1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 118 

BIOASSAY OF 

5-NITROACENAPHTHENE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 602-87-9 

NCI-CG-TR-118 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

5-NITROACENAPHTHENE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

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

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-1373
 

REPORT ON THE BIOASSAY OF 5-NITROACENAPHTHENE
 

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
 

5-nitroacenaphthene 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 5-nitroacenaphthene was conducted by
 

Mason Research Institute, Worcester, Massachusetts, initially under
 

direct contract to the NCI and currently under a subcontract to
 

Tracer 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. R. W. Fleischman (3), Dr. D. W. Hayden (3), Dr. A. S. Krishna Murthy (3), Dr. D.
 

S. Wyand (3), and Dr. Yoon (3) at the Mason Research Institute, the
 

pathology narratives were written by Dr. D. S. Wyand (3), and the
 

diagnoses included in this report represent the interpretation of
 

these pathologists. 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), and Mr.
 

R. M. Helfand (6), using methods selected for the Carcinogenesis
 

Testing Program by Dr. J. J. Gart (8).
 

111
 
This report was prepared at METREK, a Division of The MITRE Corporation (6) under the direction of the NCI. Those responsible for
 

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 Dr. B. Fuller (6), chemist Dr. N. Zimmerman
 

(6), and 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.	 Tracor 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.	 Now with 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.
 

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of 5-nitroacenaphthene for possible carcinogenicity
 

was conducted using Fischer 344 rats and B6C3F1 mice. 5-Nitroacenaphthene was administered in the feed, at either of two concentrations,
 

to groups of 50 male and 50 female animals of each species. For male
 

and female rats, the high and low dietary concentrations of 5-nitroacenaphthene were 0.24 and 0.12 percent, respectively. The high and
 

low time-weighted average concentrations for mice were 0.12 and 0.06
 

percent, respectively, for males and 0.12 and 0.05 percent, respectively, for females. After a 78-week dosing period, observation of
 

surviving rats continued for up to 22 weeks and observation of the
 

mice continued for 18 weeks.
 

For the chronic rat bioassay, 49 male and 50 female rats were
 

placed on test as high dose controls, and 50 rats of each sex served
 

as low dose controls. For the mice, 50 males and 50 females were
 

placed on test as controls.
 

Accelerated mortality was observed in all dosed groups except
 

the low dose female mice. There was a positive association between
 

mortality and dietary concentration of 5-nitroacenaphthene for both
 

sexes of both species. Early deaths were most apparent among high
 

dose male mice; half of the animals in this group were dead by week
 

20 and insufficient male mice survived to be at risk from late-


developing tumors.
 

Among rats, the incidence of malignant tumors of the ear canal
 

(incidences of ceruminous carcinomas and squamous-cell carcinomas
 

were combined) was significant at each dose level in each sex. Among
 

both dosed groups of female rats, the incidence of clitoral gland
 

carcinoma and the incidence of mammary adenocarcinoma were each significant. A significant incidence of alveolar/bronchiolar carcinoma
 

was observed in low dose rat groups of each sex.
 

Among female mice, the incidence of hepatocellular carcinoma was
 

significant at each dose level. The combined incidence of granulosacell tumors, luteomas, and tubular-cell adenomas of the ovary was significant in the high dose female mouse group.
 

Under the conditions of this bioassay, 5-nitroacenaphthene was
 

carcinogenic to Fischer 344 rats, causing increased incidences of
 

malignant tumors of the ear canal and lung in both sexes, and of the
 

clitoral gland and mammary gland in females. 5-Nitroacenaphthene was
 

also carcinogenic to female but not male B6C3F1 mice, causing carcinomas of the liver and ovarian tumors.
 

vii
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 3
 

A. Chemicals 3
 

B. Dietary Preparation 3
 

C. An imaIs 4
 

D. Animal Maintenance 4
 

E. Selection of Initial Concentrations 7
 

F. Experimental Design 9
 

G. Clinical and Histopathologic Examinations 12
 

H. Data Recording and Statistical Analyses 14
 

III. CHRONIC TESTING RESULTS: RATS 19
 

A. Body Weights and Clinical Observations 19
 

B. Survival 19
 

C. Pathology 23
 

D. Statistical Analyses of Results 24
 

IV. CHRONIC TESTING RESULTS: MICE 42
 

A. Body Weights and Clinical Observations 42
 

B. Survival 42
 

C. Pathology 45
 

D. Statistical Analyses of Results 47
 

V. DISCUSSION 54
 

VI. BIBLIOGRAPHY 58
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH 5-NITROACENAPHTHENE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH 5-NITROACENAPHTHENE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 5-NITROACENAPHTHENE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 5-NITROACENAPHTHENE D-l
 

ix
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF 5-NITROACENAPHTHENE 2 

2 GROWTH CURVES FOR 5-NITROACENAPHTHENE CHRONIC 

STUDY RATS 20 

SURVIVAL COMPARISONS OF 5-NITROACENAPHTHENE 

CHRONIC STUDY RATS 21 

GROWTH CURVES FOR 5-NITROACENAPHTHENE CHRONIC 

STUDY MICE 43 

SURVIVAL COMPARISONS OF 5-NITROACENAPHTHENE 

CHRONIC STUDY MICE 44 

LIST OF TABLES 

Table Number Page 

DESIGN SUMMARY FOR FISCHER 344 RATS— 

5-NITROACENAPHTHENE FEEDING EXPERIMENT 10 

DESIGN SUMMARY FOR B6C3F1 MICE—5-NITROACENAPHTHENE FEEDING EXPERIMENT 11 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN MALE 

RATS TREATED WITH 5-NITROACENAPHTHENE 25 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN FEMALE 

RATS TREATED WITH 5-NITROACENAPHTHENE 31 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN MALE 

MICE TREATED WITH 5-NITROACENAPHTHENE 48 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN FEMALE 

MICE TREATED WITH 5-NITROACENAPHTHENE 50 
LIST OF TABLES (Concluded)
 

Table Number 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH 5-NITROACENAPHTHENE A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH 5-NITROACENAPHTHENE A-8 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH 5-NITROACENAPHTHENE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH 5-NITROACENAPHTHENE B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH 5-NITROACENAPHTHENE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 5-NITROACENAPHTHENE C-ll 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH 5-NITROACENAPHTHENE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 5-NITROACENAPHTHENE D-8 

XI
 

I. INTRODUCTION
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for 5-nitroacenaphthene (Figure 1) (NCI No. C01967), an
 

*
 

aromatic nitro compound, is 1,2-dihydro-5-nitro-acenaphthylene. It
 

is also known as 5-nitroacenaphthylene.
 

5-Nitroacenaphthene has never had any known commercial application in the United States and is apparently produced in this country
 

solely for research purposes (Urso, 1977). The compound is, however,
 

used in Japan as a captive intermediate in the synthesis of naphthalimide dyes; 95 percent of these dyes find application as fluorescent
 

whitening agents in laundry detergents while the remainder are used
 

to dye paper (Urso, 1977).
 

Exposure to 5-nitroacenaphthene in the United States is presently
 

restricted to experimentalists engaged in laboratory research.
 

5-Nitroacenaphthene exhibited significant mutagenic activity in
 

two highly sensitive histidine-requiring tester strains of Salmonella
 

typhimurium (The Ames Test using the frame-shift-mutagen sensitive
 

strain TA 98 and the base-pair-substitution-mutagen sensitive strain
 

TA 100) (Yahagi et al., 1975). Reversion to histidine independence
 

occurred both in the presence and absence of microsomal activation,
 

with the largest number of revertent colonies (more than 100 times
 

the number expected as a result of spontaneous reversion) observed
 

in strain TA 98- in the presence of rat liver microsomes.
 

*
 

The CAS registry number is 602-87-9.
 
FIGURE 1

 

CHEMICAL STRUCTURE OF 5-NITROACEIMAPHTHENE 
II. MATERIALS AND METHODS
 

A. Chemicals
 

One batch of 5-nitroacenaphthene was purchased from Carroll Products, Wood River Junction, Rhode Island by the NCI for Mason Research
 

Institute, Worcester, Massachusetts. Spectroscopic analyses performed
 

at Mason Research Institute confirmed the identity of the compound.
 

The melting point of 98° to 100°C suggested a compound of fairly high
 

purity due to its narrow range and proximity to the literature value
 

of 103° to 104°C (Grasselli and Ritchey, 1975).
 

Throughout this report the term 5-nitroacenaphthene is used to
 

represent this material.
 

B. Dietary Preparation
 

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

5-Nitroacenaphthene was administered to the dosed animals as a component of the diet.
 

Proper amounts of the chemical were removed from the stock bottle
 

under a fume hood. The compound was ground to a powder and mixed with
 

an aliquot of meal using a mortar and pestle. Once visual homogeneity
 

was attained, the mixture was placed into a 6 kg capacity Patterson-


Kelley twin-shell stainless steel V-blender with the remainder of the
 

meal. After 20 minutes of blending, the mixtures were placed in
 

double plastic bags and stored in the dark at 4°C. The mixture was
 

used for only one week.
 

3
 
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. High dose and all control rats were obtained from
 

Charles River Breeding Laboratories, Inc., Wilmington, Massachusetts.
 

Low dose rats were obtained from ARS/Sprague-Dawley, Madison, Wisconsin, as were high and low dose mice. Control mice were obtained from
 

Charles River Breeding Laboratories, Inc. Dosed and control animals
 

for both 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 two 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 sex and species.
 

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 13 months of study, high dose and high dose control
 

rats were housed in galvanized- or stainless-steel wire-mesh cages
 
suspended above newspapers. Low dose rats and their controls were
 

housed in galvanized- or stainless-steel wire-mesh cages during quarantine and for the first 7 months of study. Newspapers under cages
 

were replaced daily and cages and racks washed weekly. For the remainder of the study, all rats were maintained in suspended polycarbonate cages equipped with disposable nonwoven fiber filter sheets.
 

Clean bedding and cages were provided twice weekly. High dose rats
 

and their controls were provided with corncob bedding (Bed-o-Cobs ,
 

The Andersons Cob Division, Maumee, Ohio) while they were in polycar­

®
 bonate cages. A second type of corncob bedding (SAN-I-CEL , Paxton
 

Processing Company, Paxton, Illinois) was supplied to low dose rats
 

for 7 months and low dose control rats for 8 months after they were
 

placed in polycarbonate cages. Bed-o-Cobs was used for the remainder of the study. Stainless-steel cage racks were cleaned once every
 

two weeks, and disposable filters were replaced at that time.
 

Mice were housed by sex in polycarbonate 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. Both types of lids
 

were from Lab Products, Inc., Garfield, New Jersey. Nonwoven fiber
 

filter bonnets were used over cage lids. Mice were housed ten per
 

cage for the first 18 months of study and five per cage thereafter.
 

Cages, lids, filters, and bedding were provided three times per week
 
when cage populations were ten and twice per week when cage populations were reduced to five. Ab-sorb-dri hardwood chip bedding ,
 

(Wilner Wood Products Company, Norway, Maine) was used through the
 

first 7 months of study followed by SAN-I-CEL for the next 12
 

months. After 19 months on study, Bed-o-Cobs was utilized as
 

bedding. Reusable filter bonnets and pipe racks were sanitized
 

every two weeks throughout the study.
 

Water was available ad libitum for both species from 250 ml
 

water bottles equipped with rubber stoppers and stainless steel
 

sipper tubes. Bottles were replaced twice weekly and, for rats only,
 

refilled as needed between changes. '
 

Wayne Lab-Blox was supplied ad libitum throughout the entire
 

test. Pelleted Wayne Lab-Blox was supplied during the quarantine
 

period and final observation period. During the chemical administration period, all dosed animals were fed Wayne Lab-Blox meal containing the appropriate concentration of 5-nitroacenaphthene, while
 

control animals had untreated meal available. Alpine aluminum feed
 

cups (Curtin Matheson Scientific, Inc., Woburn, Massachusetts) containing stainless steel baffles were used to distribute powdered feed
 

to high dose rats and their controls for the first 11 months of the
 

study. These cups were used for all mice and for low dose rats and
 

their controls during the entire study. During the last 7 months of
 

chemical administration, high dose rats and their controls were fed
 

from stainless steel gangstyle feed hoppers (Scientific Cages, Inc.,
 
Bryan, Texas). During the observation period following chemical
 

administration, rats were fed pellets on the cage floor and mice were
 

fed pellets from a wire bar hopper incorporated into the cage lid.
 

Food hoppers were changed on the same schedule as were cages. Food
 

was replenished daily in Alpine® feed cups.
 

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

containing 3-amino-4-ethoxyacetanilide (17026-81-2); 4-nitroanthranilic acid (619-17-0); l-amino-2-methylanthraquinone (82-28-0); and 5­

nitro-o-anisidine (99-59-2). Control rats were in a room with other
 

rats receiving diets containing amitrole (61-82-5); 2-methyl-l-nitroanthraquinone (129-15-7); and 3-nitro-p-acetophenetide (1777-84-0).
 

Dosed and control mice were 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; l-amino-2­

methylanthraquinone (82-28-0); 2,4-diaminoanisole sulfate (615-05-4);
 

APC (8003-03-0); 5-nitro-o-anisidine .(99-59-2); 4-nitroanthranilic
 

acid (619-17-0); 1-nitronaphthalene (86-57-7); and 3-nitro-p-acetophenetide (1777-84-0).
 

E.	 Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

5-nitroacenaphthene for administration to dosed animals in the
 

*
 

CAS registry numbers are given in parentheses.
 
chronic studies, 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. 5-Nitroacenaphthene was incorporated into the basal laboratory diet and supplied
 

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

groups in concentrations of 0.03, 0.06, 0.12, 0.24, and 0.48 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 a period of 4 weeks, followed by a 2-week observation period during which all animals were fed the basal diet.
 

The highest concentration causing no deaths, no compound-related
 

gross abnormalities, and no mean group body weight depression in
 

excess of 10 percent relative to controls was selected as the high
 

concentration utilized for the rat and mouse chronic bioassays.
 

There were no rat deaths during the 6-week subchronic study.
 

Mean body weight depression occurred in only one group of male rats,
 

those receiving 0.48 percent, the highest concentration. At this
 

concentration the mean body weight depression was in excess of 10
 

percent. Female rats experienced no mean group body weight depression. At 0.24 and 0.48 percent male rats were noted at gross necropsy
 

to have enlarged submaxillary lymph nodes and white foci on external
 

renal surfaces. Females exhibited no abnormalities. The high concentration selected for male and female rats in the chronic study was
 

0.24 percent.
 

8
 
There were three mouse deaths during the 6-week subchronic study,
 

all in the male group receiving the highest concentration of 5-nitroacenaphthene. Mean group body weight depression was in excess of 10
 

percent at dosages of 0.48 percent for male mice, and 0.24 and 0.48
 

percent for female mice. The mice receiving 0.48 percent 5-nitroacenaphthene exhibited chronic convulsions, prostration, emaciation, and
 

rapid respiration. Upon necropsy, discolored spleens and yellow kidneys and livers were noted. The high concentration selected for male
 

and female mice in the chronic study was 0.12 percent.
 

F. Experimental Design
 

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

of treated and untreated observation periods, and the 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. Low dose rats were started on
 

test approximately 7 months prior to initiation of the high dose rats.
 

The control rats for both dosed groups were approximately 1 month
 

younger than the respective dosed groups and were, therefore, included in the bioassay approximately 1 month after their corresponding dosed group. The dietary concentrations of 5-nitroacenaphthene
 

administered were 0.24 and 0.12 percent. Throughout this report
 

those rats receiving the former concentration are referred to as the
 

high dose groups, while those rats receiving the latter concentration
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

5-NITROACENAPHTHENE FEEDING EXPERIMENT
 

INITIAL 5-NITROACE­ OBSERVATION PERIOD 

GROUP NAPHTHENE TREATED UNTREATED 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) 

MALE 

LOW DOSE CONTROL 50 0 0 108 

HIGH DOSE CONTROL 49	 0 0 109
 

LOW DOSE 50	 0.12 78
 

0 22
 

HIGH DOSEb 50	 0.24 70
 

FEMALE
 

LOW DOSE CONTROL 50 0 0 108
 

HIGH DOSE CONTROL 50 0 0 110
 

LOW DOSE 50 0.12 78
 

0 22
 

HIGH DOSE 50 0.24 78
 

0 9
 

Concentrations given in percentages of feed.
 

This group was terminated in week 70, before the period of chemical
 

administration was to be completed.
 

10
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

5-NITROACENA,PHTHENE FEEDING EXPERIMENT
 

INITIAL 5-NITROACEGROUP NAPHTHENE 

SIZE CONCENTRATION3 

MALE
 

CONTROL 50	 0
 

LOW DOSE 50	 0.06
 

0
 

HIGH DOSE 50	 0.12
 

0
 

FEMALE
 

CONTROL 50	 0
 

LOW DOSE 50	 0.06
 

0.03
 

0
 

HIGH DOSE 50	 0.12
 

0
 

a
 Concentrations given in percentages
 

b
 

Timp-wpi pht-pd averafrt? ronrenl-rat- ion
 

OBSERVATION PERIOD
 

TREATED UNTREATED
 

(WEEKS) (WEEKS)
 

0 96
 

78
 

18
 

78
 

18
 

0 97
 

51
 

27
 

18
 

78
 

18
 

of feed.
 

2 (concentration X
 

TIME-WEIGHTED
 

AVERAGE
 

CONCENTRATION
 

0
 

0.06
 

0.12
 

0
 

0.05
 

0.12
 

weeks received)
 

2(weeks receiving chemical)
 

11
 
are referred to as the low dose groups. Except for the high dose
 

male group, which was terminated in week 70, the dosed rats received
 

feed containing 5-nitroacenaphthene for a period of 78 weeks, followed
 

by an observation period of up to 22 weeks.
 

