National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 144 

1978 

BIOASSAY OF 

2-AMINOANTHRAQUINONE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 117-79-3 

NCI-CG-TR-144 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

2-AMINOANTHRAQUINONE
 

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) 78-1399
 

REPORT ON THE BIOASSAY OF 2-AMINOANTHRAQUINONE
 

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
 

2-aminoanthraquinone 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 2-aminoanthraquinone was conducted by
 

Mason Research Institute, Worcester, Massachusetts, initially under
 

direct contract to the NCI and currently under a subcontract to Tracor Jitco, Inc., prime contractor for the NCI Carcinogenesis Testing
 

Program.
 

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

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

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

by Mr. G. Wade (3) and Ms. E. Zepp (3). Chemical analyses were performed by Mason Research Institute (3) and Midwest Research Institute
 

(4) and the analytical results were reviewed by Dr. N. Zimmerman (5).
 

Histopathologic examinations were performed by Dr. R. W. Fleischman (3), Dr. D. W. Hayden (3), Dr. A. S. Krishna Murthy (3), and Dr.
 

Yoon (3) at the Mason Research Institute, the pathology narratives
 

were written by Dr. A. S. Krishna Murthy (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 (6).
 

111
 
Compilation of individual animal survival, pathology, and summary tables was performed by EG&G Mason Research Institute (7); the
 

statistical analysis was performed by Mr. W. W. Belew (5,8) and Mr.
 

R. M. Helfand (5), using methods selected for the Carcinogenesis
 

Testing Program by Dr. J. J. Gart (9).
 

This report was prepared at METREK, a Division of The MITRE Corporation (5) under the direction of the NCI. Those responsible for
 

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

(5), task leader Dr. M. R. Kornreich (5,10), senior biologist Ms. P.
 

Walker (5), biochemist Dr. B. Fuller (5), and technical editor Ms.
 

P. A. Miller (5). 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 (1,10), Dr.
 

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

T. W. Orme (1), Dr. R. A. Squire (1,11), 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.	 Now with 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.	 Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

5.	 Th MITR e E Corporation, METREK Division, 1820 Dolley Madison
 

Boulevard, McLean, Virginia.
 

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

Maryland.
 

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

Rockville, Maryland.
 

8.	 Now with the Solar Energy Research Institute, Cole Boulevard,
 

Golden, Colorado.
 

IV
 
9.	 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.
 

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

Washington, B.C.
 

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of 2-aminoanthraquinone for possible carcinogenicity
 

was conducted using Fischer 344 rats and B6C3F1 mice. 2-Aminoanthraquinone was administered in the feed, at either of two concentrations
 

(except for female rats), to groups of 50 male and 50 female animals
 

of each species. The time-weighted average dietary concentrations
 

used in the chronic bioassay were 0.69 and 0.35 percent for high and
 

low dose male rats, respectively, 0.2 percent for the treated female
 

rats, and 1.0 and 0.5 percent, respectively, for high and low dose
 

mice of both sexes. After a 78-week period of chemical administration (80 weeks for high dose mice), observation of the rats continued
 

for up to an additional 32 weeks and observation of the mice continued
 

for up to an additional 16 weeks.
 

In both species adequate numbers of animals in all groups, except
 

the treated female rats, survived sufficiently long to be at risk from
 

late-developing tumors. The survival among treated female rats was
 

poor and, as a result, no conclusions could be made regarding the carcinogenicity of the compound in these animals.
 

When male rats having either hepatocellular carcinomas or neoplastic nodules of the liver were combined and the resulting tumor
 

incidences were analyzed statistically, there was a significant positive association between dosage and the incidences of these combined
 

neoplasms. Hepatocellular carcinomas were observed at significantly
 

higher incidences when dosed mice were compared to controls. There
 

was a significantly higher incidence of malignant hematopoietic
 

lymphomas in high dose female mice when compared to controls.
 

Under the conditions of this bioassay, dietary administration of
 

2-aminoanthraquinone was carcinogenic in male Fischer 344 rats, causing a combination of hepatocellular carcinomas and neoplastic nodules
 

of the liver. The compound was also carcinogenic in B6C3F1 mice,
 

causing hepatocellular carcinomas in both sexes and malignant hematopoietic lymphomas in females.
 

vii
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 4
 

A. Chemicals 4
 

B. Dietary Preparation 5
 

C. Animals 6
 

D. Animal Maintenance 7
 

E. Selection of Initial Concentrations 8
 

F. Experimental Design 10
 

G. Clinical and Histopathologic Examinations 13
 

H. Data Recording and Statistical Analyses 15
 

III. CHRONIC TESTING RESULTS: RATS 20
 

A. Body Weights and Clinical Observations 20
 

B. Survival 20
 

C. Pathology 23
 

D. Statistical Analyses of Results 25
 

IV. CHRONIC TESTING RESULTS: MICE 33
 

A. Body Weights and Clinical Observations 33
 

B. Survival 33
 

C. Pathology 36
 

D. Statistical Analyses of Results 37
 

V. DISCUSSION 44
 

VI. BIBLIOGRAPHY 46
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH 2-AMINOANTHRAQUINONE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH 2-AMINOANTHRAQUINONE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 2-AMINOANTHRAQUINONE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 2-AMINOANTHRAQUINONE D-l
 

ix
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF 2-AMINOANTHRAQUINONE 2 

2 GROWTH CURVES FOR 2-AMINOANTHRAQUINONE 

CHRONIC STUDY RATS 21 

SURVIVAL COMPARISONS OF 2-AMINOANTHRAQUINONE CHRONIC STUDY RATS 22 

GROWTH CURVES FOR 2-AMINOANTHRAQUINONE 

CHRONIC STUDY MICE 34 

SURVIVAL COMPARISONS OF 2-AMINOANTHRAQUINONE CHRONIC STUDY MICE 35 

LIST OF TABLES 

Table Number Page 

DESIGN SUMMARY FOR FISCHER 344 RATS— 

2-AMINOANTHRAQUINONE FEEDING EXPERIMENT 11 

DESIGN SUMMARY FOR B6C3F1 MICE—2-AMINOANTHRAQUINONE FEEDING EXPERIMENT 12 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN MALE 

RATS TREATED WITH 2-AMINOANTHRAQUINONE 26 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH 2-AMINOANTHRAQUINONE 29 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

2-AMINOANTHRAQUINONE 38 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH 2-AMINOANTHRAQUINONE 40 
LIST OF TABLES (Concluded)
 

Table Number 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH 2-AMINOANTHRAQUINONE A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH 2-AMINOANTHRAQUINONE A-7 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH 2-AMINOANTHRAQUINONE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH 2-AMINOANTHRAQUINONE B-7 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH 2-AMINOANTHRAQUINONE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 

2-AMINOANTHRAQUINONE C-10 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH 2-AMINOANTHRAQUINONE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 

2-AMINOANTHRAQUINONE D-9 

XI
 

I. INTRODUCTION
 

2-Aminoanthraquinone (Figure 1) (NCI No. C01876), an intermediate
 

in the synthesis of anthraquinone dyes, was selected for bioassay by
 

the National Cancer Institute in an attempt to determine which chemicals may be responsible for the increased incidence of bladder cancer
 

observed among workers in the dye manufacturing industry (Wynder et
 

al., 1963; Anthony and Thomas, 1970). Aromatic amines are one of
 

several classes of chemicals thought to contribute to the increased
 

cancer risk in this industry (Wynder et al., 1963). The structural
 

relationship of 2-aminoanthraquinone to two documented carcinogens,
 

2-naphthylamine (Occupational Safety and Health Administration, 1973)
 

and 2-anthramine (Griswald et al., 1968), was also a factor in its
 

selection.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is 2-amino-9,10-anthracenedi.one.* It
 

is also commonly referred to as AAQ.
 

Although the unsubstituted aminoanthraquinones have no importance
 

as dyes in and of themselves, introduction of the amino group into the
 

anthraquinone nucleus provides the basis for innumerable anthraquinone
 

dye intermediates (Cofrancesco, 1963). 2-Aminoanthraquinone per se is
 

a direct precursor of five dyes and one pigment that are produced commercially in the United States. These include Colour Index Vat Blues
 

k
 

The CAS registry number is 117-79-3.
 
FIGURE 1

 

CHEMICAL STRUCTURE OF 2-AMINOANTHRAQUINONE 
4, 6, 12 and 24; Vat Yellow 1; and Pigment Blue 22 (Society of Dyers
 

and Colourists, 1971; as cited in Urso, 1977).
 

Recent production statistics for 2-aminoanthraquinone are considered proprietary and are therefore not available. However, 2.01 x
 

10 pounds of the compound and its salts were produced in 1971, the
 

latest year for which such data were reported (U.S. Tariff Commission, 1973; as cited in Urso, 1977). Imports of 2-aminoanthraquinone
 

through principal U.S. customs districts are quite substantial and
 

amounted to 3.57 x 10 pounds in 1974 (U.S. International Trade
 

Commission, 1976; as cited in Urso, 1977).
 

Since 2-aminoanthraquinone appears to be used on a commercial
 

scale exclusively by the dye industry, the potential for exposure to
 

the compound is greatest for workers at dye manufacturing facilities.
 
II. MATERIALS AND METHODS
 

A. Chemicals
 

One batch of 2-aminoanthraquinone was purchased from J. T. Baker
 

Chemical Company (Lot #1-8041) and a second batch was purchased from
 

American Cyanamid. The second batch did not bear a lot number when
 

it was received at Mason Research Institute. The batch received from
 

Baker was used only for the first 8 weeks of the chronic bioassay of
 

male rats (female rats and mice of both sexes were not yet on test).
 

After week 8 of the male rat bioassay, only the American Cyanamid
 

batch was used for dosed feed preparation. Dosed female rats and all
 

dosed mice received only the American Cyanamid chemical.
 

The initial chemical analyses of both batches were performed at
 

Mason Research Institute. Five years later a sample of the American
 

Cyanamid batch was analyzed by Midwest Research Institute. The melting points determined at Mason Research Institute were 215° to 235°C
 

for the American Cyanamid batch. The chemical decomposed at 235°C.
 

When the American Cyanamid batch was tested at Midwest Research
 

Institute, a melting point range of 255° to 292°C was observed,
 

with decomposition noted at 292°C. The deviation of these observed
 

melting point ranges from the melting point range of 303° to 306°C
 

reported in the literature (Pollock and Stevens, 1965), suggests that
 

either the chemicals tested were of very low purity or that attempts
 

to determine the melting point were unsuccessful because decomposition occurred before the melting point was reached. The ultraviolet
 

4
 
spectra for the compound purchased from American Cyanamid and the
 

literature reference spectra (Sadtler Standard Spectra,a) are shown
 

below:
 

American 

Cyanamid 

(Analyzed by 

Midwest) Reference 

Solvent: Methanol Methanol 

445 445
 

297 297
 

279
 

240 240
 

The ultraviolet spectra of the American Cyanamid sample in methanol determined at Midwest Research Institute included all three of
 

the peaks found in the reference spectrum, confirming identity of the
 

compound, and also had a peak at 279 nm. The extra peak indicated
 

the presence of an impurity. The ultraviolet spectra of both batches
 

determined in chloroform at Mason Research Institute do not agree with
 

the reference spectra.
 

The infrared spectrum, determined at Midwest Research Institute
 

for the American Cyanamid batch, was consistent with the literature
 

spectrum (Sadtler Standard Spectra,b).
 

Throughout this report the term 2-aminoanthraquinone is used to
 

represent these materials.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals
 

consisted of Wayne Lab-Blox

(f& 

 (Allied Mills, Inc., Chicago, Illinos).
 

5
 
2-Aminoanthraquinone was administered to the dosed animals as a component of the diet. The chemical was mixed in the feed in a 6 kg
 

capacity Patterson-Kelley standard model stainless steel twin-shell
 

V-blender. After 20 minutes of blending, the mixtures were placed
 

in double plastic bags and stored in the dark at 4°C. Mixtures were
 

prepared weekly and stored for not longer than 1 week.
 

C. Animals
 

Two animal species, rats and mice, were used in the chronic
 

carcinogenicity bioassay. Fischer 344 rats and B6C3F1 mice were
 

obtained through contracts of the Division of Cancer Treatment, National Cancer Institute. All rats and their controls were supplied
 

by Laboratory Supply Company, Inc., Indianapolis, Indiana. The high
 

dose and high dose control mice and some of the low dose mice (male
 

and female) were supplied by Charles River Breeding Laboratories,
 

Inc., Wilmington, Massachusetts. The remaining low dose mice and
 

all the low dose control mice were supplied by ARS/Sprague-Dawley,
 

Madison, Wisconsin. Each group of animals to be dosed was received
 

in a shipment separate from their controls.
 

Upon arrival, a sample of animals was examined for parasites and
 

other signs of disease. The remaining animals were quarantined 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, rats were housed in galvanized- or
 

stainless-steel wire-mesh cages suspended above newspapers. Newspapers under cages were replaced daily and cages and racks washed
 

weekly. For the remainder of the study, rats were housed in suspended polycarbonate cages equipped with disposable nonwoven fiber
 

filter sheets. Clean bedding and cages were provided twice weekly.
 

Stainless steel cage racks were cleaned once every two weeks, and
 

disposable filters were replaced at that time.
 

Mice were housed by sex in shoe box type polycarbonate cages fitted with perforated stainless steel lids and nonwoven fiber filters.
 

All mice were housed either five or ten per cage. Cages, lids, and
 

bedding were provided two to three times per week. 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. Alpine aluminum feed cups (Curtin Matheson Scientific, Inc.,
 

Woburn, Massachusetts) containing stainless steel baffles or stainless steel gangstyle hoppers were used to distribute powdered feed.
 

Pelleted Wayne Lab-Blox was supplied during the quarantine period
 

and final observation period. Mice were fed pellets from a wire bar
 

hopper incorporated into the cage lid, while rats received pellets
 

on the cage floor.
 

All rats were housed in a room in which other rats were receiving
 

&
 

diets containing 5-nitro-o-toluidine (99-55-8); hydrazobenzene (530­

50-7); 3-amino-9-ethylcarbazole hydrochloride; 6-nitrobenzimidazole
 

(94-52-0); 1-nitronaphthalene (86-57-7); and APC (8003-03-0).
 

All dosed mice in this study were housed with their respective
 

diet controls.
 

E.	 Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

2-aminoanthraquinone for administration to dosed animals in the
 

chronic studies, subchronic toxicity tests were conducted with both
 

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

groups, each consisting of five males and five females. 2-Aminoanthraquinone was incorporated into the basal laboratory diet and
 

* .
 CAS registry numbers are given in parentheses.
 
supplied ad libitum to three of the four groups of each species in
 

concentrations of 1.0, 3.0, and 5.0 percent. The fourth 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 laboratory diet.
 

