National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 133 

1979 

BIOASSAY OF 

3-NITRO-p-ACETOPHENETIDE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 89-25-8 

NCI-CG-TR-133 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

3-NITRO-p-ACETOPHENETIDE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20205
 

US. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
 

Public Health Service
 

National Institutes of Health
 

NIH Publication No. 79-1388
 

REPORT ON THE BIOASSAY OF 3-NITRO-p-ACETOPHENETIDE
 

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-nitro-p-acetophenetide conducted for the Carcinogenesis Testing
 

Program, Division of Cancer Cause and Prevention, National Cancer
 

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

This is one of a series of experiments designed to determine whether
 

selected chemicals have the capacity to produce cancer in animals.
 

Negative results, in which the test animals do not have a significantly greater incidence of cancer than control animals, do not
 

necessarily mean the test chemical is not a carcinogen because
 

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 a potential
 

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

carcinogens requires a wider analysis.
 

CONTRIBUTORS: This bioassay of 3-nitro-p-acetophenetide was conducted
 

by Mason Research Institute, Worcester, Massachusetts, initially under
 

direct contract to the NCI and currently under a subcontract to Tracor
 

Jitco, Inc., prime contractor for the NCI Carcinogenesis Testing Program.
 

The experimental design was determined by the NCI Project Officers, Dr. J. H. Weisburger (1,2) and Dr. E. K. Weisburger (1). The
 

principal investigators for the contract were Dr. E. Smith (3) and
 

Dr. A. Handler (3). Animal treatment and observation were supervised
 

by Mr. G. Wade (3) and Ms. E. Zepp (3).
 

Histopathologic examinations were performed by Dr. D. S. Wyand
 

(3) at the Mason Research Institute, and the diagnoses included in
 

this report represent the interpretation of this pathologist. Histopathology findings and reports were reviewed by Dr. R. L. Schueler
 

(4).
 

Compilation of individual animal survival, pathology, and summary
 

tables was performed by EG&G Mason Research Institute (5); the statistical analysis was performed by Mr. W. W. Belew (6,7), using methods
 

selected for the Carcinogenesis Testing Program by Dr. J. J. Gart (8).
 

This report was prepared at METREK, a Division of The MITRE Corporation (6) under the direction of the NCI. Those responsible for
 

iii
 
this report at METREK are the project coordinator, Dr. L. W. Thomas
 

(6), task leader Dr. M. R. Kornreich (6,9), senior biologist Ms. P.
 

Walker (6), biochemist Mr. S. C. Drill (6), chemist Dr. N. Zimmerman
 

(6), and the technical editor Ms. P. A. Miller (6). The final report
 

was reviewed by members of the participating organizations.
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

the results, and reporting the findings: Dr. K. C. Chu (1), Dr. C.
 

Cueto, Jr. (1), Dr. J. F. Douglas (1), Dr. D. G. Goodman (1,9), Dr.
 

R. A. Griesemer (1), Dr. M. H. Levitt (1), Dr. H. A. Milman (1), Dr.
 

T. W. Orme (1), Dr. R. A. Squire (1,10), Dr. S. F. Stinson (1), Dr.
 

J. M. Ward (1), and Dr. C. E. Whitmire (1).
 

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

2.	 No wit w h the Naylor Dana Institute for Disease Prevention, American Health Foundation, Hammon House Road, Valhalla, New York.
 

3.	 Mason Research Institute, 57 Union Street, Worcester, Massachusetts.
 

4.	 Tracer Jitco, Inc., 1776 East Jefferson Street, Rockville,
 

Maryland.
 

5.	 EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

6.	 The MITRE Corporation, METREK Division, 1820 Dolley Madison
 

Boulevard, McLean, Virginia.
 

7.	 No wit w h the Solar Energy Research Institute, Cole Boulevard,
 

Golden, Colorado.
 

8.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

Cause and Prevention, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
 

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

Washington, D.C.
 

10. Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

iv
 
SUMMARY
 

A bioassay for possible carcinogenicity of 3-nitro-p-acetophenetide was conducted using Fischer 344 rats and B6C3F1 mice. 3-Nitrop-acetophenetide was administered in the feed, at either of two
 

concentrations, to groups of 50 male and 50 female animals of each
 

species, with the exception of low dose male mice, of which there
 

were 49. Fifty animals of each sex and species were placed on test
 

as controls. The high and low time-weighted average dietary concentrations of 3-nitro-p-acetophenetide were, respectively, 0.36 and
 

0.18 percent for rats and 1.46 and 0.73 percent for mice. The compound was administered in the diet for 78 weeks, followed by an observation period of up to 30 weeks for rats and 20 weeks for mice.
 

There were no significant positive associations between the concentrations of 3-nitro-p-acetophenetide administered and mortality in
 

rats or mice of either sex. In addition, adequate numbers of animals
 

in all groups survived sufficiently long to be at risk from late-


developing tumors.
 

There was a statistically significant increased incidence of
 

a combination of hepatocellular carcinomas and adenomas when high
 

dose male mice were compared to controls. No other neoplasm in any
 

other dosed group occurred in significant positive increased incidences when compared to controls.
 

Under the conditions of this bioassay, dietary administration
 

of 3-nitro-p-acetophenetide was not carcinogenic in Fischer 344 rats
 

of either sex or in female B6C3F1 mice. The compound, however, was
 

considered carcinogenic in male B6C3F1 mice based on a significant
 

increase in the combined incidence of hepatocellular carcinomas and
 

hepatocellular adenomas in these animals.
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 5
 

A. Chemicals 5
 

B. Dietary Preparation 5
 

C. Animals 6
 

D. Animal Maintenance 6
 

E. Selection of Initial Concentrations 9
 

F. Experimental Design 10
 

G. Clinical and Histopathologic Examinations 12
 

H. Data Recording and Statistical Analyses 15
 

III. CHRONIC TESTING RESULTS: RATS 21
 

A. Body Weights and Clinical Observations 21
 

B. Survival 21
 

C. Pathology 24
 

D. Statistical Analyses of Results 24
 

IV. CHRONIC TESTING RESULTS: MICE 33
 

A. Body Weights and Clinical Observations 33
 

B. Survival 33
 

C. Pathology 36
 

D. Statistical Analyses of Results 38
 

V. DISCUSSION 45
 

VI. BIBLIOGRAPHY 47
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 3-NITRO-p-


ACETOPHENETIDE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 3-NITRO-p-


ACETOPHENETIDE D-l
 

vii
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF 3-NITRO-p-ACETOPHENETIDE 2 

2 GROWTH CURVES FOR

CHRONIC STUDY RATS 

 3-NITRO-p-ACETOPHENETIDE 

22 

SURVIVAL COMPARISONS OF

TIDE CHRONIC STUDY RATS 

 3-NITRO-p-ACETOPHENE23 

GROWTH CURVES FOR

CHRONIC STUDY MICE 

 3-NITRO-p-ACETOPHENETIDE 

34 

SURVIVAL COMPARISONS OF

TIDE CHRONIC STUDY MICE 

 3-NITRO-p-ACETOPHENE35 

LIST OF TABLES 

Table Number 

DESIGN SUMMARY FOR FISCHER 344 RATS—3-NITROp-ACETOPHENETIDE FEEDING EXPERIMENT 11 

DESIGN SUMMARY FOR B6C3F1 MICE—3-NITRO-pACETOPHENETIDE FEEDING EXPERIMENT 12 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

3-NITRO-p-ACETOPHENETIDE 25 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH 3-NITRO-p-ACETOPHENETIDE 28 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN MALE 

MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE 39 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH 3-NITRO-p-ACETOPHENETIDE 41 

Vlll 
LIST OF TABLES (Concluded) 

Table Number Page 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE 

RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE A-7 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH 3-NITRO-pACETOPHENETIDE B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH 3-NITROp-ACETOPHENETIDE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 3-NITROp-ACETOPHENETIDE C-8 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH 3-NITROp-ACETOPHENETIDE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 3-NITROp-ACETOPHENETIDE D-6 

IX
 

I. INTRODUCTION
 

3-Nitro-p-acetophenetide (Figure 1) (NCI No. C01978), a derivative of the analgesic phenacetin, was selected for bioassay by the
 

National Cancer Institute because of the suspected renal pelvic carcinogenicity of the parent compound (juusela, 1973; Weisburger, 1977;
 

Odashima, 1978).
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is N-(4-ethoxyphenyl)-3'-nitroacetamide. It is also called 3'-nitro-p-acetophenetide (or 3'-nitro-pacetophenetidin); and 4-acetamino-2-nitrophenetole.
 

3-Nitro-p-acetophenetide can be produced by the nitration of
 

phenacetin, and this synthesis has been exploited for the detection
 

of the latter (Oelschlaeger and Welsch, 1964).
 

Specific production data for 3-nitro-p-acetophenetide are not
 

available; however, 1970 appears to have been the last year in which
 

commercial production (in excess of 1000 pounds or $1000 in value
 

annually) of the compound was reported in the United States (U.S.
 

Tariff Commission, 1972; as cited in Urso, 1977).
 

The potential for exposure to 3-nitro-p-acetophenetide is appar- .
 

ently limited to researchers and workers engaged in the synthesis of
 

this compound.
 

No information on the human toxicological properties of 3-nitrop-acetophenetide is currently available; however, the para aminophenols
 

*
 

The CAS registry number is 1777-84-0.
 
CH2CH3 

FIGURE 1 

CHEMICAL STRUCTURE OF 3-NITRO-p-ACETOPHENETIDE 
as a class have been implicated in the production of a variety of
 

effects on the hematopoietic system of man (Barr and Penna, 1971).
 

II. MATERIALS AND METHODS
 

A. Chemicals
 

3-Nitro-p-acetophenetide was purchased from Carroll Products,
 

Wood River Junction, Rhode Island. Chemical analysis was performed
 

by Mason Research Institute, Worcester, Massachusetts. The experimentally determined range in the melting point was 116° to 118°C.
 

Thin-layer chromatography utilizing ultraviolet light for detection
 

showed only one spot. Ultraviolet analyses showed peaks at 245 and
 

353 nm. The compound was identified as 3-nitro-p-acetophenetide with
 

a purity of 99 percent based on high pressure liquid chromatography
 

analysis conducted three weeks after the chronic studies were
 

terminated.
 

Throughout this report, the term 3-nitro-p-acetophenetide is used
 

to represent this chemical.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals consisted of Wayne Lab-Blox® (Allied Mills, Inc., Chicago, Illinois).
 

3-Nitro-p-acetophenetide was administered to the dosed animals as a
 

component of the diet.
 

The chemical was removed from its amber glass container, sifted,
 

weighed out in proper amounts under an exhaust hood, and mixed in a
 

mortar and pestle with an aliquot of the ground feed. Once visual
 

homogeneity was attained, the mixture was placed into a 6 kg capacity
 

Patterson-Kelley twin-shell V-blender along with the remainder of the
 

meal. The blender was sealed and operated for 20 minutes. Prepared
 

diets were placed in double plastic bags and stored in the dark at
 

5
 
4°C. The mixtures were prepared weekly and discarded 14 days after
 

formulation.
 

C. Animals
 

Two animal species, rats and mice, were used in the carcinogenicity bioassay. Fischer 344 rats and B6C3F1 mice were obtained through
 

contracts of the Division of Cancer Treatment, National Cancer Institute. Rats and mice were supplied by Charles River Breeding Laboratories, Inc., Wilmington, Massachusetts. Dosed and control animals
 

for each species were received in separate shipments. Upon arrival,
 

a sample of animals was examined for parasites and other signs of
 

disease. The remaining animals were quarantined by species for 2
 

weeks prior to initiation of test. Animals were assigned to groups
 

and distributed among cages so that average body weight per cage was
 

approximately equal for a given species and sex.
 

D. Animal Maintenance
 

All animals were housed by species in rooms having a temperature
 

®
 

range of 23° to 34°C. Incoming air was filtered through Tri-Dek
 

®
 15/40 denier Dacron filters (Tri-Dim Filter Corp., Hawthorne, New
 

Jersey) providing six changes of room air per hour. Fluorescent
 

lighting was provided on a 12-hour-daily cycle.
 

Rats were housed five per cage by sex. During quarantine and
 

for the first 7 months of study, rats were kept in galvanized- or
 

stainless-steel wire-mesh cages (Fenco Cage Products, Boston, Massachusetts) suspended above newspapers. Newspapers under cages were
 
replaced daily and cages and racks washed weekly. For the duration
 

of the study, all rats were held in suspended polycarbonate cages
 

(Lab Products, Inc., Garfield, New Jersey) equipped with disposable
 

nonwoven fiber filter sheets. Clean corncob bedding and cages were
 

provided twice weekly. Stainless steel cage racks (Fenco Cage Products) were cleaned once every 2 weeks, and disposable filters were
 

replaced at that time.
 

Mice were housed by sex in polycarbonate shoe box type cages.
 

During quarantine and periods of chemical administration, cages were
 

fitted with perforated stainless steel lids. During the final observation period, stainless steel wire bar lids were used. Cages and
 

both types of lids were from Lab Products, Inc. Nonwoven fiber filter bonnets were used over cage lids. Dosed mice were housed ten per
 

cage for the first 17 months and five per cage thereafter. Control
 

mice were changed from ten to five per cage after 14 months. Clean
 

cages, lids, and bedding were provided three times per week when cage
 

populations were ten and twice per week when cage populations were
 

reduced to five. Hardwood chip bedding was used for the first 6
 

months of study and corncob bedding was used for the remainder of
 

the study. Reusable filter bonnets and pipe racks were sanitized
 

every 2 weeks throughout the study.
 