The dosed and control mice were all approximately 6 weeks old
 

at the time they were placed on test. The dosed mice shared the same
 

median date of birth and were, therefore, started on test at the same
 

time. Control mice were approximately 3 weeks younger than the dosed
 

mice and, as a result, were started on test approximately 3 weeks
 

after the dosed groups. For both males and females, the initial
 

dietary concentrations of 5-nitroacenaphthene administered were 0.12
 

and 0.06 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. The concentration received by female low
 

dose mice was reduced to 0.03 percent after 51 weeks because of excessive mortality. After the 78-week period of chemical administration,
 

the dosed mouse groups were observed for up to 18 additional weeks.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. Body weights were recorded twice weekly for the first
 

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
 

12
 
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,
 

or 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, brain,
 

ear, testis, prostate, uterus, mammary gland, and ovary.
 

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
 

13
 
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
 

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
 

14
 
equality and used Tarone's (1975) extensions of Cox's methods when
 

testing a dose-related trend for mice. 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
 

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
 

15
 
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
 

for mice. 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
 

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
 

16
 
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; Tarone's extension to testing
 

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

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

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

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


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared 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
 

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
 

17
 
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.
 

18
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Severe compound-related mean group body weight depression was
 

evident in male rats after week 20. Compound-related,mean body
 

weight depression, although not as extreme, was also apparent in the
 

female rats (Figure 2). Fluctuations in the growth curve may be due
 

to mortality; as the size of the group diminishes, the mean body
 

weight may be subject to wide variations.
 

Subcutaneous and/or cutaneous masses were observed in 4 low dose
 

control males, 4 low dose control females, 2 high dose control males,
 

and 10 high dose control females. Four low dose males, 10 low dose
 

females, and 4 high dose females had lesions or masses in the ear
 

region. One high dose male developed a growth on the head and died
 

shortly thereafter. One high dose female developed several subcutaneous vaginal masses and one low dose control female had a firm nodule on the nose. One case of exopthalmia and one of severe alopecia
 

were observed among high dose control females.
 

B. Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and 5-nitroacenaphthene-dosed groups are shown in
 

Figure 3. For both male and female rats the Cox tests indicated that
 

each dosed group had a significantly elevated mortality compared to
 

its control group.
 

19
 
MEAN BODY WEIGHT (GRAMS) MEAN BODY WEIGHT (GR

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For males five rats were sacrificed from the high dose control
 

group in week 78 and five from the low dose control group in week 80.
 

Survival was high in all groups through the first 45 weeks of the
 

study, after which accelerated mortality was observed in the dosed
 

groups. In the high dose group the median survival was 61 weeks,
 

with the last animal dying in week 70. In the high dose group there
 

were not adequate numbers of rats at risk long enough for meaningful
 

statistical analyses of late-developing tumors. Survival was slightly
 

better in the low dose group, with a median survival of 83 weeks.
 

Thirty-six of 49 (73 percent) high dose control and 38/50 (76 percent)
 

low dose control males survived on test at least 100 weeks.
 

For females five rats were sacrificed from the high dose control
 

group in week 78 and five from the low dose control group in week 80.
 

Survival was good in all groups through week 33, after which accelerated mortality was observed in the high dose group. In the high dose
 

group the median survival was 56 weeks, with the last animal dying by
 

week 88. In the high dose group there were not adequate numbers of
 

rats at risk to permit meaningful statistical analyses of late-


developing tumors. Survival was slightly better in the low dose
 

group, with a median survival of 76 weeks, but no survivors after
 

week 100. Forty-three of 50 (86 percent) high dose control and 39/50
 

(78 percent) low dose control females survived on test at least 100
 

weeks.
 

22
 
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).
 

The incidence of mammary gland adenocarcinoma was elevated in
 

both dosed female groups (5/49 [10 percent] low dose and 5/48 [10
 

percent] high dose) when compared to controls (0/99); however, the
 

incidence of mammary fibroadenoma was less in dosed groups than in
 

controls. Morphologically the malignant mammary tumors were papillary adenocarcinomas which showed evidence of marked secretory activity (intracytoplasmic vacuoles and eosinophilic material in tumor
 

spaces and glandular lumens). Cells grew in single layers of cuboidal epithelium or multiple layers of more pleomorphic cells which
 

obliterated glandular patterns. Mitotic figures were numerous.
 

The incidence of clitoral gland carcinoma was elevated in dosed
 

females (0/99 controls, 6/49 [12 percent] low dose and 5/48 [10 percent] high dose). This neoplasm was distinguished from squamous and
 

sebaceous-type carcinomas by location and histologically, by the
 

presence of prominent intracytoplasmic eosinophilic granules.
 

In low and high dose rats of both sexes, there was a marked increase in incidence of ceruminous or squamous-cell carcinoma arising
 

in the region of the external ear canal (0/96 control males, 21/43
 

[49 percent] low dose males, 20/47 [43 percent] high dose males, 0/99
 

23
 
control females, 27/^-9 [55 percent] low dose females, 35/48 [73 percent] high dose females). These tumors probably arose from the auditory sebaceous glands (Zymbal's gland) in most instances. Ceruminous
 

carcinoma usually contained some squamous epithelial cells although a
 

few tumors containing only sebaceous cells were seen. The squamouscell carcinomas were mostly composed of pleomorphic squamous epithelium with little keratinization.
 

Increased incidences of alveolar/bronchiolar adenomas or alveolar/
 

bronchiolar carcinomas were observed in dosed rats (1/96 [1 percent]
 

control males, 7/41 [17 percent] low dose males, 3/47 [6 percent] high
 

dose males, 1/99 [1 percent] control females, 8/48 [17 percent] low
 

dose females, 3/48 [6 percent] high dose females).
 

There was a variety of spontaneous inflammatory and degenerative
 

lesions seen which commonly occur in aging Fischer 344 rats. These
 

were not considered to be compound-related.
 

It is concluded from this pathologic examination that 5-nitroacenaphthene was carcinogenic to Fischer 344 rats, inducing carcinomas of the ear canal and lung in both sexes and carcinomas of the
 

cliteral and mammary glands in females.
 

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
 

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

24
 
TABLE 3
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN MALE RATS TREATED WITH 5-NITROACENAPHTHENEa

»e
 

TOPOGRAPHY : MORPHOLOGY
 

Skin and Subcutaneous Tissue: Fibroma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

ho Lung: Alveplar/Bronchiolar
 

t-n Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar /Bronchiolar Adenoma
 

or Alveolar/Bronchiolar Carcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE HIGH DOSE LOW 

CONTROL CONTROL DOSE 

2/48(0.04) 3/48(0.06) 6/39(0.15) ___ 

N.S. 

3.692 

0.705 

35.610 

99 95 77 

0/48(0.00) 1/48(0.02) 5/37(0.14) 

P = 0.013 

Infinite 

1.642 

Infinite 

109 77 

0/48(0.00) 1/48(0.02) 7/38(0.18) 

P = 0.002 

_— Infinite 

2.462 

Infinite 

109 75 

HIGH
 

DOSE
 

1/40(0.03)
 

N.S.
 

0.400
 

0.008
 

4.739
 

62
 

0/40(0.00)
 

N.S.
 

0.000
 

0.000
 

22.284
 

3/45(0.07)
 

N.S.
 

3.200
 

0.268
 

164.119
 

49
 
___ 

_ __ 

TABLE 3 (CONTINUED)
 

TOPOGRAPHY: MORPHOLOGY
 

Kidney: Tubular-Cell Adenoma or
 

Tubular-Cell Adenocarcinoma^1
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

N3	 Pituitary: Adenoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma or Pheochromocytoma, Malignant
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW
 

DOSE
 

4/39(0.10)
 

P = 0.037
 

Infinite
 

1.144
 

Infinite
 

90
 

0/28(0.00)
 

N.S.
 

0.000
 

0.000
 

26.911
 

2/34(0.06)
 

N.S.
 

0.276
 

0.031
 

1.186
 

76
 

HIGH
 

DOSE
 

0/39(0.00)
 

N.S.
 

0/29(0.00)
 

P = 0.004(N)
 

0.000
 

0.000
 

0.487
 

0/36(0.00)
 

P = 0.008(N)
 

0.000
 

0.000
 

0.565
 

LOW DOSE
 

CONTROL
 

0/48(0.00)
 

1/41(0.02)
 

108
 

10/47(0.21)
 

99
 

HIGH DOSE
 

CONTROL
 

0/48(0.00)
 

9/38(0.24)
 

85
 

8/47(0.17)
 

107
 
___ 

TOPOGRAPHY : MORPHOLOGY
 

Preputial Gland: Carcinoma NOS
 

P Values
 

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
 

Ear Canal or Skin of Ear : Squamous-


Cell Carcinomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

2/48(0.04) 0/48(0.00)
 

—_ ___
 

104
 

45/47(0.96) 42/47(0.89)
 

—_
 

80 78
 

1/48(0.02) 0/48(0.00)
 

—_ ___
 

.
 

99
 

LOW
 

DOSE
 

3/39(0.08)
 

N.S.
 

1.846
 

0.222
 

21.132
 

79
 

19/33(0.58)
 

P < O.OOl(N)
 

0.601
 

0.526
 

0.787
 

71
 

7/39(0.18)
 

P = 0.014
 

8.615
 

1.178
 

376.472
 

68
 

HIGH
 

DOSE
 

0/37(0.00)
 

N.S.
 

_—
 

2/37(0.05)
 

P < O.OOl(N)
 

0.060
 

0.010
 

0.192
 

58
 

2/40(0.05)
 

N.S.
 

Infinite
 

0.356
 

Infinite
 

58
 
_ _ _ 

___ 

___ 

___ 

___ 

__ _ 

_ __ 

___ 

00 

TABLE 3 (CONTINUED)
 

TOPOGRAPHY: MORPHOLOGY
 

Ear Canal: Ceruminous Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ear Canal or Skin of Ear: Squamous-


Cell Carcinoma or Ceruminous Car­ cinoma*3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Body Cavities : Mesothelioma NOS or
 

Mesothelioma , Malignant^*
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE
 

CONTROL
 

0/48(0.00)
 

1/48(0.02)
 

99
 

1/48(0.02)
 

__—
 

108
 

HIGH DOSE
 

CONTROL
 

0/48(0.00)
 

0/48(0.00)
 

2/48(0.04)
 

«__
 

106
 

LOW
 

DOSE
 

14/42(0.33)
 

P < 0.001
 

Infinite
 

5.006
 

Infinite
 

38
 

21/42(0.50)
 

P < 0.001
 

24.000
 

4.200
 

946.120
 

38
 

4/39(0.10)
 

N.S.
 

4.923
 

0.513
 

235.669
 

79
 

HIGH
 

DOSE
 

18/47(0.38)
 

P < 0.001
 

Infinite
 

5.895
 

Infinite
 

47
 

20/47(0.43)
 

P < 0.001
 

Infinite
 

6.614
 

Infinite
 

47
 

2/40(0.05)
 

N.S.
 

1.200
 

0.090
 

15.868
 

66
 
TOPOGRAPHY rMORPHOLOGY
 

Thyroid: Papillary Adenoma or
 

Follicular-Cell Adenoma1*
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Papillary Adenocarcinoma 

VO or Follicular-Cell Carcinoma13 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Parathyroid: Adenoma NOS
 

c


P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

LOW DOSE HIGH DOSE


CONTROL CONTROL


0/39(0.00) 0/48(0.00)


0/39(0.00) 0/48(0.00)


0/23(0.00) 1/28(0.04)


109


 LOW
 

 DOSE
 

 4/33(0.12)
 

P = 0.040
 

Infinite
 

1.108
 

Infinite
 

79
 

 4/33(0.12)
 

P = 0.040
 

Infinite
 

1.108
 

Infinite
 

75
 

 1/20(0.05)
 

N.S.
 

Infinite
 

0.063
 

Infinite
 

 91
 

HIGH
 

DOSE
 

0/38(0.00)
 

N.S.
 

1/38(0.03)
 

N.S.
 

Infinite
 

0.068
 

Infinite
 

2/19(0.11)
 

N.S.
 

2.947
 

0.164
 

163.981
 

68 

62 
TABLE 3 (CONCLUDED)
 

TOPOGRAPHY :MORPHOLOGY 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Pancreatic Islets: Islet-Cell Adenoma 3/45(0.07) 0/46(0.00) 0/31(0.00) 1/35(0.03) 

P Values0 _—_ N.S. N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

__._ 0.000 

0.000 

2.375 

Infinite 

0.071 

Infinite 

Weeks to First Observed Tumor 85 62 

Hematopoietic System:

Malignant Lymphoma^ 

 Leukemia or 

6/48(0.13) 6/48(0.13) 2/39(0.05) 0/40(0.00) 

OJ 

o 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

N.S. 

0.410 

0.042 

2.138 

P = 0.023(N) 

0.000 

0.000 

0.745 

Weeks to First Observed Tumor 98 93 96 

Treated groups received doses of 0.12 or 0.24 percent in feed.
 

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

"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. A negative designation (N) indicates a lower incidence
 

in the treated group than in the control group.
 

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

B

These analyses were based solely upon animals surviving at least 52 weeks, except for sites
 

where the first tumor of interest was observed earlier than 52 weeks, where the analyses were
 

based upon all animals that survived until or past the date that the first tumor was observed.
 
___ 

TABLE 4
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN FEMALE RATS TREATED WITH 5-NITROACENAPHTHENEa>e
 

LOW DOSE HIGH DOSE LOW
 

TOPOGRAPHY rMORPHOLOGY CONTROL CONTROL DOSE
 

Skin: Squamous-Cell Carcinoma, Basal-Cell
 

Carcinoma, or Sebaceous
 

Adenocarcinomab 0/49(0.00) 1/50(0.02) 0/46(0.00)
 

P Values0 N.S.
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 110
 

Subcutaneous Tissue: Fibroma

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

 0/49(0.00) 1/50(0.02) 

___ 

3/46(0.07) 

N.S. 

Infinite 

0.641 

Infinite 

Weeks to First Observed Tumor 102 75 

Lung: Alveolar/Bronchiolar Carcinoma

P Values0 

 0/49(0.00) 0/50(0.00) 4/46(0.09) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Infinite 

0.989 

Infinite 

Weeks to First Observed Tumor 65 

HIGH
 

DOSE
 

3/31(0.10)
 

N.S.
 

4.839
 

0.408
 

245.211
 

85
 

2/31(0.06)
 

N.S.
 

3.226
 

0.174
 

183.956
 

68
 

1/31(0.03)
 

N.S.
 

Infinite
 

0.086
 

Infinite
 

87
 
TABLE 4 (CONTINUED)
 

LOW DOSE HIGH DOSE
 

TOPOGRAPHY : MORPHOLOGY CONTROL CONTROL
 

Lung: Alveolar/Bronchiolar Adenoma
 

or Alveolar/Bronchiolar Carcinoma 0/49(0.00) 1/50(0.02)
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 110
 

Hematopoietic System: Leukemia or
 

UJ Malignant Lymphoma^ 7/49(0.14) 5/50(0.10)
 ho
 

P Values0
 

Relative Risk (Control)
 _—
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 106 104
 

Pituitary: Adenoma NOS or Chromophobe
 

Adenoma 18/44(0.41) 17/40(0.43)
 

P Values0
 

Relative Risk (Control) —_
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 90 78
 

LOW
 

DOSE
 

8/46(0.17)
 

P = 0.002
 

Infinite
 

2.437
 

Infinite
 

55
 

1/46(0.02)
 

P = 0.036(N)
 

0.152
 

0.003
 

1.116
 

83
 

5/42(0.12)
 

P - 0.002(N)
 

0.291
 

0.094
 

0.727
 

60
 

HIGH
 

DOSE
 

3/31(0.10)
 

N.S.
 

4.839
 

0.408
 

245.211
 

53
 

0/30(0.00)
 

N.S.
 

0.000
 

0.000
 

1.298
 

0/28(0.00)
 

P < O.OOl(N)
 

0.000
 

0.000
 

0.257
 
___ _ __ 

TABLE 4 (CONTINUED) 

TOPOGRAPHY : MORPHOLOGY
 

Adrenal: Pheochromocytoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Carcinoma
 

UJ P Values0 

u> 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Carcinoma
 

or Follicular-Cell Adenoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE
 

CONTROL
 

2/49(0.04)
 

108
 

1/40(0.03)
 

_—
 

106
 

1/40(0.02)
 

106
 

HIGH DOSE
 

CONTROL
 

3/49(0.06)
 

109
 

1/45(0.02)
 

109
 

1/45(0.02)
 

109
 

LOW
 

DOSE
 

1/45(0.02)
 

N.S.
 

0.544
 

0.009
 

10.083
 

88
 

3/43(0.07)
 

N.S.
 

2.791
 

0.236
 

142.985
 

77
 

4/44(0.09)
 

N.S.
 

3.636
 

0.381
 

174.735
 

51
 

HIGH
 

DOSE
 

0/31(0.00)
 

N.S.
 

0.000
 

0.000
 

2.587
 

2/27(0.07)
 

N.S.
 

3.333
 

0.181
 

188.994
 

60
 

3/27(0.11)
 

N.S.
 

5.000
 

0.424
 

251.837
 

59
 
___ 

___ 

TOPOGRAPHY : MORPHOLOGY
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma or
 

Papillary Cystadenocarcinoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED)
 

LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

3/40(0.08) 2/45(0.04)
 

108 110
 

0/49(0.00) 0/50(0.00)
 

_— _—_
 

0/49(0.00) 0/50(0.00)
 

_ —
 

LOW
 

DOSE
 

0/43(0.00)
 

N.S.
 