The highest concentration causing no deaths, no compound-related
 

gross abnormalities, and no mean group body weight depression in
 

excess of 20 percent relative to controls was selected as the high
 

concentration utilized for the rat and mouse chronic bioassays.
 

No deaths were recorded for any dosed rat group. Mean body
 

weight depression was approximately 6 and 10 percent for males receiving chemical concentrations of 1.0 and 3.0 percent, respectively,
 

and 18 and 19 percent for females receiving the same respective
 

concentrations. The high concentration selected for use in the rat
 

chronic bioassay was 2.0 percent for both males and females.
 

The only death recorded among dosed mice was one male receiving
 

5.0 percent 2-aminoanthraquinone. Mean body weight depression was
 

approximately 8 percent for males receiving concentrations of 1.0
 

and 3.0 percent, respectively, and 14 and 18 percent for females
 

receiving the same respective concentrations. The high concentration
 

ultimately selected for use in the mouse chronic bioassay was 1.0
 

percent for both males and females.
 
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.
 

There were 10 rat groups (male and female) originally included
 

in this research. As five of these groups consisted of too few animals for meaningful statistical analysis for bioassay purposes and/
 

or included gonadectomized and ovariectomized rats, only the 5 remaining rat groups were considered when evaluating this bioassay.
 

Dosed male rats were approximately 6 weeks old when chemical
 

administration was initiated and they shared the same median date
 

of birth. Control males were approximately 3 weeks older than the
 

dosed males. The initial concentrations of 2-aminoanthraquinone
 

administered to male rats were 2.0 and 1.0 percent, respectively.
 

Throughout this report those males initially receiving 2.0 percent
 

are referred to as the high dose group and those initially receiving 1.0 percent are referred to as the low dose group. After having
 

received 2-aminoanthraquinone at the indicated concentrations for 10
 

weeks, the dosages were reduced to 0.5 and 0.25 percent for the high
 

and low dose groups, respectively. These concentrations were maintained for the remainder of the chemical administration period. The
 

dosed group of female rats was approximately 6 weeks old when they
 

10
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS 

2-AMINOANTHRAQUINONE FEEDING EXPERIMENT 

TIME-WEIGHTED 

AVERAGE 

INITIAL 2-AMINOANTHRA­ OBSERVATION PERIOD CONCENTRATION 

GROUP QUINONE TREATED UNTREATED OVER A 78-WEEK 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) PERIODb 

MALE 

CONTROL 50 0 0 107 0 

LOW DOSE 50 1.0 10 0.35 

0.25 68 

0 28 

HIGH DOSE 50 2.0 10 0.69 

0.5 68 

0 28 

FEMALE 

CONTROL 25 0 0 109 0 

DOSED 50 0.2 78 0.2 

0 32 

a
 Concentrations in percentages in feed .
 

S (concentration X weeks received)
 

Ti trip— w<=> i oh t-prl flvpi •fltrp f nnrpnfraf i nn =
 78 weeks
 

11
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

2-AMINOANTHRAQUINONE FEEDING EXPERIMENT
 

INITIAL 2-AMINOANTHRA­ OBSERVATION PERIOD 

GROUP QUINONE TREATED UNTREATED 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) 

MALE 

LOW DOSE CONTROL 50 0 0 93 

HIGH DOSE CONTROL 50 0 0 96 

LOW DOSE 49 0.5 78 

0 16 

HIGH DOSE 50 1.0 80 

0 15 

FEMALE 

LOW DOSE CONTROL 50 0 0 94 

HIGH DOSE CONTROL 50 0 0 96 

LOW DOSE 50 0.5 78 

0 16 

HIGH DOSE 50 1.0 80 

0 16 

Concentrations in percentages in feed. 

12
 
were included in the bioassay. The control females were also approximately 6 weeks old when they were included in the bioassay, 11 weeks
 

after compound administration had been initiated for the dosed females.
 

The dosed females received 0.2 percent 2-aminoanthraquinone for the
 

78-week period of compound administration. Dosed male rats were
 

observed for an additional 28 weeks after compound administration
 

ceased and dosed female rats were observed for an additional 32 weeks.
 

The concentrations utilized for both male and female mice were
 

1.0 and 0.5 percent. Throughout this report those mice receiving a
 

dietary 2-aminoanthraquinone concentration of 1.0 percent are referred to as the high dose groups while those receiving 0.5 percent
 

are referred to as the low dose groups. The low dose mice were
 

approximately 6 weeks old when they were first administered the
 

compound. At that time the low dose control mice were approximately
 

3 weeks older. The high dose mice were also approximately 6 weeks
 

old when they first received the compound, which was approximately 8
 

months after the low dose mice had been started on test. The high
 

dose control mice were approximately 3 weeks younger than the high
 

dose mice. The high dose mice were dosed for 80 weeks while the low
 

dose mice received the chemical for 78 weeks. An observation period
 

of up to 16 weeks followed the period of compound administration.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. Body weights were recorded twice weekly for the first
 

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

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

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

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

bioassay and for three consecutive days each month thereafter. The
 

presence of tissue masses and lesions was determined by monthly observation and palpation of each animal.
 

A necropsy was performed on each animal regardless of whether it
 

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

bioassay. The animals were euthanized by carbon dioxide inhalation,
 

and were immediately necropsied. The histopathologic examination
 

consisted of gross and microscopic examination of major tissues,
 

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

possible, from animals found dead.
 

Tissues were preserved in 10 percent buffered formalin, embedded
 

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

to microscopic examination. An occasional section was subjected to
 

special staining techniques for more definitive diagnosis.
 

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

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

urinary bladder, pituitary, adrenal, thyroid, parathyroid, brain, ear,
 

testis, prostate, Zymbal's gland, mammary gland, uterus, and ovary.
 

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

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

preclude histopathologic interpretation. Thus, the number of animals
 

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

animals that were placed on experiment in each group.
 

H. Data Recording and Statistical Analyses
 

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

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

as recommended by the International Union Against Cancer (Berenblum,
 

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

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

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

15
 
missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect
 

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

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

testing a dose-related trend. One-tailed P-values have been reported
 

for all tests except the departure from linearity test, which is only
 

reported when its two-tailed P-value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

specific anatomic site (numerator) to the number of animals in which
 

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

histologically. However, when macroscopic examination was required
 

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

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

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

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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
 

16
 
with those for a control group, a correction to ensure an overall
 

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

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

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

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

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

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

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

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

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

was found. Once this reduced set of data was obtained, the standard
 

17
 
procedures for analyses of the incidence of tumors (Fisher exact
 

tests, Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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

during which animals died naturally or were sacrificed was entered
 

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

these curves were used for two groups; 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.
 

18
 
The lower and upper limits of the confidence interval of the
 

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

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

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

would be within the interval calculated from the experiment. When
 

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

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

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

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

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

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

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

detected under the conditions of this test.
 

19
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Slight mean body weight depression was observed when dosed males
 

were compared with control males. There was a more distinct difference between the mean body weights of the dosed and control females
 

(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.
 

One low dose male rat developed a palpable mass on the ventral
 

surface and one control male had a crusted lesion on the dorsolateral
 

surface. No other clinical abnormalities were observed in dosed or
 

control rats of either sex.
 

B. Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and 2-aminoanthraquinone-dosed groups are shown in
 

Figure 3. For male rats, there was no significant association between
 

dose and mortality. For female rats a significant positive association between chemical administration and mortality was observed.
 

For males ten rats were sacrificed from the control group in
 

week 29 and five rats were sacrificed from each dosed and control
 

group in week 78. Adequate number of males were at risk from late-


developing tumors with 70 percent (35/50) of the high dose, 64 percent (32/50) of the low dose, and 54 percent (27/50) of the controls
 

surviving on test until the termination of the study.
 

20
 
TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR 2-AMINOANTHRAQUINOIME CHRONIC STUDY RATS 

21 
TIME ON TEST (WEEKS) 

FIGURE 3 

SURVIVAL COMPARISONS Of 2-AMINOANTHRAQUINONE CHRONIC STUDY RATS 

22 
For females five rats were sacrificed from the dosed group and
 

five from the control group in week 78. Survival in the dosed group
 

declined steadily from week 40 on. After one year, 70 percent
 

(35/50) of the dosed rats and 92 percent (23/25) of the control rats
 

were alive on test. By the termination of the study, however, only
 

14 percent (7/50) of the dosed rats and 48 percent (12/25) of the
 

control rats remained alive. Survival of female rats was not adequate to permit analysis of late-developing tumors.
 

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 hepatic neoplasms in male rats fed the two concentrations of 2-aminoanthraquinone appeared to be compound-related.
 

Hepatocellular carcinomas occurred in 8/49 (16 percent) low dose,
 

5/46 (11 percent) high dose and 0/46 control male rats. Hepatocellular carcinoma was found in 1/42 (2 percent) dosed females but in
 

none of the controls. Neoplastic nodules were also commonly observed
 

in dosed male rats.
 

Morphology of the hepatocellular carcinoma and neoplastic nodules
 

was similar to that described by Squire and Levitt (1975). Hepatocellular carcinoma involved one or many lobes of the liver. The tumor
 

appeared well-differentiated, but the normal lobular architecture was
 

distorted. Liver plates were two cells in thickness. There was a
 

23
 
pleomorphism in the size of transformed hepatocytes. Cytoplasm of the
 

cells was either acidophilic or basophilic. In some tumors, cytoplasm
 

of the cells was vacuolated and suggested fatty infiltration. Nuclei
 

were hyperchromatic and nucleoli prominent. Mitotic figures were
 

numerous. Areas of necrosis and clusters of inflammatory cells were
 

found in a few tumors. The tumor had metastasized to the lung in
 

one control female rat, but in none of the dosed rats.
 

Neoplastic nodules were smaller, demarcated, and compressed the
 

normal liver in areas. Cells in these nodules were large. Cytoplasm
 

of the cells was basophilic and the nuclei were large. A few mitotic
 

figures were present.
 

2-Aminoanthraquinone is considered to be a hepatocarcinogen since
 

there was an increase in the incidence of hepatocellular carcinomas
 

and neoplastic nodules of the liver in male Fischer 344 rats fed the
 

two concentrations of the chemical.
 

One low dose male rat and 16/42 (38 percent) of the female rats
 

originally included in the bioassay had foreign body granulomatous
 

nephritis (FBGN) of the kidney and died in less than 25 weeks. This
 

lesion differed from the renal changes normally seen in aging Fischer
 

344 rats, in that the lumen of cortical tubules were plugged with crystals, and there was a granulomatous reaction and fibrosis. The lesions
 

resembled those described by Baker et al. (1975) as foreign body granulomatous nephritis (FBGN). In those female rats fed 0.2 percent 2-aminoanthraquinone which survived as long as the respective controls, the
 

incidence of neoplasms was low.
 

24
 
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
 

tumors were observed in at least one of the control or 2-aminoanthraquinone-dosed groups and where such tumors were observed in at least
 

5 percent of the group. Because of the sacrifice of 10 male control
 

rats in week 29, the analyses for males were based solely on those
 

rats surviving at least 30 weeks.
 

For males the Cochran-Armitage test showed a significant (P <
 

0.001) positive association between dose and the combined incidence
 

of hepatocellular carcinomas or neoplastic nodules of the liver.
 

The departure from linear trend was also significant (P = 0.006)
 

due to the comparable incidence rates in both dosed groups. These
 

positive results were supported by significant (P < 0.001) Fisher
 

exact tests when either dosed group was compared to the control.
 

When hepatocellular carcinomas alone were considered, the Fisher
 

exact test showed a significantly (P = 0.005) higher incidence in
 

the low dose than in the control. The high dose comparison had a
 

probability level of P = 0.048, a marginal result which was not
 

significant under the Bonferroni criterion. Based on these statistical findings the administration of 2-aminoanthraquinone was associated with the increased incidence of hepatocellular neoplasms in
 

male rats.
 

25
 
TABLE 3
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH 2-AMINOANTHRAQUINONEa»f
 

S3
 

ON
 

HIGH
 

DOSE
 

5/45(0.11)
 

P = 0.048
 

Infinite
 

1.020
 

Infinite
 

78
 

18/45(0.40)
 

P < 0.001
 

Infinite
 

4.667
 

Infinite
 

78
 

3/42(0.07)
 

P = 0.002(N)
 

0.190
 

0.038
 

0.635
 

106
 

TOPOGRAPHY: MORPHOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma or
 

Neoplastic Nodule"
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Adenoma NOS or Chromophobe
 

Adenoma'3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/36(0.00)
 

N.S.
 

P = 0.018
 

__.«
 

0/36(0.00)
 

P < 0.001
 

P = 0.006
 

12/32(0.38)
 

P - O.OOl(N)
 

101
 

LOW
 

DOSE
 

8/41(0.20)
 

P = 0.005
 

Infinite
 

2.032
 

Infinite
 

78
 

18/41(0.44)
 

P < 0.001
 

Infinite
 

5.134
 

Infinite
 

78
 

4/38(0.11)
 

P = 0.008(N)
 

0.281
 

0.074
 

0.824
 

106
 
___ 

TABLE

TOPOGRAPHY: MORPHOLOGY
 

Adrenal: Pheochromocytoma or Pheochromocytoma, Malignant
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pancreatic Islets: Islet-Cell Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

3 (CONTINUED) 

CONTROL
 

6/35(0.17)
 

N.S.
 

107
 

1/35(0.03)
 

P = 0.044
 

107
 

2/32(0.06)
 

N.S.
 

__«.
 

107
 

LOW
 

DOSE
 

2/40(0.05)
 

N.S.
 

0.292
 

0.030
 

1.511
 

106
 

0/39(0.00)
 

N.S.
 

0.000
 

0.000
 

16.661
 

1/39(0.03)
 

N.S.
 

0.410
 

0.007
 

7.536
 

106
 

HIGH
 

DOSE
 

5/44(0.11)
 

N.S.
 

0.663
 

0.175
 

2.398
 

104
 

5/39(0.13)
 

N.S.
 

4.487
 

0.539
 

206.175
 

104
 

1/44(0.02)
 

N.S.
 

0.364
 

0.006
 

6.707
 

106
 
___ 

TABLE 3 (CONCLUDED) 

LOW HIGH
 

TOPOGRAPHY :MORPHOLOGY CONTROL DOSE DOSE
 

Testis: Interstitial-Cell Tumor 33/35(0.94) 36/39(0.92) 38/45(0.84)
 

P Values0 N.S. N.S. N.S.
 

