morphology^
Control
Hematopoietic System: Malignant Lymphoma,
Undif f erentiated Leukemia, or
Lymphocytic Leukemia^
20/50 (40)
P Valuesc»d
P = 0.021 (N)
Relative Risk (Control)f
Lower Limit
Upper Limit
KJ
Weeks to First Observed Tumor
18/50
(36)
21/50
P Valuesc»d
P = 0.014 (N)
Relative Risk (Control)f
(42)
21/50
N.S.
1.000
0.603
1.659
Lower Limit
Upper Limit
79
84
(20)
0.500
0.235
0.996
100
84
79
10/50
P = 0.024 (N)
N.S.
0.900
0.516
1.561
Hematopoietic System: All Neoplasmsb
Weeks to First Observed Tumor
High
Dps e
Dose
(42)
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
Tqp_ograj>hy:
Morphology
Liver: Hepatocellular Carcinomab
P Valuesc>d
Dose
High
Dose
1/49 (2)
1/50 (2)
2/50 (4)
N.S.
N.S.
N.S.
Control
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
N>
S3
All Sites: Hemangiosarcoma^
P Valuesc»d
0.980
0.013
75.404
104
105
1/50 (2)
1/50 (2)
3/50 (6)
N.S.
N.S.
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
—
1.960
0.106
113.310
1.000
0.013
76.970
104
105
3.000
0.251
154.270
101
Table F2.
Analyses of the Incidence of Primary Tumors in Female Mice
Administered 3-Nitropropionic Acid by Gavage3
(continued)
Low
Topography;
Morphology
All Sites: Sarcoma of All Kindsb
P Valuesc»d
Control
2/50 (4)
4/50 (8)
N.S.
N.S.
Relative Risk (Control)f
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Pituitary: Chromophobe Adenoma
P Valuesc'd
2.000
0.301
21.316
104
96
High
Dose
6/50
(12)
N.S.
3.000
0.569
29.254
98
2/43 (5)
4/48 (8)
1/42 (2)
N.S.
N.S.
N.S.
1.792
0.272
19.046
Relative Risk (Control) ^
Lower Limit
Upper Limit
Weeks to First Observed Tumor
Dose
104
105
0.512
0.009
9.452
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.
,_•
NJ
*­
e
The probability level for departure from linear trend is given when P < 0.05 for any comparison.
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 Clear­
inghouse is to advise on the National Cancer Institute's
bioassay program to identify and evaluate chemical carcino­
gens in the environment to which humans may be exposed. The
members of the Clearinghouse have been drawn from academia,
industry, organized labor, public interest groups, State
health officials, and quasi-public health and research
organizations. Members have been selected on the basis of
their experience in carcinogenesis or related fields and,
collectively, provide expertise in organic chemistry, bio­
chemistry, 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 carcino­
genic) 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 hepato­
ceullular carcinomas for the purposes of obtaining a stat­
istically 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