0.000
 

0.000
 

1.538
 

5/46(0.11)
 

P = 0.024
 

Infinite
 

1.345
 

Infinite
 

60
 

6/46(0.13)
 

P = 0.011
 

Infinite
 

1.706
 

Infinite
 

60
 

HIGH
 

DOSE
 

0/27(0.00)
 

N.S.
 

0.000
 

0.000
 

5.519
 

5/31(0.16)
 

P = 0.007
 

Infinite
 

2.044
 

Infinite
 

65
 

5/31(0.16)
 

P = 0.007
 

Infinite
 

2.044
 

Infinite
 

65
 
___ 

TOPOGRAPHY : MORPHOLOGY
 

Mammary Gland: Fibroadenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Clitoral Gland: Carcinoma NOS 

u> P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Uterus: Endometrial Stromal Polyp
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

16/49(0.33) 19/50(0.38)
 

_—_
 

80 107
 

0/49(0.00) 0/50(0.00)
 

_—
 

12/49(0.24) 10/50(0.20)
 

80 78
 

LOW
 

DOSE
 

12/46(0.26)
 

N.S.
 

0.799
 

0.389
 

1.593
 

78
 

6/46(0.13)
 

P = 0.011
 

Infinite
 

1.706
 

Infinite
 

75
 

1/45(0.02)
 

P = O.OOl(N)
 

0.091
 

0.002
 

0.573
 

60
 

HIGH
 

DOSE
 

4/31(0.13)
 

P = 0.013(N)
 

0.340
 

0.092
 

0.902
 

74
 

5/39(0.13)
 

P = 0.014
 

Infinite
 

1.620
 

Infinite
 

40
 

1/26(0.04)
 

N.S.
 

0.192
 

0.005
 

1.227
 

77
 
TOPOGRAPHY: MORPHOLOGY
 

Ear Canal: Squamous-Cell Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ear Canal: Ceruminous Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ear Canal: Squamous-Cell Carcinoma,
 

Ceruminous Carcinoma or
 

Carcinosarcoma"3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED)
 

LOW DOSE HIGH DOSE


CONTROL CONTROL


0/49(0.00) 0/50(0.00)


0/49(0.00) 0/50(0.00)


0/49(0.00) 0/50(0.00)


 LOW
 

 DOSE
 

 3/49(0.06)
 

N.S.
 

Infinite
 

0.602
 

Infinite
 

67
 

 25/49(0.51)
 

P < 0.001
 

Infinite
 

8.233
 

Infinite
 

44
 

 27/49(0.55)
 

P < 0.001
 

Infinite
 

8.947
 

Infinite
 

44
 

HIGH
 

DOSE
 

9/46(0.20)
 

P = 0.001
 

Infinite
 

2.860
 

Infinite
 

34
 

26/47(0.55)
 

P < 0.001
 

Infinite
 

9.148
 

Infinite
 

33
 

35/47(0.74)
 

P < 0.001
 

Infinite
 

12.674
 

Infinite
 

33
 
TABLE 4 (CONCLUDED)
 

a

Treated groups received doses of 0.12 or 0.24 percent in feed. 

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

C

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. A negative designation (N) indicates a lower incidence 

in the treated group than in the control group. 

d

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

analyses were based solely upon animals surviving at least 52 weeks, except for sites
 

where the first tumor of interest was observed earlier than 52 weeks, where the analyses were
 

based upon all animals that survived until or past the date that the first tumor was observed.
 

OJ 

-J 
tumors were observed in at least one of the control or 5-nitroacenaphthene-dosed groups and where such tumors were observed in at
 

least 5 percent of the group. Due to high early mortality these
 

analyses have been based solely upon those rats surviving at least
 

52 weeks or, in the event that the tumor of interest was observed
 

earlier, at least as long as the time at which the first tumor of
 

interest was observed. Since all of the high dose males and all but
 

4/50 (8 percent) of the high dose females were dead after week 74,
 

the statistical tests involving the high dose groups are not meaningful for late-developing tumors. Additionally, statistical results
 

involving the low dose groups must be qualified since the low dose
 

rats came from a different supplier.
 

In both male and female rats large numbers of ceruminous carcinomas and squamous-cell carcinomas of the ear canal or skin of the
 

ear were observed. When the incidences were combined so that the
 

numerator represented rats of that sex with either one of these
 

tumors, then both for males and for females the Fisher exact tests
 

comparing both the high dose and low dose groups to their respective
 

controls were significant (P < 0.001). These tumors occurred in
 

3/334 (1 percent) of the male and 0/336 female Fischer 344 rats in
 

the historical controls observed at Mason Research Institute for
 

the NCI Carcinogenesis Testing Program. By comparison, these tumors
 

were observed in 20/47 (43 percent) of the high dose and 21/42 (50
 

percent) of the low dose males surviving on test at least 38 weeks,
 

38
 
and in 35/47 (74 percent) of the high dose and 27/49 (55 percent) of
 

the low dose females surviving on test at least 33 weeks. Based on
 

these statistical results, the administration of 5-nitroacenaphthene
 

was associated with an increased incidence of ceruminous carcinomas
 

and squamous-cell carcinomas of the ear canal or skin of the ear in
 

both male and female rats.
 

In female rats, the Fisher exact test for the incidence of clitoral gland carcinoma was significant when comparing both the high
 

dose to the high dose control (P = 0.014) and the low dose to the low
 

dose control (P = 0.011). Similarly, in female rats, the incidence
 

of mammary adenocarcinomas was significant when comparing both the
 

low dose (P = 0.024) and high dose (P = 0.007) to their respective
 

controls. In historical control females, 0/336 and 4/336 (1 percent)
 

of the untreated females had the clitoral or mammary tumor, respectively. Based on these statistical results, the administration of
 

5-nitroacenaphthene was associated with the increased incidence of
 

clitoral gland carcinomas and mammary adenocarcinomas in female
 

rats.
 

Significantly increased incidences of alveolar/bronchiolar neoplasms were noted with the Fisher exact tests for both low dose male
 

(P = 0.002) and low dose female (P = 0.002) rats. In historical
 

control data compiled by this laboratory for the NCI Carcinogenesis
 

Testing Program 5/334 (1 percent) of the male and 4/336 (1 percent)
 

of the female Fischer 344 rats had either an alveolar/bronchiolar
 

39
 
adenoma or an alveolar/bronchiolar carcinoma—compared to 7/38 (18
 

percent) low dose males and 8/46 (17 percent) low dose females in
 

this study. The absence of significant results in the high dose
 

groups probably reflected the poor survival in those high dose
 

groups. The earliest observed tumor was in week 49 in the high dose
 

males and week 53 in the high dose females, compared to week 109 in
 

the high dose control males and week 110 in the high dose control
 

females. Based on these statistical results, the administration of
 

5-nitroacenaphthene was associated with the development of alveolar/
 

bronchiolar neoplasms in male and female Fischer 344 rats.
 

In male rats the incidence of kidney tubular-cell adenomas or
 

tubular-cell adenocarcinomas was increased in the low dose group
 

over that in the control. The Fisher exact test yielded a value of
 

P = 0.037, a marginal result which was not significant under the
 

Bonferroni criterion. Similarly, the incidences of thyroid adenomas
 

and the incidences of thyroid carcinomas in males, plus the incidences
 

of leukemias or malignant lymphomas in females were not significant
 

under the Bonferroni criterion.
 

For interstitial-cell tumors of the testis, pituitary adenomas,
 

and adrenal pheochromocytomas in males and for pituitary adenomas,
 

mammary fibroadenomas, and endometrial stromal polyps in females, the
 

possibility of a negative association between chemical administration
 

and incidence was indicated. These results must be discounted, however, due to the poor survival of the dosed rats.
 

40
 
In summary, the statistical findings were that the administration
 

of 5-nitroacenaphthene was associated with the increased incidence
 

of carcinomas of the ear canal or skin of the ear and of alveolar/
 

bronchiolar neoplasms in both male and female rats and with clitoral
 

gland carcinomas and mammary gland adenocarcinomas in female rats.
 

41
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Compound-related mean body weight depression was apparent in
 

male mice but not in females (Figure 4). Fluctuations in the growth
 

curve may be due to mortality; as the size of the group diminishes,
 

the mean body weight may be subject to wide variations.
 

No abnormal clinical signs were recorded.
 

B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and 5-nitroacenaphthene-dosed groups are shown in
 

Figure 5. For male mice, the Tarone test for positive association
 

between increased dosage and accelerated mortality was significant
 

(P < 0.001). For female mice, the Cox test indicated a significantly
 

(P < 0.001) greater mortality in the high dose than in the control
 

group.
 

Five male mice were sacrificed from the control group in week 79.
 

Survival was very poor in the male high dose group with a median survival of 20 weeks: 10 died in week 13, 26 in week 20, and 6 in week
 

42. Only 3 mice survived past week 42. These deaths were not related
 

to tumor incidence since the one tumor found in a high dose male occurred in a mouse which survived until the end of the study. In the
 

low dose group 14 deaths were observed in week 50. All 14 of these
 

low dose males had a fatty metamorphosis of the liver; ten had
 

42
 
50- -50 

40­ — 40 

C/5 

V cc 

55 30— -30 

£20H -20 

o 

m 

CONTROL 

10— LOW DOSE -10 

MALE MICE HIGH DOSE 

i I 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

50­ -50 

40­ -40 

cc 

2 30­ -30 

>20- -20 

O 

CO 

CONTROL 

10— LOW DOSE -10 

HIGH DOSE 

FEMALE MICE 

I ' I I I 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR 5-NITROACENAPHTHENE CHRONIC STUDY MICE 

43 
PROBABILITY OF SURVIVAL PROBABILITY OF SURV

C CD O O C 

<D (0 J* O5 00 c3 C3 KJ -^ er 

D O 3 0 O 

0— 1 1 1 1 1 , 1 L_ o — i i 1 I 1 1 

1 -n - | tn m C 

r~ 

H

1

i 

m 33 

1 CJ1 < s- m O 

r~ ^ 1 n •" o J 0 m 

3

i £ "- CO 

^ O o 

30 

f V)

 

0 ­

V> 

O j 1 •

 —4 ^ 

-" d £- r _ tn Ul g 

1 

Z m m 

H -n 0 . O 30 — Z 2 

O £^> _i 1 

> d S m O 30 CO m CO 01 _ mrl

m m —| o H o 

Z m —. ,—, • 

J

• > s 5 "" m m 

m H 55 " I W •• C/3 f 

m — . *z S ~~ 1 01 m •• 

O 1 

I 1

30 f

O 

•

1

>r ~ 

Z r r- i r- CO 

§

o — j 1 § - 1 \ 

C; j ^JT75 

j 

\ •• I ' O | *" 

1 1 5 I r~ ") • 

x r n o S_ 5 § 0 

o — m 01 H tn O g 2 D 

O § D O /-, ^

 w m

 0 S 0 

~ m TM r 

N) — 1 ro — O 1 o 1 1 | 1

1 

1 1 1 ' 1 r- 

C3 O C i D O O C3 

D O o C3 Kj ji. O) bo cD '<3 ro J* "c 7> 
calcification of the renal papilla. Twenty-eight low dose males survived on test until the termination of the study. In the control
 

group survival was good, with 43/50 (86 percent) of the mice surviving
 

on test until the termination of the study. Only in the low dose and
 

control groups were adequate numbers of male mice at risk from late-


developing tumors.
 

Among females five mice were sacrificed from the control group in
 

week 79. The 24 deaths (48 percent) which occurred during week 50
 

in the high dose group were apparently due to toxicity, but only a few
 

additional deaths were observed after that week. At the termination
 

of the experiment 36 percent (18/50) of the high dose, 76 percent
 

(38/50) of the low dose, and 72 percent (36/50) of the control mice
 

were still alive on test. Adequate numbers of dosed and control females survived long enough to be at risk from late-developing tumors.
 

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).
 

The incidence of hepatocellular carcinoma was elevated in both
 

low dose (23/47 [49 percent]) and high dose (18/45 [40 percent])
 

females compared with female controls (2/47 [4 percent]). There
 

were no hepatocellular carcinomas in high dose males, although they
 

occurred in both control and low dose animals. The lesions were
 

typical of hepatocellular carcinoma in mice as described in the
 

45
 
literature (Reuber, 1975). The full range of lesions described by
 

Reuber as hepatocellular carcinoma was seen (i.e., from well-


differentiated to anaplastic). Four hepatocellular carcinomas in
 

treated female mice and one in a control female mouse metastasized
 

to the lung.
 

There was an increased incidence of ovarian neoplasms in dosed
 

female mice. Granulosa-cell tumors were found in 2/41 (5 percent)
 

low dose and 2/39 (5 percent) high dose mice. The incidence of
 

tubular-cell adenomas appeared to be dose-related, occurring in 2/41
 

(5 percent) low dose and 4/39 (10 percent) high dose mice. One luteoma was observed in the high dose group. None of these neoplasms
 

was observed in control female mice.
 

There were other tumors and many inflammatory and degenerative
 

lesions reported which are commonly seen in aging mice. Most of
 

these lesions were unrelated to the administration of the test compound, appearing with similar or lesser frequency in the dosed mice
 

than in the controls. Fatty metamorphosis of the liver and calcification of renal papilla were, however, compound-related lesions in
 

both sexes.
 

Based upon this pathologic examination, 5-nitroacenaphthene was
 

carcinogenic in female B6C3F1 mice, inducing hepatocellular carcinomas and ovarian tumors.
 

46
 
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 5-nitroacenaphthene-dosed groups and where such tumors were observed in at least
 

5 percent of the group. The results for high dose male mice have
 

been omitted since early mortality was so high that statistical
 

analyses of late-developing tumors were meaningless for that group.
 

Because of the high early mortality in both sexes, statistical analyses were based upon those mice surviving at least 52 weeks. All
 

of the statistical results for mice must be qualified since the
 

control mice were from a different supplier than the dosed mice.
 

For females the incidences of hepatocellular carcinoma were significant (P < 0.001) when comparing the dosed groups to the controls
 

using the Cochran-Armitage test. These Cochran-Armitage results were
 

supported by significant (P < 0.001) Fisher exact test results for
 

the comparison of both high dose and low dose to control. In historical data compiled by this laboratory for the Carcinogenesis Testing
 

Program, 12/250 (5 percent) of the untreated female B6C3F1 mice had
 

one of these tumors, compared to the 2/47 (4 percent) observed in
 

these control females. Based on these results the statistical
 

conclusion is that the administration of 5-nitroacenaphthene was
 

47
 
TABLE 5
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN MALE MICE TREATED WITH 5-NITROACENAPHTHENEa

»e
 

TOPOGRAPHY:MORPHOLOGY
 

Lung: Alveolar/Bronchiolar Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

-P- Lung: Alveolar/Bronchiolar Adenoma or
 00
 

Alveolar/Bronchiolar Carcinoma^
 

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
 

CONTROL
 

5/50(0.10)
 

95
 

5/50(0.10)
 

95
 

5/50(0.10)
 

75
 

LOW
 

DOSE
 

0/31(0.00)
 

N.S.
 

0.000
 

0.000
 

1.258
 

4/45(0.09)
 

N.S.
 

0.889
 

0.187
 

3.868
 

50
 

3/33(0.09)
 

N.S.
 

0.909
 

0.149
 

4.312
 

96
 
TABLE 5 (CONCLUDED)
 

LOW 

TOPOGRAPHY; MORPHOLOGY CONTROL DOSE 

Liver: Hepatocellular Carcinomab 12/50(0.24) 7/31(0.23) 

P Values0 N.S. 

Relative Risk (Control)d 0.941 

Lower Limit 0.348 

Upper Limit 2.273 

Weeks to First Observed Tumor 95 95 

o^ 

Low dose group received a dose of 0.06 percent in feed.
 

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

°The probability level for the Fisher exact test for the comparison of the 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. A negative designation (N) indicates a lower incidence
 

in the treated group than in the control group.
 

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

These analyses were based solely upon animals surviving at least 52 weeks, except for sites
 

where the first tumor of interest was observed earlier than 52 weeks, where the analyses were
 

based upon all animals that survived until or past the date that the first tumor was observed.
 
___ 

TABLE 6
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN FEMALE MICE TREATED WITH 5-NTTROACENAPHTHENE'

a,e
 

TOPOGRAPHY : MORPHOLOGY
 

Lung: Alveolar/Bronchiolar Adenoma or
 

Alveolar/Bronchiolar Carcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 tn
 

o
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma
 

P Values0
 

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.
 

96
 

7/47(0.15)
 

N.S.
 

83
 

2/47(0.04)
 

P < 0.001
 

94
 

LOW
 

DOSE
 

3/47(0.06)
 

N.S.
 

1.468
 

0.176
 

16.917
 

92
 

5/47(0.11)
 

N.S.
 

0.714
 

0.192
 

2.423
 

92
 

23/47(0.49)
 

P < 0.001
 

11.500
 

3.116
 

94.161
 

92
 

HIGH
 

DOSE
 

2/19(0.11)
 

N.S.
 

2.421
 

0.184
 

30.577
 

96
 

4/46(0.09)
 

N.S.
 

0.584
 

0.134
 

2.132
 

25
 

18/19(0.95)
 

P < 0.001
 

22.263
 

7.102
 

44.718
 

87
 
___ 

___ 

TABLE 6 (CONTINUED) 

TOPOGRAPHY: MORPHOLOGY CONTROL
 

Pituitary: Adenoma NOS 5/43(0.12)
 

P Values0 P - 0.028(N)
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 95
 

Ovary: Tubular-Cell Adenoma 0/45(0.00)
 

P Values0 P = 0.001
 

Relative Risk (Control) _——
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ovary: Granulosa-Cell Tumor, Luteoma,
 

or Tubular-Cell Adenoma^ 0/45(0.00)
 

P Values0 P - 0.003
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW
 

DOSE
 

0/39(0.00)
 

P = 0.035(N)
 

0.000
 

0.000
 

0.867
 

2/41(0.05)
 

N.S.
 