Relative Risk (Control) 0.979 0.896
 

Lower Limit 0.886 0.812
 

Upper Limit 1.115 1.077
 

Weeks to First Observed Tumor 78 78 78
 

Treated groups received time-weighted average doses of 34.6 or 69.2 ppm in feed.
 

NJ Number of tumor-bearing animals/number of animals examined at site (proportion).
 00
 

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.
 «
 

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

group when P < 0.05.
 

These analyses were based solely upon animals surviving at least 30 weeks.
 
TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH 2-AMINOANTHRAQUINONE*
 

TOPOGRAPHY: MORPHOLOGY
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma^3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

K3 Liver: Hepatocellular Carcinoma or
 

Neoplastic Nodule*3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Adenoma NOS or Chromophobe
 

Adenoma*5
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

2/23(0.09)
 

106
 

2/23(0.09)
 

106
 

8/21(0.38)
 

78
 

DOSED
 

2/43(0.05)
 

N.S.
 

0.535
 

0.042
 

7.038
 

106
 

1/42(0.02)
 

N.S.
 

0.274
 

0.005
 

5.023
 

106
 

6/30(0.20)
 

N.S.
 

0.525
 

0.182
 

1.477
 

78
 
___ 

TOPOGRAPHY: MORPHOLOGY
 

Adrenal: Cortical Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma or
 

Pheochromocytoma, Malignant
 

u> P Values0 o 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma*3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

CONTROL
 

0/23(0.00)
 

3/23(0.13)
 

109
 

3/21(0.14)
 

109
 

DOSED
 

2/39(0.05)
 

N.S.
 

Infinite
 

0.180
 

Infinite
 

67
 

3/39(0.08)
 

N.S.
 

0.590
 

0.087
 

4.106
 

76
 

2/29(0.07)
 

N.S.
 

0.483
 

0.044
 

3.871
 

50
 
__ _ 

TABLE 4 (CONCLUDED)
 

TOPOGRAPHY; MORPHOLOGY CONTROL DOSED
 

Mammary Gland: Adenocarcinoma NOS
 

or Papillary Cystadenoma NOSb 2/23(0.09) 0/43(0.00)
 

P Values0 N.S.
 

Relative Risk (Control) 0.000
 

Lower Limit 0.107
 

Upper Limit 1.794
 

Weeks to First Observed Tumor 98
 

Mammary Gland: Fibroadenoma 4/23(0.17) 3/43(0.07)
 

P Values0 N.S.
 

Relative Risk (Control) 0.401
 

Lower Limit 0.065
 

Upper Limit 2.194
 

Weeks to First Observed Tumor 109 64
 

o
 

Treated groups received concentrations of 0.2 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.
 
Also in male rats the Cochran-Armitage test indicated a significant (P = 0.044) positive association between dose and the incidence
 

of thyroid C-cell adenomas. The Fisher exact tests, however, were
 

not significant.
 

For male rats the possibility of a negative association between
 

chemical administration and tumor incidence was observed for the combined incidence of pituitary adenomas NOS and chromophobe adenomas.
 

This may have been due, however, to the greater than expected incidence of these tumors in the control group (12/41 or 29 percent)
 

when compared to the historical control incidence for these tumors
 

35/334 (11 percent) in data on untreated male Fischer 344 rats
 

collected by this laboratory for the NCI Carcinogenesis Testing
 

Program.
 

32
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Slight mean body weight depression was observed when low dose
 

males were compared with low dose control males. There was a more
 

distinct difference between the mean body weights of the low dose female and low dose control females. Distinct differences in mean body
 

weights were not apparent when high dose mice of either sex were compared to their high dose controls (Figure 4). It may be noted that
 

for both sexes the mean body weights of the two control groups which
 

came from different suppliers were appreciably different.
 

No clinical abnormalities were observed in dosed or control mice
 

of either sex.
 

B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and 2-aminoanthraquinone-dosed groups are shown in
 

Figure 5. The Tarone test for positive association between dosage
 

and mortality was not significant for either male or female mice.
 

Five male mice were sacrificed from the high dose control in
 

week 49 and five more in week 78. Additionally, five males were
 

sacrificed from the high dose group in week 78, plus five from the
 

low dose control in week 80. At the termination of the study, 86
 

percent (43/50) of the high dose, 94 percent (46/49) of the low dose,
 

78 percent (39/50) of the high dose control and 82 percent (41/50) of
 

33
 
TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR 2-AMINOANTHRAQUINONE CHRONIC STUDY MICE 

34 
TIME ON TEST (WEEKS) 

FIGURES 

SURVIVAL COMPARISONS OF 2-AMINOANTHRAQUINONE CHRONIC STUDY MICE 

35 
the low dose control mice were alive on test. Adequate numbers of
 

males were at risk from late-developing tumors.
 

Five female mice from the high dose control were sacrificed in
 

week 49 with five more sacrificed in week 78. Additionally, five
 

females were sacrificed from the high dose group in week 78 and five
 

from the low dose control in week 80. With 76 percent (38/50) of the
 

high dose, 88 percent (44/50) of the low dose, 76 percent (38/50) of
 

the high dose control, and 78 percent (39/50) of the low dose control
 

mice surviving on test until the termination of the study, adequate
 

numbers of females survived sufficiently long 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).
 

A dose-related increase in the incidence of hepatocellular
 

carcinomas was found in both sexes of mice receiving 2-aminoanthraquinone. In male mice, this tumor occurred in 20/47 (43 percent) of the low dose group, 36/49 (73 percent) of the high dose
 

group, and in 18/94 (19 percent) of the controls. In female mice,
 

this tumor was present in 5/47 (11 percent) of the low dose group,
 

12/47 (26 percent) of the high dose group, and 5/96 (5 percent)
 

of the controls.
 

Morphology of the hepatocellular carcinoma in these mice was
 

similar to that described in Butler and Newberne (1975). The tumor
 

36
 
had replaced a part or a whole lobe of the liver. The normal architecture was distorted with the tumor cells arranged in abnormal
 

trabecular patterns. Cytoplasm of the cells was eosinophilic and
 

vacuolated in areas. A pleomorphism in nuclear size was evident.
 

Nuclei were vesicular, and mitotic figures were present. Inclusions
 

were seen in some of the nuclei. Metastases to other organs were not
 

found.
 

An increased incidence of malignant lymphomas was observed in
 

high dose female mice when compared to controls. These lesions were
 

found in 12/49 (24 percent) of the high dose females and 7/97 (7 percent) of the controls. The incidences of these lesions in the low
 

dose and control groups were similar.
 

The dosed mice had the usual spectrum of nonneoplastic lesions,
 

and none appeared to be compound-related.
 

2-Aminoanthraquinone is considered to be a hepatocarcinogen, as
 

there was a dose-related increase in the incidence of hepatocellular
 

carcinomas in B6C3F1 mice receiving this chemical. Administration
 

of the chemical was also associated with an increased incidence of
 

malignant lymphomas in high dose female B6C3F1 mice.
 

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
 

37
 
00 

TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH 2-AMINOANTHRAQUINONE*
 

LOW DOSE HIGH DOSE LOW
 

TOPOGRAPHY : MORPHOLOGY CONTROL CONTROL DOSE
 

Lung: Alveolar/Bronchiolar Carcinoma 2/46(0.04) 5/49(0.10) 4/45(0.09)
 

P Values0 N.S.
 

Relative Risk (Control) 2.044
 

Lower Limit 0.310
 

Upper Limit 21.695
 

Weeks to First Observed Tumor 93 96 94
 

Lung: Alveolar/Bronchiolar Adenoma or
 

Alveolar/Bronchiolar Carcinoma^ 7/46(0.15) 10/49(0.20) 8/45(0.18)
 

P Values0 —_
 N.S.
 

Relative Risk (Control) 1.168
 

Lower Limit 0.404
 

Upper Limit 3.468
 

Weeks to First Observed Turner 80 96 94
 

Hematopoietic System: Malignant
 

Lymphoma'3 2/46(0.04) 5/49(0.10) 0/48(0.00)
 

P Values0 N.S.
 

Relative Risk (Control) 0.000
 

Lower Limit 0.000
 

Upper Limit 3.236
 

Weeks to First Observed Tumor 93 96
 

HIGH
 

DOSE
 

5/50(0.10)
 

N.S.
 

0.980
 

0.240
 

3.999
 

85
 

11/50(0.22)
 

N.S.
 

1.078
 

0.459
 

2.570
 

85
 

2/50(0.04)
 

N.S.
 

0.392
 

0.039
 

2.266
 

78
 
TABLE 5 (CONCLUDED)
 

TOPOGRAPHY : MORPHOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma or
 

Hepatocellular Adenoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE
 

CONTROL
 

12/46(0.26)
 

93
 

12/46(0.26)
 

93
 

HIGH DOSE
 

CONTROL
 

6/48(0.13)
 

78
 

8/48(0.17)
 

78
 

LOW
 

DOSE
 

20/47(0.43)
 

N.S.
 

1.631
 

0.867
 

3.192
 

94
 

20/47(0.43)
 

N.S.
 

1.631
 

0.867
 

3.192
 

94
 

HIGH
 

DOSE
 

36/49(0.73)
 

P < 0.001
 

5.878
 

2.843
 

14.070
 

78
 

36/49(0.73)
 

P < 0.001
 

4.408
 

2.353
 

8.925
 

78
 

Treated groups received doses of 0.5 or 1.0 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.
 
TABLE 6 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT 

SPECIFIC SITES IN FEMALE MICE TREATED WITH 2-AMINOANTHRAQUINONEc 

TOPOGRAPHY: MORPHOLOGY 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Lung: Alveolar /Bronchiolar Adenoma or 

Alveolar/Bronchiolar Carcinoma" 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

1/46(0.02) ___ 

___ 

3/50(0.06) ___ 

___ 

2/48(0.04) 

N.S. 

1.917 

0.103 

110.703 

3/48(0.06) 

N.S. 

1.042 

0.146 

7.419 

Weeks to First Observed Tumor 94 78 94 95 

° Hematopoietic System: Malignant 

Lymphoma" 

P Values0 

5/47(0.11) 2/50(0.04) 6/48(0.13) 

N.S. 

12/49(0.24) 

P = 0.003 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

1.175 

0.321 

4.548 

6.122 

1.464 

53.856 

Weeks to First Observed Tumor 80 96 83 82 

Liver: Hepatocellular Carcinoma" 

P Values0 

4/46(0.09) 1/50(0.02) 5/47(0.11) 

N.S. 

12/47(0.26) 

P = 0.001 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

1.223 

0.281 

5.803 

12.766 

2.015 

530.576 

Weeks to First Observed Tumor 94 96 94 78 
TABLE 6 (CONCLUDED)
 

LOW DOSE HIGH DOSE LOW HIGH 

TOPOGRAPHY: MORPHOLOGY CONTROL CONTROL DOSE DOSE 

Pituitary: Adenoma NOS or Chromophobe 

Adenoma^ 4/37(0.11) 3/42(0.07) 4/42(0.10) 3/36(0.08) 

P Values0 N.S. N.S. ___ 

Relative Risk (Control) 0.881 1.167 

Lower Limit 0.176 0.165 

Upper Limit 4.417 8.184 

Weeks to First Observed Tumor 80 96 94 96 

Treated groups received doses of 0.5 or 1.0 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.
 
tumors were observed in at least one of the control or 2-aminoanthraquinone-dosed groups and where such tumors were observed in at least
 

5 percent of the group. Cochran-Armitage tests were not used in
 

these analyses since the high dose and the high dose control groups
 

came from a different supplier and were started at a different time
 

from the low dose and the low dose control groups.
 

Hepatocellular carcinomas were observed in a large number of
 

male and female mice. In male mice a significant (P < 0.001) Fisher
 

exact test result was obtained for the comparison of the high dose
 

group to the high dose control. Similarly, in female mice the Fisher
 

exact test was significant (P = 0.001) for the comparison of high
 

dose to the high dose controls. Based on these statistical findings
 

the administration of 2-aminoanthraquinone was associated with the
 

increased incidence of hepatocellular carcinomas in both male and
 

female mice.
 

For female mice, the incidence of malignant lymphoma was
 

increased in the high dose group relative to that in the high dose
 

control group, with a significant (P = 0.003) comparison using the
 

Fisher exact test. Incidences of 24 percent (12/49) and 4 percent
 

(2/50) were observed in the high dose and high dose control groups,
 

respectively, compared to the historical incidence of a maximum of
 

12 percent (43/350) in untreated female B6C3F1 mice observed at Mason
 

Research Institute for the NCI Carcinogenesis Testing Program. Based
 

on these findings, there was a possibility that the administration of
 

42
 
2-aminoanthraquinone was associated with the development of malignant
 

lymphomas in female mice.
 

43
 
V. DISCUSSION
 

There were no significant positive associations between the
 

dietary concentrations of 2-aminoanthraquinone and mortality in male
 

rats or in mice of either sex; there was a significant positive association, however, between dose and mortality in female rats. Adequate
 

numbers of animals in all groups, except the dosed female rats, survived sufficiently long to be at risk from late-developing tumors.
 

When male rats having either hepatocellular carcinomas or neoplastic nodules of the liver were combined and the resulting incidences
 

(i.e., 0/46, 18/49 [37 percent], and 18/46 [39 percent] in the control,
 

low dose, and high dose, respectively) were analyzed statistically,
 

there was a significant positive association between dosage and the
 

incidences of these combined neoplasms. Both the high dose to control
 

and the low dose to control Fisher exact comparisons supported this
 

finding. When those males having hepatocellular carcinomas alone
 

were evaluated, the low dose to control Fisher exact comparison was
 

significant; however, the high dose to control comparison was not.
 

Dietary exposure to the compound resulted in nephrotoxicity in female
 

rats, as the majority of the females examined had foreign body granulomatous nephritis.
 

Hepatocellular carcinomas were observed at higher incidences in
 

dosed mice when compared to controls (i.e., 12/46 [26 percent], 6/48
 

[13 percent], 20/47 [43 percent], and 36/49 [73 percent] in the low
 

44
 
dose control, high dose control, low dose, and high dose males, respectively, and 4/46 [9 percent], 1/50 [2 percent], 5/47 [11 percent]
 

and 12/47 [26 percent] in the low dose control, high dose control,
 

low dose, and high dose females). For both sexes the high dose to
 

control Fisher exact comparisons were significant. For female mice
 

there was a significantly higher incidence of malignant hematopoietic
 

lymphomas in the high dose group when compared to the high dose control. In addition, the incidence observed in the high dose females
 

in this bioassay (i.e., 12/49 or 24 percent) is twice the historical
 

incidence observed in control female mice maintained at Mason Research
 

Institute during the NCI Carcinogenesis Testing Program.
 