Tap water was available from 250 ml water bottles equipped with
 

rubber stoppers and stainless steel sipper tubes. Bottles were
 

replaced twice weekly and, for rats only, water was supplied as
 

needed between changes. Food and water were available ad libitum.
 

7
 
Pelleted Wayne Lab-Blox \ 

 was supplied to rats during the quarantine period and to both rats and mice during the final observation
 

period. During the 78-week dosing period, all dosed animals were
 

(R)
 supplied with Wayne Lab-Blox meal containing the appropriate concentration of 3-nitro-p-acetophenetide. Control animals had untreated
 

(R)
 

meal available. Meal was dispensed from Alpine aluminum feed cups
 

(Curtin Matheson Scientific, Inc., Woburn, Massachusetts) containing
 

stainless steel baffles. Rats received pelleted feed on the cage
 

floor. Mice were fed pellets from a hopper incorporated into the wire
 

bar cage lids. Food hoppers were changed on the same schedule as were
 

(R)
 cages. Food was replenished daily in Alpinew

 feed cups.
 

All dosed and control rats were housed in a room with other
 

st
 

rats receiving diets containing amitrole (61-82-5) and 2-methyl-lnitroanthraquinone (129-15-7).
 

Dosed mice were housed in a room with other mice receiving diets
 

containing amitrole (61-82-5); N,N-dimethyl-p-nitrosoaniline (138-89­

6); 2,5-toluenediamine sulfate (6369-59-1); 2,4-dinitrotoluene (121­

14-2); 2-aminoanthraquinone (117-79-3); 3-amino-4-ethoxyacetanilide
 

(17026-81-2); 3-amino-9-ethylcarbazole hydrochloride; APC (8003-03-0);
 

l-amino-2-methylanthraquinone (82-28-0); 5-nitro-o-anisidine (99-59­

2); 4-nitroanthranilic acid (619-17-0); 1-nitronaphthalene (86-57-7);
 

5-nitroacenaphthene (602-87-9); and 2,4-diaminoanisole sulfate (615­

05-4). Control mice were housed in a room with other mice receiving
 

*
 CAS registry numbers are given in parentheses.
 
diets containing 2-methyl-l-nitroanthraquinone (129-15-7); p-cresidine (120-71-8); 4-chloro-m-phenylenediamine (5131-60-2); acetylaminofluorene (53-96-3); and fenaminosulf (140-56-7).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

3-nitro-p-acetophenetide for administration to dosed animals in the
 

chronic study, subchronic toxicity studies were conducted with both
 

rats and mice. Animals of each species were distributed among six
 

groups, each consisting of five males and five females. 3-Nitro-pacetophenetide was incorporated into the laboratory diet and supplied
 

ad libitum to five of the six rat groups and five of the six mouse
 

groups in concentrations of 0.05, 0.1, 0.2, 0.4, and 0.8 percent.
 

The sixth group of each species served as a control group, receiving
 

only the basal laboratory diet. The dosed dietary preparations were
 

administered for 7 weeks, followed by a 1-week observation period
 

during which animals were fed the basal diet. Survivors were sacrificed at the end of the test, and gross necropsies were performed.
 

The highest concentration causing no deaths, no compound-related
 

gross abnormalities, and no mean body weight depression in excess of
 

10 percent relative to controls during the 8-week subchronic test was
 

selected as the high concentration utilized for the chronic bioassays
 

of both species.
 

All animals survived until the end of test. Mean body weight
 

depression in female rats was 2.4, 6.0, and 15.0 percent at dietary
 
concentrations of 0.05, 0.1, and 0.2 percent, respectively. Mean
 

body weight depression in male rats were 18.4, 8.3, and 19.4 percent
 

at dietary concentrations of 0.05, 0.1, and 0.2 percent, respectively.
 

Enlarged submaxillary glands were noted in mice receiving concentrations of 0.8 percent. Mean body weight depression in female
 

mice was 6.3 and 8.2 percent at dietary concentrations of 0.4 and
 

0.8 percent, respectively. Mean body weight depression in male mice
 

was 2.6 percent at a dietary concentration of 0.4 percent and no mean
 

body weight depression was observed at a dietary concentration of 0.8
 

percent.
 

The high concentrations chosen for the chronic study were 0.1
 

percent for rats and 0.4 percent for mice.
 

F. Experimental Design
 

The experimental design parameters for the chronic study (species, sex, group size, concentrations administered, duration of
 

treated and untreated observation periods, and time-weighted average
 

concentrations) are summarized in Tables 1 and 2.
 

The dosed and control rats were all approximately 6 weeks old
 

at the time they were placed on test. The initial concentrations of
 

3-nitro-p-acetophenetide utilized were 0.1 and 0.05 percent. Throughout this report those rats initially receiving the former concentration are referred to as the high dose rat groups, while those initially
 

receiving the latter concentration are referred to as the low dose
 

rat groups. In week 10, concentrations were raised to 0.4 and 0.2
 

10
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

3-NITRO-p-ACETOPHENETIDE FEEDING EXPERIMENT
 

MALE 

CONTROL

LOW DOSE

INITIAL 

GROUP 

SIZE 

 50 

 50 

HIGH DOSE 50 

FEMALE 

CONTROL

LOW DOSE

 50 

 50 

HIGH DOSE 50 

0 

3-NITRO-p-


ACETOPHENETIDE
 

CONCENTRATION
 

(PERCENT)
 

0
 

0.05
 

0.2
 

0
 

0.1
 

0.4
 

0
 

0
 

0.05
 

0.2
 

0
 

0.1
 

0.4
 

0
 

OBSERVATION PERIOD TIME-WEIGHTED 

TREATED UNTREATED AVERAGE 

(WEEKS) (WEEKS) CONCENTRATION3 

0 108 0
 

9 0.18
 

69
 

28
 

9 0.36
 

69
 

29
 

0 109 0
 

9 0.18
 

69
 

29
 

9 0.36
 

69
 

30
 

£( concentration X weeks received)
 

11
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

3-NITRO-p-ACETOPHENETIDE FEEDING EXPERIMENT
 

INITIAL 

GROUP 

SIZE 

MALE 

CONTROL 50 

LOW DOSE 49 

HIGH DOSE 50 

FEMALE 

CONTROL 50 

LOW DOSE 50 

HIGH DOSE 50 

3-NITRO-p-


ACETOPHENETIDE
 

CONCENTRATION
 

(PERCENT)
 

0
 

0.2
 

0.8
 

0
 

0.4
 

1.6
 

0
 

0
 

0.2
 

0.8
 

0
 

0.4
 

1.6
 

0
 

OBSERVATION PERIOD

TREATED UNTREATED

(WEEKS) (WEEKS)

 TIME -WEIGHTED 

 AVERAGE 

CONCENTRATION3 

0 98 0 

9

69 

20 

 0.73 

9

69 

20 

 1.46 

0 98 0 

9

69 

20 

 0.73 

9

69 

20 

 1.46 

a S (concentration X weeks received)
 Ti Tn<a—wf* i ah t-f>d av(*r;ia*a rrmr£>n1~T,at-inn
 

2(weeks receiving chemical)
 

12
 
percent for the rat groups because of insufficient mean body weight
 

depression. These concentrations were not changed for the remainder
 

of the 78-week dosing period. Five males and five females from the
 

high dose and control groups were sacrificed and necropsied at the
 

end of the compound administration phase according to existing protocol. The remaining rats were observed for an additional period of
 

up to 30 weeks after compound administration ceased.
 

All mice were approximately 6 weeks old at the time they were
 

placed on test. The initial concentrations of 3-nitro-p-acetophenetide utilized were 0.4 and 0.2 percent. Throughout this report those
 

mice initially receiving the former concentration are referred to as
 

the high dose groups, while those initially receiving the latter concentration are referred to as the low dose groups. In week 10, concentrations were raised to 1.6 and 0.8 percent for the high and low
 

dose mouse groups, respectively, because of insufficient mean body
 

weight depression. These concentrations were not changed for the
 

remainder of the 78-week dosing period. Five low dose male mice were
 

sacrificed and necropsied in week 14, and five males and five females
 

from the high dose and control groups were sacrificed and necropsied
 

at the end of the compound administration phase, according to existing
 

protocol. All remaining mice were observed for an additional period
 

of 20 weeks after compound administration ceased.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. Body weights were recorded twice weekly for the first
 

13
 
12 weeks of the study and at monthly intervals thereafter. From the
 

first day, all animals were inspected twice daily for mortality.
 

Food consumption, for two cages from each group, was monitored for
 

seven consecutive days once a month for the first nine months of
 

the bioassay and for three consecutive days each month thereafter.
 

The presence of tissue masses and lesions was determined by monthly
 

observation and palpation of each animal.
 

A necropsy was performed on each animal regardless of whether it
 

died, was killed when moribund, or was sacrificed at the end of the
 

bioassay. The animals were euthanized by carbon dioxide inhalation,
 

and were immediately necropsied. The histopathologic examination
 

consisted of gross and microscopic examination of major tissues,
 

organs, and gross lesions taken from sacrificed animals and, whenever
 

possible, from animals found dead.
 

Tissues were preserved in 10 percent buffered formalin, embedded
 

in paraffin, sectioned, and stained with hematoxylin and eosin prior
 

to microscopic examination. An occasional section was subjected to
 

special staining techniques for more definitive diagnosis.
 

Slides were prepared from the following tissues: skin, subcutaneous tissue, lungs and bronchi, trachea, bone marrow, spleen, lymph
 

nodes, thymus, heart, salivary gland, liver, gallbladder (mice), pancreas, esophagus, stomach, small intestine, large intestine, kidney,
 

urinary bladder, pituitary, adrenal, thyroid, parathyroid, testis,
 

prostate, brain, uterus, mammary gland, and ovary.
 

14
 
A few tissues were not examined for some animals, particularly
 

for those that died early. Also, some animals were missing, cannibalized, or judged to be in such an advanced state of autolysis as to
 

preclude histopathologic interpretation. Thus, the number of animals
 

for which particular organs, tissues, or lesions were examined microscopically varies and does not necessarily represent the number of
 

animals that were placed on experiment in each group.
 

H. Data Recording and Statistical Analyses
 

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

(Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design, clinical observations, survival, body weight, and individual pathologic results,
 

as recommended by the International Union Against Cancer (Berenblum,
 

1969). Data tables were generated for verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques described in this section. Those analyses of the experimental results
 

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

in the form of graphs. Animals were statistically censored as of the
 

15
 
time that they died of other than natural causes or were found to be
 

missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect
 

on survival used the method of Cox (1972) when testing two groups for
 

equality and used Tarone's (1975) extensions of Cox's methods when
 

testing 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 was examined (denominator). In most instances, the denominators included only those animals for which that site was examined
 

histologically. However, when macroscopic examination was required
 

to detect lesions prior to histologic sampling (e.g., skin or mammary
 

tumors), or when lesions could have appeared at multiple sites (e.g.,
 

lymphomas), the denominators consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a significantly higher proportion of tumors than did the control animals.
 

As a part of these analyses, the one-tailed Fisher exact test (Cox,
 

1970, pp. 48-52) was used to compare the tumor incidence of a control
 

16
 
group to that of a group of treated animals at each dose level. When
 

results for a number of treated groups, k, are compared simultaneously
 

with those for a control group, a correction to ensure an overall
 

significance level of 0.05 may be made. The Bonferroni inequality
 

(Miller, 1966, pp. 6-10) requires that the P-value for any comparison
 

be less than or equal to 0.05/k. In cases where this correction was
 

used, it is discussed in the narrative section. It is not, however,
 

presented in the tables, where the Fisher exact P-values are shown.
 

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

continuity correction (Armitage, 1971, pp. 362-365), was also used
 

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

determined if the slope of the dose-response curve is different from
 

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

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

tumor was observed were excluded by basing the statistical tests on
 

animals that survived at least 52 weeks, unless a tumor was found at
 

the anatomic site of interest before week 52. When such an early
 

tumor was found, comparisons were based exclusively on animals that
 

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

17
 
found. Once this reduced set of data was obtained, the standard procedures for analyses of the incidence of tumors (Fisher exact tests,
 

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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

during which animals died naturally or were sacrificed was entered as
 

the time point of tumor observation. Cox's methods of comparing
 

these curves were used for two groups; Tarone1

s extension to testing
 

for linear trend was used for three groups. The statistical tests for
 

the incidence of tumors which used life-table methods were one-tailed
 

and, unless otherwise noted, in the direction of a positive dose
 

relationship. Significant departures from linearity (P < 0.05, two-


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared to its control was calculated from
 

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

is defined as p /p where p is the true binomial probability of the
 

t c t
 

incidence of a specific type of tumor in a treated group of animals
 

and p is the true probability of the spontaneous incidence of the
 

c
 

same type of tumor in a control group. The hypothesis of equality
 

between the true proportion of a specific tumor in a treated group
 

and the proportion in a control group corresponds to a relative risk
 

18
 
of unity. Values in excess of unity represent the condition of a
 

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical analyses. The interpretation of the limits is that in approximately 95
 

percent of a large number of identical experiments, the true ratio
 

of the risk in a treated group of animals to that in a control group
 

would be within the interval calculated from the experiment. When
 

the lower limit of the confidence interval is greater than one, it
 

can be inferred that a statistically significant result (a P < 0.025
 

one-tailed test when the control incidence is not zero, P < 0.050
 

when the control incidence is zero) has occurred. When the lower
 

limit is less than unity but the upper limit is greater than unity,
 

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

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

detected under the conditions of this test.
 