Infinite
 

0.326
 

Infinite
 

96
 

4/41(0.10)
 

P = 0.048
 

Infinite
 

1.022
 

Infinite
 

96
 

HIGH
 

DOSE
 

0/18(0.00)
 

N.S.
 

0.000
 

0.000
 

1.803
 

4/17(0.24)
 

P = 0.004
 

Infinite
 

2.497
 

Infinite
 

96
 

7/37(0.19)
 

P = 0.003
 

Infinite
 

2.376
 

Infinite
 

36
 
TABLE 6 (CONCLUDED)
 

Treated groups received time-weighted average doses of 0.05 or 0.12 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.
 

e

These analyses were based solely upon animals surviving at least 52 weeks, except for sites
 

where the first tumor of interest was observed earlier than 52 weeks, where the analyses were
 

based upon all animals that survived until or past the date that the first tumor was observed.
 
associated with the incidence of hepatocellular carcinomas in female
 

B6C3F1 mice.
 

For females the Cochran-Armitage test indicated a significant
 

(P = 0.003) positive association between dose and the combined incidences of granulosa-cell tumors, luteomas, or tubular-cell adenomas
 

of the ovary. The Fisher exact test comparing high dose to control
 

was also significant (P = 0.003) under the Bonferroni criterion.
 

The historical control data compiled by this laboratory indicated
 

that 2/350 (0.57 percent) of the untreated female B6C3F1 mice had
 

these tumors as compared to the absence of these tumors in the control females of this bioassay. Based upon these results the administration of 5-nitroacenaphthene was associated with the increased
 

incidence of ovarian neoplasms.
 

In female mice, the Cochran-Armitage test indicated a significant negative association between dose and the incidence of pituitary
 

adenomas NOS. The Fisher exact tests, however, were not significant
 

when the Bonferroni criterion was applied.
 

In summary, the statistical results indicate that the administration of 5-nitroacenaphthene was associated with the increased incidence of hepatocellular carcinomas and ovarian neoplasms in female
 

B6C3F1 mice under the conditions of this bioassay. No statistical
 

inferences could be made with respect to male mice due to the elevated
 

mortality among high dose males.
 

53
 
V. DISCUSSION
 

In spite of deficiencies in the conduct of the experiment (some
 

controls improperly matched; the maximum tolerated doses exceeded),
 

sufficient evidence was obtained to establish the carcinogenicity of
 

5-nitroacenaphthene.
 

The concentration of 5-nitroacenaphthene in the diet was associated with decreased survival in both species. Toxicity of 5-nitroacenaphthene at the dosages used in this experiment was indicated by
 

accelerated mortality in all dosed groups and by depressed growth
 

among dosed rats of both sexes and dosed male mice. Although mortality among dosed rats may have been related to tumor development, the
 

very high rate of early deaths among male mice could not have been
 

due to cancer since no tumors were found in high dose male mice that
 

died before termination of the study. During week 50, 24/50 (48 percent) high dose female mice died, presumably due to toxicity of the
 

test chemical. Survival in low dose and control mouse groups of both
 

sexes was considered sufficient to provide an adequate number of mice
 

at risk from late-developing tumors. Because of accelerated mortality
 

in both species, statistical analyses were based on those animals surviving at least 52 weeks or until the first tumor at that site was
 

observed in any group of that sex and species.
 

Dietary administration of 5-nitroacenaphthene was related to an
 

increased incidence of several neoplasms in rats. The incidences of
 

54
 
malignant tumors of the ear canal (incidences of ceruminous carcinomas and squamous-cell carcinomas were combined so that the numerator
 

represented rats with either one of these tumors) were statistically
 

significant at each dose level in each sex. Tumors were found as
 

early as week 33. Among female rats, the incidences of clitoral gland
 

carcinoma and the incidences of mammary adenocarcinomas were each significant in both the high dose group and the low dose group. Significant incidences of alveolar/bronchiolar carcinomas were observed in
 

low dose rat groups of each sex, but not in high dose groups. It is
 

possible that high dose rats did not survive long enough to be at risk
 

from these tumors. Hepatocellular carcinomas in four dosed female mice
 

and one control female mouse metastasized to the lung.
 

Ovarian tumors of several histologic types were observed at
 

increased incidences in dosed female mice. Although only tubular-


cell adenomas were significant alone, when incidences of granulosacell tumors, luteomas, or tubular-cell adenomas were combined, there
 

was a significant positive association between dosage level and tumor
 

incidence. The number of high dose female mice having at least one
 

of these tumors was significantly greater than the number of control
 

mice with at least one of these ovarian tumors.
 

There was a significant positive association between dietary concentration of 5-nitroacenaphthene and the incidence of hepatocellular
 

carcinomas in female mice. The incidences of these tumors were statistically significant for each dosed group.
 

55
 
In an earlier study conducted in Tokyo, Japan, 5-nitroacenaphthene was found to be carcinogenic in female Wistar rats following
 

dietary administration of approximately 200 mg/day (1 percent of the
 

diet) for a period of 4 months (Terasawa, 1974; Takemura et al.,
 

1974). Malignancies were observed in all surviving animals and
 

included rhabdomyosarcoma of the lower abdominal wall, sebaceous-cell
 

carcinoma of the ear duct, intraductal carcinoma of the breast, and
 

adenocarcinoma of the small intestine. In a similar experiment using
 

female Syrian golden hamsters, cholangiomas were the only tumors
 

observed in 54 percent of surviving hamsters fed a diet containing 1
 

percent 5-nitroacenaphthene for a period of 6 months (Terasawa, 1974;
 

Takemura et al., 1974). No tumors were observed in male animals of
 

either species. These indications of carcinogenicity in a different
 

strain of rats and in a third rodent species provide supporting evidence for the carcinogenicity of this compound.
 

The numbers of high dose male B6C3F1 mice surviving long enough
 

to be at risk from late-developing tumors did not constitute an adequate test for carcinogenicity; however, survival and dosage among
 

low dose male mice was adequate. On this basis, no evidence was provided by this bioassay for the carcinogenicity of 5-nitroacenaphthene
 

in male B6C3F1 mice.
 

Under the conditions of this bioassay, 5-nitroacenaphthene was
 

carcinogenic to Fischer 344 rats, causing increased incidences of
 

malignant tumors of the ear canal in both sexes, alveolar/bronchiolar
 

56
 
carcinoma in both sexes, cliteral gland carcinoma in females, and
 

mammary adenocarcinomas in females. 5-Nitroacenaphthene was also carcinogenic to female B6C3F1 mice, causing hepatocellular carcinomas and
 

ovarian tumors.
 

57
 
VI. BIBLIOGRAPHY
 

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

J. Wiley & Sons, New York, 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, Volumes 76-85, 1972-1976. American
 

Chemical Society, Washington, B.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.
 

Grasselli, J.G. and W.M. Ritchey, Atlas of Spectral Data and Physical
 

Constants for Organic Compounds, 2nd edition, Volume II. Chemical Rubber Company Press, Cleveland, Ohio, 1975.
 

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

Observations." Journal of the American Statistical Association
 

_53_: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.
 

Reuber, M.D., "Transplantation Studies." Pathology of Mouse Hepatic
 

Neoplasia, W.H. Butler and P.M. Newberne, editors. Elsevier
 

Scientific Publishing Company, Amsterdam, 1975.
 

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

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

Ferric Oxide." Cancer Research 32:1073-1079, 1972.
 

58
 
Takemura, N.S. Hashida, and M. Terasawa "Carcinogenic Action of
 

5-Nitroacenaphthene." Br. J. Cancer 30:481-483, 1974.
 

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

62:679-682, 1975.
 

Terasawa, M., "Carcinogenic Action of 5-Nitroacenaphthene." Tokyo
 

Jikeikai Ika Daigaku Zasshi 89(4), 1974.
 

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

Industries Center. Stanford Research Institute, Menlo Park,
 

California. Personal communication, 1977.
 

Yahagi, T., H. Shimizu, M. Nagao, N. Takemura, and T. Sugimira, "Mutagenicity of 5-Nitroacenaphthene in Salmonella." Gann 66:581-582,
 

1975.
 

59
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH 5-NITROACENAPHTHENE
 

TABLE A1

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH 5-NITROACENAPHTHENE 

LOW DOSE CCNTPOL(UNTR ) 

HIGH DOSE CONTROL(UNTE ) L OW DOSE HIG H DOSE 

01-0070 01-0118 01­0065 01-0107 

ANIMALS INITIALLY IN STUDY 50 350 50 50 

ANIMALS HISSING 1 

ANIMALS NECROPSIED 48 48 43 47 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 1(8 48 43 47 

INTEGUMENTAR Y SYSTEM 

*SKIN (48) (48) (43) (47)
 

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

SQUAMOUS CELL CARCINOMA 1 (2%) 1 (2%)
 

3ASAL-CELL TUMOR 1 (2%)
 BASAL-CELL CARCINOMA 1 (2%)
 

SEBACEOUS ADENOCAP.CINOMA 1 (2X)
 

FIBROMA 4 (9%)
 

*SUBCUT TISSUE (48) (48) (43) (47)
 

SARCOMA, NOS 1 (2%)
 

FIBROMA 2 (4*) 3 (6%) 2 (5X) 1 (2X)
 

FI3ROSARCOMA 1 (2%)
 

LSIOMYOSARCOMA 1 (2%)
 

PESPISATOR Y SYSTEM 

SLUSG (48) (48) (41) (47)
 

CARCINOMA, NOS, METASTATIC 1 (2%)
 

3QOAHODS CELL CARCINOMA 1 (2%)
 

SQUAMOUS CELL CARCINOMA, METASTA 2 (5%) 1 (2%)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 2 (554) 3 (6%)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 1 (2X) 5 (12X)
 

CSRUMINOUS CARCINOMA, METASTATIC 1 (2%)
 

PHEOCHROMOCYTOMA, SETASTATIC 1 (2%)
 

OSTEOSAHCOMA, METASTATIC 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

*MULTIPL S ORGAN S (48) (Ha) (43) (47) 

MALIGNAN T LYMPHOKA , NOS 1 (2%) 

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

____ MYELCMONOCYTIC_I.EUKEMI A ______________ 5_j.10%l. _________ i*_lSJ] ___________ 2__{5X) _________________ 

* NUMBE R O F ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

a 50 ANIBALS WERE INITIALLY IN STUDY BUT ONE HAS DELETED WHEN FOUND TO BE A FEMALE ANIMA L IN A
 

MALE GROUP.
 

A-3
 
TABLE Al (CONTINUED) 

. LOW DOSE CONTROL (BNTH>. 01-0070 

HIGH DOSE 

CONTROL (UNTR) 

01.-0118 

LOW DOSE 

01-0065 

_ 

HIGH DOSE 

01-0107 

•SPLEEN 

OSTEOSARCOMA, SETASTATIC 

(«8) 

1 (2X) 

(48) (38) (47) 

»B£DIASTINAL L. NODii 

ALVEOLAR/BRONCHIOLAR CA, HETASTA 

(42) (44) (23) 

1 <»*) 

(16) 

CIRCULATORY SYSTEM 

*PUL(10NiRY ARTERY 

SARCOMA, NOS 

C»8) (48) (43) 

1 (2X) 

(»7) 

DIGESTIVE SYSTEM 

•SALIVAR Y BLAN D 

ADENOCARCINOHA,

SARCOMA, NOS 

 NOS 

(46) (47) 

1 (2X) 

1 (2X) 

(38) (44) 

•SUBNAXILLAR Y GLAN D 

SARCOMA, NOS 

(46) (47) (38)

1 (3X) 

(44) 

• LIViiR 

NSOPLASTIC NODUL£ 

HEPATOC2LLULAR CARCINOM A 

(48) 

1 (2X) 

2 (4X) 

(48) 

1 (2X) 

(40) 

1 (3X) 

(46) 

• ILEUli 

SARCOMA, NOS 

(45) (46) 

1 (2X) 

(34) (39) 

•COLON 

ADJ3NOCARCINOMA, NOS 

(44) (46) (32) (38) 

1 (3X) 

URINAR Y SYSTEM 

•KIDNS Y 

TUBULAR-CELL

TUBULAR-CELL

SARCOMA , NOS 

 ADBNOM A 

 ADENOCAHCINOM A 

(48) (48) (43) 

3 (7X) 

1 (2X) 

(46) 

1 (2X) 

•U5INAE Y BLADDER 

TRANSITIONAL-CEL L PAPILLOM A 

(46) 

1 (2S) 

(43) (37) (45) 

ENDOCRINE SYST2M 

• PITUITARY

Ap.EN2S4x_NQS

 (41)

 1_A2%1

 (38) 

9_J24X)._ 

(31) (33) 

* NU1BER

* NU.1BSR

 OF

 OF

 ANIMAL S

 ANI.1ALS

 HITH TISSUE

 NSCEOPSIID 

 EXAMINE D MICROSCOPICALLY 

A-4 
TABLE A1 (CONTINUED)
 

LOW DOSE CONTROL (UNTR) 

01-0070 

HIGH DOSE CONTROL (UNTR) 

01-0118 

EPENDIMOMA 

tADRENAL 

CORTICAL ADENOMA 

PHEOCHROMOCYTOMA 

PHEOCHROflOCYTOMA, MALIGNANT 

GANGLIONEUROMA 

(147) 

1

10

1

 (2%) 

 (21%) 

 (2%) 

(47) 

7

1

 (15%) 

 (2%) 

*THYROID 

PAPILLARY ADENOMA 

PAPILLARY ADENOCABCINOHA 

FOL1ICULAR-CELL ADENOMA 

FOLLICULAR-CELL CARCINOMA 

C-CELL CARCINOMA 

(39) (48) 

1 (2%) 

#PARATHYBOID 

ADENOMA, NOS 

(23) (28) 

1 (4%) 

tPANCREATIC ISLETS 

ISLET-CELL ADSNOMA 

(45) 

3 (7%) 

(46) 

REPRODUCTIVE SYSTEM
 

*MAHMARY GLAND (U8) (48)
 

PAPILLARY ADENOCASCINOMA 1 (2S)
 

FIBROADENOMA 1 (2%)
 

*PRfiPUTIAL GLAND (48) (48)
 

CARCINOMA, NOS 2 (4%)
 

#TESTIS (47) (47)
 

INTERSTITIAL-CELL TUMOR 45 (96%) 42 (89%)
 

NERVOUS SYSTEM
 

#BPAIN (47) (48)
 

GLIOMA, NOS 1 (2%)
 

SPECIAL SENSE ORGANS
 

*EYS/LJCRIMAL GLAND (48) (48)
 

ADENOMA, NOS
 

•EAR	 CANAL (48) (48)
 

SQUAMOUS CELL CARCINOMA
 

CSRUMIJIOUS CARCINOMA
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-5
 

LOW DOSE
 

01-0065
 

1 (3%)
 

(37)
 

2 (5«)
 

(37)
 

2 (5%)
 

2 (5%)
 

2 (5%)
 

2 (5%)
 

(22)
 

1 (5»)
 

(35)
 

(43)
 

(43)
 

3 (7%)
 

(37)
 

19 (51%)
 

(37)
 

(43)
 

1 (2%)
 

(43)
 

7 (16 JS)
 

14 133%1
 

HIGH DOSE
 

01-0107
 

(42)
 

(44)
 

1 (2«)
 

(22)
 

2 (9%)
 

(42)
 

1 (2%)
 

(47)
 

(47)
 

(44)
 

2 (5%)
 

(44)
 

(47)
 

(47)
 

2 (4X)
 

18 _[38%)_
 
TABLE Al (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTROL (UNTR ) CONTROL(UNTR ) L OW DOSE HIGH DOSE 

01-0070 01-011 8 01-0065 01-0107 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*BOOY CAVITIES (US ) (t8) (U3 ) (17)

 

MESOTHELI0.1A, NOS 1 (2X) 2 (H%) 

llESOTHELIOMA , MALIGNAN T 2 (14%) I* (9X)

 

ALL OTHER SYSTEMS
 

DIAPHRAGM
 

ADENOCAHCINOMA, NOS
 

ANIMA L DISPOSITION SUMMAR Y
 

ANIMALS INITIALLY IN STUDY 50 50 50 50
 

NATURAL DEATHS 5 6 23 19
 

SORIBUND SACRIFICE 5 8 27 31
 

SCHEDULED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 31 30
 

ANIMAL MISSING 1
 

ANIMA L DELETED/WRONG SEX 1
 

JL!MCLUDES_AUTQLYZED_ANI!1ALS _____________________________________________________________________ _ 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE R OF ANIMALS NECROPSIED
 

A-6 
_
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PHIMARY TUHOES
 

TOTAL ANIMALS WITH BENIGN TUM08S
 

TOTAL EENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORS*
 TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT


TOTAL UNCERTAIN TUMORS


TABLE Al (CONCLUDED)
 

LOW DOSE
 CONTROL (UNTR)
 

01-0070
 

K5
 

81
 

45
 

66
 

10
 

13
 

2
 3


 2
 

2
 

HIGH DOSE
 CONTROL (UNTR)
 

01-0118
 

It
 

80
 

i«3
 

62
 

17
 

18
 

1
 1


LOW DOSE HIGH DOSE
 

01-0065 01-0107
 

39 30
 

87 36
 

25 7
 

UO 9
 

31 25
 U7 25
 

3	 .
 