In a previous report, no evidence was provided for the carcinogenicity of 2-aminoanthraquinone in female Sprague-Dawley rats following administration of a total of 1000 mg of the compound. The animals
 

were intubated with 10 equal doses over a period of 30 days, the first
 

dose administered when they were 40 days old. Final observations in
 

this study were completed approximately 9 months after the first dose
 

was given (Griswald et al., 1968).
 

Under the conditions of this bioassay, dietary administration of
 

2-aminoanthraquinone was carcinogenic in male Fischer 344 rats, causing a combination of hepatocellular carcinomas and neoplastic nodules
 

of the liver. The compound was also carcinogenic in B6C3F1 mice,
 

causing hepatocellular carcinomas in both sexes, and it was associated
 

with malignant lymphomas in females. No evidence was provided for the
 

carcinogenicity of 2-aminoanthraquinone in female rats.
 

45
 
VI. BIBLIOGRAPHY
 

Anthony, H.M., and G.M. Thomas, "Tumors of the Urinary Bladder: An
 

Analysis of the Occupations of 1,030 Patients in Leeds, England."
 

Journal of the National Cancer Institute 45:879-895, 1970.
 

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

J. Wiley & Sons, New York, 1971.
 

Baker, J.R., E.R. Smith, Y.H. Yoon, G.G. Wade, H. Rosenkrantz, and B.
 

Schmall, "Nephrotoxic Effect of 2-Aminoanthraquinone in Fischer
 

Rats." Journal of Toxicology and Environmental Health 1:1-11,
 

1975.
 

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

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

Union Against Cancer, Geneva, 1969.
 

Butler, W.H. and P.M. Newberne, editors, Pathology of Mouse Hepatic
 

Neoplasia. Elsevier Scientific Publishing Company, Amsterdam,
 

1975.
 

Chemical Abstracts Service, The Chemical Abstracts Service (CAS)
 

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

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

Cofrancesco, A.J., "Anthraquinone and Related Quinoid Dyes." Kirkothmer Encyclopedia of Chemical Technology. Second edition,
 

Volume 2, p. 447, 1963.
 

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.
 

Griswald, D.P., Jr., A.E. Casey, E.K. Weisburger, and J.H. Weisburger,
 

"The Carcinogenicity of Multiple Intragastric Doses of Aromatic
 

and Heterocyclic Nitro or Amino Derivatives in Young Female
 

Sprague-Dawley Rats." Cancer Research 28:924-933, 1968.
 

46
 
Kaplan, E.L., and P. Meier, "Nonparametric Estimation from Incomplete
 

Observations." Journal of the American Statistical Association
 

_53_: 45 7-481, 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.
 

Occupational Safety and Health Administration, "Occupational Safety
 

and Health Standards 1910.93c. Carcinogens." Federal Register
 

38_:974, 1973.
 

Pollock, J.R.A. and R. Stevens, editors. Dictionary of Organic Compounds , 4th edition. Oxford University Press, New York, New
 

York, p. 83, 1965.
 

Sadtler Standard Spectra,a. Sadtler Research Laboratories, Philadelphia, Pennsylvania. UV No. 7792.
 

Sadtler Standard Spectra,b. Sadtler Research Laboratories, Philadelphia, Pennsylvania. IR No. 19527.
 

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.
 

Society of Dyers and Colourists. Colour Index, 3rd edition, Volume 4.
 

Yorkshire, England, 1971.
 

Squire, R.A. and M.H. Levitt, "Report of a Workshop on Classification
 

of Specific Hepatocellular Lesions in Rats." Cancer Research 35:
 

3214-3223, 1975.
 

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

62_:679-682, 1975.
 

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

Industries Center, Stanford Research Institute, Menlo Park, California. Personal communications, 1977.
 

U.S.	 International Trade Commission, Import os f Benzenoid Chemicals
 

and Products, 1974. USITC Publication 762, U.S. Government
 

Printing Office, Washington, B.C., 1976.
 

47
 
U.S.	 Tariff Commission, Synthetic Organic Chemicals, U.S. Production
 

and Sales 1971. USTC Publication 614, U.S. Government Printing
 

Office, Washington, D.C., 1973.
 

Wynder, E.L., J. Onderdonk, and N. Mantel, "An Epidemiological Investigation of Cancer of the Blader." Cancer 16:1388-1407, 1963.
 

48
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH 2-AMINOANTHRAQUINONE
 

TABLE Al

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH 2-AMINOANTHRAQUlNONE 

LOW DOSE HIGH DOSE CONTROL(UNTR ) 

01-0037 01-0031 01-0032 

ANIMAL S INITIALL Y IN STUD? 50 50 50 

AHIHALS NECROPSIED 46 49 47 

ANIMAL S EXAMINE D HISTOPATHOLOGICAL! Y ** U6 49 46 

INTZGUNENIABY SYS1EM 

*SKIN 

FIBROSARCOMA 

(46) (49) (47) 

1 (2%) 

*SUBCUT TISSUE 

FIBROMA 

LEIOMYOSARCOMA 

(46) (49) 

1

1

 (2%) 

 (2X) 

(47) 

RESPIRATORY SYSTEM 

tTFACHEA (45) (49) (45) 

ADENOCARCI NOHA , NOS, METASTATIC 1 (2X) 

tLUNG (46) (49) (46) 

ADENOCAHCmCHA. NOS, METAS1ATIC 1 (2X) 

ALVEOLAR/BRONCHIOLAR ADENOM A 1 (2X) 

ALVEOLAR/BRONCHIOLAR CARCINOM A 1 <2X) 

C-CELL CARCINOHA , METASTATIC 1 (2X) 

PHEOCHROMOCYTOMA , METASTATIC 2 (4X) 

HEKATCPOIETIC SYSTEM 

*BUL1IFLE CRGAKS (46) (49) (47) 

MALIGNAN T LYBEHOMA , NO S 

HALIG.LYMPHOMA , HISTIOCYTIC TYSI 1 (2X) 

UNDIFFERENTIATED LEUKEMI A 1 (2X) 

MONOCYTIC LEUKEMIA 1 (2X) 

*IY»PH SCEE (38) (43) (40) 

ADENOCARCINOMA , NOS, METAS1ATIC 1 (3X) 

CIBCOLATCB Y SYSTE M 

* NUMBER CF ANIMAL S HITH TISSBH EXAMINE D MICROSCOPICALLY 

* NUMBER Cf ANIMAL S NECROPSIED 

**EXCLODES PARTIALLY AUTOLYZED ANIMALS 

A-3 
TABLE Al (CONTINUED) 

CONTBOL(UNTB ) LOW DOSE 

01-0037 01-0031 

DIGESTIV E SYSTEM 

#IIVE R (16) (49) 

NEOPLASTIC NOtULE 10 (20X) 

HEPATOCELLULA B CARCINOM A 8 (16X) 

»STOKAC H (45) (47)

 

PAPILLCMA , NOS 

ISBAIL I MESTINI (43) (47)

 

LEIOMYCM A 

atUCEENU M (43) (47) 

LEIOMYCM A 

URINAR Y SYSTE M 

NON E 

ENDOCRIN E SYSTEM 

IPITUITAR Y (41) (46) 

ACENCFA , NC S 2 <5X ) 1 (2*) CHBOMOPHOEE ADENOMA 10 (2i*%) 3 (1%) 

fADRENA L (43) (48) 

ADENOCARC I NCMA , NOS, METASTATI C 1 (2X) PHEOCFHCHOCYTCM A 6 (14X) 2 (4X) PHEOCHROMOCYTOMA , MALIGNAN T 

#THYROI D (45) (46) 

ACEkCPA , NCS 1 (2X) 1 <2X) ACENOCARCIKCMA , NOS 2 (H%1 C-CELL ADENOM A 1 (2X) C-CELL CARCINOM A 1 <2X) 

IPANCEEATIC ISLETS (42) (47) 

ISLET-CELL AEENOM A 2 (5)!) 1 (2X) 

BEPRCDUCT3V E SYSTE M 

*RAHfAB Y GLAN C (46) (49)

 

ACENOCABCI!,CMAJ_HO^ 

* NUMBE R CF ANIMAL S WITH TISSUB IXAtlNE D MICROSCOPICALLY 

* NUMBE R CF ANIMAL S NEC80PSIEE 

HIGH DOSE 

01 -0032 

(46) 

1-1 (28X) 

5 (11«) 

(40) 

1 (2X) 

(15)

1 (2X) 

(4S) 

1 (2*) 

(43) 

3 <7X) 

(45) 

:i (7X) 

2 (4X) 

(40) 

1 (3X) 

b (13X) 

(45) 

I (2X) 

(47) 

1 12XL ___ _ 

A-4 
TABLE Al (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE 

01-0037 01-0031 01-0032 

FIBBCAtENCKA 1 (2X) 

IPBOSTATE (45) (47) (44) 

PABAGAKGIICMA , NOS 1 (2%) 

ITESTIS (45) (47) (46) 

INTEBSTITIAL-CH.L TUMOR 33 (73*) 36 (77*) 38 (83%) 

NERVOU S SYSTEM 

tBRAI N (44) (48) (46) 

ASTRCCYTOMA 1 (2*) 

SPECIAL SEliSE CBGAN S 

NONE 

MUSCULOSKELETA L SYSTEM 

NON E 

EOCY CAVITIES 

*BCDY CAVITIE S (46) (49) (47) MESOTHELIOMA, NOS 1 (2*) 

All CTHEB SYSTEM S 

NON E 

ANIMA L CISPOSITION SUMMAR Y 

ANIRAL S IMTIALL Y IN STUDY 50 50 50 

NATURA I DEATHS 6 3 6 

MCBIBUN t SACRIFICE 2 10 4 

SCHEDULED SACBIEICE 15 5 5 

ACCIDENTALL Y KILIED 

TERMINA L SACRIFICE 27 32 35 

ANIMA L MISSING 

a_ISCIUEES_AUTCLYZI D ANIMALS. 

I NUMBE R CF ANIMAI S WITH TISSUE EX8BINE D MICBO SCOP ICALIY 

* NUMBE K CI ANIMAL S NECBOPSIED 

A-5 
TABLE Al (CONCLUDED) 

CONTBOL(UNTB) LOW DOSE HIGH DOSE 

01-0037 01-0031 01-0032 

TUI-CF SUHHJRY
 

TOTAL AUMAL S WITH PRIMARY ItWOHS* 3U 39 42 

TCTAL PRIMAR Y TUMORS 61 68 79 

TOTAL ANIBAL S KITH BENIG N TOBCRS 33 37 39 

TOTAL EENIGN TUMORS 55 46 5<» 

TOTAL ABIBAL S SI TH BALIGBANT TUMORS 5 11 10 

TOTAL BALIGNAN T TUBOBS 5 11 12 

TOTAL AKIBAL S 8ITH SECONDARY TOMCES* 1 1 2 

TCTAL SECONDAR Y TUSO8S 4 1 2 

TOTAL AKIMAL S BITH TUMORS UNCEBTAINEENIGN OR MALIGNANT 1 11 13 

TCTAL UNCIBTAI N TUMOHS 1 11 13 

TOTAL AMBAL S WITH TOBORS ONCBHTAINPRIBAH Y OB METASTATIC

 

TOTAL DNCERTAIN TOMOES

 

* PRIMAR Y 1UMORS: ALL TOBORS EXCEPT SECONDARY TUMORS 

* SECCNEAB Y TUBCBS : METASTATIC 1UBOBS OB TUM08S INVASIVE INTO AN ADJACENT ORGAN 

A-6 
TABLE A2 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH 2-AMINOANTHRAQUINONE 

COHTBOL(UHTR) DOSED 

02-0084 02-0078 

ANIBAI S IMTIALLY III STUDY 25 50 

ANIHilS NECROPSIED 23 43 

ANIMALS EXAMINE D HISTOPATHOLOGICAL!,Y ** 23 43 

INTIGOHESTARY SYSTEH
 

• SKI N (23) (43) 

SQUAHCD S CELL CARCINOMA 1 (2%) 

SEBACECUS ADENOCARCINOMA 1 (4X) 

HESPIBATCBY SYSTEH 

• LON G (23) (42) 

ALVECIAE/BBCHCHIOLAB ADENOM A 1 (4%) 

ALVEOLAR/BRONCHIOUB CABCIHOHA 1 (2X) 

HEBATCPOIETIC SYSTIH 

*PULTIPL E CBGANS (23) (43) 

ONDIFJIRENTIATI D LEUKEHI A 2 (9X) 

BYEIOHCKOCYTIC IEOKEMIA 2 (5X) 

ISPIEEK (23) (39) 

SARCOfA , DCS 1 (3X) 

CIBCUIATCBY SYS1EM 

NON E 

DIGESTIVE SYSTIH 

tLIVEB (23) (42) 

NEOPLASTIC NOEOLE 2 (9%) 

HEPATOCELLULAR CARCINOMA 1 (2%) 

URINAB Y SYSTEH 

I NOHEER CF ANIHAI S WITH TISSUE EXAMINE D BICBO SCOPICALLY * NUMBE R Cf ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

A-7
 
TABLE A2 (CONTINUED) 

CONTBOL(DNTH) DOSED 

02-008« 02-0078 

ENECCRINE SYSTEM
 

tEITUITABY (21) (30) 

ADENCPA , NOS 1 (5X) K (13X) 

CHRONCPHOBE ADENOMA 7 ( 33X) 2 (7X) 

(ADRENAL (23) (39) CCRTICAL ADENOM A 2 (5X) 

PhECCtBOMOCYTCMA 2 (9X) 3 <8X) 

PHEOCHROMOCYTOMA. MALIGNAN T 1 (IX) 

tlHYBOIC (21) (29) 

C-CELI ADENCM A 2 (10X) 1 (3X) 

C-CELL CARCINCB A 1 (5*) 1 <3X) 

KTHYBOI E FCLLICLI (21) (29) 

PAPILLAR Y CYSliDEHOCABCINOHA.MOS 1 (5X) 

IPAFATHJROI D (9) (11) 

ADEHCRA , NCS 1 (7X) 

REPRODUCTIV E SYSTBB 

*PABBAB Y GLAK t

ACENCCARC I KCBS , NOS

INFILTRATIN G DDCT CA

FIBROADENOM A

RCISCHA

 (23)

 2

 1

 U

 (9X) 

 (US) 

 (17X)

 (U3 ) 

3 (7X) 

IUTERU S

fNDOHf^BIA L S1BOMAL POLYP

 (23)

 6 (26X)

 (33) 

1 (3X) 