19
 

III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Mean body weight depression was apparent in both male and female
 

dosed rats when compared to controls. Mean body weight depression
 

was observed after 15 weeks in males and after 8 weeks in females
 

(Figure 2).
 

Subcutaneous and/or cutaneous growths developed in one high dose
 

male, one low dose male, one low dose female, four control males, and
 

five control females. Exudate in the conjunctival sac of one high dose
 

female and in the ear of one low dose female was observed. Alopecia
 

was recorded in one low dose female. No other clinical abnormalities
 

were observed.
 

B. Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and 3-nitro-p-acetophenetide-dosed groups are shown in
 

Figure 3. For both male and female rats the Tarone test for positive
 

association between dosage and mortality was not significant.
 

For males five animals from the high dose and five from the control group were sacrificed in weeks 79 or 80. Sufficient numbers of
 

male rats were at risk from late-developing tumors, with 80 percent
 

(40/50) of the high dose, 84 percent (42/50) of the low dose, and
 

68 percent (34/50) of the control group surviving on test until the
 

termination of the study.
 

21
 
750 750 

— 600 

-450 

— 300 

— 150 

TIM EON TEST (WEEKS) 

120 

750­ •750 

600­ -600 

DC 

e? 

I- 450­ X 

o 

UJ 

§300-

CO 

-450 

-300 

150 — -150 

FEMALE RATS 

I 

15 

I 

30 

I 

45 

I 

60 

\ 

75 

TIME ON TEST (WEEKS) 

90 

I 

105 120 

FIGURE 2 

GROWTH CURVES FOR 3-NITRO-p-ACETOPHENETIDE CHRONIC STUDY RATS 

22 
1.0- -1.0 

0.8­ -0.8 

0.6­ -0.6 

CO 0.4­ -0.4 

co 

O 

cc 

o. 

CONTROL 

0.2­ -0.2 

LOW DOSE 

MALE RATS HIGH DOSE 

0.0­ \ I ' I ' I -0.0 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

-1.0 

-0.8 

0.6­ -0.6 

CO 0.4­ -0.4 

CO 

O 

CC 

Q­ CONTROL 

0.2­ LOW DOSE -0.2 

FEMALE RATS HIGH DOSE 

0.0­ ~r -1 1 1 1 r —T~ T -0.0 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

FIGURES 

SURVIVAL COMPARISONS OF 3-NITRO-p-ACETOPHENETIDE CHRONIC STUDY RATS 

23 
For females five animals from the high dose and five from the
 

control group were sacrificed in week 80. Sufficient numbers of
 

female rats were at risk from late-developing tumors, with 78 percent
 

(39/50) of the high dose, 94 percent (47/50) of the low dose, and 70
 

percent (35/50) of the control animals surviving on test until the
 

end of the study.
 

C. Pathology
 

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).
 

Tumors were found in comparable numbers in control and dosed
 

animals and were not considered to be related to compound administration. There were instances in this study, as noted in the summary
 

tables, where neoplastic lesions occurred only in dosed animals or
 

with increased frequency when compared to the control group. However,
 

the nature and incidence of these lesions are similar to those known
 

to occur spontaneously in aged Fischer 344 rats.
 

A variety of inflammatory and degenerative lesions, which commonly occur in aging rats of this strain, was seen. None of these
 

lesions was considered to be compound-induced.
 

Based upon the results of this pathologic examination, 3-nitrop-acetophenetide is not carcinogenic in Fischer 344 rats.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

rats are summarized in Tables 3 and 4. The analysis is included for
 

24
 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE*
 

TOPOGRAPHY: MORPHOLOGY
 

Pituitary: Adenoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromo cy toma
 

P Values0
 

to Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pancreatic Islets: Islet-Cell Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

1/41(0.02)
 

N.S.
 

108
 

10/47(0.21)
 

P = O.OOl(N)
 

P = 0.045
 

99
 

3/45(0.07)
 

N.S.
 

85
 

LOW
 

DOSE
 

4/46(0.09)
 

N.S.
 

3.565
 

0.373
 

171.546
 

105
 

1/49(0.02)
 

P = 0.003(N)
 

0.096
 

0.002
 

0.633
 

105
 

2/46(0.04)
 

N.S.
 

0.652
 

0.057
 

5.426
 

106
 

HIGH
 

DOSE
 

0/44(0.00)
 

N.S.
 

0.000
 

0.000
 

17.344
 

1/48(0.02)
 

P = 0.003(N)
 

0.098
 

0.002
 

0.646
 

107
 

1/45(0.02)
 

N.S.
 

0.333
 

0.007
 

3.964
 

107
 
___ 

TOPOGRAPHY: MORPHOLOGY
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma°
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma or
 

Neoplastic Nodule"
 

O> P Values0 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Testis: Interstitial-Cell Tumor
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

CONTROL
 

6/48(0.13)
 

P = 0.026(N)
 

98
 

3/48(0.06)
 

P = 0.035(N)
 

99
 

45/47(0.96)
 

N.S.
 

80
 

LOW
 

DOSE
 

2/50(0.04)
 

N.S.
 

0.320
 

0.033
 

1.687
 

106
 

0/50(0.00)
 

N.S.
 

0.000
 

0.000
 

1.596
 

47/50(0.94)
 

N.S.
 

0.982
 

0.911
 

1.082
 

65
 

HIGH
 

DOSE
 

1/50(0.02)
 

N.S.
 

0.160
 

0.004
 

1.249
 

104
 

0/49(0.00)
 

N.S.
 

0.000
 

0.000
 

1.628
 

39/49(0.80)
 

N.S.
 

0.831
 

0.765
 

0.987
 

80 
TABLE 3 (CONCLUDED)
 

Treated groups received time-weighted average doses of 0.18 or 0.36 percent in feed.
 

Number of tumor-bearing animals/number of animals examined at site (proportion).
 Q


The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in
 

the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability
 

level for the Fisher exact test for the comparison of a treated group with the control group is
 

given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 o
 

The probability level of the test for departure from linear trend is given beneath the control
 

group when P < 0.05.
 
___ 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE3
 

TOPOGRAPHY: MORPHOLOGY 

Pituitary: Adenoma NOS 

P Values0 

Departure from Linear Trend 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Thyroid: C-Cell Adenoma 

to P Values0 

oo
 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Thyroid: C-Cell Adenoma or 

C-Cell Carcinoma*5 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

CONTROL 

18/44(0.41) 

P < O.OOl(N) 

P = 0.020 

90 

2/40(0.05) 

N.S. 

108 

3/40(0.08) 

N.S. 

108 

LOW 

DOSE 

3/45(0.07) 

P < O.OOl(N) 

0.163 

0.033 

0.506 

106 

2/47(0.04) 

N.S. 

0.851 

0.064 

11.301 

107 

2/47(0.04) 

N.S. 

0.567 

0.050 

4.717 

107 

HIGH 

DOSE 

2/37(0.05) 

P < O.OOl(N) 

0.132 

0.016 

0.499 

108 

0/45(0.00) 

N.S. 

0.000 

0.000 

2.995 

3/45(0.07) 

N.S. 

0.889 

0.126 

6.296 

107 
TOPOGRAPHY: MORPHOLOGY
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma'3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma or
 

Neoplastic Nodule^3
 

vO P Values0 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma
 

P Values
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

CONTROL
 

7/49(0.14)
 

P = 0.014(N)
 

106
 

2/49(0.04)
 

N.S.
 

-.__
 

93
 

16/49(0.33)
 

P < O.OOl(N)
 

P = 0.002
 

80
 

LOW
 

DOSE
 

2/50(0.04)
 

N.S.
 

0.280
 

0.030
 

1.383
 

93
 

0/50(0.00)
 

N.S.
 

0.000
 

0.000
 

3.313
 

0/50(0.00)
 

P < O.OOl(N)
 

0.000
 

0.000
 

0.194
 

HIGH
 

DOSE
 

1/49(0.02)
 

P = 0.030(N)
 

0.143
 

0.003
 

1.051
 

92
 

3/48(0.06)
 

N.S.
 

1.531
 

0.183
 

17.665
 

108
 

1/49(0.02)
 

P < O.OOl(N)
 

0.063
 

0.002
 

0.375
 

108
 
TABLE 4 (CONCLUDED)
 

LOW HIGH
 

TOPOGRAPHY:MORPHOLOGY CONTROL DOSE DOSE
 

Uterus: Endometrial Stromal Polyp 12/49(0.24) 0/47(0.00) 0/41(0.00)
 

P Values0 P < O.OOl(N) P < O.OOl(N) P < O.OOl(N)
 

Departure from Linear Trend P = 0.015
 

Relative Risk (Control) 0.000 0.000
 

Lower Limit 0.000 0.000
 

Upper Limit 0.284 0.324
 

Weeks to First Observed Tumor 80
 

Treated groups received time-weighted average doses of 0.18 or 0.36 percent in feed.
 

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

°The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in
 

the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability
 

level for the Fisher exact test for the comparison of a treated group with the control group is
 

given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 f\
 

The probability level of the test for departure from linear trend is given beneath the control
 

group when P < 0.05.
 
every type of malignant tumor in either sex where at least two such
 

tumors were observed in at least one of the control or 3-nitro-pacetophenetide-dosed groups and where such tumors were observed in
 

at least 5 percent of the group.
 

None of the statistical tests for any site in rats of either
 

sex indicated a significant positive association between the administration of 3-nitro-p-acetophenetide and an increased tumor incidence.
 

Thus, at the dose levels used in this experiment, there were no statistically significant results indicating that 3-nitro-p-acetophenetide was a carcinogen in Fischer 344 rats.
 

The Cochran-Armitage and Fisher exact tests indicated the possibility of a negative association between administration and the incidence of adrenal pheochromocytomas in the male, and the incidence of
 

pituitary adenomas NOS, of mammary fibroadenomas, and of endometrial
 

stromal polyps in the female rats.
 

The possibility of a negative association between dosage and
 

incidence was also noted for liver tumors in males and for either
 

leukemia or malignant lymphomas in both males and females. For these
 

tumors, however, the Cochran-Armitage test and the Fisher exact tests
 

were not significant under the Bonferroni criterion.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

risk have been estimated and entered in the tables based upon the
 

observed tumor incidence rates. In many of the intervals shown in
 

31
 
Tables 3 and 4, the value one is included; this indicates the absence
 

of statistically significant results. It should also be noted that
 

many of the confidence intervals have an upper limit greater than
 

one, indicating the theoretical possibility of tumor induction in
 

rats by 3-nitro-p-acetophenetide that could not be established under
 

the conditions of this test.
 

32
 
IV. CHRONIC TESTING RESULTS: MICE
 

A.	 Body Weights and Clinical Observations
 

Mean body weight depression relative to controls was apparent in
 •
 

both male and female dosed mice (Figure 4).
 

No clinical abnormalities were observed in mice of any group.
 

Salivary glands were not enlarged.
 

B.	 Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and 3-nitro-p-acetophenetide-dosed groups are shown in
 

Figure 5. For both male and female mice the Tarone test for positive
 

association between dosage and mortality was not significant.
 

For males five mice from the low dose group were sacrificed in
 

week 14, five from the high dose group in week 80, and five from the
 

control group in week 78. Sufficient numbers of male mice were at
 

risk from late-developing tumors, with 74 percent (37/50) of the
 

high dose, 82 percent (40/49) of the low dose, and 74 percent (37/50)
 

of the control mice surviving on test until the termination of the
 

study.
 

For females five mice from the high dose group were sacrificed
 

in week 80 and five from the control group in week 78. Sufficient
 

numbers of female mice were at risk from late-developing tumors, with
 

80 percent (40/50) of the high dose, 82 percent (41/50) of the low
 

dose3 and 70 percent (35/50) of the control mice surviving on test
 

until the end of the study.
 

33
 
—50 

T 

60 75 

TIME ON TEST (WEEKS) 

O 

50­ -50 

to 

-40 

-30 

-20 

10 

3

FEMALE MICE 

I 1 ' 

 15 

I

30

 I I ' I ' I 

 45 60 75 

TIME ON TEST (WEEKS) 

I

90 

' 

LOW DOSE 

HIGH DOSE 

I « 

105 12 

-10 

FIGURE 4 

GROWTH CURVES FOR 3-NITRO-p-ACETOPHENETIDE CHRONIC STUDY MICE 
c3 O P p p = 0 P 0 

c3 k> *• bi bo 3 D 'co -f^ b) 

i i i i i i i i > , 1 1 1 , 

o — o — 

C/S 

Tl 

30 m 

S 1

m 

- 

J» Ol |— w 01 ~~ m 1 ^ [-• ^ 

0 O 

O o J m 

s m 1• i •o 

2 8- J 8­ 

o f 

o 1 Tl 1 w H ^,_ d ft­

"^ m 1 m 1 0 

OZ3 ° •z. 