3
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR M2TASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

A-7
 
TABLE A2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH 5-NITROACENAPHTHENE 

cmmwn, 02-0070 SffiMSfcuiTH] 

02-0118 

! LOW DOSE 

02-0065 

HIGH DOSE 

02-0107 

ANIMAL S INITIALLY IN STUDY 50 50 50 50 ANIMAL S NECROPSIES 4 9 50 49 48 ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* * 4 9 50 43 48 

INTI3UMENTAE Y SYSTEM 

*SKIN (49) (50) (49) (48)
 

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

SQUAMOUS CELL CARCINOMA 1 (2X)
 

BASAI-C2LL CARCINOMA 1 (2%)
 

SEBACEOUS ADENOMA 1 (2X)
 

SIBACSOUS ADENOCAHCINOtfA 2 (4X)
 

*SUBCUT TISSUE (49) (50) (49) (48)
 SEBACEOUS ADSNOMA 1 (2X)
 FIBROMA 1 (2%) 3 (6X) 2 (4X)
 FI3BOSARCOMA 1 (2X)
 

RESPIRATORY SYSTEM
 

*LUNG (49) (50) (48) (48)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (2)8) 1 (2%) 1 (2X)
 

ALVEOLAR/BSONCHIOLAR ADENOMA 1 (2%) 4 (8X) 2 (4X)
 

ALVEOLAH/BRONCHIOLAR CARCINOMA 4 (8X) 1 (2X)
 

HEHATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS (49) (50) (49) (48)
 

MALIGNANT LYMPHOHA, NOS 2 (4%)
 

'^DIFFERENTIATED LEUKEMIA 1 (2%)
 

lYELCaONOCYTIC LEUKZMIA 5 (10X) 3 ((,%)
 

*SPL2EN (48) (48) (48) (47)
 

UNDIFFERENTIATED LEUKEMIA 1 (2%)
 

• LIVER (49) (50) (48) (47)
 

UNCIFFERENTIATED LEUKEMIA 1 (2%)
 

CIRCULATORY SYSTEM
 

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

* NUMBE R OF ANIMALS NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-8 
TABLE A2 (CONTINUED)
 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) CONTROL (UNTR)


02-0070 02-0118
 

DIGESTIVE SYSTEM
 

»MOUTH (49) (50)
 

SQUAMOUS CELL PAPILLOMA
 

#PAROTID GLAND (49) (50)
 ADENOMA, NOS
 

#LIVER (49) (50)
 NEOPLASTIC NODULE 2 (4X)
 

HEPATOCELLULAR CARCINOMA 1 (2X)
 

IILSUM (49) (48)
 

LEIOMYOSARCOMA 1 (2X)
 

URINARY SYSTEM
 

IKIDN2Y (49) (50)
 CARCINOMA, NOS 1 (2X)
 TUBULAR-CELL ADENOMA
 

SARCOMA, NOS
 

ENDOCRINE SYSTEM 

*PITUITARY 

ADENOMA, NOS 

CHROMOPHOBE ADENOMA 

(44) 

18 (41X) 

(40) 

17 (43%) 

•ADRENAL 

CORTICAL ADENOMA 

CORTICAL CARCINOMA 

PHEOCHROMOCYTOMA 

(49) 

1 (2%) 

2 (45) 

(49) 

1 (2X) 

3 (6X) 

*ADR£NAL MEDULLA 

GANGLIONJJUROMA 

(49) (49) 

1 (2*) 

•THYROID 

FOLLICULAR-CELL ADcNOMA 

FOLLICULAR-CELL CARCINOMA 

C-CELL ADENOMA 

C-CELL CARCINOMA 

(40) 

1 (3X) 

2 (5X) 

1 (3») 

(45) 

1 (2X) 

1 <2X) 

1 (2X) 

*PANCREATIC ISLETS 

J3LETz£SLL_4ESNOM4

(47) 

 1_I2X.)

(48) 

 2_i4X) 

* NUMBEP OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-9
 

 LOW DOSE
 

02-0065
 

(49)
 

1 (2%)
 

(45)
 

1 (2K)
 

(48)
 

(48)
 

(48)
 

(44)
 

4 (9X)
 

1 <2X)
 

(47)
 

1 (2X)
 

(47)
 

(45)
 

1 (2*)
 3 (7*)
 

(47)
 

HIGH DOSE
 

02-0107
 

(48)
 

(44)
 

(47)
 

(47)
 

(48)
 

1 (2X)
 

1 <2X)
 

(43)
 

(48)
 

1 (2X)
 

(48)
 

(40)
 

1 (3X)
 

2 (5X)
 

(47)
 
TABLE A2 (CONTINUED)
 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) CONTROL (UNTR)
 

02-0070 02-0118
 

i-EPROCUCTlVE SYSTEM
 

• MAMMAR Y	 GLAND . (49) (50)
 ADENOMA, NOS 2 (4S)
 

ADENOCARCINOHA, NOS
 

PAPILLARY CYSTADENOCAHCINOMA.NOS
 

FIBFOADENOMA 16 (33%) 19 (38X)
 

*CLITORAL GLAND (49) (50)
 CARCINOMA, NOS
 

SgUAMOUS CELL PAPILL0.1A 1 (2X)
 SIUAHOUS CELL CARCINOMA
 

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

IUTERUS (49) (50)
 

ADENOCARCINOHA, NOS 1 (2X)
 LSIOMYOSARCOMA 1 (2%)
 

ENDOMETRIAL STHOHAL POLYP 12 (24X) 10 (20%)
 rNDO«2IElAL STROBAL SARCOMA 1 (2X)
 

*UTERUS/£,NDOM2TRI(Jt! (49) (50)
 AD2SOCARCINOMA, NOS 2 (4*)
 

40VAHY (47) (49)
 

3RANrjLOSA-CSLL TUMOR 1 (2%)
 

MJRVOU S SYSTEM
 

•BRAIN	 (49) (50)
 

OLIGODiiNDROGLIOHA 1 (2X)
 

SPTCIAL SENSE ORGANS
 

*TA<! CANAL (49) (50)
 S3UAMOUS CELL CARCINOMA
 

CiPUfllNOUS CARCINOHA
 

CARCINOSAECOMA
 

tiUSCULOSKELSTAL SYSTEM
 

« NU132R OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* N'J:',B5R OF ANIMAL S NECROPS1ED
 

A-10
 

LOW DOSE
 

02-0065
 

(49)
 

5 (10X)
 

1 (2X)
 

12 (24X)
 

(49)
 

6 (12X)
 

1 (2X)
 

(47)
 

1 (2X)
 

(47)
 

(45)
 

(48)
 

(49)


3 (6X)
 

25	 (51*)
 

1 (2X)
 

HIGH DOSE
 

02-0107
 

(48)
 

5 (10X)
 

4 (8X)
 

(48)
 

5 (10X)
 

1 (2X)
 

(42)
 

1 (2X)
 

(42)
 

(45)
 

(47)
 

(48)
 

9 (19*)
 

26 (54X)
 
BODY CAVITIES
 

•PERITONEUM
 

MESOTHELIOHA, NOS
 

ALL OTHER SYSTEMS
 

SITE UNKNOWN
 

SO.UAMOUS CELL CARCINOMA
 

ANIMAL DISPOSITION SUMMARY
 

ANIUALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDUL3D SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

* INCLUDES AUTOLYZYD ANIMALS
 

TUMOR SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 

TOTAL B3NIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORS!


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT


TOTAL UNCERTAIN TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

TABLE A2 (CONCLUDED) 

LOW DOSE CONTROL (UNTR ) 

02-0070 

1 (2)5)
 

50
 

3
 

7
 

5
 

35
 

U5
 

73
 

37
 

5U
 

13
 

16
 

3


 3


HIGH DOSE CONTROL(UNTR )

02-0118

(50)
 

50
 

5
 

3
 

5
 

37
 

38
 

73
 

35
 

59
 

12
 

13
 

1


 1


 1
 

1
 

 LOU DOS 2 HIGH DOSE 

 02-0065 02-0107 

(U9) (UB) 

50 50 

9 11 

U1 39 

1*6 U5 

81 67 

214 10 

32 13 

39 414 

U9 5U 

1 

1 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

A-ll
 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH 5-NITROACENAPHTHENE
 

TABLE Bl
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH 5-NITROACENAFHTHENE
 

CONTROL(UMTH) LOW DOSE

05-0070 05-006U

 HIGH DOSE 

 05-0065 

ANIMALS INITIALLY IN STUDY 50

ANIMALS NECROPSIED 50

ANIMALS EXAMINED HI3TOPATHOLOGICALLY ** 50

 50

 19

 17

 50 

 15 

 15 

INTEGUMENTARY SYSTEM
 

NON!
 

RESPIRATORY SYSTEM
 

• LUNG (50) (16) (11)
 

'^DIFFERENTIATED CARCINOMA NETAS 1 (2*)
 HEPATOCELLULAR CAECINOMA, METAST 1 (2%)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 1 (9%)
 

ALVEOLAE/BHONCHIOLAR CARCINOMA 5 (10X)
 

HEBATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS (50) (19) (15)
 MALIGNANT LYMPHOMA, NOS 2 <"»%) 2 (IX)
 SALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 <2X) 1 (2X)
 

tSPLEEN (50) (12) (U1)
 HEHANGIOMA 1 (2X)
 

•LYMPH	 NODE (15) (31) (30)
 

MALIC. LYMPHOMA. HISTIOCYTIC TYPE 2 (1%)
 

*PEYERS PATCH (19) (11) (11)
 

MALIGNANT LYHPHOMA. MIXED TYPE 1 (2X)
 

CIRCULATORY SYSTEM
 

NOME
 

DIGESTIVE SYSTEM 

*LIVSR (50) (U6) (11)
 

HEPATOCELLULAR CARCINOMA 12 (21%) 7 (15X)
 

f NU1BES OF ANIMALS KITH TISSUE EXAMINE D MICROSCOPICALLY 

« NUMBER OF ANIMALS NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3
 
URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

• THYROID


^DENOCARCINOMA, NOS


• PANCREATIC ISLETS


ISLET-CELL ADENOMA


REPRODUCTIVE SYSTEM
 

*MAHMARY GLAND


^DIFFERENTIATED CARCINOMA


NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

»HARDZRIAN GLAND


ADENCMA, NOS


MUSCULOSKELETAL SYSTEM
 

NIN E
 

BODY CAVITIES
 

NONE
 

ALL	 OTHER SYSTEMS
 

'MULTIPLE ORGANS


• NUMBE R OF ANIMAL S WIT H TISSUE

« NUMBE R OF ANIMAL S N2CROPSIED 

TABLE Bl (CONTINUED) 

CONTROL (UNTR ) LOW DOSE HIGH DOSE 

05-0070 05-0061 05-0065 

 (40)	 (31) (13)
 

1 (3*)
 

 (16)	 (39) (38)
 

1 (2%)
 

 (50)	 (19) (15)
 

1 (2%)
 

 (50)	 (19) (15)
 

1 (2S)
 

 (50) («9) (15)
 

 EXAMINE D MICROSCOPICALL Y 

B-4
 
TABLE Bl (CONCLUDED)
 

ANIMA L DISPOSITION SUMMAR Y
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE
 ANIMAL i-ilSSING
 

3 INCLUDES AUTOLYZilD ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORS*


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY 08 HETASTATIC
 

TOTAL UNCERTAIN TUMORS
 

CONTROL (UNTR)
 

05-0070
 

50
 

2
 

5
 

HJ
 

23
 

27
 

3
 

3
 

22
 

24
 

1


 1


LOW DOSE HIGH DOSE 

05-006U 05-0065 

50 50 

16 19 6 28 

28 3 

15 1 

16 1 

5 

5 

10 1 

11 1 

1 

1 

* PPIMAF.Y TUMOHS: ALL TUMORS EXCEPT SECONDARY TUMORS
 * SECJNDARY TUMORS: M2TA3TATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-5
 
TABLE B2
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH 5-NITROACENAPHTHENE
 

CONTROL (UNTR) LOU DOSE

06-0070 06-0061

 HIGH DOSE 

 06-0065 

ANIMALS INITIALLY IN STUDY 50

ANIMALS HISSING

ANIMALS NECROPSIED U8

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 17

 50

 1

 17

 17

 50 

1 

 16 

 16 

INTEGUMENTARY SYSTEM
 

• SUBCUT TISSUE (18) (U7) («6)
 

HEMANGIOSARCOMA 1 (2X)
 

RESPIRATORY SYSTEM
 

*LUNG (16) (17) (46)
 HBPATOCELLULAR CARCINOMA, METAST 1 <2X) 3 (6X) 1 (2X)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 2 (IX) 2 (IX)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 2 (IX) 1 <2X)
 OSTEOSARCOMA, METASTATIC 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS (18) (17) (16)
 

MALIGNANT LYMPHOMA. NOS 2 (IX) 3 (6X) <» (9X)
 

LYNPHOCYTIC LEUKEMIA 1 (2X)
 

SHYTHROCYTIC LEUKEMIA 1 (2X)
 

ISPLEEN (17) (11) (U5)
 

HEMANGIOSARCOMA 1 (2X)
 

MALIGNANT LYMPHOMA. NOS 1 (2X)
 

*HILAR LYMPH NODE (36) (38) (35)
 

HEPATOCELLULAR CARCINOMA, MEIAST 1 (JX)
 

tMESENTERIC L. NODji (36) (38) (35)
 

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

IPEYERS PATCH (15) (12) (39)
 

MALIGNANT LYMPHOMA, NOS 1 (2X)
 

*THYMUS (27) (21) (18)
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

« NUMBER OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-6
 
TABLE B2 (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE 

06-0070 06-0060 06-0065 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

*LIVSR (17) (17) (145)
 

HSPATOC3LLULAR CARCINOMA 2 (<tX) 23 (U9X) 18 (10X)
 

tSTOMACH («5) (t"4) (Q3)
 

SiJUAMOUS CELL CARCINOMA 1 (2X)
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

fPITUITARY (13) (39) (39)
 

ADENOMA, NOS 5 (12%)
 

IA3RSNAL (17) (13) (HO)
 

ADENOCARCINOMA, NOS 1 (2X)
 

CORTICAL ADENOMA 1 (2X)
 

PHEOCHROMOCYTOSA 1 (3*)
 

REPRODUCTIVE SYSTEM
 

•UTERUS	 (13) (39) (12)
 

LEIOBYOMA 1 (2X)
 

IUTERUS/ENDOMETRIUM (13) (39) (12)
 

HEMANGIOMA 1 (2X)
 

HOVASY/OVIDUCT (13) (39) (12)
 

PAPILLARY ADENOMA 1 (2X)
 

KOV^RY (HI) (39)
 

LUTEOMA 1 (3X)
 

•3P.ANULOSA-CELL TOMOR 2 (5X) 2 (5X)
 

TUBULAR ADENOMA 2 (5%) 1 (10X)
 

NERVOUS SYSTEM
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

B-7
 
SPECIAL SENSE OBGANS
 

*HARDERIAN GLAND


ADENOMA, NOS


MUSCULOSKELETAL SYSTEM
 

HONE
 

BODY CAVITIES
 

NONE
 

ALL OTHE3 SYSTEMS
 

OM3NTUM
 

HSMANGIOSAECOMA


ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY


NATURAL DEATHS


MORIBUND SACRIFICE


SCHEDULED SACRIFICE


ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE


ANIMAL MISSIN3


a_INCiUDES_AUTOLY2ED_ANIMALS
 

TABLE B2 (CONTINUED) 

CONTROL(UNTR) LOB DOSE HIGH DOSE 

06-0070 06-006M 06-0065 

 (U8) (U7) (U6) 

1 (2%) 1 (2%) 

1 

 50 50 50 

6 9 21 

3 2 10 

5 

 36 38 18 

1 1 

HUMB3R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

NUMBER OF ANIMALS N2CROPSIED
 

B-8
 
TABLE B2 (CONCLUDED)
 

TUMOB SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS«
 TOTAL PRIMARY TUMORS


TOTAL ANIiiALS WITH BENIGN TUMORS


TOTAL BENIGN TUMORS


TOTAL ANIMALS WITH MALIGNANT TUMORS


TOTAL MALIGNANT TUMORS


TOTAL ANIMALS WITH SECONDARY TUMORS*


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT


TOTAL UNCERTAIN TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINPHIMAHY OR METASTATIC
 TOTAL UNCERTAIN TUHORS
 

CONTROL (UNTR)


06-0070


 18
 22


 8


 9
 

 12


 13


 2


 2
 

 LOW DOSE HIGH DOSE
 

 06-006« 06-0065
 

 29 21
 39 33
 

« 8
 

5 9
 

 29 21
 

 32 22
 

3 1
 

3 2
 

2 2
 

2 2
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

B-9
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS TREATED WITH 5-NITROACENAPHTHENE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH 5-NITROACENAPHTHENE

 

LOW DOSE CONTROL (UNTR) 

HIGH DOSE CONTROL (DNTR) LOB DOSE 

01-0070 01-0118 01­0065 

ANIMAL S INITIALLY IK STODY 50 350 50 

ANIMAL S MISSING 1 

ANIMALS NECHOPSIED us 148 U3 

ANIMAL S EXAMINED HISTOPATHOLOGICALLY** us 43 < 43 

INTEGOMENTAR Y SYSTEM 

•SUBCUT TISSUE (18) (<I8) (U3)
 

INFLAMMATION, ACUTE
 

ABSCESS, NOS
 

NECSOSIS, NOS
 

METAPLASIA, OSSEOUS 1 (2%)
 

RESPIRATORY SYSTEM
 

tTRACHE A (145) (US) (35) 

INFLAMMATION, NOS 2 

INFLAMMATION, ACUTE/CHBONIC 18 (<40%) 3 (9X) 

INFLAMMATION, CHRONIC 3 (9X) 

tLUNG/BBONCHUS (148) (148) (141) 

EHONCHIECTASIS 3 (6X) 1 (2%) 1 (2X) 

INFLAMMATION, NOS 7 (15X) 

ABSCESS, NOS 

*LUNG/BRONCHIOLE (148) (148) (141) HYPERPLASIA, EPITHELIAL 1 (2X) 

*LUNG (148) (141) CONGESTION, NOS 1 <2X) 7 (17X) EDEMA, NOS 2 (5X) HEMORRHAGE 2 (5X) INFLAMMATION, NOS 2 (5X) INFLAMMATION, FOCAL 2 <<tX) 

INFLAMMATION, INTERSTITIAL 14 (8X) 1 (2X) INFLAMMATION, NECEOTIZING 1 (2X) ABSCESS, HOS 1 (2X) PNEUMONIA, CHRONIC HURINE 1 (2X) INFLAMMATION, CHRONIC NECROTIZIN 1 (2X) 

I NUMBER OF ANIMALS WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMALS NECP.OPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

» 50 ANIMALS HERE INITIALLY IN STUDY BUT ONE HAS DELETED WHEN FOUND TO BE A FEMALE

GROUP. 