NERVCUS SYSTEM
 

KCNE
 

SPECIAL SINSE CEGAN S
 

*ZYMEAL' S GLAN D (23) (t3) 

SCUABCU S CELL CARCINOMA 1 (2X) 

MUSCOIOSKE1ETAI SYSTEM 

NONE 

» NUMBE R CF ANIMAL S WITH TISSDE EXAMINE D MICROSCOPICALLY 

* NUHBE R CI ANIMAL S NECROPSIED 

A-8 
TABLE A2 (CONCLUDED) 

CONTfiOL(UNTR) DOSED 

02-008U 02-0078 

EOtY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

kCN I 

ANIMA l DISPOSITION SUMMAR Y 

8NIBALS INITIALL Y IN STUD Y 25 50 

NATURAL DEATHS 3 20 

MCRIEUN E SACRIFICE 5 18 

SCHEDULE D SACBIEICE 5 5 

ACCIDENTALL Y KIIIED 

TERMINAL SACRIFICE 12 7 

ANIMA L MISSIN G 

9 INCLUDE S AUTCLYZI C ANIMAL S 

TUBCB SUBPAE Y 

TOTAL AMMAL S WITH PHIMABY 1UMOBS* 19 17 

TOTAL PRIMAR Y TUMORS 3t 25 

TOTAL ANIMAL S KITH BENIS N TUMORS 18 14 

TOTAL EENIG N TUMORS 23 17 

TOTAL AMP.AL S WITH MALIGNAN T TUMORS 8 6 

TCTAL BALIGNAN T TUMORS 9 8 

TOTAL ANIMAL S WITH SECONDARY TUMCfSI 

TCTA L SECONDAR Y TUMOH S 

TOTAL AMMAL S WITH TUMORS HNCERTAIN ­

EENIGN OR MALIGNAN T 2 

TOTAL UNCERTAI N TUMORS 2 

TOTA L AUKAL S WITH TUMORS UNCER1AINPBIKAR Y CH METASTATIC

 

TOTAL UNCERTAI N TUHORS 

* PRIMAR Y TUBCBS: ALL TUMORS EXCEPT SECONDAR Y TUMORS 

i SECONDAB Y TUMCRS : METASTATIC TUHORS OR TUMOBS INVASIVE INTO AN ADJACENT ORGA N 

A-9 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH 2-AMINOANTHRAQUINONE
 

TABLE Bl

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH 2-AMINOANTHRAQUINONE 

HIGH DOSE LOW DOSE 

CONTROL(UNTR) CONTHOL(UNTR ) LO U DOSE 

05-0118 05-0030 05-0032 

ANIMAL S INITIALL Y I N STUDY 50 50 49 ANIMAL S MISSING 1 

ANIMAI S KECROESIEr 49 46 48 ANIMAI S EXAMINE E HISTOPATHOL CGICALLY ** 49 46 47 

IKTIGU8IMABY SYSTEM 

*SUECUT 1ISSDE (U9) (46) (48)

 LEIOMYOSARCOM A

 

RESPIRATOR Y SYSTE M 

*IUNG <K9 ) (46) (45) 

ACENOCARCI hCHA , NOS. METASTSIIC 1 (2%) 

HEPATOCELLULA R CARCINOMA , METAST 1 (2%) 1 (2X) ALVEOLAR/BRONCHIOLAR ADENOM A 5 (IDS) 5 (11X) 4 (9X) 

ALVEOLAR/BRONCHIOLA R CARCINOM A 5 (10«) 2 (US) 4 (98) 

HEMATOPOIETIC SYSTE M 

*FULTIELt OBGAN S (U9 ) (46) (48) 

MALIGNA M LYMPHOMA , NO S 1 (2%)

 

MALIG . LYMPHOMA , HISTIOCYTIC TYP E 3 (6X) 1 (2%) 

tSPLEEN (49) (46) (46) HEBANGICSARCGM A 1 (2%)

 

MALIG-IYMPFCMA , HISTIOCYTIC TYIE 1 (2%)

 

*IYMP H NCt E (12) (34) (36) BALIG .LYMPHOMA , HISTIOCYTIC TYP E 1 (2%)

 

IPEYERS PATC H (49) (46) (47)

 MALIG.1YMPHCHA , HISTIOCYTIC TYP I

 

CIRCOIATOF Y SYSTE M 

NONE 

I NOMEE B OF ANIMAL S WITH TISSUE EXAKINE D MICROSCOPICALL Y 

* NUMBE R CI ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

HIG H DOSE 

05-0112 

50 

50 

50 

(50) 

1 (2%) 

(50) 

6 (12X ) 

b (10X) 

(50) 

(49) 

1 (2%) 

(42) 

(47) 

1 (2%) 

B-3 
TABLEBl (CONTINUED) 

HIGH DOSE CONTROL (UNTK )

05-0118

 LOW DOSE CONTROL (U NIB)

 05-0030

 LCH DCSE

 05-0032

 HIGH COSE 

 05-0112 

CICESTIVE SYSTE H 

(SALIVAR Y GLAH C

CABCINCSABCCM A

#IIVER

HEPATOCELLULAR ADENOMA

HEPATOCELLULA R CARCINOM A

HEMANGIOSARCOMA . UNC PRIB CB Mil

 (48)

 (48)

 2 (4X) 

6 (13*)

 1 (2!!) 

 (37)

 (46)

 12 (26%}

 (47)

 1 (2*) 

 (47 )

 20 (43*)

 (50) 

 (49) 

 36 (73!l) 

URINAR Y SYSTEB 

f KIDNE Y

TUBULAR-CEL L AIENOM A

 (49) (46) (47)

 1 (2*) 

 (50) 

INCOCRINE SYSTEB 

*EITUI1AEY

AEEKCI'A, NOS

tADBENA L

PHECCHRCBCCYTOM A

 (40)

 (44)

 1 (2«)

 (39)

 1 (3X)

 (44)

 (42)

 1 (2X) 

 (46)

 (42) 

 (38) 

1 (3$) 

#THYFOI D

AEENCCABCINCMA , NOS

 (45) (44)

 2 (5X) 

 (44) (46) 

REERCDUCTIVE SYSTEM 

*1ES1IS

IMEKS1IIIAL-CILL TUMOR

SEBINOBS/DYSGERBINOM A

 (48) (46)

 1 (2%) 

 (46) (49) 

2 (4X) 

NIBVOUS SYSTEB 

NON E 

SPECIAL SENSE OBCAN S 

*HARDERIAt v GLAND (49) (46) (4H) (50) 

» NUHBFR

* NUMBE R

 CE AHIBAL S

 CE ANIMAL S

 WITH TISSUE

 NECROPSIED 

 EXAMINE D MICHOSCOP ICALIY 

B-4 
TABLE Bl (CONTINUED) 

HIGH DOSE LOW DOSE 

COHIHOLtUNTR)

05-0118 CONTROL (UNTR) 05-0030 ICH DOSE 05-0032

 HIGH EOSH 

 05-0112 

PAPILLAR Y ACB1.CM A 1 (2X) 

PAPILLAB Y CYSTJDENOBA, NOS 1 (2%) 

MUSCOIOSKEIEIA I SYSTEM 

NONE 

BODY CAVITIES 

liCNE 

A L I CTHE B SYSTIM S 

NCN I 

SNIMBl DISPOSITION SUMMAR Y 

ANISAL S IMTIALL Y IB STUDY 50 50 50 50 

NATORA I DEATHS 1 3 1 

MCSIEUN E SACRIFICE 1 1 

SCHEEDIE E SACBIFICB 10 5 5 

ACCIDENTALL Y KILLE D 3 

TERMINA L SACRIFICE 39 11 46 43 

ANIMA L MISSIN G 1 

* NUBEE R CF ANIMAL S WITH TISSUE EXAMINE D MIC BO SCOP 1C ALL Y 

* I.UMBER Cf ANIMAL S NECROPSIED 

B-5 
TABLE Bl (CONCLUDED) 

HIGH DOSE LOW DOSE 

CONTROL (UNTB) CONTROL (UNTB) LCH ECSE HIGH COSE 

05-0 118 05-0030 05-0032 05-0112 

TUMO B SUMMAR Y 

TOTAL AMMAL S WITH PBISABY TUMORS * 22 19 28 12 

TOTAL PRIMAR Y TUMORS 26 26 32 5t 

TOTAL AMMAL S WIT H 8 BENIG N TUMORS 6 6 10 

TOTAL EENIC N TUMORS 8 7 6 10 

TOTAL AMCAL S HUH MALIGNAN T 1UMCRS 15 16 22 38 

TOTAL PALIGl.AliT TUMORS 17 19 26 uu 

TOTAL AMHAI S BITH SECONDARY TOMCES* 1 1 1 

TOTAL SECONEAR Y TUMORS 1 1 1 

TOTAL AMMAL S WIT H TUMORS UNCERTAIN ­

EENIGN 08 MALIGNAN T 

TOTAL UNCERTAI N TUMORS 

TOTAL AMMAL S KITH TUMORS UNCERTSIN ­

PRIHAR Y CR METASTATIC 1 

TOTAL UNCERTAI N TUMORS 1 

* PRIMAR Y TUMORS : ALL TUMORS EXCEP T SECONDAR Y TUMOBS 

I SECONDAR Y TUMORS : METASTATIC TUMORS OR TUMORS INVASIV E INTO AN ADJACENT ORGA N 

B-6 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH 2 AMINOANTHRAQUINONE 

HIGH DOSE ~ LOW DOSE 

CONTROL(UNTR) CONTROL (UNTR) 

06-0118 06-0030 

ANIMAL S INITIALL Y III STUD Y 50 50 

ANIMAL S NECROPSIED 50 «7 

ANIHAI S EXAMINE E HISTOPATHOLOGICALL I ** 5 0 47 

INTEGUHINTJR Y SYSTE M 

*SUECUT TISSUE (50) (147)

 

FIBROSARCOMA 

RESPIRATCB Y SYSTEM 

»IUN G (50) («6)

 

CABCIbCMA , NOS, METASTATIC 1 (2X)

 ALVECIAR/BBCNCHIOIA R ADENCM A 2 (4X)

 

ALVEOIAR/BRCNCHIOLA R CARCINOM A 1 ( 2%) 1 (2X)

 

HEHATCPCIETIC SYSTE M 

*MULTIPLI ORGAN S (50) (U7) 

MAUGHAM LYMPHOMA. NOS 2 (US) 2 («) 

NALIG.LYMPBOMA , HISTIOCYTIC TYP E 2 («X) 

•SPLEEN (49) (15)

 HEMANGIOSARCOM A 1 (2%)

 

ILYMP H NODE (44) (27)

 

MALIG . LYMPHOMA , HISTIOCYTIC TYP I 

tllVEF (50) (1*6) MAIIG.IYMPHOHA , HISTIOCYTIC TYPE 1 (2X)

 

tPEYERS PATC H (US ) (1*5)

 

MAIIG . LYMPHOMA , HISTIOCYTIC TYP I 

CIRCDIATCB Y SYSTE M 1 

i NONE 

I NUMEE R OF ANIMAL S WIIH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBER CE ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

L OU DOSE 

06-0032 

50 

1(8 

48 

(48) 

2 <4X ) 

(48) 

2 (4*) 

(43) 

1 <2X) 

4 (8X) 

(48) 

(39) 

(47) 

(48) 

1 (2X) 

« 

HIGH DOSE 

06-0112 

50 

49 

49 

(49) 

(48) 

2 (4X) 

1 (2*) 

(49) 

2 (4X) 

8 (16X ) 

(46) 

(36) 

1 (3X) 

(47) 

(39) 

1 OX) 

!*­

B-7 
DIGESTIVE SYSTEM 

tllVER 

CARCIKHA , NOS, METASTATIC 

HEPATOCELLDLA B CARCINOM A 

tSTOMAC H 

SGOAMCU S CILL PAPILLOMA 

URINAR Y SYSTE M 

NONE 

ENDOCRIN E SYSTEM 

tEITOITABY 

A CE (.CCA, NCS 

CHHOHOPHOEE ADENOM A 

tTHYROI D 

FCLLICULAB-CELL ADENOMA 

REPRODUCTIV E SYSTEM 

*BABKAH Y GLAN C 

FIBRCACENCB A 

*UIERUS 

ACEKCBA , NCS 

ADENOCARCINOMA , NOS 

LEIOMYOSARCOM A 

ENDOMETRIA L STROMAL POLYP 

tUTERUS/ENrOMETBIU M 

CABCIICMA,NC S 

tOVARY/OVIDUCT 

PAPILLARY AEENOM A 

INTRADOCTA L PAPILLOMA 

NERVOU S SYSTE M 

« NDMEE R OF ANIMAL S WITH TISSUE

» NUMBE R CI ANIMAL S NECROPSIED 

TABLE B2 (CONTINUED) 

HIGH DOSE LOW DOSE 

CONTROL (UNTB) CONTROL (UNTB) 

06-0118 06-0030 

(50) (46) 

1 UX) 

1 (2X) 4 (9X) 

(49) (45) 

(42) (37) 

1 (2X) 3 <8X) 

2 <5X> 1 (3X) 

(44) (44) 

(50) (47) 

1 (2X) 

(47) (43) 

1 (2X) 

(47) (43) 

1 (2X) 

(47) (43) 

1 (2X) 1 UX) 

1 (2X) 

 EXAMINE D MICROSCOPICALLY 

LOW DCSE 

06-0032 

(47) 

5 (11X) 

(48) 

1 (2X) 

(42) 

3 (7X) 

1 (2%) 

(42) 

(48) 

(43) 

1 (2X) 

(43) 

(48) 

HIGH COSE 

06-0112 

(47) 

12 (26 X) 

(46) 

(36) 

3 <8X) 

(42) 

1 (2X) 

(49) 

(46) 

1 <2X ) 

1 (2X) 

(46) 

(46) 

B-8 
TABLE B2 (CONTINUED) 

HIGH DOSE

CONTROLfUNTB )

06-0118

 LOW DOSE 

 CONTROL (U NIB)

 06-0030

 LCB DOSE

 06-0032

 HIGH COSE 

 06-0112 

SPECIA L SEKSE CBGABS 

*HAHDERIA K GLAN t

PAPIIIABY ADESCMA

 (50)

 1 (2X) 

 (47) (48) (49) 

MUSCULOSKELETA L SYSTEM 

NONE 

EOEY CAVITIES 

*ABDCHIHA L CAVIT Y

HEMAtiGIOSABCOB A

 (50) (47)

 1 (2*) 

 (48) (49) 