-\ 

?? §5 TM m 

w o> _ 

I

r 

H 01 ^ 

m

i m 

1 en m — H -J _ VI 

Ul — Ul 

0 i 

m J 

_ O

• 

p

SbIi 

30 

O 1 

S _ ~Z to 1 F; 

5 °~ 

1 j^ S* _

• «E 1 f"* \ 

a 

g 

1 

I r

0 

- I r o ^ 

O —

o 8- 01 x ^ 5 m 0 2 3 0 § I

 

O en ^ 0 g 0

m s

 r - K ' s

ro — 

S- i 1 1 I i 1 1 1 o 1 1 1 1 1 1 

<:D O O p O —» c3 O O p 

c3 N} ^ O) 00 O t3 N> '•*»• O) 
C. Pathology
 

Histopathologic findings on neoplasms in mice are summarized in
 

Appendix B (Tables Bl and B2); findings on nonneoplastic lesions are
 

summarized in Appendix D (Tables Dl and D2).
 

There was a dose-related increase in the incidence of hepatocellular carcinomas observed in the dosed male mice as shown below:
 

MALES FEMALES 

Low High Low High 

Control Dose Dose Control Dose Dose 

No. of Animals with
 

Livers Examined
 

Histopathologically (45) (41) (43) (45) (47) (49)
 

Hepatocellular
 

Carcinoma
 

Hepatocellular
 

Adenoma 6 14 2 0
 

Adenoma NOS 0 0 0 2
 

Hepatocellular carcinomas tended to lose their trabecular pattern
 

and grew in solid sheets, papillary arrangements, or less commonly in
 

adenomatous patterns with considerable variation in cytology. There
 

was usually marked variation in cell size and cytoplasmic vacuolation
 

and hyaline bodies were common findings. There were often bizarre
 

hyperchromatic giant nuclei and multinucleate hepatocytes. Mitoses
 

•
 

were urually numerous with occasional abnormal mitotic figures. Bile
 

ducts were usually absent. Carcinomas often compressed the adjacent
 

36
 
normal parenchyma and invasion was difficult to demonstrate. Some
 

tumors contained cystic and blood-filled spaces and areas of necrosis.
 

Metastases were not detected in the dosed mice in this study.
 

Hepatocellular adenomas were roughly spherical, expanding unencapsulated tumors which compressed adjacent hepatic cords. They had
 

a more trabecular pattern than carcinomas, being composed of closely
 

packed cords of liver cells and sinusoids. Normal mitoses were
 

present. Adenomas also had some cellular pleomorphism but to a
 

lesser degree than carcinomas. There were areas of fatty change and
 

cytoplasmic hyaline bodies. Bile ducts and triads were occasionally
 

seen in adenomas. Generally, adenomas were smaller than carcinomas,
 

with the majority occupying less than the width of a liver lobe.
 

The incidence of hepatic tumors was not significantly elevated
 

in dosed female mice as compared with controls; thus the hepatic tumors observed in females were not considered to be compound-related.
 

None of the other tumors noted in either sex were considered to be
 

compound-related.
 

In a few mice the diagnosis of telangiectasis was made. This
 

consisted of a focal area of dilated vascular spaces bounded usually
 

by a single layer of hepatocytes and lined by somewhat enlarged endothelial cells.
 

A variety of other tumors was observed in both dosed and control
 

mice. The incidence and distribution of these neoplasms were similar
 

37
 
to spontaneously occurring neoplasms in aged B6C3F1 mice and were not
 

considered to be compound-related.
 

A variety of inflammatory and degenerative lesions which commonly occur in aging mice of this strain was seen. These nonneoplastic lesions were not considered to be compound-induced.
 

Based upon the results of this pathologic examination, there was
 

an increased incidence in liver neoplasms in male B6C3F1 mice fed
 

3-nitro-p-acetophenetide.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 5 and 6. The analysis is included for
 

every type of malignant tumor in either sex where at least two such
 

tumors were observed in at least one of the control or 3-nitro-pacetophenetide-dosed groups and where such tumors were observed in
 

at least 5 percent of the group. Due to the sacrifice of five low
 

dose males in week 14, the analyses in Table 5 include only those
 

male mice surviving at least 15 weeks.
 

For male mice when incidences were combined so that the numerator
 

represented males with either a hepatocellular carcinoma or a hepatocellular adenoma, the Cochran-Armitage test indicated a significant
 

(P = 0.002) positive association between dose and incidence. The
 

Fisher exact test comparing high dose to control was also significant
 

(P = 0.002). In historical control data collected by this laboratory
 

for the NCI Carcinogenesis Testing Program 51/350 (15 percent) of the
 

38
 
___ 

TABLE 5
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

a,f
 SPECIFIC SITES IN MALE MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE


TOPOGRAPHY: MORPHOLOGY
 

Hematopoietic System: Malignant Lymphoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar Adenoma or
 OJ Alveolar/Bronchiolar Carcinoma*'
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

2/46(0.04)
 

N.S.
 

98
 

11/45(0.24)
 

N.S.
 

P = 0.032
 

78
 

4/45(0.09)
 

N.S.
 

93
 

LOW
 

DOSE
 

4/41(0.10)
 

N.S.
 

2.244
 

0.340
 

23.727
 

98
 

2/41(0.05)
 

P = O.Oll(N)
 

0.200
 

0.023
 

0.843
 

98
 

8/41(0.20)
 

N.S.
 

2.195
 

0.639
 

9.244
 

98
 

HIGH
 

DOSE
 

4/45(0.09)
 

N.S.
 

2.044
 

0.310
 

21.695
 

98
 

6/43(0.14)
 

N.S.
 

0.571
 

0.190
 

1.525
 

98
 

9/43(0.21)
 

N.S.
 

2.355
 

0.715
 

9.722
 

98
 
TABLE 5 (CONCLUDED)
 

LOW HIGH 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE DOSE 

Liver: Hepatocellular Carcinoma or 

Hepatocellular Adenoma" 10/45(0.22) 13/41(0.32) 23/43(0.53) 

P Values P = 0.002 N.S. P = 0.002 ___ 

Relative Risk (Control) 1.427 2.407 

Lower Limit 0.651 1.269 

Upper Limit 3.212 4.829 

Weeks to First Observed Tumor 93 98 80 

Treated groups received time-weighted average doses of 0.73 or 1.46 percent in feed.
 

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

G

The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in
 

the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability
 

level for the Fisher exact test for the comparison of a treated group with the control group is
 

given beneath the incidence of tumors in the treated group when P < 0.05; ^otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 

S

The probability level of the test for departure from linear trend is given beneath the control
 

group when P < 0.05.
 

These analyses were based solely upon animals surviving at least 15 weeks.
 
TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDEa
 

TOPOGRAPHY : MORPHOLOGY 

Lung: Alveolar/Bronchiolar Adenoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Hematopoietic System: Leukemia or 

Malignant Lymphoma" 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Liver: Hepatocellular Carcinoma or 

Hepatocellular Adenoma^ 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

LOW 

CONTROL DOSE 

1/45(0.02) 4/47(0.09) 

N.S. N.S. 

3.830 

0.399 

184.347 

98 97 

12/46(0.26) 7/47(0.15) 

N.S. N.S. 

0.571 

0.209 

1.426 

95 80 

4/45(0.09) 3/47(0.06) 

N.S. N.S. 

0.718 

0.111 

4.010 

78 97 

HIGH 

DOSE 

2/49(0.04) 

N.S. 

1.837 

0.099 

106.150 

98 

6/49(0.12) 

N.S. 

0.469 

0.158 

1.232 

98 

1/49(0.02) 

N.S. 

0.230 

0.005 

2.209 

98 
TABLE 6 (CONCLUDED) 

TOPOGRAPHY: MORPHOLOGY CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Stomach: Squamous-Cell Papilloma 

P Values0 

3/42(0.07) 

P = 0.031(N) 

0/44(0.00) 

N.S. 

0/47(0.00) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 98 

0.000 

0.000 

1.580 

0.000 

0.000 

1.482 

Pituitary: Adenoma NOS 

P Values0 

6/37(0.16) 

P = 0.004(N) 

1

P

/41(0.02) 

= 0.040(N) 

0

P

/38(0.00) 

= 0.012(N) 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

0.150 

0.003 

1.159 

0.000 

0.000 

0.602 

Weeks to First Observed Tumor 98 97 

treated groups received time-weighted average doses of 0.73 or 1.46 percent in feed.
 

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

The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in
 

the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability
 

level for the Fisher exact test for the comparison of a treated group with the control group is
 

given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
untreated male B6C3F1 mice had one of these tumors, compared to the
 

23/43 (53 percent) observed in the high dose group in this bioassay.
 

Based on these statistical results, the administration of 3­

nitro-p-acethophenetide was associated with an elevated incidence of
 

liver neoplasms in male mice under the conditions of this experiment.
 

Both the Cochran-Armitage test and the high dose Fisher exact
 

comparison indicated a significant negative association between compound administration and the incidence of pituitary adenomas NOS in
 

female mice. The low dose Fisher exact test was not significant under
 

the Bonferroni criterion. For alveolar/bronchiolar neoplasms in the
 

males and for squamous-cell papillomas of the stomach in females the
 

possibility of a negative association was indicated; for both cases,
 

however, the results of the Cochran-Armitage test did not agree with
 

those of the Fisher exact test as to the significance of the results.
 

43
 

V. DISCUSSION
 

There were no significant positive associations between the
 

concentrations of 3-nitro-p-acetophenetide administered and mortality
 

in rats or mice of either sex. In addition, adequate numbers of animals in all groups survived sufficiently long to be at risk from
 

late-developing tumors.
 

In rats of both sexes there were no significant positive associations between the administration of 3-nitro-p-acetophenetide and
 

the incidence of any tumor.
 

In dosed male mice when compared to controls there were increased incidences of hepatocellular carcinomas (i.e., 4/45 [9 percent] , 8/41 [20 percent], and 9/43 [21 percent] in the control, low
 

dose, and high dose groups) and hepatocellular adenomas (i.e., 6/45
 

[13 percent], 5/41 [12 percent], and 14/43 [33 percent] in the control, low dose, and high dose groups). When the incidences of male
 

mice having either of these neoplasms were combined, the resulting
 

incidences (i.e., 10/45 [22 percent], 13/41 [32 percent], and 23/43
 

[53 percent] in the control, low dose, and high dose groups, respectively) indicated a significant positive association between concentration administered and tumor incidence. This finding was supported
 

by a significant high dose to control Fisher exact comparison.
 

Under the conditions of this bioassay, dietary administration
 

of 3-nitro-p-acetophenetide was not carcinogenic in Fischer 344 rats
 

of either sex or in female B6C3F1 mice. The compound, however, was
 

45
 
considered carcinogenic in male B6C3F1 mice based on a significant
 

increase in the combined incidence of hepatocellular carcinomas and
 

hepatocellular adenomas in these animals.
 
VI. BIBLIOGRAPHY
 

Armitage, P., Statistical Methods in Medical Research, Chapter 14.
 

J. Wiley & Sons, New York, 1971.
 

Barr, W.H., and R.P. Penna, "Internal Analgesics." Handbook of Non-


Prescription Drugs, 1971 edition. G.B. Griffenhagen and L.L.
 

Hawkins, editors. American Pharmaceutical Association, Washington, D.C., 1971.
 *
 

Berenblum, I., editor, Carcinogenicity Testing. International Union
 

Against Cancer, Technical Report Series, Vol. 2. International
 

Union Against Cancer, Geneva, 1969.
 

Chemical Abstracts Service. The Chemical Abstracts Service (CAS)
 

Ninth Collective Index, Voluumes 76-85, 1972-1976. American
 

Chemical Society, Washington, D.C., 1977.
 

Cox, D.R., Analysis of Binary Data, Chapters 4 and 5. Methuen and
 

Co., Ltd., London, 1970.
 

Cox, D.R., "Regression Models and Life-Tables." Journal of the Royal
 

Statistical Society, Series "B" 34:187-220, 1972.
 

Gart, J.J., "The Comparison of Proportions: A Review of Significance
 

Tests, Confidence Limits, and Adjustments for Stratification."
 

International Statistical Institute Review 39:148-169, 1971.
 

Juusela, H. , "Carcinoma of the Renal Pelvis and its Relationship to
 

Analgesic Abuse." Annales Chirurgiae et Gynaecologiae Fenniae
 

62:386-390, 1973.
 

Kaplan, E.L., and P. Meier, "Nonparametric Estimation from Incomplete
 

Observations." Journal of the American Statistical Association
 

_^3:457-481, 1958.
 

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

"Carcinogenesis Bioassay Data System." Computers and Biomedical
 

Research 7:230-248, 1974.
 

Miller, R.G., Simultaneous Statistical Inference. McGraw-Hill Book
 

Co., New York, 1966.
 

Odashima, S. , Chief, National Institute of Hygienic Sciences, Department of Chemical Pathology, Tokyo, Japan. Personal communication, May 1978.
 

47
 
Oelschlaeger, H., and C. Welsch, "Detection of Phenacetin by Nitration."
 

Archives of Pharmacology 297(10):608-617, 1964; Chemical Abstracts
 

£_2, 3885g.
 