C-3
 

HIGH DOSE 

01-0107 

50 

U7 

147 

(147)
 

1 <2X)
 

2 (IX)
 

1 (2X)
 

(147) 

3 (6X) 

(U7 ) 

1 (2X) 

(47) 

(147) 

1 (2X) 

 ANIMA L IN A MALE 
TABLE Cl (CONTINUED)
 

LOW DOSE CONTROL(UNTH) 

01-0070 

HIGH DOSE CONTROL(UNTR) 

01-0118 

PERIVASCULITIS 

HYPERPLASIA. EPITHELIAL 

HYPEHPLASIA, FOCAL 

HYPERPLASIA, ALVEOLAR EPITHELIUM 

1 (2X) 1 (2X) 

1 (2X) 

*LUNG/ALVEOLI 

MINERALIZATION 

CALCIFICATION. NOS 

HEMATOPOIF.TIC SYSTEM
 

*BON! HARROW
 

HYPERPLASIA, NOS
 

MYELOFIBROSIS 1 <2X)
 

HYPERPLASIA, HEMATOPOIETIC 1 <2X)
 

HYPERPLASIA. GRANULOCYTIC 1 (2X)
 

HYPERPLASIA, MEGAKARYOCYTIC 1 (2X)
 

«SPLEEN (148) (18)
 

FIBROSIS (2X)
 

NECROSIS, DIFFUSE
 

HEMOSIDEROSIS 1 (2X)
 

ATROPHY, NOS
 

LYMPHOID DEPLETION
 

HYPERPLASIA. HEMATOPOIETIC 9 (19X)
 

HYPERPLASIA, ERYTHROID 10 (21X)
 

EBYTHROPOIESIS
 

ULYNFH NODE
 

HEMORRHAGE 1 (2X)
 

ATBOPHY, NOS
 

PLASMACYTOSIS 1 (2X)
 

HYPE8PLASIA, LYMPHOID 3 (7X)
 

tMANDIBULAR L. NODE
 

DILATATION, NOS 1 (2X)
 

HYPEHPLASIA. NOS 1 (2X)
 

HYPERPLASIA, PLASMA CELL
 

CIRCOIATORY SYSTEM
 

IHSART
 

MINERALIZATION
 

I NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

« NUMBER OF ANIMALS NECROPSIED
 

C-4
 

LOU DOSE 

01-0065 

(2X)
 

(HI)
 

' (2X)
 

1 (2X)
 

(39)
 

1 (3X)
 

(38)
 

1 (3X)
 

1 (3X)
 

1 (3X)
 

(23)
 

2 (9X)
 

(23)
 

1 (4X)
 

(U1)
 

a (iox)
 

HIGH DOSE
 

01-0107
 

1 (2X)
 

C»3)
 

1 (2X)
 

10 (21X)
 

7 (15X)
 

1 (2X)
 

(16)
 

(16)
 
FIBEGSIS, DIFFUSE
 

NECROSIS. NOS
 

tMYOCARDIUM
 

MINERALIZATION
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, ACUTE/CHRONIC
 

FIBROSIS
 

FIBROSIS, FOCAL
 

FIBROSIS, DIFFUSE
 

DEGENERATION, NOS
 

NECROSIS, NOS
 

NECROSIS, FOCAL
 

ICARDIAC VALVE
 

INFLAMMATION, ACUTE/CHRONIC
 

*BLOOD VESSEL
 

MINERALIZATION
 

MEDIAL CALCIFICATION
 

*ARTEHY
 

MINERALIZATION
 

*AORTA
 

3EDIAL CALCIFICATION
 

*CORONARY ARTBRY
 

PERIVASCULITIS
 

*PULMONARY ARTERY
 

MINERALIZATION
 

*PASCREATIC ARTERY,
 

MINERALIZATION
 

ARTERIOSCLEROSIS, NOS
 

HYPERTROPHY, NOS
 

HYPERPLASIA, NOS
 

*RENAL ARTEHY
 

INFLAMMATION PROLIFEHATIVE
 

NSCROSIS, NOS
 

*TESTICULAR ARTERY
 

DEGENERATION^ EOSIJOPHILIC
 

*	 NUMBER OF ANIMALS HITH TISSUE
 

*	 NUMBER OF ANIMALS NECROPSIED
 

TABLE Cl (CONTINUED) 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) CONTROL (UNTR)
 

01-0070 01-0118
 

1	 (2X)
 

(IB) (18) 

2 (»X) 

3 (6X) 

2 (UX) 

23

12

 (U8X) 

 (25%) 

1 (2X) 

(18)	 (18)
 

1 (2%)
 

(18) (US)
 

(48) (18)
 

(18) (18)
 

(18)	 (18)
 

1 <2X)
 

(18)	 (18)
 

11 (23%)
 

(48) (18)
 

(18) (18)
 

(18) (18)
 

EXAMINED MICROSCOPICALLY
 

C-5
 

LOW DOSE
 

0 1-0065
 

1 (10X)
 

(11)
 

1 (2X)
 

1 <2X)
 

1 (2X)
 

1 <2X)
 

1 (2X)
 

5 (12X)
 

1 (2X)
 1	 (2%)
 

(11)
 

(13)
 

1 (2X)
 

1 [2X)
 

(13)
 

2 (5X)
 

(13)
 

1 (2%)
 

(13)
 

(13)
 

5 (12X)
 

(13)
 

1 <2X)
 

1 (2X)
 

1 (2S)
 

1 (2X)
 

(13)
 

1 (2X)
 

1 (2X)
 

(13)
 

1 12X1
 

HIGH DOSE
 

01-0107
 

(17)
 

(17)
 

(17)
 

(17)
 

(17)
 

(17)
 

(17)
 

(17)
 

(17)
 

(47)
 
TABLE Cl (CONTINUFD) 

LOW DCSE CONTRJL (UNTK) 

01-0070 

HIGH DOSE CONTROL (UNTR) 

01-0118 

LOW DOSE 

0 ,-0065 

HIGH DC "E 

01-0107 

DIGESTIVE 'YSTEH 

•SALIVARY GLAND 

ATYPIA, NOT 

IIYPERPLASIA, PLAShA CELL 

(16) (' 7) (38) 

1 (3%) 

(11) 

1 (2%) 

•LIVER T

 FLAMMATION, NECROTIZING 

r'IBSrSIS 

FIBPOSIS SEPTAI LIVER 

NECROSIS, NOS 

NECROSIS, FOCAL 

' JSTMORTEH CHANGE 

.1ETAJ10KPH03IS FATTY 

BASOPHILIC ~YTO CHANGE 

HYPERPIASI , FOCAL 

ANGIECTASIS 

ESYTHROPCIESIS 

(18) 

1 (2%) 

8 (17%) 

4 (8*) 

3 (17%) 

2 (4%) 

1 (2%) 

(18) 

2

2

15

1

 (1%) 

 (1%) 

 ,31%) 

 :2%) 

(10) 

1 (3%) 

1 (3%; 

1 (ji) 

2 (5%) 

(5?, 2 (5%, 

(16) 

1 (2%) 

*LIV£R/CENTRILOBULAT( 

DiGENERATION, EC TNOPHILIC 

NECROSIS, NOS 

(;8) 

2 ,4%) 

(18) 

1 (2-,) 

(10) ('16) 

•LIVER/HEPATOCYTES 

NECROSIS, NOS 

(48) (18) (40) 

1 (3%) 

(46) 

*BILE DUCT 

INFLAMMATIOH, NOS 

HYPERPLASIA, NOS 

(48) 

5 (13%) 

(18) 

3

13

 (6%) 

 (90%) 

(1") (17) 

•PANCREAS 

THROMBOSIS, NOS 

INFLAMMATION, NOS 

INFLAMMATION, ACUTE/CHROKIC 

INFLAMMATION, CHRONIC 

PERIARTERITIS 

ATROPHY, FOCAL 

(15) 

6 (13%) 

1 (2%) 

1 (2%) 

(16) 

17 (37%) 

(35) 

1 (3%) 

1 (3%) 

3 (9%) 

(1.) 

•PANCREATIC ACINUS 

ATROPHY, NOS HYPEHELASIA, FOCAL 

US) (16) 

1 (2%) 

(35) 

1 (3%) 

(42) 

•ESOPHAGUS 

MINERALIZATION 

DYSPLASIA^ HOS _ . 

(45) (15) 

1 -L2S1-

(36' 

1 ,3!:; 

(17) 

t NUM3EE OF ANIMALS WITH TISSUE EXAMINED MICROSCCFINALLY 

« NUMBER Or ANIMALS NECfoPSIED 

-C-6 
#ESOPHAGEAL MUSCULARI
 

MINERALIZATION
 

•STOMACH
 

MINERALIZATION
 

EPIDERMAL INCLUSION CYST
 

INFLAMMATION, NOS
 

INFLAMMATION, ACUTE/CHRONIC
 

CALCIFICATION, NOS
 

HYPERPLASIA, BASAL CELL
 

HYPERKERATOSIS
 

ACANTHOSIS
 

tGASTRIC MUCOSA
 

CALCIFICATION, NOS
 

CALCIFICATION, FOCAL
 

#PEYERS PATCH
 

HYPERPLASIA, N'jS
 

HYPERPLASIA, '.(ETICULUM CELL
 

#ILEUM
 

INFLAMMATION, NOS
 

HYPERPLASIA, LYMPHOID
 

tCOLON
 

NEMATODIASIS
 

PARASITISM
 

URINAR Y SYSTEM
 

tKIDNEY
 

MINERALIZATION
 

HYDRONEPHROSIS
 

CYST, NOS
 

GLOMEROLONEPHRITIS, NOS
 

INFLAMMATION, ACUTE/CHRONIC
 

SLOMERULONEPHHITIS, SUBACUTE
 

INFLAMMATION, CHRONIC
 

FIBROSIS, DIFFUSE
 

PERIAHTERITIS
 

NEPHROSIS, NOS
 

IKIDNEY/CORTEX
 

SISTj. HOS
 

t NUMBE R OF ANIMAL S WIT H TISSUE

* NUMBE R OF ANIMAL S NECROPSIE D 

TABLE Cl (CONTINUED) 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) CONTROL (UNTR)
 

01-0070 01-01 18
 

(15) (15)
 

(18) (18)
 

1 (2S)
 1 (2X)
 

1 (2X)
 

2 (IX)
 

2 (1%)
 

(18) (18)
 

(15) (16)
 

12 (26%)
 1 (2%)
 

(15) (16)
 

2 (1*)
 1 (2%)
 

(11) (16)
 

1	 (9%)
 

3 (7%)
 

(18) (18)
 

3 (6%) I' 7 (98X)
 

1 (2%)
 

6 (13X)
 

11 (85*)
 

(18) (18)
 

 EXAMIN£ D MICROSCOPICALL Y 

C-7 

LOW r
 01-0065
 

(36)
 

1
 

(35)
 

«
 

1
 

1
 

(35)
 

2
 

(31)
 

(31)
 

(32)
 

6
 

(«?;


3
 

1
 

2
 

2
 

8
 

1
 

31
 

(13)
 

2
 

(3X)
 

(11%)
 

(3%)
 

(3*)
 

(6%)
 

(19X)
 

(7%)


<2X)
 

C5%)
 

(5%)
 

(19X)
 

(2X)
 

(7 IS)
 

J5X)L
 

HIGH DOSE
 

01-0107
 

(17)
 

(11)
 

(11)
 

1 (2X)
 1 (2%)
 

(39)
 

(39)
 

(38,
 

(16)
 

1 (2X)
 

!)3 (93*)
 

(16)
 
TABLE Cl (CONTINUED) 

LOW DOSE HIGH DOSE CONTROL (DNTR) CONTROL (UNTR) LOR DOSE HIGH DOSE 

01-0070 01-0118 01-0065 01-0107 

tKIDNEY/PELVIS (48) (48) (43) (46) 

MINERALIZATION 1 (2X) 

HYPEBPLASIA, EPITHELIAL 1 (2%) 

•UKINAHY BLADDER 

HYPERPLASIA, EPITHELIAL (16) (43) 

1 (2X) 

(37) 

1 (3%) 

(45) 

METAPLASIA, SQUAMOOS 1 (2X) 

ENDOCRINE SYSTEM 

IPITUITARY (41) (38) (31) (33) 

HYPERPLASIA, SOS 1 (3X) 

HYPERPLASIA, FOCAL 3 <7X) 2 (5X) 

fADRENAL (<*7) (47) (37) (42) 

METAMORPHOSIS FATTY 

HYPERPLASIA, KODULAR 

1 (2%) 

2 (5X) ANGIECTASIS 3 (6X) 

IADRENAL CORTEX (47) (47) (37) (t2) 

HYPERPLASIA, FOCAL 1 (2X) 

fADRENAL MEDULLA (47) (47) (37) (42) ' 

HYPERPLASIA, NODULAR 1 (2X> 

HYPEHPLASIA, NOS 1 (3X) HYPERPLASIA, FOCAL 4 (9X) 2 (5X) 

iTHYROID (39) (48) (37) (44) 

FOLLICOLAR CYST, NOS 1 (3%) 

HYPERPLASIA, C-CELL 1 (3X) 3 (6X) 

*PARATHYROID (23) (28) (22) (22) 

CALCIFICATION, NOS 

HYPEHPLASIA, NOS 1 (4<) 

1

11

 (5%) 

 (SOX) 

tPANCREATIC ISLETS (45) (46) (35) (42) 

HYPERPLASIA, NOS 1 <2X) 1 (3X) 

HYPERPLASIA, FOCAL 1 (2X) 

REPRODUCTIVE SYSTEM 

*MAMMARY GLAND (48) (»8) (43) (47) 

GAtACTOCELE „ . , . — 1 -UXX- _. 2_I1S1- ' „ . 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

C-8 
TABLE Cl (CONTINUED) 

LOW DOSE CONTROL (UNTR)

01-0070

 HIGH DOSE CONTROL (UNTR) 

 01-0118 

LOW DOSE 01-0065 HIGH DOSE 01-0107 

HYPERPLASIA. NOS 

•PREPOTIAL GLAND ABSCESS, NOS 

•PROSTATE INFLAMMATION, NOS 

INFLAMMATION, FOCAL INFLAMMATION, NECBOTIZING INFLAMMATION, ACUTE INFLAMMATION, ACUTE FOCAL 

INFLAMMATION, ACUTE/CHRONIC 

•SEMINAL VESICLE 

ATROPHY, NOS 

»TESTIS MINERALIZATION 

PERIARTERITIS DEGENERATION, NOS 

ATROPHY, NOS 

HYPERPLASIA, INTERSTITIAL CELL 

ITESTIS/TUBULE DEGENERATION, NOS 

CALCIFICATION, FOCAL 

3 (6%)

(48)

(43)

1 <2X) 

6 (14X) 

8 (19X) 

2 (5X) 

(48)

2 <4X) 

(47)

39 (83X) 

1 (2X)

(47)

 4 (8X) 

 (48) 

 (44) 

17 (39X) 

 (48) 

 (47) 

1 (2X) 

6 (13X) 

3 (6X) 

 (47) 

(43)

1 (2X) 

(35)

1 (3X) 2 (6X) 

2 (6X) 

(43)

(37)

1 <3X) 

8 (22X) 

4 (11X) 

(37)

 (47) 

 (43) 

 (47) 

 (44) 

 (44) 

5 (11X) 

2 (5X) 

NERVOUS SYSTEM 

N3NE 

SPECIAL SENSE OBGANS 

•EYE/CORNEA INFLAMMATION, ACUTE/CHRONIC 

•EYE/RETINA DEGENERATION. NOS 

(48)

(48)

 (48) 

 (48) 

(43)

1 <2X) 

(43)

1 <2X) 

 (47) 

 (47) 

HOSCUIOSKEIETAL SYSTEM 

NONE 

BODY CAVITIES 

* NUHBEF

* NUMBER

 OF ANIMALS

 OF ANIMALS

 WITH TISSUE

 NECROPSIED 

 EXAMINE D MICROSCOPICALLY 

C-9 
TABLE Cl (CONCLUDED) 

LOW DOSE HIGH DOSE CONTROL (UNTR ) CONTROL(UNTR ) LOW DOSE HIGH DOSE 

01-0070 01-0118 01-0065 01-0107 

ALL OTHER SYSTEMS
 

'MULTIPLE ORGANS (U8) (48) (43) (47)
 

POSTMORTEM CHANGE 2 (5%)
 