All OTHER SYSTJMS 

SITE DNKNOH N 

SARCCBA, DCS 1 

ANIMA L DISPOSITION SUMMAR Y 

ANIMALS INITIALLY IN STUDY

SATUBAI DEAtHS

MCBIEUN E SACBIFICE

SCHEDDLE D SACBIEICE

ACCIDENTALI Y HLIED 

TERMINA L SACRIFICE

ANIMA L MISSING 

a_INCIUEiS.AUICLYZt E ANiaAL 

 50

 2 

 10

 38

 50

5 

1

 5

 39

 50

2 

4 

 44

 50 

7 

5 

 38 

I NUMEE B

* NUMBE B

 C? ASIBAL S WITH TISSUE EXJUINE D

 Cf ANIMAL S NECROPSIED 

 BICBO SCOPICALLY 

B-9 
TABLE B2 (CONCLUDED) 

HIGH DOSE 

CONTROL (UNTR) 

06-0118 

LOW DOSE 

CONTROL (UNTR ) 

06-0030 

ICII DOSE 

06-0032 

HIG H tOSE 

06-0112 

TUMO B SUMMAR Y 

TOTAL ANIMAL S WITH PBIHABY

TCTA L FBIMAB Y TUMOBS

 TUMORS * 10

 11

 17

 21

 16 

 22 

29 

33 

TOTAL AMBAL S HITH BENIG N

TOTAL EENIG N TUMORS

 7 TUBCBS

7 

7 

8 

7 

8 

TCTSL AMFAL S KITH MALIGNAN T TUBCBS

TOTAL PALIGNAN T TUMOBS

 t

 1

 11

 13

 12 

 1<t 

25 

27 

TOTA L &MKAL S WITH SECONDAR Y

TCTAL SECONCAB Y TUMOBS

 TUHCBS * 1 

2 

TOTAL AMMAL S WITH TUMORS

EENIG N O R MALIGNAN T 

TCTAL UNCERTAI N TUMOBS 

 UNCEBTAIN ­

TCTAL AMMAL S WITH TUMOBS UNCERTJIN -

PRIflAB? CB METASTATIC 

TOTAL DNCEBTAI N TUMOBS 

* FBIMAB Y TUMORS : ALL TUMORS EXCEP T SECONDAR Y TUMOBS 

* SECONDAR Y TUMCBS : METASTATIC TUSOBS OB TUMOBS INVASIV E INTO AN ADJACE M ORGA N 

B-IO 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 2-AMINOANTHRAQUINONE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH 2-AMINOANTHRAQUINONE 

CONTROL (UNTH) LOW DOSE 

01-0037 01-0031 

ANIMAI S 1MTIAHY IN STUDY 50 50 

ANIMAL S NECROPSIED 46 49 

ANIMALS EXAMINE D HISTOPAT HOLOGICAILY ** 46 49 

INTEGUMENTAR Y SYSTEM 

*SKIN 

EPIDERMA L JNC1USION

INFLAMMATION , NECRO

FIBROSIS 

 CYST 

TIZING 

(46) <«9) 

1

1

1

 (2*> (2X) 

 (2%) 

*SUBCUT TISSUE 

FIBROSIS 

(46) (49) 

1 (2*) 

8ESPIHATCB Y SYSTEM 

#TRACHE A (45) (49) INFIABPATICN, SOS 9 (20X) 7 (14X) 

INFLAMMATION , ACHTE 1 (2X) 

INFLAMMATION , ACUTE/CHRONIC 

INFLAMMATION , CHRONIC 10 (22%) 8 (16%) 

*LUNG/BRONCHU S (46) (49) 

BRONCHIECTASIS 

INFLAMMATION . NOS 2 <4X) 

INFLAMMATION , FOCAL 1 (2X) 

INFLAMMATION , CHRONIC 8 (17X) 6 (12X) 

• BRONCHIA L MUCOU S GLA (46) (49) ABSCESS, NOS 1 (2%) 

NECRCSIS , NCS 1 (2X) 

HYPERPLASIA , ADENOMATOU S 1 (2X) 

tLUNG/BRCNCHICL E (16) (49) INFLA P fATICN , NOS 1 (2%) INFLAMMATION , FOCAL 1 (2%) 

tlUN G (46) (49) 

BIJERAIIZATJCN 2 (4X1 

* BOHEEB CF ANIMAL S WITH TISSUE EXJMNE D MICROSCOPICALLY 

* NUMBE R CI ANIMAL S NECBOPSIEC 

**EXCLDDES PARTIALLY AUTOLYZED ANIMALS
 

HIGH DOSE 

01-0032 

50 

47 

46 

(47) 

(47) 

(45) 

2 (4X) 

21 (47«) 

1 (2X| 

(46) 

2 (4J) 

(46) 

<t6) 

1 (2*) 

(46) 

C-3
 
ATEIF.C1ASIS 

CONGESTION, NOS 

EDEMA , NOS 

INFtAHMATION , NOS 

INFLAMMATION , FOCAL 

INFLAMMATION , INTEBSTITIAL 

INFLAMMATION , SDPPUBATIV E 

PNEUMONIA , CHBONIC MUBIN E 

INFLAMMATION , CHRONIC 

PERIVASCULITIS 

HYPERPLASIA , ADENOMATOU S 

IIUNG/ALVIOLI 

INFLACPATICN , NOS 

INFLABPATICB , FOCAL 

FIBROSIS 

HEBATOPOIET1C SYS1IB 

tSELEES


THBOHBCSIS, NCS
 FIBROSIS


INFARCT, HEAIED

RETICOLOCYTOSIS

HYPERPLASIA, HEMATOPOIETIC 

HYPERPLASIA , EEYTHROID

HYPERPLASIA , RETICDLUM CELL

HEMATOPOIESIS 

ERYTHROPOIESIS 

MYELOPOIESIS 

• ZYHPH NCE E

IkFLAPBATICN , NOS

HYPEBPLASIA , NOS

PLASMACYTOSI S 

HYPERPLASIA , RETICOLUM CELI

* SUBMANDIBULA R L.NODI

HIMORBHAG E 

tHEDIASTINA L L. NODE

PIASBACYTOSI S

tBENAL LYMPH NOCE

CCNGJS1IOJL._JJOS 

* NUMBER CF AKIMAL S WITH TISSUI * NUMBER CI ANIMAL S

 NSCBOPSIEE 

TABLECl (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE 

0 1-0037 01-0031 01-0032 

1 <2X) 1 (2X)

 1 (2X) 1 (2X)

 1 <2X )

 1 (2X) 1 (2X)

 

3 (756) 7 (11%)

1 (2X) 5 (10X) 

1 (2X)

 

1 (2X)

 

1 (2X)

 

5 1 <8X) I (2X) (

 11") 1 (2X) 

(46) (19) (H>) 

2 (IX) 

1 (2X) 

1 (2X) 

 (16)	 (18)

 

1 (2X)
 1 (2X)
 

1 (2X) 

1 ( 2X) 

1 (2«) 

 12 (26%) 10 (21X) (2X) 

8 ( 17X) 

3 (6X) 

8 (17X) 1 (2X) 

6 (13X) 1 (2X) 

 (38) (13) (10) 

1 (3X) 

1 (3X) 1 <3X) 

1 (2X) 

3 (8X) 

 (38) (13) (40) 

(2X> 

 (38) (13) (10) 

1 (3X) 

 (38) (13) (10) 

1 

 IXA6INE D MICBOSCOPICALLY 

C-4 
CIRCUIATOB Y SYSTEM 

*IYMEHATIC VESSELS 

INFLABFATICN . NOS 

• HEART 

FIBROSIS, FOCAL 

FIBROSIS, DIFFUSE 

PERIARTERITIS 

tMYOCARCID M 

INFLABBATICN , NOS 

INFLAMMATION , INTERSTITIAL 

INFlAMfATICN , ACOTE/CHRONIC 

INFLAMMATION , CHRONIC FOCAL 

FIBROSIS 

FIBROSIS, FOCAL 

*ACRTA 

INFLAMMATION , CHRONIC FOCAL 

PERIARTERITI S 

*COSCNARY ABTEBY 

HIMCRBFAG E 

*PULBONAB Y ABIIR Y 

HYPERTRCPHY, NCS 

DIGESTIVE SYSTEM 

tSAIIVAR Y GLAS t 

HEMOBRHAG E 

tPAROTID GLAN D 

HYPERPLASIA , AtENOMATOU S 

tllVEB 

FIBROSIS 

DEGENERATION , SOS 

NECROSIS, FOCAL 

NECROSIS, COAGDLATIVE 

HETAMORPHOSI S PATTY 

* NUMBE R CE 1NIBALS WITH TISSUE

* NUMBE R CE ASIMAL S NECROPSIED 

TABLE Cl (CONTINUED) 

CONTBOL(DNTB) 

0 1-0037 

(46) 

1 (2X) 

(46) 

(46) 

3 <2X ) 

22 (48X) 

3 <7X) 

7 (15X) 

(46) 

1 (2X) 

(46) 

(46) 

1 (2X) 

(38) 

(38) 

(46) 

3 (7X) 

1 <2X ) 

1 (2X) 

LOW DOSE 

01-0031 

(49) 

(49) 

(49) 

33 (67*) 

3 (6X) 

18 (37X) 

(49) 

1 <2X) 

(49) 

(49) 

(44) 

1 (2X) 

(44) 

1 <2X) 

(49) 

1 (2X) 

6 (12X) 

31 (63X) 

 EXAMIHE D MICBOSCOPICALLY 

HIGH DOSE 

01-003! 

(47) 

(46) 

2 (4t) 

1 (2X) 

1 (2«) 

(46) 

4 (9X) 

1 (2X) 

2 <4S) 

4 (9S) 

(47) 

(47) 

1 (2*) 

(47) 

(46) 

(46) 

(46) 

5 (11X) 

1 (2X) 

26 (57X) 

C-5 
TABLE Cl (CONTINUED) 

CONTBOL(UNTB) LOW DOSE HIGH DOSE 

01-0037 01-0031 01 -0032 

HYPERPLASIA , FOCAL 23 (SOX) 25 (51S) 27 (59X) 

HYPERPLASIA, DIFFUSE 1 <2X) 

ANGIECTASIS 1 (2X) 

ILIVER/PIFIPCBTAL (46) (49) (46) 

FIBROSIS 1 <2X) 

*BILE DUCT (46) (49) (47) 

INFLABBATICN. NOS 

HYPERPLASIA, NOS 

6 (13X) 

32 (70X) 

2

13

 (4X) 

 (27X) 5 (13X) 

HYPERPLASIA, FOCAL 1 <2X ) 7 (15X) 

IPANCBEA S (42) (47) (45) 

INFLABBATICN , NOS 10 (24X) 20 (43X) 2 (4X) 

INFLAMMATION , FOCAL 1 <2X) 

INFLAMMATION , ACUTE/CHHONIC 7 (16X) 

HYPERPLASIA , INTHADOCTA L 1 (2X) 1 (2X) 

*PANCREATIC DUCT (42) (47) (45) 

HYPERPLASIA , NOS 2 (4X) 

tPANCBEATIC ACINUS (42) (47) (45) 

ATBCPHY , NOS 4 (10X) 1 (2X) 2 (4X) 

tSTGBACH (45) (47) (46) 

EPIDEBBAL INCLUSION CYST 1 (2X) 

ULCER, NOS 2 (U%) 

INFLAMMATION, FOCAL 2 (4%) 1 (2X) 

HYPERPLASIA , NOS 6 (13X) 

HYPERPLASIA, EPITHELIAL 

HYPERPLASIA , FOCAL 

1

3

 <2X) 

 (6X) 1 (2X) 

HYPERKERATOSIS 

ACANTHOSIS 

1

1

 (2X) 

 (2X) 

tPEYFRS PATCH <t3) (47) (45) 

INFLABBATIC N FBOLIFERATI VE 1 (2X) 

HYPERPLASIA , NOS 7 (16X) 4 (9X) 

HYPERPLASIA, LYMPHOID 1 (2X) 

«COLCN (43) (46) (44) 

INFLABKATICN , NOS 1 (2X) 

INFLAMMATION , ACUTE 1 (2X) 

NEMATCD1&SIS 3 (7X) 4 (9X) 7 (16X) 

PARASITISM 2 (4X) 

* NOBBEH CF ABIBAL S HITH TISSUE EXAMINE D MICBOSCOPICALLY 

* NUMBER Cl ANIMAL S NECROPSIED 

C-6 
TABLE Cl (CONTINUED) 

COHTHOL(UNTB) LOW DOSE HIGH DOSE 

01-0037 01-0031 01-0032 

URINAR Y SYSTEM 

*KIDNE Y (46) (49) (46) 

CONGESTION. NOS 1 <2X) 

GLOMERULONEFHRITIS, NOS 33 (12%) 38 (78X) 37 (SOX) 

GLOMEROLONEPHRITIS, FOCAL 3 (7*) IN PL ASH AT ION, INTERSTITIAL 1 (2%) 1 <2X) 2 («*) 

INFLAMMATION . CHRONIC FOCAL 1 (2*) 

GRANULOMA, FOREIGN BODY 1 (2X) 

FIBROSIS 1 <2X) 

NBPHROSIS, NOS 7 (14X1 1 (2X) 

tUEINAPY BLACIEB (42) (17) (46) 

HEHORHHAGl 1 <2X) 

INFLAMMATION , NOS 1 (2%) 

HYPERPLASIA. EPITHELIAL <7X) 1 (2X) 1 (2X) ? 