Saffiotti, U., R. Montesano, A.R. Sellakumar, F. Cefis, and D.G.
 

Kaufman, "Respiratory Tract Careinogenesis in Hamsters Induced
 

by Different Numbers of Administration of Benzo (a) Pyrene and
 

Ferrix Oxide." Cancer Research 32; 1073-1079, 1972.
 

Tarone, R.E., "Tests for Trend in Life-Table Analysis." Biometrika
 

£2_:679-682, 1975.
 

Urso, S., Research Analyst, Chemical-Environmental Program, Chemical
 

Industries Center, Stanford Research Institute, Menlo Park,
 

California. Personal communications, 1977.
 

U.S.	 Tariff Commission, Synthetic Organic Chemicals, U.S. Production
 

and Sales, 1970. USTC Publication 479, U.S. Government Printing
 

Office, Washington, D.C., 1972.
 

Weisburger, E.W., Chief, Carcinogen and Toxicology Branch, National
 

Cancer Institute, National Institutes of Health, Bethesda,
 

Maryland. Personal communication, August 5, 1977.
 

48
 
Review of the Bioassay of 3-Nitro-p-Acetophenetide*
 

for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DREW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

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

academia, industry, organized labor, public interest groups,
 State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

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

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

on the bioassay of 3-Nitro-p-Acetophenetide for carcinogenicity,
 

The reviewer agreed with the conclusion in the report
 

that the compound was not carcinogenic, under the conditions
 

of test, in treated rats or female mice but did induce a
 

statistically significant incidence of liver tumors in
 

treated male mice. After a brief description of the experi­ mental design, the reviewer noted several flaws that detracted
 

from the study, e.g., analyses were not done to confirm the
 

identification of the 3-Nitro-p-Acetophenetide or on its
 

stability or concentration in the diet and the study was
 

conducted in a room in which other compounds were under
 

test. The reviewer opined that the flaws were sufficient to
 

cast doubt on the validity of the bioassay. He also said
 

that the "very slight response" in only one sex of one
 

species was of considerable concern.
 

A discussion ensued as to whether the study flaws were
 

sufficiently serious as to invalidate the conclusions drawn
 

from the bioassay. A Program staff member noted that the
 

49
 
3-Nitro-p-Acetophenetide was procured from the manufacturer
 

of the chemical. A Subgroup member said that the structure
 

of the test material would indicate it to be unstable. It
 

was agreed that further consideration of the report should
 

be deferred until a sample of the reference 3-Nitro-p-
 Acetophenetide could be analyzed for identity and stability.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

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

Louise Strong, University of Texas Health Sciences Center
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

GOVERNMENT PRINTING OFFICE 1979-2812173221 

50 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE
 

TABLE Al

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE 

CONTROL (UNTR) LOW DOSE HIGH DOSE
 

01-0070 01-0073 01-0074
 

ANIMALS INITIALLY IN STUDY 50 50 50
 ANIMALS MISSING 1
 ANIMALS NECROPSIED 48 50 50
 ANIMALS EXAMINED HISTOPATHOLOGICALLY** 48 50 49
 

INTEGUMENTARY SYSTEM
 

*SKIN (48) (50) (50)
 

SQUAMOUS CELL PAPILLOMA 1 (2X)
 

SQUAHOUS CELL CARCINOMA 1 <2X)
 BASAL-CELL CARCINOMA 1 (2X)
 KERATOACANTHOMA 1 (2X) 1 (2X)
 

*SUBCUT TISSUE (18) {50} (50)
 ADNEXAL ADENOMA 1 (2X)
 FIBROMA 2 (4*) 1 (2X)
 NEUROFIBROHA 1 (2X>
 

BESPIBATORY SYSTEM
 

*LUNG (48) (50) (49)
 CARCINOMA, SOS, METASTATIC 1 (2X)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 2 <4X)
 OSTEOSARCOMA, METASTATIC 1 <2X)
 

HEMATOPOIETIC SYSTEM
 

*MDLTIPLE ORGANS (48) (50) (50)
 LEUKEMIA, NOS 1 <2X)
 MYELOMONOCYTIC LEBKEMIA 5 (10X) 2 (4X)
 

*SKIN (48) (50) (50)
 MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2X)
 

#SPLEEN (48) (50) (49)
 OSTEOSARCOMA, METASTATIC 1 (2X)
 

CIRCULATORY SYSTEM
 

NON£
 

# NUMBE R OF ANIMALS WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

**EXl,LUDES PARTIALLY AUTOLYZED ANIMALS 

A-3 
TABLE Al (CONTINUED)
 

CONTROL (UNTR) LOW DOSE HIGH DOSE
 

01-0070 01-0073 01-0074
 

DIGESTIVE SYSTEM
 

tLIVER (48) (50) (49)
 

NEOPLASTIC NODULE 1 (2%)
 

HEPATOCELLULAR CARCINOMA 2 (4%)
 

ISTOMACH (48) (49) (49)
 

SQUAMOUS CELL PAPILLOMA 1 (2X) 1 (2X)
 

#SMALL INTESTINE (45) (49) (48)
 

CYSTADENOCAPCINOBA, NOS 1 (2X)
 

L3IOMYOSABCOMA 1 (2X) 1 (2X)
 

USINARY SYSTEM
 

tKIDNEY (48) (50) (49)
 

TRANSITIONAL-CELL CARCINOMA 1 (2X)
 

HSMANSIOSAHCOMA 1 (2X)
 

*KIDNEY/PELVIS (48) (50) (49)
 

TRANSITIONAL-CELL CARCINOMA 1 (2%)
 

#UBINARY BLADDER (46) (50) (17)
 

TRANSITIONAL-CELL PAPILLOMA 1 (2*)
 

ENDOCRINE SYSTEM
 

fPITUITARY (41) (46) (44)
 

ADENOMA, NOS 1 (2X) 4 (9X)
 

#ADRENAL (47) (49) (48)
 

CORTICAL ADENOMA 1 (2X)
 

PHEOCHEOMOCYTOMA 10 (21%) 1 (2X) 1 (2X)
 

GANGLIONEUROMA 1 (2%)
 

ITHYROID (39) (45) (47)
 

C-CELL ADENOMA 2 (4S) 2 (4X)
 

*PAHCEEATIC ISLETS (45) (46) (45)
 

ISLET-CELL ADENOMA 3 (7X) 2 (4X) 1 (2X)
 

REPRODUCTIVE SYSTEM
 

*MAMMAEY GLAND (48) (50) (50)
 

____ PAPILLASI_ADENOCAECINOMA
 

* NDMBEE OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-4
 
TABLE Al (CONTINUED) 

CONTROL (UNTR)

01-0070

 LOW DOSE

 01-0073

FIBROADENOMA 1 (2%) 

*PENIS (U8) (50)

SQUAMOUS CELL CARCINOMA 1 (2%) 

•PBEPUTIAL GLAND (18) (50)

CABCINOMA.NOS 2 (IS) 

tTESTIS (47) (50)

INTERSTITIAL-CELL TUMOR 15 (9631) 17 (91X)

NERVOUS SYSTEM
 

*CE3EBRAL HEMISPHERE (U7) (50)


OLIGODENDROGLIOMA


tCEREBELLUM (17) (50)


ASTROCYTOMA 1 (2%)
 

SPECIAL SENSE ORGANS
 

NJN E
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*BODY CAVITIES (18) (50)
 MESOTHELIOMA, NOS 1 (2%) 1 (2%)


*ABDOMINAL CAVITY (18) (50)


SARCOMA, NOS 1 (2%)
 

ALL OTHER SYSTEMS
 

NINE
 

* NUMBER Or ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

 HIGH DOSE 

 01-0071 

 (50)
 

 (50)
 

 (19)
 

 39 (80X)
 

 (19)
 

1 (2%)
 

 (19)
 

 (50)
 

2 (U»)
 

 (50)
 

A-5
 
TABLE Al (CONCLUDED)
 

(;ONTROL (UNTR) LOW DOSE HIGH DOSE
 

01-0070 01-0073 01-0074
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 NATURAL DEATHS 5 3 3
 

MORIBUND SACRIFICE 5 5 2
 

SCHEDULED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 34 42 10
 ANIMAL MISSING 1
 

3 INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TOHORS* 45 49 42
 TOTAL PRIMARY TUMORS 81 71 53
 

TOTAL ANIMALS WITH BENIGN TUMORS US 49 41
 

TOTAL BENIGN TUMORS 66 62 46
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 10 8 5
 TOTAL MALIGNANT TUMORS 13 8 5
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2
 TOTAL SECONDARY TUMORS 3
 

TOTAL ANIMALS WITH TUMORS UNCEBTAINBENIGN OR MALIGNANT 2 1 2
 TOTAL UNCERTAIN TUMORS 2 1 2
 

TOTAL ANIHALS WITH TUMORS ONCERTAINPRMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

A-6
 
TABLE A2
 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE
 

CONTROL (UNTR) LOH DOSE 02-0070 02-0073 HIGH DOSE 02-0074 

ANIMALS IHITIALLY IN STUDY 50

ANIMALS NECROPSIED U9

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 49

 50

 50

 50

 50 

 49 

 49 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE (49) (50) («9)
 TRICHOEPITHELIOBA 1 (2%)
 FIBROMA 1 (2*)
 

RESPIRATORY SYSTEM
 

ILONG (49) (50) (49)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS (49) (50) (49)
 

MALIGNANT LYMPHOMA, NOS 2 (4X)
 

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

MYELOflONOCYTIC LEUKEMIA 5 (10X) 1 (2X)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

ULIVER (49) (50) (48)
 NEOPLASTIC NODULE 2 (4X) 1 (2X)
 HEPATOCELLULAR CARCINOMA 1 (2X) 2 (4X)
 

URINARY SYSTEM
 

tKIDNEY (49) (50) (48)
 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBS* OF ANTMALS NBCSOPSIED
 **EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

A-7
 
TUBULAR-CELL ADENOCABCINOMA
 

tKIDNEY/PELVIS
 

TRANSITIONAL-CELL CARCINOMA
 

ENDOCRINE SYSTEH
 

tPITHITARY
 

ADENOMA, NOS
 

tADRENAL
 

CORTICAL CARCINOMA
 

PHEOCHROMOCYTOMA
 

#THYROID
 

FOLLICDLAR-CELL CARCINOMA
 

C-CELL ADENOMA
 

C-CELL CARCINOMA
 

tPANCREATIC ISLETS
 

ISLET-CELL ADENOHA
 

ISLET-CELL CARCINOMA
 

REPRODUCTIVE SYSTEM
 

*HAHMARY GLAND
 

ADENOMA, NOS
 

ADENOCARCINOMA, SOS
 

FIBROADENOBA
 

*CLITORIS
 

ADENOMA, NOS
 

•CLITOBAL GLAND
 

ADENOMA, NOS
 

*VAGINA
 

LEIOHYOSARCOMA
 

tUTERUS
 

LEIOMYOSARCOMA
 

ENDOBETHIAL STROMAL POLYP
 

tUTERUS/ENDOHETRIUM
 

ADENOCARCINOHA. NOS
 

TABLE A2 (CONTINUED)
 

CONTROL (DNTR)
 

02-0070
 

(49)
 

(44)
 

18 (41X)
 

(49)
 

1 (2X)
 

2 (4X)
 

(40)
 

1 (3X)
 

2 (5X)
 

1 <3X)
 

(47)
 

1 (2*)
 

(49)
 

2 (4X)
 

16 (33X)
 

(49)
 

(49)
 

1 (2X)
 

(49)
 

(49)
 

1 <2X)
 

12 (24X)
 

(49)
 

2 14X.1
 

LOW DOSE HIGH DOSE
 

02-0073 02-0074
 

1	 <2X)
 

(50)	 (48)
 

1 (2X)
 

(45)	 (37)
 

3 <7X) 2 (5X)
 

(49)	 (48)
 

(47)	 (45)
 

2	 (4X)
 

3 <7X)
 

(48)	 (46)
 

1 (2X)
 

(50)	 (49)
 

2	 (4X)
 

1 <2X)
 

(50)	 (49)
 

1 (2X)
 

(50)	 (49)
 

(50)	 (49)
 

1 <2X)
 

(47)	 (41)
 

(47)	 (41)
 

1 (2X)
 

t	 NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSIED
 

A-8
 
NERVOUS SYSTEM
 

JBRAIN


OLIGODENDROGLIOMA


SPECIAL SENSE ORGANS
 

*EYELID


SQUAMOUS CELL CARCINOMA


MUSCULOSKEIETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*PERITONEBB


MESOTHELIOMA, NOS


ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY


NATURAL DEATH3


MORIBUND SACRIFICE


SCHEDULED SACRIFICE


ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE


ANIMAL MISSING
 

8 INCLUDES AUIQLYZED ANIMALS
 

TABLE A2 (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE 

02-0070 02-0073 02-007U 

 (49) (50) (47) 

1 (2S) 

 (49) (50) (49) 

1 (2X) 

 (49) (50) (49) 

1 (2X) 

 50 50 50 

3 3 5 

7 1 

5 5 

 35 47 39 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-9
 
TABLE A2 (CONCLUDED)
 