ADIPOSE TISSUE
 

INFLAMMATION. ACUTE/CHRONIC
 

OflSNTBM
 

NECROSIS, FAT
 

SPECIAL MORPHOLOGY SUMMAR Y
 

ANIMAL MISSING/NO NECROPSY
 

AUTO/NECROPSY/HISTO ?ERF
 

AUTOLYSIS/NO NECROPSY
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

c-io
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH 5-NITROACENAPHTHENE

 

LOW DOSE HIGH DOSE CONTROL (UNTR ) CONTROL (HNTR ) LOW DOSE HIGH DOSE 

02-0070 02-0118 02-0065 02-0107 

ANIMALS INITIALLY IN STUDY 50 50 50 50
 

ANIMALS NECROPSIED 49 50 49 48
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 49 50 48 48
 

INTEGUMENTARY SYSTEM
 

*SKIN (1*9) (50) (19) (48)
 

EPIDERMAL INCLUSION CYST 1 <2X)
 

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

NECROSIS, NOS 1 (2X)
 

*SDBCOT TISSUE (U9) (50) (49) (48)
 

MINERALIZATION 1 (2%)
 

ABSCESS, NOS 1 (2X)
 

RESPIRATORY SYSTEM
 

tTRACHEA (49) (49) (48) (47)
 

INFLAMMATION, ACUTE 1 (2%)
 

INFLAMMATION, ACUTE/CHRONIC 15 (31%) 4 <8X)
 

INFLAMMATION, CHRONIC 8 (17X) 7 (15*)
 

•LUNG/BRONCHUS	 (49) (50) (48) (48)
 

BRONCHIECTASIS 2 (4X)
 

INFLAMMATION, NOS 3 <6X)
 

INFLAMMATION, ACUTE/CHRONIC 1 (2%)
 

HYPERPLASIA, PAPILLARY 1 (2X)
 

ILUNG (49) (50) (48) (48)
 

INFLAMMATION, FOCAL 2 (4S)
 

INFLAMMATION, INTERSTITIAL 2 (4%) 6 (12X) 1 (2%)
 

INFLAMMATION, SUPPURATIVE 1 (2X)
 

PNEUMONIA, CHRONIC MURINE 1 <2X)
 HYPERPLASIA, EPITHELIAL 1 <2X)
 

HYPERPLASIA, FOCAL 1 <2X)
 

HYPERPLASIA, ALVEOLAR EPITHELIUM 1 <2X)
 

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

12*1
 

» NUMBE R OF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY » NUMBER OF ANIMALS NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-l 
__ _ _____ ____ _ ____ _ _________________________ _ _

TABLE C2 (CONTINUED) 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) CONTROL (UNTR) LOW DOSE HIGH DOSE 

02-0070 02-0118 02-0065 02-0107 

HEMATOPOIETIC SYSTEM 

• BONE MARRO B (46) (46) (47) (»3) 

OSTEOSCLSROSIS 1 (2X) 1 (2X) 

HYPEEPLASIA, NOS 1 (2X) 

*SPLEEN (48) (48) (48) (»7) 

HEMOSIDSROSIS 12 (25X) 2 («XJ 

ATROPHY, NOS 1 (9X) 

HYPEHPLASIA, HEMATOPOIETIC 1 (2X) 25 (52X) 

HYPERPLASIA, EEYTHEOID 19 (40X) 

HYPEHPLASIA, RETICULUM CELL 1 (2X) 

SSYTHEOPOIESIS 3 (6X) 

ISPLENIC CAPSULS (48) (48) (48) (17) 

H£MORRHAGIC CYST 1 (2X) 

iLYMPH NODJi (42) (47) (42) (3<t) 

PLASMACYTOSIS . 1 (2X) 

HYPESPLASIA, LYMPHOID 4 (9X) 

*MANDIBULAR L. NODlJ (42) (47) (42) (31) 

HYPERPLASIA, PLASMA CELL 2 (5X) 

*MEDIASTINAL L.NODE (42) (47) (42) (3») 

HYPERPLASIA, NOS 1 (2X) 

IPASCREATIC L.NODE (42) (47) (42) (31) 

HYPERPLASIA, PLASMA CELL . ___-.__— — _____•—— ­.———_ — —__•••———— — —— 1 (3X) _——.•—————• 

CIRCULATORY SYSTEM
 

fHEART (49) (50) (48) («8)
 

FIBROSIS, FOCAL 1 (2X)
 

FIBROSIS, DIFFUSE 1 (2X)
 

*MYOCARDIUM (49) (50) (48) (»8)
 

INFLAMMATION, NOS 1 (2X)
 

INFLAMMATION, INTERSTITIAL 2 (4X) 23 (46X)
 

INFLAMMATION, ACUTE FOCAL 1 (2X)
 

INFLAMMATION, ACUTE/CHRONIC 1 (2X) 1 (2X)
 

FIBROSIS 15 (30X)
 

FIBEOSIS, FOCAL 2 (4X)
 

IIS1!^_NQS 1_J2S1
 

« NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIHALS NECROPSIED
 

C-12
 
TABLE C2 (CONTINUED)
 

LOU DOSE HIGH DOSE CONTROL (UNTR) CONTROL (UNTR) 

02-0070 02-0118 

CALCIFICATION, FOCAL 

IENDOCARDIUM (49) (50) INFLAMMATION, NOS 1 (2X) 

ICARDIAC VALVE (i»9) (50) INFLAMBATION, ACUTB/CHRONIC 1 <2X) 

*AORTA (49) (50) MEDIAL CALCIFICATION 

*COHONART ARTERY (49) (50) INFLAMMATION. ACOTE/CHHONIC 

MEDIAL CALCIFICATION 

*PULMONARY ARTERY (19) (50) HINERALIZATION 9 (18X) 

*MESENTERIC ARTERY (49) (50) MEDIAL CALCIFICATION 

DIGESTIVE SYSTEM
 

»LIVER (19) (50)
 INFLAMMATION. ACOTE FOCAL
 

DEGENERATION, EOSI NOPHILIC 2 (IX)
 NECROSIS, FOCAL 3 (6X) 2 (4X)
 METAMORPHOSIS FATTY 1 (8X) 6 (12X)
 HYPERPLASIA, FOCAL 29 (59X) 38 (76X)
 

AHGIECTASIS 1 <2X)
 HYPERPLASIA, ERYTHROID 1 (2X)
 

HYPEHPLASIA, RETICULUM CELL
 

HEMATOPOIESIS 2 (UX)
 

ILIVSR/CENTRILOBULAR (49) (50)
 

CONGESTION, CHRONIC PASSIVE
 

*LIVER/HEPATOCYT2S (19) (50)
 NECROSIS, NOS
 

*BILE DDCT (49) (50)
 INFLAMMATION, NOS 1 (2X>
 

HYPERPLASIA, NOS 5 (10X) 32 (6 IX)
 HYPERPLASIA, FOCAL 1 <2X) 1 (2X)
 

#PANCREAS (K7) (KB)
 _ TflROM^OSIS,. NOS
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NSCROPSIE0
 

C-13
 

LOW DOSE
 

02-0065
 

(48)
 

1 (2X)
 

(i»8)
 

09)
 

1 <2X)
 

(19)
 

1 (2X)
 

(19)
 

(19)


1 (2X)
 

(18)
 

2 (IX)
 

1 (2X)
 

1 (2X)
 

(18)
 

1 (2X)
 

(18)
 

1 (2X)
 

(19)
 

(17)
 

1 IISL .
 

HIGH DOSE
 

02-0107
 

1 (2X)
 

(48)
 

(48)
 

(48)
 

1 (2X)
 

(48)
 

1 (2X)
 

(48)
 

(48)
 

1 (2X)
 

(47)
 

1 (2X)
 

1 (2X)
 

(47)
 

(47)
 

(18)
 

(47)
 
TABLE C2 (CONTINUED)
 

LOW DOSE HIGH DOSE CONTROL (UNT.c) CONTROL (UNTR) 

02-0070 02-0118 

INFLAMM.'.TION, NOS 6 (13X) 

INFLAMMATION, ACUTE/CHRONIC 4 (9X) 

ATROPHY, NOS 1 (2%) 

•STJKACH (49) (48) 

INFLAMMATION, NOS 1 (2X) 

ULCER, FOCAL 1 (2%) CALCIFICATION, NOS 

ACANTHOSIS 2 (4%) 

tPEYERS PATCH (49) (48) 

HYPERPLASIA, NOS 15 (31X) 

tCOLON (44) '.46) 

NEMATODIASIS 2 <5X> 

PARASITISM 2 (i»X) 

URINARY SYSTEM
 

tKIDNEY (49) (50)
 

MINERALIZATION 1 (2%)
 

CYST, NOS
 

GLCMEaULONEPhtUTIS, NOS 43 (86X)
 

INFLAMMATION, SUPPURATIVE
 

INFLAMMATION, ACUTE/CHRONIC
 

jLOMERULONEPHRITIS, SUBACUTE
 

FIBROSIS, DIFFUSE 1 (2X)
 

NEPHROSIS, NOS 34 (69X)
 

«KIDNEY/TU3ULE (49) (50)
 

CALCIFICATION, NOS
 

tKIDNEY/PELVIS (49) (50)
 HYPERPLASIA, FOCAL
 

ENDOCRINE SYSTEM
 

•PITUITARY	 (44) (40)
 PERIVASCULITIS 1 (3X)
 HYPEBPLASIA, NOS 1 (2X)
 HYPERPLASIA, FOCAL 2 (5X) 3 (8X)
 

*AOSLNJU (49) (19)
 

METAMORPHOSIS FATTY 3 16X1 1 12X1
 

•' NUN3ER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NSCROPSIED
 

C-I4
 

LOW DOSE

02- 06i

 HIGH DOSE 

 02-0107 

1 (2X) 

1 (2X) 

(48) (46) 

1 (2X) 1 (2X) 

(48) (47) 

(48)

4 (8X)

 (45) 

1 <2X) 

(48)

2 (4%) 

2 (4X) 

1 (2X)

15 (31X)

29 (60X)

(48)

(48)

1 (2X) 

 (48) 

1 (2X) 

1 (2X) 

9 (19X) 

 35 (73X) 

 (48) 

1 (2%) 

 (48) 

(44) (43) 

(47) (48) 
TABI EC2 (CONTIN' I
 

LOW COSE HIGH DOS':
 CONTROL (UN R) CONTROL IUNT R)
 

02-0070 02-0 111,
 

•ADRENAL	 CORTEX (.19) (19)
 

CYS", NOS
 

METAMORPHOSIS FATTY 3 (6X)
 

HYPERPLASIA, FOCAL 1 (2%)
 

ANGIECTASIS
 

•ADRENAL MEDULLA (19) ;u9)
 HYPERPLASIA, NODULAR 3 (6%)
 

HYPERPLASIA, NOS (2%)
 HYPERPLASIA, FOCAL i (2V 3 (6%;
 

•THYROID	 (tu) (15)
 

CYSTIC FOLLICLES 1 (2*)
 

HYPERPLASIA, C-CELL 1 '2")
 

y ,
 

HYPERPLASIA, NOS
 

•PARATHYROID	 (29)
 

REPRODUCTIVE SYSTEM
 

*MAMMAR Y GLAND (49) (50)
 

GALACTOCELE 9 (18%) 16 (32%)
 

CYSTIC DUCTS
 

INFLAMMATION, ACUTE 'i 1 2%)
 

HYPERPLASIA, NOS ."3 (47X) I (16%)
 

HYPEBPLASIA, FOCAL 2 (4%)
 

*CL!TOFAL GLAND (19; (50)
 

ABSCESS, NOS
 

(. j)
 

INFLAMMATION, ACUTE/CHRONIC 1 (2X)
 

"VAGINA	 (50)
 

•UTERUS	 (19) (30)
 

HYDRONETRA 6 (12X)
 HEK.iTOMA, NOS
 

HYP! TPLASIA, ADENJMAT US 1 (2X)
 

• CEPVIX UTEi.I (19) (50)
 

: IFLAMMATION, ACUTE/CHRONIC 2 lIX)
 

hi'F5RPLASIA, BASAL CELL 1 (2%)
 

ACANTHOSIS 1 (2%)
 

tUTERl'S/ENDOMETRIUM (19) (50)
 

INFLAKMiTION^NQS
 

» NUMBER OF ANIMALS WITH TISSUE EXAMINED MlCtvOSCO^ ^.CALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-l"
 

LOH DO i
 

02- 0( 5
 

(17)
 

1 (2%)
 1 (2X>
 

1 (2X)
 

(17)
 

(15)
 

1 (2S)
 

.23)
 

1 (4X)
 

(19)
 

1 (2%)
 

2 (IX)
 

(19)
 

1 (2X)
 

(49)
 

(17)
 

1 (2%)
 

1 (2X)
 

1 (2X)
 

(17)
 

(47)
 

i!IGH DOSE
 

02-0107
 

(48)
 

1 (IX)
 

(18)
 

(10)
 

(23)
 

(18)
 

(48)
 

C'8)
 

(42)
 

1 (2X)
 

(42,
 

(12)
 
TABLE C2 (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTROL (UNTR) CONTROL (UNTR) LOW DOSE HIGH DOSE 

02-0070 02-0118 02-0065 02-0107 

INFLAMMATION, SUPPURATIVE 1 (2%) 1 (2%) 

INFLAMMATION, ACUT3 23 (47%) 2 (4X) 

HYPESPLASIA, NOS 5 (10%) 6 (12%) 

HYPEEPLASIA, CYSTIC 5 (10%) 3 (6%) 

HYPERPLASIA, ADENOMATOUS 1 (2%) 

(OVARY/OVIDUCT (49) (50) (47) (42) 

INFLAMMATION, NOS 10 (20%) 

INFLAMMATION, SUPPURATIVE 2 (4%) 

INFLAMMATION, ACUTE 1 (2%) 

tOVARY (47) (49) (45) (45) 

CYST, NOS 2 (4%) 8 (16X) 1 (2%) 

SSRVOUS SYSTEM 

•BSAIN/MiNISGES (49) (50) (48) (47) 

HEMORRHAGE 1 (2X) 

SPECIAL SENS2 ORGANS 

*SYS (49) (50) (49) (48) 

INFLAMMATION, NOS 1 (2%) 

SYNECHIA, NOS 1 (2%) 

CATARACT 1 (2%) 1 (2%) 

VASCULAHIZATION 1 (2%} 

'EY3/CORNEA (49) (50) (49) (48) 

INFLAMMATION, ACUTE 1 (2%) 

ULCER, ACUTE 1 (2%) 

INFLAMMATION, ACUTE DIFFUSE 1 (2%) 

INFLAMMATION ACUTE AND CHRONIC 1 (2X) 

INFLAMMATION, CHRUNIC 1 (2%) 

'EYS/RETINA (49) (50) (49) (48) 

DEGENERATION, NOS 1 (2%) 

ATROPHY, NOS 1 (2X) 

*EYS/CRYSTALLINi; LENS (49) (50) (49) (48) 

CALCIFICATION , FOCAL 1 (2X) 

*HARDERIAN GLAND (49) (50) (49) (48) 

HYPJ.RPLASIA. NOS 1 _[2%]_ 

» NUM3ER OF ANIMALS 'WITH TISSUE EXAMINED MICROSCOPICALLY * NUMBE R OF ANIMALS NBCROPSIED 

e-16 
MOSCULOSKELSTAL SYSTEM 

*STBRNUB

OSTEOPETBOSIS

*HUSCLE OF NECK

HEMORRHAG E

BOOT CmtlBS 

*nEDI»STttTOH

PERIABTERITIS

*P£RICARDIUH


INFUBMATION, ACUTE/CHRONIC


ALL OTHER SYSTEMS
 

ADIPOSE TISSUE
 

INFLAMMATION, ACUTE/CHRONIC
 INFLAMMATION, CHRONIC


OMINTUB
 

MINERALIZATION


NECROSIS, FAT


SPECIAL HOSPHOLOGY SUMMARY
 

A01TO/NECROPSY/NO HISTO


AUTOLYSIS/NO NECROPSY


TABLE C2 (CONCLUDED) 

LOW DOSE HIGH DOSE 

CONTROL (UNTR) CONTROL(UMTR)

02-0070 02-0118

 (U9 ) (50)

 1 (2X) 

 (49) (50)

 (09) (SO)

 1 (2%) 

 (49) (50)


 3
 2
 

 1.
 

1
 

1


 LOW DOSE HIGH DOSE 

 02-0065 02-0107 

 (49) (U8 ) 

 (49) (48) 

1 (2X) 

 (49) (US) 

 (49)	 (48)
 

1 (2X)
 

1
 

1 2
 

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

C-17
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE TREATED WITH 5-NITROACENAPHTHENE
 

TABLED!