ENEOCEIN E SYSTID 

IEITHITABY (41) (46) <43) 

HYPERPLASIA , NOS 3 (7X) 

HYPEBPLASIA, FOCAL 6 (14X) 

HYPERPLASIA , C HROHOPH QBE-CELL 2 (5X) 

IADRENA L (43) (48) (45)

BBTAMCBrHOSIS FATTY 3 (6X) 2 (4x;i 

HYPEHPLASIA , NODULAR 1 <2X) 

fACBENS L CCSTIX (43) (48) (45) 

NCDDLI 2 (4X) 

NECROSIS, ICCAl 1 <2X) 

HYPERTROPHY , FOCAL 1 (2X) 

HYPERPLASIA , NODULA R 1 (2X) 

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

HYPERPLASIA, FOCAL 1 (2X) 

(ADRENA L EEDULL A (43) (48) (45) 

NECROSIS, NOS 1 <2X) 

CALCIFICATION, NOS 1 (2X) 

HYPERPLASIA , NODULAR 1 (2X) 

HYPERPLASIA, NOS 6 (1UX) 3 (6X) 

HYPERPLASIA , FOCAL 3 (7X.I 

tTHYROIE (45) (46) (40)

 

INFLABtATICN . fOCAL 

* NUMBE R CF ANIMAL S WITH TISSUE EXAHNE D MICROSCOPICALLY 

» NUMBER CI ANIMALS NECHOPSIED 

C-7 
TABLE Cl (CONTINUED) 

— — -' — -.-.I •- -r .- -. I — -.—•—— ____»—« 

CONTROL (UNTB) LOW DOSE HIGH DOSE 

01-0037 01-0031 01-0032 

HYPEBPIASIA , AtENOMATOD S 1 (2X) 1 (3%) 

HYPEBPIASIA, C-CELL 1 (2X) 2 (*X) 

IPABATHYBCI D (32) (28) (32)

 

HYPERPLASIA , NOS 1 (IX)

 

tPANCBEATIC ISLETS (12) (17) (15) 

HYPEBPLASIA, NOS 2 <5X) 3 (6X) 

HYPEBPIASIA , FOCAL 1 <2X) 

REPRODUCTIV E SYSTEM 

*MABMAB Y GLAN t (16) (19) (17) 

GALACTCCELI 2 (IX) 

HYPERPLASIA , NOS 5 (11X) 3 (6X) 1 <2X) 

*PBEEUTIAL GLAN C (16) (19) (17) 

ABSCESS, NCS 1 <2X ) 1 (2%) 

HYPERPLASIA , NOS 1 (2X) 

tPROSTATE (15) (17) (41) 

INFLAfHATICN , NOS 21 (17X) 21 (15X) 5 (11%) 

INFLAMMATION , FOCAL 3 (7X) 1 <2X) 1 <2X) 

INFLAMHATION , INTERSTITIAL 1 (2X) 

INFLAMMATION , ACUTZ 3 (7X) 

INFLAMMATION , ACUTE FOCAL 3 (7%) 

INFLAMMATION , ICUTE/CHBONIC a (9%) 

HYPERPLASIA , NOS 1 <2X) 

HYPERPLASIA , FOCAL 5 (11%) 3 (6X) 

HYPEBPLASIA , PAPILLAR Y 2 (IX) 

METAPLASIA , SQUAMOUS 5 (11%) 1 <9X) 2 (5%) 

I1ESTIS (US) (17) (16)

MINEBALIZATIC N 18 (38X) 2 (IX) 

DEGENEBATION, NOS 32 (70X) 

A1BOPHY, NOS 2 (IX) 29 (62X) 3 (7%) 

ASPEBHATO GENES IS 1 (2%) 

HYPERPLASIA , INTERSTITIAL CELL 19 (12X) 30 (6«X) 10 (22X) 

ITESTIS/1UBULH (15) (17) (16) 

DEGENIBATICN , NOS 6 (13X) 2 (IX) 1 (2X) 

NEBVCU S SYSTEM 

*CEBHBBU K (11) (18) (16) 

HgsqEss, lie? , 1 (2%) 

* NOHBE B Cl ANIMAL S WITH TISSUE EXAMINE D HICBOSCOP 1CALLY 

* KUBBEB 01 ASIMALS NECROPSIED 
TABLE Cl (CONCLUDED) 

COSTOOLfDHTR) LOW DOSE HIGH DOSE 

01-0037 01-0031 01-0032 

SPECIAL SEKSE CBGAN S 

NONE 

MUSCULOSKELETA L SYSTEM 

• CARTILAGE,HG S (46) (19) (ll) 

CYST, NOS 1 (2*) 

BOEY CAVITIES 

NON E 

All OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOG Y SUMMAR Y 

AUTO/NECROPSY/HISIO PERF

AUTC/MCRCr£Y/H O HISTO

 1 1 

1 

AUTCLYSIS/N C NICSOPSY 1 1 3 

i NUMBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER Cf ANIMAL S NECROPSIED 

C-7 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH 2-AMINOANTHRAQUINONE 

CONTBOL(UNTB) DOSED 

02-0084 02-0078 

ANIBJLS INITIALL Y IN STDDY 25 50 

ANIBAI S KECBOPSIE E 23 13 

ANIBALS IXAMNE r HISTOPATHOLOGICALLY ** 23 13 

INTEGUBENTABY SYS1IB
 

NOME
 

RESPIBATOBY SYSTEB 

*LABYN X (23) (i*3) 

INFLABBATIC N ACDTI AN D CHBONIC 1 («X) 

INFLABBATION, CHBONIC 3 (13X) 

ITHACHF.A (5) (36) 

IKFLA f PATICN , NOS 1 (3X) 

tLONG/EBONCHOS (23) (U2) 

INFLAtKATICN , NOS 

HUNG (23) 

BINEBALIZATION 1 (2X) 

INFLABBATICN , INTERSTITIAL 3 (13X) 12 (29X) 

INFLABBATICN , NECROTIZING 1 (2X) 

PNEOBONIA, CHBOHIC SOBINE 8 (35%) 

GRANULOMA , FOREIGN BODY 1 (IK) 

CALCIFICATION, FOCAL 1 (IX) 

HYPERPLASIA , EPITHELIAL 1 (IX) « (10X) 

*IUNG/ALVECLI (23) (t2) 

niNFBALIZATIC N 1 (2X) 

FIBROSIS, BOCAI 1 (2X) 

HZBATCPOIBTIC SYSTEB 

IEONE HABBO H (22) (35) 

BYPIBFLASI J...H IB.ATOEO.IJTIC .__J_J5S1_ 

t NUHBEB Cf ANIHAL S KITH TISSUE EXJHINE D HICHOSCOPICALLY 

» BDBBEB CI ANIMAL S NECBOPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

c-io 
ISPLEE N 

HEBATCtA, NCS
 

HEBCSIDEROSIS
 

HYPERPLASIA, HEMATOPOIETIC
 

HYPERPLASIA, EBYTHROID
 

HEMATOPOIESIS
 

*IYMP H NCt £ 

IKFLAKHATICN , NO S 

HETICUICCYTOSIS 

LYMPHOCYTOSIS 

PLASMJCYTOSIS 

CIRCUIATOBY SYS1SB
 

*(YCCABEIU B 

MINERALIZATIO N 

INFL4HKATICN , INTERSTITIAL 

FIBROSI S 

DEGENERATION , NOS 

*ACRTA 

BINERftLIZATIO N 

DIGESTIVE SYSTJM
 

ILIVER 

CCNGESTICN , CHBONIC PASSIV E 

FIBROSIS, FOCAL 

CHOLSNGIOFIEROSIS 

FIBROSIS SEPTAL LIVES 

METAMORPHOSI S FATTY 

BASOPHILIC CYTO CHANGE 

HYPERPLASIA ,

*BILE DUCT 

INFLAJtATICN ,

HYPERPIASIA ,

fEANCREA E 

INFLAPfATICN ,

IS10MACH 

 FOCAL 

 NOS 

 NCS 

 NO S 

TABLE C2 (CONTINUED) 

DOSED 

02-0081 02-0078 

(23) (39)
 1 (IX)
 

2 (9X) 18 (16X)
 

3	 (13X) 7 (18X)
 

7 (18X)
 3 (13X)
 

(21) (29) 

1 (3X) 

2 (7X) 

2 (7X) 

1 (3X) 

(23) (U1 ) 

1 <2X) 

1 (4X) 9 (22J) 

9 (22X) 

U (17X) 

(23) (13) 

1 (2%) 

(23) (12) 

1 (UX) 

1 (2X) 

1 (US) 

1 <2X) 

2 (9X) 1 (2X) 

(4 (17*) 

3 (13X) 8 (19X) 

(23) (13) 

1 (2X) 

2 (9«) 9 (21X) 

(22) (31) 

2 (6X) 

(23) (31) 

5_J15X1__ 

* BDMBE R Cf ANIMAL S WITH TISSUE EXAMINE D MICfiOSCOP ICALLY 

* NUMBE R CT ANIMAL S NECROPSIED 

C-l 
IkFLAPfATICN , NO S 

HECROSIS, NOS 

HYPERPLASIA , E»SAL

HYPERKERATOSIS 

ACANTHOSIS 

tGASTRIC MUCOSA 

MINERALIZATIO N 

• GASTRIC MUSCULARI S 

MINERALIZATIO N 

tPEYEH S PATCH 

HYPERPLASIA , NOS 

tCOLON 

PA8ASI1ISM 

*HECTU M 

HYPERKERATOSI S 

URINAR Y SYSTE M 

*KIDNE Y 

BINEEAIIZATION 

 CEIL 

GLOME8ULONEPHBITIS , NOS 

PYELONEPHRITIS , ACUTE 

PYELONEPHRITIS , CHRONIC 

GRANULOMA , FOREIGN BODY 

FIBROSIS, DIFFUSE 

NEPHROSIS, NOS 

GLOMERULOSCLEROSIS, NOS 

CALCIFICATION , FOCAL 

PIGMENTATION , NOS 

METAPLASIA , NOS 

REGENERATION, NOS 

tKIDNEY/TUBUL F 

MINERALIZATIO N 

CYST, NCS 

INFLAMMATION , NOS 

NEPHROSIS , NOS 

NECROSIS, NOS 

TABLE C2 (CONTINUED) 

CONTROL(UNTR ) 

02-0084 

(23) 

(23) 

(23) 

U (17X) 

(22 ) 

2 (9%) 

(23) 

(23) 

4 (17X) 

1 (4X) 

1 (4X ) 

10 (»3X) 

1 <4X) 

(23) 

1 (KX) 

DOSED 

02-0078 

1 (3*) 

2 (6X) 

1 (12X) 

1 <3X) 

3 (9X) 

(31)

6 (181) 

(34) 

4 (12X) 

(33) 

2 (6X) 

(30) 

(43) 

1 (2X) 

(43) 

3 <7X) 

24 (56X) 

16 (37X) 

19 (44X) 

2 (5J) 

H (9X) 

4 <9X) 

1 (2X) 

(43) 

1 <2X) 

1 (2X) 

1 (2X) 

2 (5X) 

f NUMBE R CI ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBER CF ANIMAL S NECROPSIED 

C-I2 
TABLE C2 (CONTINUED) 

COHTBOL(UNTH) DOSED 

02-0084 02-0078 

tURINABY E1ADDEB (22) (34) 

HYPERPLASIA , JflTHELIAL <3X) 

ENDOCRIN E SYSTE M 

*PItOITAR Y (21) (30) 

HEMCBEHAGIC CYST 1 (5*)

 

HYPERPLASIA , FOCAL 

tADRENA I CEDULL A (23) (39) 

HYPERPLASIA , NODULiR 1 (3X) 

HYPERF1ASIA, FOCAL 1 (3X) 

#THYBOIC (21) (29) 

HYPEBPIASIA , C-CELL 3 (14X) 1 (3X) 

tPARSTHYROI D (9) (14) 

HYPERPLASIA , NCS 2 (14X) 

REPRODUCTIV E SYSTEM 

"PAflBAB Y GLAN t (23) (43) 

GALACTCCEL I 1 (4X) 1 <2X) 

HYPEBPIASIA , NOS 1 (4X) 9 (21X) 

LACTATION 9 ( 39X) 

* VAGIN A (23) (43) 

INFLAKMTICN , NOS 1 <2X) 

*U1ERUS (23) (33) 

PYOKETB A 3 (13X) 

HYPEBPLASIA, ADENOMATOUS 1 (3X) 

tUTEBUS/HSCCHITBIU H (23) (33) 

INFIAt^ATICN , NOS 1 (IX) 4 (12X) 

INFIAHPATICN , SUPPURATIV E 1 <3X) 

INFLAMMATION , NECROTIZING 1 <3X) 

INFLAMMATION , CHRONIC 1 («X) 

HYPERPLASIA , NOS 1 (IX) 1 (3X) 

HYPERPLASIA , CYSTIC 1 (4X) 

ICVABY/CVItUCI (23) (33) 

* HUBEE F CF ANIMAL S WITH TISSUE EXAflHE D MICROSCOPICALLY 

» NUMBE R CE ANIMAL S NECROPSIED 

C-I3 
TABLE C2 (CONCLUDED) 

CONTROL (UNTR )

02-0081

 DOSED 

 02-0078 

IKFIAffATICN ,

INFLAMMATION ,

ABSCESS. SOS

 SDEPURiTIVB

 ACUTE 1

 1

 (M) 

(14%) 

1 (3)1) 

tCVAEY

MINIBALIZATIO N

CYST. NOS

INFLAMMATION , NOS

 (22)

 3 (14X)

 (34) 

1 (3%) 

1 (3X) 

1 (3X) 

NJBVOUS SYSTE M 

tBRAI N (23) (36) 

HYDROCEPHALUS , NOS 1 (4X) 

HEMOFBhAG E 1 (I*) 

CALCIFICATION, FOCAL 1 (tX) 

SPBCIA L SENS E ORGAN S 

NON E 

HUSCULOSKEIETA I SYSTEM 

NONE 

BODY CAVITIES 

*PLEOBA (23) (13) 

NEC8OSIS, NOS 1 (28) 

A LL OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOG Y SUMMAR Y 

AUTO/NECBOPSY/BISTO PERF 1 

AUTCLYSIS/hC NBCBOPSY 2 7 

I NOMEE R CF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALL Y 

* MJHBER CF ANIMAL S NECROPSIED 

C-14 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 2-AMINOANTHRAQUINONE
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH 2-AMINOANTHRAQUINONE

 

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

05-0118 05-0030 05-0032 

SNIMAI S IMTIAII Y IN STUDY 50 50 49 

ANIMAL S MISSIN G 1 

ANIMAL S NECROPSIE D 49 46 48 

ANIHAI S EXAMINE D HI STOPATHOLOGICALL Y ff 49 46 47 

INTEGUMENTAR Y SYS1EM 

*SKI« (49) (46) (48) 

INFLAMMATION , NOS 1 (2») 

INFLAMMATION , FOCAL 3 (6%) 1 (21) 

INFLAMMATION , NECROTIZING 1 (2X ) 

GRANULOMA , PYOGENIC 1 (2X) 

RESPIRATOR Y SYSTE M 

• TRACHE A (48) (46) (45) 

INFLACHATICN , NO S 

tlUNG/BRCNCHU S (49) (46) (45) 

INFLACPATICN , FOCAL 1 (2%) 1 (2%) 2 (4S) 

IIUNG/BRCNCHICL H (49) (46) (45) 

IKFLAP.KATICN , FOCAL 1 (2% ) 

ILUN G (49 ) (46) (45) 

EWHYSIBA , NO S 1 (2X) 

HEMORRHAG E 1 (2%) 2 (4X) 

INFLAMMATION , FOCAL 1 (2X) 

INFLAMMATION , INTERSTITIA L 10 (20%) 7 (15JS) 12 (21%) 

HYPERPLASIA , EPITHELIA L 1 (2%) 

HIUNG/AIVECL I (49) (46) (45) 

INFLACHTICN , NO S 1 (2%) 

HEPATCPCIE1IC SYS1E M 

•SPLEE N (49) (46) (46) 

. . INFLAPKATICN , NOS J J2XL. .... 