(:ONTROL (ONTRJ I LOW DOSE HIGH DOSE
 

02-0070 02-0073 02-0074
 

TUMOS SUMMARY
 

TOTAL ANIMALS WITH P.RIMARY TUMORS* <»5 12 15
 TOTAL PRIMARY TUMORS 73 12 17
 

TOTAL ANIMALS WITH BENIGN TUMORS 37 5 6
 TOTAL EENIGN TUMORS 51 5 7
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 13 7 9
 TOTAL MALIGNANT TUMORS 16 7 9
 

TOTAL ANIMALS WITH SECONDARY TUBORS*
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUHORS UNCERTAINBENIGN OR MALIGNANT 3 1
 

TOTAL UNCERTAIN TUMORS 3 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR BETASTATIC
 TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SSCONDARY TUMORS
 

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

A-10
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE
 

TABLE Bl

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE 

~~ >J '"­ —TM—__.—,_,«.— 

COHTROL(ONTB) LOR DOSE HIGH DOSE 

05-0077 05-0075 05-0076 

ANIMAL S INITIALLY IN STDDY 50 950 50 ANIMAL S HISSING 1 

ANIMALS NECROPSIBD H6 46 45 ANIMALS EXAMINE D HISTOPATHOLOGICALLY **45 41 43 

INTEGUMENTAR Y SYSTSH 

NON E 

RESPIRATOR Y SYSTEM 

*L0NG (45) (41) (43) HEPATOCELLOLAE CARCIHOHA, HETAST 1 (2X) ALVEOLAR/BRONCHIOLAH ADENOHA 7 (16X) 2 (5«) 6 (14X) 

ALVEOLAB/BRONCHIOLAR CARCINOMA 4 (9«) 

HTMATOPOIETIC SYSTEM 

*NULTIPLE ORGAN S (46) (46) (45) MALIGNAN T LYMPHOHA. NOS 1 (2X) MALIG.LYBPHOMA , HISTIOCYTIC TYPE 1 (2X) 

iSPLE?N t«5) (41) (43) HEMANGIOSAECOM A 1 (2%) MALIG.LYMPHOMA , HISTIOCYTIC TYPE 1 (2«) 2 (5X) 

*B?NDIBOLA R 1. NODE (35) (36) (35) MALIG.LYMPHOMA , HISTIOCYTIC TYPE 1 <3X) 

IMFSENTSRIC L. NODE (35) (36) (35) MALIGNAN T LYMPHOMA , NOS 1 (3*) 

fEENA L LYMPH NODE (35) (36) (35) MALIGNAN T LYMPHOMA , NOS 1 (3X) 

tPEYESS PATCH (43) (41) (43) MALIGNAN T LYMPHOMA . NOS 2 (5X) 

CIEC'ILATORY SYSTEM 

NOSE 

* ND1BER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* NU1BEB OF ANIMAL S NECROPSIED 

»*EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

S> SO ANIMAL S WERE INITIALLY IN THE STDDY, BUT ONE ANIMA L WAS FOUN D TO BE A FEMALE 

IN A MALE GROUP . 

B-3 
TABLE Bl (CONTINUED) 

CONTHOL(UNTR ) LOW DOSE HIGH DOSE 

05-0077 05-0075 05-0076 

DIGESTIVE SYSTEM 

tLIVEP (K5 ) (<»1 ) (U3) 

ADENOMA , NOS 1 (2J) 

HEPATOCELLOLiR ADEHOMA 6 (13*) 5 (12*) 1 <• (33%) 

HEPATOCELLOLAR CARCINOMA U (9X) 8 (20X) 9 (21X ) 

HEBANGIOSAKCOMA 1 (2X) 

• STOMACH (U2) (141) (13) 

SQUANOU S CELL PAPILLOBA 1 (2X) 

URINAR Y SYSTEM 

NONE 

ENDOCPINF SYSTEM 

NON E 

REPRODUCTIV E SYSTEM 

NONE 

NERVOU S SYSTEM 

NON F 

SPFCIAL SENSE ORGAN S 

*HARDSRIAN GLAND (U6) («6) (US) 

"APILLAP Y ADENOMA 1 (2X) 

*EAS CANA L (U6 ) (U6) (45) 

SQUAMOU S CELL CARCINOMA 1 (251) 

MUSCULOSKELETA L SYSTEM 

NON E 

BOtY CAVITIES 

NOK E 

* SOMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECBOPSIED 

B-4 
TABLE Bl (CONCLUDED) 

CONTROL (UNTB ) LOW DOSE HIGH DOSE 

05-0077 05-0075 05-0076 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

NATURAL DEATHS 7 1
 

MORIBUND SACRIFICE 1
 

SCHEDULED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 37 10 37
 

ANIMAL HISSING 1
 

ANIMAL DELETED(WRONG SEX) 1
 

3 INCLUDES AOTOLYZED ANIMALS
 

TUBOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TOMORS* 21 20 29
 

TOTAL PRIMARY TUMORS 25 22 3«
 

TOTAL ANIMALS WITH BENIGN TUMORS 13 8 20
 

TOTAL BENIGN TUMORS 1* 8 21
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 9 13 12
 

TOTAL MALIGNANT TUMORS 11 14 13
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 1
 

TOTAL SECONDARY TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCEBTAINPRIMAPY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TOMORS OE TUHORS INVASIVE INTO AN ADJACENT ORGAN
 

B-5
 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE 

CONTROL(UNTR ) LOW DOSE 

06-0077 06-0075 

ANIMAL S INITIALL Y IN STUDY 50 50 

ANIMAL S NECBOPSIED 16 17 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 16 t7 

INTEGUMENTAP Y SYSTEM 

*SKI N (16) (17) 

FIBBOSA°COM A 2 (IX) 

*SUBCUT TISSUE (16) (17) 

HEMANGIOM A 

RFSPIPATOE Y SYSTSM 

#LUSG (15) (17) 

ALVEOLAR/BPONCHIOLAR ADENOMA 1 (2X) 1 (9X) 

HEMATOPOIETIC SYSTEM 

*«ULTIPLF ORGAN S (16) (17) 

MALIGNANT LYBPHOMA, NOS 3 (7X) 5 (11X) 

1ALIG.LYNPHOHA , DNDIFFER-T YPE 1 <2X ) 

MALIG.LYMPHOSA , HISTIOCYTIC TYPE 6 (13%) 1 (2X) 

LYMPHOCYTIC LEUKEMIA 1 (2X) 

tSPLEBN (13) (17) 

MALIGNANT LYBPHOMA, NOS 

tLIVER (15) (17) 

1ALIG.LYMPHOMA , HISTIOCYTIC TYPE 1 (2X) 

#SMAL L INTESTINE (13) (11) 

MALIG.LYMPHOSA, HISTIOCYTIC TYPE 1 (2X) 

tPEYESS PATCH (13) (11) MALIG.LYMPHOBA , HISTIOCYTIC TYPE 1 (2X) 

*KIDNE Y (13) (16) _ MAL.IGNANT_LYSPjJOHAi_NOS_ 

* NUHBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* HUMBEP OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

HIGH DOSE 

06-0076 

50 

19 

19 

(19) 

(19) 

1 (2X) 

(19) 

2 (IX) 

(19) 

2 (IX) 

1 (2X) 

(18) 

1 (2X) 

(19) 

(18) 

(18) 

(19)

1_I2S1 

B-6 
TABLE B2 (CONTINUED) 

COHTROL(UNTR) LOW DOSE HIGH DOSE 

06-0077 06-0075 06-0076 

tTHYBUS (27) (32) (36)
 

MALIGNANT LYBPHOMA, NOS 1 (3X)
 

CIRCULATORY SYSTEB
 

NONE
 

DIGESTIV E SYSTEH 

#LIVER (45) (17) (U9) 

ADENOMA , NOS 1 (2%) 2 (IX) 

HEPATOCELLOLAR ADENOM A 2 (IX) 1 (2X) 

HEPATOCELLULAR CARCINOMA 2 (4X) 2 (IX) 1 (2X) 

tSTOMACH (42) (44) (17) 

SQUAMOU S CELL PAPILLOSA 3 (7X) 

URINAR Y SYSTEM 

NOSE 

ENEOCEIN E SYSTEH 

tPITUITARY (37) (41) (38)
 

ADENOMA , NOS 6 (16X) 1 (2X)
 

#ADRENA L (45) (47)
 

CORTICAL ADENOM A 1 <2X)
 

tTHYROI D (30) (45) (42) 

ADENOCARCISOMA , NOS 1 (2%) 

iPANCREATIC ISLETS (44) (48)
 

ISLET-CELL ADENOM A 1 (2X)
 

REPRODUCTIVE SYSTEH
 

*BSMBAHI GLAND (46) (47) (49)
 

ADENOCARCINOMA. NOS 1-J2S1 1-J2SL.
 

* NU«BEP OF ANIMALS WITH TISSDE EXABINED BICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

B-7
 
TABLE B2 (CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE 

06-0077 06-0075 06-0076 

tUTERUS (43) (47) (42) 

HEMANGIOfl A 2 (5*) 

#OVAH Y (41) (45) (47) 

LUTEOM A 1 (2*) 1 (2X) 

NERVOU S SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

*HAHDFRIAN GLAND (46) (47) (49) 

PAPILLARY CYSTADENOHA, NOS 1 (2X) 

MUSCULOSKELETAL SYSTEM 

NONE 

BOEY CAVITIES 

NONE 

ALL OTHFB SYSTEMS 

NONF 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 50 50 50 

NATURAL DEATHS 8 9 5 

MORIBUND SACRIFICE 2 

SCHEDULED SACRIFICE 5 5 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 35 41 40 

ANIMSL MISSING 

* NUMBE R OF ANIMAL S HITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUHBE F OF ANIMAL S NECROPSIED 
TABLE B2 (CONCLUDED)
 

CONTROL(UNTR ) LOW DOSE HIGH DOSE 

06-0077 06-0075 06-0076 

TU«O P SUMMAR Y 

TOTAL ANIMAL S WITH PRIMAR Y TDHORS* 22 17 15 

TOTA L PRIMAR Y TUMORS 32 18 17 

TOTAL ANIMALS HITH BENIGN TDMOBS 13 7 8 

TOTAL BENIGN TDMORS 15 7 9 

TOTAL ANIMAL S WITH MALIGNAN T TUHOBS 16 10 7 

TOTAL MALIGNAN T TUMORS 17 11 8 

TOTAL ANIMALS HITH SECONDARY TUMOBS#
 

TOTAL SECONDARY TOMORS
 

TOTAL ANIMALS WITH TUMORS ONCERTilN-


BSNIGN OH MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCEBTAINPPIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUBORS EXCEPT SECONDARY TUMORS
 

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

B-9
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 3-NITRO-p-ACETOPHENETIDE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH 3-NITRO-p-ACETOPHENETIDE 

CONTROL (UNTR ) 

01-0070 

LOW

01­

 DOSE 

0073 

HIGH DOSE 

01-0074 

ANIMALS INITIALLY IN STUDY 50 

ANIMALS MISSING 1 

ANIHALS NECROPSIED 48 

ANIMALS EXAHINED HISTOPATHOLOGICALLY** 48 

50 

50 

50 

50 

50 

49 

INTEGUMENTARY SYSTEM
 

*SKIN (48) (50) (50)
 

DILATATION, NOS 1 (2%)
 

HEMATOMA, NOS 1 (2%)
 

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

BESPIHATOEY SYSTEM
 

*TRACHEA (15) (49) (48)
 

INFLAMMATION, ACUTE/CHRONIC 18 (405C)
 

•LUNG/BRONCHUS	 (48) (50) (49)
 

ERONCHIECTASIS 3 (6X)
 

*LUNG (18) (50) (49)
 

CONGESTION, NOS 1 (2%)
 

INFLAMHATION, FOCAL 2 (4X)
 

ABSCESS, NOS 1 (2%)
 

PNEUMONIA, CHRONIC MUBINE 1 (2X)
 

GRANULOMA, NOS 1 (2X)
 HYPERPLASIA, FOCAL 1 (2%)
 

HYPERPLASIA, ALVEOLAR EPITHELIUM 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

tBONE MARROW (48) (19) (47) 

MYELOFIBROSIS 1 (2») HYPERPLASIA, HEMATOPOIETIC 1 (2%) HYPERPLASIA, GRANULOCYTIC 1 (2%) 

HYPEHPLASIA, MEGAKARYOCYTIC 1 (2%) 

#SPLEEN (48) (50) (49) 

CONG_gSTj:ONj,._NOS 

I NUMBER OF ANIMALS WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-3
 
TABLECl (CONTINUED) 

CONTROL (UNTR) LOH DOSE HIGH DOSE
 

01-0070 01-0073 01-0074
 

ERYTHEOPOIESIS	 1 (2%) 1 (2%)
 

•MANDIBULAR	 L. NODE (42) (40) (46)
 

DILATATION, NOS 1 (2X)
 HYPERPL4SIA, NOS 1 (2X)
 

•MEDIASTINAL L.NODE (42) (40) (46)
 

HYPERPLASIA, NOS 2 (5X)
 

CIRCULATORY SYSTEM
 

•HEART	 (48) (50) (49)
 FIBROSIS, FOCAL 11 (23X)
 

FIBROSIS, DIFFUSE 1 (2X)
 