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH 5-NITROACENAPHTHENE 

HIGH DOSE
 

05-0065
 

50
 

45
 

45
 

(45)
 

(45)
 

(44)
 

(44)
 

2 <5X)
 

1 (2X)
 

(43) 

1 (2X) 

11 (26X) 

(41) 

10 (24X) 

(41) 

(30)

.__l_iiSl 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY**
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

ABSCESS, NOS
 

INFLAMMATION, CHRONIC FOCAL
 

*SUBCUT TISSUE
 

NECROSIS, FAT
 

RESPIRATORY SYSTEM
 

fLUNG/BRONCHIOLE
 

INFLAMMATION, NOS
 

PESIVASCULITIS
 

*LUHG
 

CONGESTION, NOS
 

HEMORRHAGE
 

PNEUMONIA, ASPIRATION
 

HYPERPLASIA, ALVEOLAR EPITHELIUM
 

HEMATOPOIETIC SYSTEM
 

CBONi; MARRO W 

HYPERPLASIA, HEMATOPOIETIC
 

HYPOPLASIA, HEMATOPOIETIC
 

*SPL3EN
 

ATROPHY, NOS
 

HYPE3PLASIA, LYMPHOID
 

tSPLENIC FOLLICLES
 

HYP£RPLASIA, NOS
 

• LYMP H NODii 

HYPERPLASIAt.NOS 

CONTROL (UNTR)
 

05-0070
 

50
 

50
 

50
 

(50)
 

2 (4X)
 

(50)
 

1 (2X)
 

(50)


1 (2X)
 

1 (2X)
 

(50)


2 (4X)
 

2 <<4X)
 

(48) 

150) 

1 (2X) 

(50) 

2 (4X) 

(45) 

LOW DOSE
 

05-0064
 

50
 

49
 

47
 

(49)
 

1 <2X)
 

(49)
 

 (46)
 

 (46)
 

(11) 

(12) 

(31) 

» NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NiJCFOPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

0-3 
HYPERPLASIA, LYMPHO'O
 

KCEBVICAL LYMPH NODE
 

HYPERPLASIA, NOS
 

IABDOMINAL LYMl'H NODK
 

HYPERPLASIA, NOS
 

IMESENTERIC L. NODI)
 

LYMPHOID DEPLETION
 

HYPERPLASIA. NOS
 

HYPERPLASIA. RETICULUM CELL
 

CIRCULATOBY SYSTEM
 

tHEART
 

INFLAMMATION, SUPPURATIVE
 

•AORTIC VALVE
 

INFLAMMATION, ACUTE/CH/'ONIC
 

DIGESTIVE SYSTEM
 

ISALIVARY GLAND
 

PERIVASCULITIS
 

*LIVER
 

CONGESTION, DOS
 

CONGESTION, PASSIVE
 

NECROSIS, FOCAL
 

METAMORPHOSIS FATTY
 

HEIATOCYTOMEGALY
 

DEPLETION
 

HYPERPLASIA, NODULAR
 

HYPERPLASIA, FOCAL
 

HYPERPLASIA, DIFFUSE
 

tLIVER/CENTRILOBULAR
 

NECROSIS, NOS
 

*LIVER/KUPFFER CELL
 

HYPERPLASIA, NOS '
 

((PANCREAS
 

_TNFLJK«A.TigN,. FOCA.L
 

« NUMBER OF ANIMALS WITH TISSUE

* NUMBE R OF ANIMAL S NiCROPSISD 

TABLE Dl (CONTINUED) 

CONTROL (UNTR) ' 1W DOSE
 

05-0070 05-0061
 

(U5) (31)
 

(15) (31)
 

(15) (31)
 

1	 <2X)
 

(19) (16)
 

(19)	 (16)
 1 (2X)
 

(19)	 (1U)
 1 (2X)
 

(50) I1- 6)
 

1 (2%)
 

2 (IX) 15 (35*)
 

2 (IX)
 

1 (2X)
 2 (IX)
 1 (2X)
 1 (2X)
 

(50)	 (16)
 

1 (21)
 

(50)	 (16)
 

1 (2X)
 

(16)	 (39)
 

1 .12JL
 

 EXAMINE D MICROSCOPICALLY 

0-4 

HIGH DOSE
 

05-0065
 

1 <3X)
 

(30)
 

1 (3X)
 

(30)
 

1 <3X)
 

(30)
 

1 (3X)
 

2 (7X)
 

(11)
 

1 (2%)
 

(11)
 

(12)
 

(11)
 

1 (2X)
 

1 (2X)
 

16 (36X)
 

',
 

(11)
 

l«1)
 

(38)
 
INFLAMMATION, CHRONIC
 

((ESOPHAGUS
 

HYPERKERATOSIS
 

ISTOMACH
 

ULCER, ACUTE
 

HYPERKERATOSIS
 

tGASTRIC MUCOSA
 

INFLAMMATION, FOCAL
 

•PEYERS	 PATCH
 

HYPERPLAS1A, NOS
 

KCOLON
 

3HANULOMA, NOS
 

HYPOPLASIA, NOS
 

URINARY SYSTEM
 

•KIDNEY
 

EMBRYONAL DUCT CYST
 

HYDRONEPHROS1S
 

CONGESTION, NOS
 

INFLAMMATION, INTERSTITIAL
 

FIBROSIS
 

FIBROSIS, FOCAL
 

N2PHP.OPATHY
 

METAMORPHOSIS FATTY
 

CALCIFICATION, NOS
 

fKIDNEY/CORTSX
 

SCAR
 

DEGENERATION, NOS
 

#KIDNEY/MEDULLA
 

DEGENERATION, NOS
 

CALCIFICATION, NOS
 

tRENAL PAPILLA
 

METAMORPHOSIS FATTY
 

CALCIFICATION, NOS
 

•KIDNEY/TUBULE
 

DILiTiTION^NOS
 

TABLE Dl (CONTINUED) 

CONTROL (UNTR )

05-0070

(49)
 

(49)
 

1 (2%)
 

(49)
 

(46)
 

1 (2%)
 

(K9)
 

3 (6X)
 

(149)
 

(49)
 

 LOW DOSE 

 05-0064 

(10)
 

(43)
 

2 (5X)
 

(43)
 

(41)
 

(39)
 

(46)
 

1 (2X)
 

(t6)
 

1 (2X)
 26 (6IX)
 

(46)
 

HIGH DOSE 

05-0065 

1 (3X)
 

(43)
 

4 (9X)
 

(43)
 

4 (9X)
 

(43)
 

(44)
 

(43)
 

1 (2X)
 

(44)
 

2 (5X)
 

1 (2X)
 

5 (11X)
 

1 (2X)
 

2 (5X)
 

5 (11X)
 

1 (2X)
 

(44)
 

8 (18X)
 

1 (2X)
 

(44)
 

1 (2X)
 

1 (2X)
 

(44)
 

(9X)
 

(44)
 

* NUMBER OF ANIMALS «ITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIES
 

D-5
 
FIBROSIS
 DEGENERATION, NOS


CALCIFICATION, NOS


• KIDNEY/PELVIS
 DILATATION, NOS


HYPE8PLASIA, EPITHELIAL


tURINARY BLADDER
 INFLAMMATION, ACUTE
 HYPERPLASIA, EPITHELIAL


ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTSM
 

ITESTIS
 ATPOPHY, NOS


ASPEHMATOGENESIS


tTESTIS/TBBULE


DEGENERATION, NOS


HOLTIHUCLEATE GIANT-CELL


NEEVOUS SYSTEM
 

*BRAIN/(1ENING£S
 HEMORRHAGE


fCEREBROH
 HEMORRHAGE


• CEREBELLUM
 HEMORRHAGE


SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

TABLE Dl (CONTINUED)
 

CONTROL (UNTR)


05-0070


 (1(9)


 (U7)


 1(2%)
 

(50)


 (50)


 (50)


 (50)


 (50)


 LOH DOSE HIGH DOSE
 

 05-006U 05-0065
 

3 (7*)
 21 (U8X)
 

2 (<tX)
 

 (16)	 (4<l)
 1 (2*)
 

2 (5X)
 

 (UU) (00)
 1 (2%)
 

 (<45) (Ul<)
 

2 (5X)
 

1 (2J)
 

 (<*5) (ait)
 

1 (2X)
 

« <9X)
 

 (13)	 (13)
 1 (2»)
 

 (U3) (U3)
 1 (2%)
 

 (43)	 (13)
 1 (2X)
 

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

D-6
 
TABLE DI (CONCLUDED)
 

CONTROL (UNTR) LOU DOSE HIGH DOSE
 

05-0070 05-006U 05-0065
 

BODY CAVITIES
 

NOSE
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 12 7 2
 

AUTO/NECROPSY/NO HISTO 2
 

AUTOLYSIS/NO NECROPSY 1 5
 

* NU13ER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-7
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH 5-NITROACENAPHTHENE

 

ANIMALS INITIAL!.! IN STUDY
 

ANIMALS HISSING
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY**
 

INTEGUMENTARY SYSTEM
 

NON3
 

RESPIRATORY SYSTEM
 

•LUNG/BRONCHIOLE
 

INFLAMMATION. NOS
 

*LUNG
 

INFLAMMATION, INTERSTITIAL
 

HEMATOPOIF.TIC SYSTEM
 

1BCNE MARBOH
 

MYELOFI3ROSIS
 

tSPLEZN
 

HYPERPLASIA, PLASMA CELL
 

HYPERPLASIA, LYMPHOID
 

SRYTHROPOIESIS
 

•SPLENIC	 FOLLICLES
 

HYPERPLASIA, NOS
 

iLYMPH NODE
 

INFLAMMATION, NOS
 

HYPERPLASIA, NOS
 

HYPERPLASIA, PLASMA CELL
 

•ABDOMINAL	 LYMPH NODE
 

PLASMACYTOSIS
 

CIRCULATORY SYSTEM
 

tMYOCARDIUM
 

1CONTROL (UNTR)
 

06-0070
 

50
 

18
 

17
 

(146)
 

1 (2%)
 

(<46)
 

1 (2%)
 

(46)
 

1 (2%)
 

(U7)
 

1 (2X)
 

(17)
 

3 (6%)
 

(36)
 

1 (3K)
 

1 (3X)
 

1 (354)
 

(36)
 

1 (3X)
 

(<414)
 

LOW DOSE HIGH DOSE 

06-00614 06-0065 

50 50 

1 1 

147 146 

«7 146 

(147) (146) 

(147) (146) 

(146) (14U) 

(14«) (U5) 

1 (2«) 

1 (2X) 

(1414) (U5) 

(38) (35) 

1 (3X) 

(38) (35) 

(15) (146) 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE R OF ANIrtAL S NSCROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-8 
TABLE D2 (CONTINUED) 

CONTROL (HNTR )

06-0070 LOW DOSE 06-0064

 HIGH DOSE 

 06-0065 

DIGESTIVE SYSTEM 

*SALIVARY GLAND

PERIVASCULITIS

PERIVASCULAP. CUFFING

 (15)

 3

 1

 (7%) 

 (2X) 

 (46) (15) 

tLIVER

INFLAMMATION, ACUTE FOCAL

INFLAMMATION. ACUTE/CHRONIC

NECROSIS, FOCAL

.1ETAMORPHOSIS FATTY

HYPERPLASIA, NODULAR

HYPERPLASTIC NODULE

HYPERPLASIA, FOCAL

 (47)

 1 (2%) 

1 (2X) 

2 (U%) 

 (47)

 1 (2X)

 1 (2X) 

1 (2X) 

1 (2%) 

 (45) 

 24 (53%) 

*SIL£ DUCT 

INFLAMMATION, ACUTE/CHRONIC 

(18) 

4 (8%) 

(47) (46) 

*PANCREAS 

INFLAMMATION, NOS 

INFLAMMATION, INTERSTITIAL 

PERIARTERITIS 

(43) 

1 (2X) 

1 <2X) 

1 (2X) 

(42) 

1 (2%) 

(37) 

#PASCREATIC ACINUS 

ATROPHY, NOS 

(43) 

1 (2X) 

(42) (37) 

#STOMACH 

ULCER. FOCAL 

(45) 

1 (2X) 

(44) (43) 

1PEY2RS PATCH 

HYPERPLASIA, NOS 

(45) 

1 (2JS) 

(42) (39) 

URINARY SYSTEM 

tKIONEY

GLONERULONEPHRITIS, NOS

3LOMSEULOSEPHRITIS, FOCAL

INFLAMMATION. INTERSTITIAL

GLOSERULONEPHRITIS, MEMBRANOUS

PYELONGPHF.ITIS, ACUTE/CHRONIC

GLOMEBULONEPHHITIS, CHRONIC

INFLAMMATION, CHRONIC FOCAL

PERIART3RITIS

 (45)

 3 (7X) 

2 (4X) 

1 (2X)

 2 (4X) 

1 (2%) 

1 (2«) 

 (46)

 4

 1

 (9X) 

 (2X) 

 (45) 

1 (2X) 

• KIDNEY/CORTEX

DILiTATIQNx_NOS

 (45) (46)

 ;

 (45) 

l_ 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS SSCROPSIED 

0-9 
INFLAMMATION, INTERSTITIAL
 

SCAS
 

fRENAL PAPILLA
 

CALCIFICATION, NOS
 

•KIDNEY/TUBULE
 

CALCIFICATION. NOS
 

•URINARY	 BLADDER
 

INFLAMMATION, CHRONIC FOCAL
 

PERIARTEHITIS
 

HYPERPLASIA, EPITHELIAL
 

ENDOCRINE SYST3M
 

• ADRENAL


HYPERPLASIA, NODULAR
 

IADRINAL CORTEX


HYPERPLASIA, NODULAR
 

HYPERPLASIA, NOS
 

*THYROID


HYPERPLASIA, FOLLICULAR-CELL


REPRODUCTIVE SYSTEM
 

tUTSRUS
 

HYDROHETRA
 

ABSCESS, NOS
 

•UTERUS/ENDOMETRIUM
 

INFLAMMATION, NOS
 

INFLAMMATION, SUPPURATIVE
 

INFLAMMATION, ACUTE
 

INFLAMMATION, ACUTE FOCAL
 

INFLAMMATION, ACUTE/CHRONIC
 

HYPERPLASIA, NOS
 

HYPERPLASIA, CYSTIC f
 

METAPLASIA, SQUAMOUS
 

fOVARY/OVIDUCT
 

TABLE D2 (CONTINUED) 

CONTROL (UNTH) 

06-0070 

(15)
 

(45)
 

1 (2X)
 

1 (2X)
 

 (47)
 

 (17)
 

 (11)
 

1 (2*)
 

(13)
 

3 (7X)
 

2 (5X)
 

(13)
 

2 <5X)
 

2 (5X)
 

6 (11*)
 

(2X)
 

(7X)
 

(2*)
 20 (K7X)
 

1 (2X)
 

(13)
 

LOW DOSE 

06-0061 

1 (2X)
 

U (9X)
 

(46)
 

(16)
 

1 (2X)
 

(it 3)
 

(13)
 

1 (2X)
 

(13)
 

12 (28*)
 

3 (7X)
 

(13)
 

(39)
 

(39)
 

21 (5<»X)
 

(39)
 

HIG H DOSE 

06-0065 

(15)
 

23 (51%)
 

(U5)
 

6 (13X)
 

1 (3X)
 

1 (3X)
 

(10)
 

(10)
 

13 (33X)
 

(10)
 

(12)
 

(12)
 

14 (33X)
 

(12)
 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

» SOMBER OF ANIMALS NECROPSIED
 

D-IO
 
TABLE D2 (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE 

06-0070 06-006U 06-0065 

ABSCESS. NOS 1 (2X) HYPERPLASIA , ADENONATOU S 2 (5%) 

IOVABY (U5) (41) (39)
 

CYST, NOS « (10X) « (10*)
 

HEMORRHAGE 1 (2X)
 

HEMORRHAGIC CYST 1 (3X)
 

INFLAMMATION, SUPPURATIVE 6 (13X)
 

INFLAMMATION, CHRONIC 1 (2X)
 

ABSCESS, CHRONIC 1 (2X)
 

FIBROSIS 1 (2X)
 

PERIART2RITIS 1 (2%)
 

CALCIFICATION, NOS 1 (2%)
 

HYPERPLASIA, CYSTIC 1 (2%)
 

METAPLASIA, OSSEOUS 1 (3X)
 

NERVOU S SYSTEM
 

I3RAIN/.12NINGES (H6) (U5) (13)
 

INFLAMMATION, ACUTE/CHRONIC 1 (2X)
 

INFLAMMATION, CHRONIC FOCAL 1 (2«)
 

SPECIAL SENSE ORGANS
 

NONE
 

WUSCULOSKEI2TAL SYST3M
 

NONE
 

BODY CAVITIES
 

•ABDOMINAL CAVITY (U8) (U7) (U6)
 

ABSCESS, NOS 1 (2X)
 

ALL OTHER SYSTEMS
 

• MULTIPLE ORGAN S («8) (17) (16) 

PERIVASCOLITIS 1 (2%) 

SPECIAL MORPHOLOGY SUMMARY
 

f NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-ll
 
TABLE D2 (CONCLUDED)
 

CONTROL (UNTB) LOW DOSE HIGH DOSE
 

06-0070 06-006H 06-0065
 

ANIMA L MISSING/NO NECROPSY 1 1
 

AUTO/NECROPSY/HISTO PER? 2
 

A'JTO/NECHOPSY/liO HISTO 1
 

AUTOLYSIS/NO NECROPSY 2 2 3
 

* NU13ER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUPI3ER OF ANIMALS NECROPSIED
 

D-12
 
Review of the Bioassay of 5-Nitroacenaphthene* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

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

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

organizations. Members have 'oeen selected on the basis of
 

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

as ad noc 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 5-Nitroacenaphthene for carcinogeni.city.
 

The reviewer agreed with the conclusion in the report
 

that 5-Nitroacenaphthene was carcinogenic, under the con­ ditions of test, in treated female mice and both sexes of
 

rats. He said that poor survival among treated male mice
 

precluded an analysis of this group. After a brief descrip­ tion of the experimental design, the reviewer commented on
 

the various tumors induced by the test material. He opined
 

that the study was adequate to support the conclusion on the
 

carcinogenicity of 5-Nitroacenaphthene. The reviewer moved
 

that the report on the bioassay of 5-Nitroacenaphthene be
 

accepted as written. The motion was approved without
 

objection.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

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

Louise Strong, University of Texas Health Sciences Center
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

61
 

•6U-S. GOVERNMEN T PRINTIN G OFFICE i 1 978—26 0'8 99/32 03
 

DHEW Publication No. (NIH) 78-1373