CALCIFICATION, NOS 1 <2X>
 

•MYOCARDIUM	 (48) (50) (49)
 INFLAMMATION, INTERSTITIAL 2 <4X)
 

INFLAMMATION, ACUTE/CHRONIC 3 (6X)
 FIBROSIS, FOCAL 2 (4X) 1 (2X)
 DEGENERATION, NOS 1 (2X) 1 (2%)
 

•CARDIAC VALVE (48) (50) (49)
 INFLAMMATION, ACOTE/CHROMC 1 (2%)
 

*CORONAHY ARTERY (48) (50) (50)
 PERIVASCULITIS 1 (2X)
 

MEDIAL CALCIFICATION 1 (2X)
 

*PULBONARY ARTERY (48) (50) (50)
 MINERALIZATION 11 (23X)
 

DIGESTIVE SYSTEM
 

•LIVER	 (48) (50) (49) INFLAMMATION, NECROTIZIN& 1 (2X) 

PERIARTERITIS 1 (2X) NECROSIS, FOCAL 8 (17X) 1 (2X) METAMORPHOSIS FATTY 4 (8X) HYPERPLASIA, FOCAL 8 (17X) ANGIECTASIS 2 (4X) SRYTHEOPOIESIS 1 (2X) 

•LIVER/CENTRILOBULAR	 (48) (50) (49) DEGENERATION, EOSINOPglLIC_ .__2_i»XJ._
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-4
 
TABLE Cl (CONTINUED) 

HIGH DOSE
 

01-0074
 

(49)
 

1 (2X)
 

(50)
 

<i (8X)
 

1 (2X)
 

(45)
 

(49)
 

(49)
 

1 (2X)
 

(48)
 

(48)
 

(45)
 

(49)
 

1 (2%)
 

42 (86%)
 

(49)
 

(47)
 

1 (2%)
 

#LIVEB/PE HI PORTAL
 

FIBROSIS, ?OCAL
 

*BILE DHCT
 

INFLAMMATION, CHRONIC FOCAL
 

METAMORPHOSIS FATTY
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

HYPEHPLASIA, DIFFUSE
 

CPANCREAS
 

INFLAMMATION, ACUTE/CHRONIC
 

PEEIARTERITIS
 

ATROPHY, FOCAL
 

tSTOMACH
 

EPIDERMAL INCLUSION CIST
 

#GASTRIC MDCOSA
 

EROSION
 

*P£YSRS PATCH
 

HYPERPLASIA, RETICULUM CELL
 

#ILEUM
 

HYPERPLASIA, LYBPHOID
 

#COLON
 

NEMATODIASIS
 

URINARY SYSTEM
 

IKIDNEY
 

GLOMERULONEPHRITIS, NOS
 

INFLAMMATION, ACUTE/CHRONIC
 

SCAR
 

NEPH80SIS, NOS
 

#KIDNEY/PELVIS
 

MINERALIZATION
 

#URINARY BLADDER
 

POLYP
 

METAPLASIA^. SQflABOUS
 

CONTROL (ONTR)
 

01-0070
 

(48)
 

(48)
 

6 (13S)
 

(45)
 

6 (13*)
 

1 (2X)
 

1 (2*)
 

(48)
 

1 (2%)
 

(48)
 

(45)
 

1 (2X)
 

(45)
 

1 (2X)
 

(44)
 

4 (9*)
 

(48)
 

3 (6X)
 1 (2*)
 

41 (85%)
 

(48)
 

1 (2X)
 

(46)
 

1 J2%1
 

LOH DOSE
 

01-0073
 

(50)
 

(50)
 

1 (2*)
 

1 (2X)
 

3 (6X)
 

1 (2*)
 

(46)
 

(49)
 

(49)
 

(49)
 

(49)
 

(42)
 

(50)
 

45 (90%)
 

(50)
 

(50)
 

* HU1BEH OF ANIMALS BITH TISSUE EXAMINE D HICHOSCOPICALLY 

* NO-1BER OF ANIKAL S NECROPSIED 

C-5 
TABLE Cl (CONTINUED)
 

HIGH DOSE
 

01-0074
 

(44)
 

(48)
 

(48)
 

(47)
 

(45)
 

(50)
 

1 (2X)
 

1 (2%)
 

(47)
 

5 (11%)
 

1 (2X)
 

(50)
 

(49)
 

4 <8X)
 

(49)
 

1 12 Si
 

ENDOCEINE SYSTEM
 

#PITUITARY
 

CONGESTION, NOS
 HYPEHPLASIA, FOCAL
 

#ADRENAL
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

*ADRENAL CORTEX
 

HYPERPLASIA, FOCAL
 

ITHYROID
 HYPERPLASIA, C-CELL
 

iPANCREATIC ISLETS
 HYPERPLASIA, FOCAL
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND
 

GALACTOCELE
 CYSTIC DUCTS
 

HYPERPLASIA, NOS
 

fPROSTATE
 INFLAMMATION, FOCAL
 

INFLAMMATION, SUPPURATIVE
 

INFLAMMATION, ACUTE
 

INFLAMHATION, ACUTE FOCAL
 INFLAMMATION, ACUTE SUPPUBATIVE
 

INFLAMMATION, ACUTE/CHRONIC
 

*SIMINAL VESICLE
 

ATHOPHY, NOS
 

tTESTIS
 

DEGENERATION, NOS
 ATROPHY, NOS
 HYPESPLASIA, INTERSTITIAL CELL
 

#TESTIS/TUBULE
 

DEGENERATION, NOS
 

CONTROL (ONTR)


01-0070


(41)


3 (7X)
 

(47)


1 (2X)
 

3 (6X)
 

(47)


1 (2X)
 

(39)


1 (3X)
 

(45)


1 (2X)
 

(48)


1 (2X)
 

3 (6X)
 

(43)


1 (2X)
 

6 (14X)
 

8 (19X)
 

2 (5X)
 

(48)


2 (4X)
 

(47)


39 (83X)
 

1 (2X)
 

(47)


 LOW DOSE
 

 01-0073
 

 (46)
 

1 (2X)
 

 (49)
 

 (49)
 

 (45)
 

 (46)
 

 (50)
 

 (48)
 

3 (6X)
 

1 (2X)
 

 (50)
 

 (50)
 

2 (4X)
 

 (50)
 

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

C-6
 
TABLE Cl (CONCLUDED) 

CALCIFICATION, NOS

CONTROL (UNTR)

01-0070

 LOW DOSE

 01-0073

 HIGH DOSE 

 01-007U 

1 (2%) 

NERVOUS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

«EYE/CORNEA

ULCER, ACUTE

 (18) (50) (50) 

1 (2%) 

MUSCULOSKELETAL SYSTEM 

NINE 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION, ACUTE/CHRONIC 2 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED

f»NIMAL MISSING/NO NECROPSY 1 

AUTO/NICROPSY/HIST 1 O PERF

AUTO/NECROPSY/NO HISTO

AUTOLYSIS/NO NECROPSY 1 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

1 

1 

1 

1 

C-7
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH 3-NITRO-p-ACETOPHENETIDE 

(:ONTROl (ONTR) LOW DOSS HIGH DOSE
 

02-0070 02-0073 02-0074
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

ANIMALS NSCROPSIF.D 49 50 49
 ANIMALS EXAMINED HISTOPATHOLOGICALLY** 49 50 49
 

INTEGUMENTARY SYSTEM
 

*SKIN (49) (50) (49)
 

EPIDERMAL INCLUSION CYST 1 (2JS)
 

EESPIHATOPY SYSTEM
 

#TEACHEA (49) (48) (47)
 

INFLAMMATION, ACUTE/CHRONIC 15 (31X)
 

iLUNG/BRONCHUS (49) (50) (49)
 

SRONCHIECTASIS 1 (2X)
 

INFLAHHATION, ACUTE/CHRONIC 1 (2%)
 

*LUNG (49) (50) (49)
 INFLAMMATION. FOCAL 2 (UX)
 

INFLAMMATION, INTERSTITIAL 2 (4X)
 

INFLAMMATION, SUPPURATIVE 1 (2X)
 GRANULOMA, NOS I (2S)
 

HYPEHPLASIA, ALVEOLAR EPITHELIUM 1 (2%)
 

HMATOPOIETIC SYSTEM
 

*BONE MABROW (46) (50) (49)
 

OSTEOSCLEROSIS 1 (2X)
 

#SPLEEN (48) (50) (48)
 

HYPSRPLASIA, HEMATOPOIETIC 1 (2X)
 

HYP2RPLASIA. RETICOL'JB CELL 1 (2%)
 

CIRCULATORY SYSTEM
 

# HEART (49) (50) (49)
 

_ FIBROSIS^ FOCA.L 1 1221 1 J.2%1
 

# NU1BER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NOHBS B OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C_ oo 
FIBROSIS, DIFFUSE
 

PEHIARTERITIS
 

NECROSIS, FOCAL
 

CALCIFICATION, NOS
 

•MYOCARDIUM
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, ACUTE/CHRONIC
 

INFLAMMATION, CHRONIC
 

FIBROSIS, FOCAL
 

tCARDIA C VALVE 

INFLAMMATION, ACUTE/CHRONIC
 

*COfONARY ARTERY
 

INFLAMMATION, CHRONIC FOCAL
 

*PULMONAHY ARTERY
 

MINERALIZATION
 

DIGESTIVE SYSTEM
 

tLIVER
 

DEGENERATION, EOSINOPHILIC
 

NECROSIS, FOCAL
 

MITANORPHOSIS FATTY
 

HYPERPLASIA, FOCAL
 

ANGIECTASIS
 

*BILE DUCT
 

HYPERPLASIA, NOS
 

HYPERPIASIA, FOCAL
 

tPANCREAS
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, ACUTE/CHRONIC
 

ATROPHY, NOS
 

•STOMACH
 

ULCER, FOCAL
 

•DUODENAL MUSCULARIS
 

INFLAMMATION, GRANULOMATOUS
 

•COLON
 

TABLE C2 (CONTINUED) 

CONTROL (UNTR) 

02-0070 

1 (2%) 

(49)
 

2 (4%)
 

1 (2%)
 

(49)
 

1 (2%)
 

(49)
 

(49)
 

9 (18%)
 

(49)
 

2 (4%)
 

3 (6%)
 

4 (8%)
 

29 (59%)
 

1 (2%)
 

(49)
 

5 (10%)
 

1 (2%)
 

(47)
 

4 (9%)
 

1 (2%)
 

(49)
 

1 (2%)
 

(49)
 

(44)
 

LOH DOSE 

02-0073 

(50)
 

1 (2%)
 

(50)
 

(50)
 

(50)
 

(50)
 

(50)
 

5 (10%)
 

(48)
 

1 (2%)
 

(50)
 

(49)
 

1 (2%)
 

(44;
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-9
 

HIGH DOSE 

02-0074 

1 (2%) 

1 <2X) 

1 (2%) 

(49) 

(49) 

(49)
 

1 (2%)
 

(49)
 

(48)
 

(49)
 

1 (2%)
 

1 (2%)
 

(46)
 

(46)
 

(48)
 

(35)
 
TABLE C2 (CONTINUED)
 

HIGH DOSE
 

02-0074
 

(48)
 

15 (31X)
 

1 (2»)
 

(37)
 

(48)
 

(48)
 

(48)
 

(49)
 

(49)
 

(41)
 

(41)
 

URINARY SYSTEM
 

*KIDNEY
 

EMBRYONAL DUCT CYST
 

MINERALIZATION
 

CYST, NOS
 

NEPHROSIS, NOS
 

CALCINOSIS, NOS
 

ENDOCRINE SYSTEM
 

#PIT'JITARY
 HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

tADRENAL
 

METAMORPHOSIS FATTY
 

#ADRENAL CORTEX
 

METAMORPHOSIS FATTY
 

HYP3PPLASIA, FOCAL
 

IADRENAL MEDULLA
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND
 

GALACTOCELE
 INFLAMMATION, ACUTE
 

HYPEHPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

*VAGINA
 

INFLAMMATION, ACUTE/CHRONIC
 

tUTERUS
 

DILATATION, NOS
 

HYDROMETRA
 

HEMORRHAGE
 

HYPERPLASIA, FOCAL
 

#CERVIX UTERI
 

_ INFLAMMATION, ACUTE/CHROMIC
 

CONTROL (UNTR)
 

02-0070
 

(49)
 

1 (2«)
 

34 (69X)
 

(44)
 

1 (2%)
 

2 (5*)
 

(49)
 

3 (6%)
 

(49)
 

3 (6%)
 

1 (2*)
 

(49)
 

1 (2*)
 

1 (2%)
 

(49)
 

9 (18%)
 

1 (2X)
 

23	 (47X)
 

2 (4*)
 

(49)
 

1 (2X)
 

(49)
 

6 (12*)
 

(49)
 

2 J4Jii
 

LOW DOSE
 

02-0073
 

(50)
 

1 (2%)
 

1 (2X)
 

21 (42X)
 

(45)
 

(49)
 

(49)
 

(«9)
 

(50)
 

(50)
 

(47)
 

1 (2X)
 

1 (2X)
 

1 (2«)
 

(47)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NOMBER OF ANIMALS NECROPSIED
 

C-10
 
TABLE C2 (CONTINUED) 

CONTROL (ONTR) LOB DOSE HIGH DOSE
 

02-0070 02-0073 02-0074
 

HYPERPLASIA, BASAL CELL 1 (2X)
 

ACANTHOSIS 1 (2%)
 

*UTERUS/ENDOMETRIUM (49) (47) (41)
 

INFLAMMATION, SUPPURATIVE 3 <6S) 3 (7S)
 

INFLAMMATION, ACDTE 23 (47X)
 

GRANULATION, TISSUE 1 (2X)
 

HYPERPLASIA, NOS 5 (10X)
 HYPEKPLASIA, CYSTIC 5 (10%)
 

tOVARY/OVIDUCT (49) (47) (41)
 

INFLAMMATION, ACOTE 1 (2X)
 

tOVARY (47) (50) (46)
 CYST, NOS 2 (UX) 1 (2X)
 ABSCESS, NOS 1 (2%)
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

*EYE (19) (50) (49)
 

SYNECHIA, NOS 1 (2X)
 

CATARACT 1 (2X)
 

*EYE/CORNEA (49) (50) (49)
 INFLAMMATION, CHRONIC 1 (2X)
 

*EYE/RETINA (49) (50) (49)
 DEGENEPATION, NOS 1 (2X)
 

*EYS/CRYSTALLINE LENS (49) (50) (49)
 

CALCIFICATION, NOS 1 (2X)
 

*LENS CAPSULE (49) (50) (49)
 

CALCIFICATION, NOS 1 (2X)
 

*EXTERNAL EAR (19) (50) (49)
 

HEMATOMA, NOS 1 (2X)
 

MUSCULOSKELETAL SYSTEM
 

»ST3RNUM (49) (50) (49)
 

____ OSTEQPETEQSIS
 

* NU1BER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-ll
 
TABLE C2 (CONCLUDED) 

CONTROL(UNTR)

02-0070

 LOW DOSE

 02-0073

 HIGH DOSE 

 02-007U 

BODY CAVITIES 

*M£DIASTINUM

PSRIARTERITIS

 (19)

 1 (2%) 

 (50) (t9) 

ALL OTHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION, ACUTE/CHRONIC

INFLAMMATION, CHRONIC

OMENTUM 

MINERALIZATION

 3 

2 

1 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED 12

AOTOLYSIS/NO NECROPSY 1

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

 22 

1 

C-12
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 3-NITRO-p-ACETOPHENETIDE
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH 3-NITRO-p-ACETOPHENETIDE

 

CONTROL(UNTR) LOW DOSE HIGH DOSE 

05-0077 05-0075 05-0076 

ANIMAL S INITIALL Y IN STUDY 50 S50 50 

ANIMAL S MISSIN G 1 

ANIMAL S NECROPSIED 46 46 45 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 45 41 43 

INTEGUMENTAR Y SYSTE M 

*SUBCUT TISSUE (46) (46) (45) HYPEHPLA3IA , FOCAL 1 (2X) HYPERKERATOSI S 1 (2X) 

RESPIRATOR Y SYSTEM 

tLUNG (45) (41) (43) ARTERIOSCLEROSIS, NOS 1 (2%) 

HEMATOPOIFTIC SYSTEM 

#SPLEBN (45) (41) (43) FIBROSIS 1 (2%) 1 (2X) HEMOSIDEROSI S 1 (2*) HYPERPLASIA , RETICULOM CELL 3 (1%) 

HYPERPLASIA , LYMPHOI D 1 (2*) HEBATOPOIESIS 1 (2%) 1 (2X) 

IKESENTERIC L. NODE (35) (36) (35) INFLAMMATION , ACUTE/CHRONIC 1 (3X) 

CIRCULATOR Y SYSTEH 

NON E 

DIGESTIVE SYSTEM 

tLIVER (45) (4T) (43) CYST, N0§ 1_I2SL 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIED **EXCLUDES

 PARTIALLY AUTOLYZED ANIMALS 

NOT!!: ANIMA L f010 WAS DELETED FBOK STUDY WHEN FOUND TO BE A FEMALE III A MALE GFOOP. 

D-3 
TABLE Dl (CONTINUED)
 

CONTROL (UNTR )

05-0077

 LOW DOSE 

05-0075 

INFLAMMATION ,

DEGENERATION ,

METAMORPHOSI S

ANGISCTASI S 

 FOCAL 

 DOS 

 FATTY 

2 (4%) 

1 (2X ) 

2 («X) 

1 <2X) 

#LIVEE/CENTEILOBULAR 

METAMORPHOSIS FATTY 

(45) (41) 

1 <2X)» 

tLIVER/PERIPOBTAL 

INFLAMMATION, NOS 

MEGALOCYTOSIS 

(45) 

1 (2X) 

1 (2*) 

(41)

*BILE DUCT 

CYST, NOS 

INFLAMMATION, NOS 

HYPERPLASIA, FOCAL 

(46) 

1 (2%) 

1 (2X) 

(46)

1 (2X) 

#STOMACH 

HYPEBPLASIA, FOCAL 

(42) 

1 (2X) 

(41)

URINAEY SYSTEM
 

• KIDNEY (45) (41) 

CALCULUS, NOS 20 (44%) 

INFLAMMATION, INTERSTITIAL 5 (11%) 

INFLAMMATION, CH80HIC 1 <2X) 

PERIVASCULITIS 2 (**) 

ARTERIOSCLEROSIS, NOS 1 (2%) 

NEPHROSIS, NOS 1 (2X) 

GLOMESULOSCLEROSIS, NOS 25 (61X) 

HYPEEPLASIA, TUBULAR CELL 2 (WX) 

*KIDNEY/TUBOLE (45) (41) 

DEGEN"PATION, NOS 1 (2%) 

METAMORPHOSIS FATTY 9 (20X) 

ENDOCRIN E SYSTEM 

NONE 

REPRODUCTIVE SYSTEM
 

*TESTIS/TUBULB (45) (41) 

* NUMBER OF ANIMALS WITH TISSDE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NEC80PSIED
 

HIGH DOSE 

05-0076 

3 (7X) 

(13)
 

 (43)
 

 (45)
 

 (43)
 

<H3) 

22 (51X) 

(43) 

(43) 

D-4
 
TABLE Dl (CONCLUDED)
 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 05-0077 05-0075 05-0076
 

NFRVOU S SYSTE M 

NON E 

SPECIAL SENSE ORGANS
 

NONE
 

MOSCOLOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*SBDOMINAL CAVITY (l»6) (16) (H5)
 STEATITTS 1(2%)
 

ALL OTHEE SYSTEMS
 

ADIPOSE TISSUE
 

INFLAMMATION, FOCAL 1
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 8 10 6
 ANIMAL MISSING/NO NECROPSY 1
 NECROPSY PERF/NO HISTO PERFORMED 5
 

SUTO/NECSOPSY/NO HISTO 1 2
 

AUTOLYSIS/NO NECROPSY It 3 U
 

» NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NDMBEE OF ANIMALS NECROPSIED
 

D-5
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH 3-NITRO-p-ACETOPHENETIDE

 

CONTROL (OKIE) LOW DOSE HIGH DOSE 

06-0077 06-0075 06-0076 

ANIMAL S INITIALL Y IN STUDY 50 50 50 

ANIMAL S NECROPSIED 46 47 49 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 46 47 49 

INTEGUMENTAR Y SYSTEM 

*SKIN (46) (47) (49) 

FI3ROSIS 1 (2«) 

FIBROSIS, FOCAL 1 (2%) 

RESPIRATOR Y SYSTEM 

*LUMG (45) (47) (49) 

INFLAMMATION , INTERSTITIAL 2 (4*) 

PERIARTERITI S 1 (2X) 

HE1ATOPOIETIC SYSTEM 

*SPLEFN (43) (47) (48) 

HYPFRPLASIA , FETICULUfl CELL 2 (5«) 

HYPEBPLASIA , LYMPHOID 4 (9%) 

HEMATOPOIESI S 1 (2%) 2 (4*) 

CISC'JLATORY SYSTE M 

#HEABT (45) (47) (48)
 

PFHIARTERITIS 1 (2X)
 

*1YOCARDIUM (45) (47) (48)
 

CALCIFICATION, FOCAL 1 (2*)
 

*PHLHONAEY ARTERY (46) (47) (49)
 

HYP3RPLASTA, NOS 1 (2X)
 

*MESENTFPIC ARTERY (46) (47) (49)
 

* NU1BEP OF ANIMAL S WITH TISSDE EXAMINE D MICROSCOPICALLY 

* NtlHBF P OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-6
 
TABLE D2 (CONTINUED) 

DIGESTIVE SYSTEH 

ILIVER 

INFLAMMATION , FOCAL 

INFLAMMATION , CHRONIC 

CYTOPLASMIC CHANGE, NOS 

BASOPHILIC CYTO CHANGE 

MEGBLOCYTOSIS 

HYPEFTROPHY , NOS 

HYPEEPLASIA , NO S 

HYPERPLASIA , FOCAL 

ANGIECTASIS 

*LIVER/PEBIPORTAL 

INFLAMMATION , NOS 

(tLIVER/KOPFFER CELL 

HYPERPLASIA, NOS 

*BILE DUCT 

INFLAMMATION , NOS 

INFLAMMATION , CHRONIC 

tSTOMAC H 

PERIARTERITI S 

URINAR Y SYSTEM 

IKIDNE Y 

INFLAMMATION , INTERSTITIAL 

PFRIVASCULITIS 

GLOMERULOSCLEROSIS , NOS 

tKIDNEY/GLOBERULU S 

AMYLOIDOSI S 

SIDESOSIS 

tKIDNEY/PELVI S 

INFLAMMATION, ACOTE/CHHONIC 

fORINAR Y BLADDER 

INFLAMMATION , ACUTE 

PERIART£RITJ5_ 

# NUMBF R OF ANIMAL S WITH TISSUE

* NUMBE R OF ANIMAL S NECROPSIED 

CONTROL (UNTR ) LOW DOSE HIGH DOSE 

06-0077 06-0075 06-0076 

(45) (47) (49) 

1 (2*) 1 (2*) 

1 (2%) 

1 (2%) 1 (2*)

 

1 (2%) 

1 (2*) 

1 (2«) 

1 (2*) 

1 <2X ) 

7 (15*) 1 (2*) 

(45) (47) (49) 

1 (2% ) 

(45) (47) (49) 

1 (2%) 

(U6 ) (47) (49) 

1 (2%) 

1 (2% ) 

(«2 ) (44) (47) 

2 (4*) 

(43) (46) <«9) 

3 <7X ) 

ft (9%) 

17 (37*) 25 (51*) 

(43) (46) (49) 

1 (2% ) 

1 (2X) 

(»3) (46) (49) 

1 (2%) 

(U1 ) (47) (46) 

1 (2%) 

1_12*L 

 EXAMINE D MICROSCOPICALL Y 

D-7 
TABLE D2 (CONTINUED) 

CONTROL (UHTB) LOW DOSE 

06-0077 06-0075 

ENEOCEIN E SYSTEH 

#THYEOID (30) (U5)

 

PERIARTFRITIS 

REPRODUCTIVE SYSTEM 

fOTEEU S (13) (17) 

DILATATION , NOS 1 <2X) 

HYDROMETR A 1 (9X) 3 (6X) 

IDTEEUS/yNDOMETRIUM (13) (1?) 

CYST, NOS 2 (5X) 

INFLAMMATION, SUPPORATIVE 7 (15X) 

INFLAMMATION , ACUTE 1 (2X) 

HYPERPLASIA , NOS 1 (2X) 

HYPERPLASIA , CYSTIC 35 (81X) 27 (57X) 

*OVAPY (11) (15) 

CYST, NOS 1 (2X) it (9X) 

HEMATOMB , NOS 

INFLAMMATION , SOPPURATIVE 2 (<*X) 

ABSCESS, NOS 2 (IX) 

INFLAMMATION , CHRONIC 1 (9X) 

NERVOUS SYSTEM 

NOSE 

SPECIAL SENSE ORGANS 

*EYE/CORNfA (16) (17)

 

ULCER, ACUTE 

MUSCULOSKELETA L SYSTEH 

*VERTEEEA
 

OSTEOSCLEROSIS 1 (2X)

 

BODY CAVITIES
 

*MESENTSRY (46) (17) 

* NUMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBER OF ANIMAL S NECEOPSIED 

HIGH DOSE 

06-0076 

(12) 

1 (2X) 

(12) 

1 (2X) 

(12) 

1 (2X) 

1 (2*) 

6 (UX) 

(17) 

1 (9X) 

1 (2X) 

(19) 

1 (2X) 

(19) 

(19)
 

2_iiSL.­

D-8 
TABLE D2 (CONCLUDED) 

CONTROL(ONTR )

06-0077

 LOW DOSE

 06-0075

 HIGH DOSE 

 06-0076 

A LL OTHFR SYSTEMS 

NON E 

SPECIAL MORPHOLOG Y SUMMAR Y 

NO LESION REPORTED

AtlTO/NECROPSY/HISTO PERF

ACJTOLYSIS/NO NECROPSY

* NUMBE R OF ANIMAL S WITH TISSUE

* NOMBE B OF ANIMAL S NECROPSIED 

1

 2

 U

 EXAMINE D MICROSCOPICALL Y 

H

 1

 3

 12 

1 

1 

D-9 


NIH Publication No. 79-1388