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

* KUHBE R CF ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

. 

HIG H DOSE 

05-0112 

50 

50 

50 

(50) 

(49) 

1 (2X) 

(SO) 

(50) 

(50) 

1 (2%) 

7 (14J) 

2 (4X) 

(50) 

(49) 

D-3 
TABLE Dl (CONTINUED) 

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

05-0118 05-0030 05-0032 05-0112 

FIBBCEI S 1 (2X) 

HYPEBPLASIS , NOS 6 (12X) 21 (46X) 10 (20X) 

RETICOLOCYTOSIS 1 <2X ) 

HYPEBPLASIA , HEMATOPOIETIC 5 (10X) 6 (13X) 6 (12X) 

HYPEBPLASIA , EEYTHROID 1 (2X) 3 (7X) 2 (4%) 

HYPERPLASIA , RETICULU M CEL L 1 (2X) 

HYPERPLASIA , LYMPHOID 1 (2X) 2 (4X) 2 (4X) 

HEHATOPOIESIS 1 (2X) 

ILYME H NCE E (42) (34) (36) (42) 

HEMORRHAG E 1 (3X) 1 <3X) 1 <2X ) 

INFLAMMATION , NOS 10 (24X) 19 (53X) 14 (33X) 

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

RETICULOCY10SIS 2 <5X ) 5 (14X) it (10X) 

LYMPHOCYTOSI S 5 (14X) 4 (10K ) 

PLASMACYTOSI S 1 (2X) 

MEGAKABYOCYTOSI S 1 (3X) 

HYPERPLASIA , HEMATOPOIETIC 1 (2%) 

HYPERPLASIA , RETICULUM CELL 2 <6X) 

HYPERPLASIA , LYMPHOI D 3 (7X) 5 (14X) 3 (7X) 

tCANDIBULA B L. NCt E (42) (34) (36) (42) 

HYPERPLASIA . EJTICULUM CELL 1 (3X)

 

1MESENTERIC L. NODE (42) (34) (36 l (42)

 

THROPBCSIS , NCS 1 (3X)

 

HEMORRHAG E 1 <3X )

 

CIRCUIATOF Y SYSTE M 

*CABCICVASCULAB SYSTE (49) (46) (48) (50) 

PEHIVASCULITI S 1 (2%) 

#HEAR T (49) (46) (46) (49) 

MINEBALIZA1IC N 1 (2%)

 

tMYOCARDIU M (49) (46) (4 6 | (49) 

INFLAPBATICd . NO S 1 <2X ) 1 (2X ) 

FIBROSIS 1 (2X) 

tENDCCARIIU M (49) (46) (46| (49)

 

H,FLAM«ATICB PfOLIFERATI VE 1 (2%)

 

*AOR1A (49 ) (46) (4 8I (50) 

t NUMBE R CF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUHBE B CJ ANIMAL S NECROPSIED 

D-4 
TABLE Dl (CONTINUFD) 

HIGH DOSE LOW DOSE 

CON180L ( UNTB ) CONTRO L (UNTB) L OW DOSE 

05-0118 05-0030 05-0032 

DIGESTIV E SYSTE M 

• SAIIVAB Y GLAN C (1*8) (37) (1*7) 

PERIVASCULA R CUFFIN G 5 (14S) 

•LIVER (48) (1*6) (1*7) 

HINERAIIZATIC N 1 <2X) INFLAMMATION , NECROTIZIN G 1 (2%) 

FIBROSIS, FOCA L 1 (2X ) NECROSIS , FOCA L 9 (19X ) 2 (U% ) 16 (3UX ) 

NECROSIS , COAGULATIV E 

NECROSIS , hEMORRHAGI C 1 (2%) 

METAMORPHOSI S FATT Y 8 (17% ) 3 (6X) CYTOPLASMI C VACUOLIZATIO N 2 (U% ) 

HYPERPLASTI C NODUL E 1 (2%) 10 (21X ) HYPERPLASIA , NOS 1 (2%) 

HYPERPLASIA , FOCAL 1 (2X) 

*GALIBlACrE B (19) («6) (48) 

INFIAKFATICN , NOS 1 (2%) 

*BILE CUC T (49) <U6 ) (1*8) 

LYMFHCCYTI C INFLAMMATOR Y INFILT R 1 (2%) HYPERPLASIA , NOS 1 (2X) 

• PANCREA S (1*7) (4H) (45) 

I1.FLAF11ATICN, NOS 1 (2X ) 3 (7X) INFLAMMATION , FOCA L 2 (5X) 

INFLAMMATION , INTERSTITIA L 1 (2%) INFLAMMATION , ACOTE/CHRONIC 1 (2X ) DEGENERATION , CYSTIC 1 (2X) HYPERPLASIA , FOCAL 2 (5%) 

• PANCREATI C CDCT (47 ) (1*4) (45) 

HYPERPLASIA , NC S 1 (2X) 

IEANCBEATI C ACINU S (1*7) (Kit) (45) 

NECROSIS , NCS 1 (2%) ATROPf-Y , FOCA L 1 (2%) 

ISTCMACH (1*8) (1*5) (46) 

INFLAFfATICN , NO S 2 (dX) 7 (15%) 

INFLAMMATION , FOCAL 2 (i*%) 3 <7X ) 

_ INFLAMMATION , ..NECROTIZING 1 ±2%l 

* NUMBE R CE ANIMAL S DITH TISSU E EXAMINE D MICROS COP 1C AL LY 

* MJHBER CE AMM&L S NECROPSIE D 

HIG H EOSE 

05-0112 

(50) 

(U9) 

5 (10X ) 

1 (2X) 

6 (125!) 

(50 ) 

(50) 

(45) 

(45) 

(45) 

(47) 

2 (UX ) 

5 (11)1) 

0-5 
TABLE Dl (CONTINUED) 

IkFLAI'fATICN , SCUTE 

HYPERPLASIA , EPITHELIAL 

HYPERPLASI A FOCAL 

HYPERPLASIA , ADENOMATOUS 

HYPERKERATOSI S 

ACANTHOSI S 

IGASTRIC MUCOSA 

HYPOPLASIA , JOCAL 

tEEYEHS PATCH 

HYPERPLASIA , NO S 

KCOLC N 

NHMATCEIASI S 

PARASITIS M 

URINAR Y SYSTEM 

«KIDNE Y 

GIOMEEDLONEPHPITIS , NO S 

GLOMERDLONEPH8ITIS , FOCAL 

INFLAMMATION , INTERSTITIAL 

HYPERPLASIA , TUBULA R CELL 

METAPLASIA , OSSEOUS 

tKIDNEY/TUBUL E 

MINEBALIZATIC N 

IKIBSEY/EF.LVI S 

INFLAKPAT I CN , ACUTE/CHRONIC 

• URINAR Y ELADDE B 

INFlAfCATICN , NO S 

HYPERPLASIA , EPITHELIA L 

ENDOCRIN E SYSTE M 

tEITUITAE Y 

HYPERPIASIA , NO S 

HYPERPLASIA , FOCAL 

tADRENA L 

.NECROSIS.. FOCA1 

t liUHEE H CF ANIMAL S KITH TISSU E

* NUMBE R CF ANIMAL S NECROPSIEE 

HIGH DOSE LOW DOSE 

CONTROL (UNTH ) CONTROL (UNTR ) LOU E'OSE 

05-0118 05-0030 05-0032 

1 <2X) 

2 (4X) 

1 (2%) 2 (4X) 

2 (4X) 

1 (2%) 1 <2X ) 

1 (2X) 1 (2X) 

(48) (45) (46) 

5 (11X) 

(49) (46) (47) 

7 (114X) 3 (6X) 

C*3) (41) (45) 

3 (7X) 

(49) (46) (47) 

2 (IX) 2 (4%) 

1 <2X ) 

16 (33%) 7 (15X) 31 (66X) 

1 <2X ) 

(49) (46 ) (47 ) 

1 (2X) 

(49) (46) (47) 

3 (7*) 

(48) (46) (47) 

1 (2X) 

4 (8%) 2 (4X) 7 (15X) 

(40) (39) (42) 

1 (3X) 

(44) (44) (46) 1 J2XJ. 

 BXAC.INE D MICROSCOPICA LIY 

HIGH COSE 

05-0112 

2 <«X ) 

5 (11% ) 

(47) 

(47) 

9 (19% ) 

(43) 

4 (9% ) 

(50) 

12 (24% ) 

1o (325!) 

1 (2X ) 

(50) 

(50) 

(48) 

7 (15% ) 

(42 ) 

1 (2% ) 

2 (5%) 

(33) 

D-6 
TABLE Dl (CONTINUED) 

HIGH DOSE CONTEOL(UNTB) 

05-0 118 

LOW DOSE 

CONTROL (ONTR) 

05-0030 

HYPEBEIASIA , NCS 3 (7X > 

• ADRENAL/CAPSUL E 

NODDL E 

HYPERPLASIA , NOS 

(44) 

3 <7X) 

(44) 

• ADRENA L CORTE X 

NCDUL F 

NECROSIS, FOC

HYPEHPLASIS ,

HYPERPLASIA ,

A L 

 NOS 

 FOCA L 

(44) (44) 

2

14

 <5X) 

 (32X) 

• THYROI D 

HYPERPLASIA ,

HYPERPLASIA ,

 FCCAL 

 PAPILLAR Y 

(15) (44) 

• PARATHYRCI D 

CYST, NCS 

(24) (17) 

1 (6*) 

• PANCREATI C ISLE

HYPERPLASIA ,

TS 

 NCS 

(47) (44) 

REPRODUCTIV E SYSTE M 

"PREPUTIA L GLAN C (49) (46) 

ABSCISS, NCS 1 (2%) 

• PROSTATE («9) (46) 

HYPESELASIA , EPITHELIAL 1 (2X) HYPERPLASIA , FOCAL 

• TESTIS (48) (46) ATRCPHY , NO S 

ATROPHY , FOCA L 

HYPERPLASIA , INTERSTITIAL CELL 

•TESTIS/TUBULE («8) (46) 

DEGENERATION , NO S 3 (7X) 

*EPIDIEYfI E (49) (46) 

OFLABPATICN , NOS 1 <2X ) 

NERVOU S SYSTEM 

• ERAI N (49) (46) PERIVASCULITI S 

* NUHEE R CE ANIMAL S WITH TISSUE EX8KINE D MICROSCOPICALLY 

* NUMBER CI ANIMAL S NECROPSIED 

LCH DCSE 

05-0032 

(46) 

5 (11X) 

(46) 

1 <2X) 

1 (2%) 

(44) 

1 (2X) 

(17) 

(45) 

(48) 

(46) 

2 <4X) 

(46) 

1 (2X) 

10 (22X) 

(46) 

(48) 

(46) 

HIGH tOSE 

05-0112 

(38) 

1 <3X > 

1 <3X) 

(38) 

(46) 

1 (2X) 

1 <2X ) 

(22 ) 

(45) 

2 (4X) 

(50) 

(43) 

1 (2X ) 

(49) 

2 (1%1 

8 (16)1) 

(49 ) 

1 (2X) 

(50) 

(49) 

1 -(2XL 

D-7 
TABLE Dl (CONCLUDED) 

HIGH DOSE COJITBOL(UNTB )

05-0118

 LOW DOSE CONTROL (0 NTS)

 05-0030

 LCW DOSE

 05-0032

 HIG H DOSE 

 05-011^ 

liECFCSIS, NOS 1 (2%) 

SPECIA L SEliSE CBGAN S 

NONE 

MUSCDLOSKELETA L SYSTEM 

NCK E 

BCD Y CAVITIE S 

*AEDOHINA L CAVIT Y

MCBCSIE, FA T

 (U9) (16)

 1 (2X) 

 (U8) (50) 

SLl OTHES SYSTEM S 

ADIEOS E USSLE 

IBFLABHATICN . ACUTE 1 

CHENTU M 

SKECSIS. EAT

CALCIEICATICN . NOS

 1 1 

1 

SPECIAL PCFPHCLCG Y SUHMAE Y 

N C LESIC N 5EECETE D

AMBA I BISSING/NO NECROPS Y

ACCIDENTAL DEATH AUTO/NECBOPSY/HO HISTO

AUTOLYSIS/NC KECBOPSY

* NDMEE E OF ANIBAL S WITH TISSUE EXAMINE D

* NUMBE B CE ANIMAL S NECROPSIED 

5 2 

1 

3 

1

 MICROSCOPICALL Y 

1 

 .! 

0-8 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH 2-AMINOANTHRAQUINONE 

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

06-0118 06-0030

ANIBAI S IUTIAIIY IN SIDDY 50 50 ANIMAL S NECEOPSIEC 50 47

ANIMAL S EXAMINE D H ISTOPAT HOLOGICA11Y ** 50 47

IN1EGCMENTAR Y SYSTE M 

*SUBCUT TISSUE (50) (47)

ABSCESS, NOS 1 (2X ) 

RESPIRATOR Y SYSTE M 

• LUNG/EEONCHU S (50) (46) INFLAPKATICN , FOCAL 1 (2%) 

IIUNG/BRCKCHICL E (50) (46)

HYPERPLASIA , NC S 1 (2X) 

*LUNG (50) ( 46 ) INFLABCATICN , INTERSTITIA L 14 (28%) 2 (4JS)

HYPERPLASIA , EPITHELIA L 

•IUNG/ALVECL I (50) (46)

EPPHYSEfii , NO S 1 <2X) 

HECATCPCIEII C SYSTE M 

IEON E MARRO W (49) (45)

HYPCPIASIA , NO S 1 (2X) 

MYELOFIEROSI S 1 (2X) SEGAKARYOCYTOSI S 

ISPLEES (49) (45)

HYPERPLASIA , NCS 9 (18X ) LYMPHOCYTOSI S 2 (4X) 

HYPERPLASIA , fiEH ATOPOIET 1C 6 (12%) 

HYPERPLASIA , ERYTHROI D 

. HYPERPLASIA ^ LYMPHOI D 2 _(4%]_ 3 (7X) 

* NUMBE R CE ANIMAL S HITH TISSU E EXAMINE D MICROSCOPICALL Y * NUMBE R Cf A M HALS NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

 LOW DOSE 

 06-0032 

 50 

 48 

 48 

 (48) 

 (48) 

 (48) 

 (18) 

5 (10X) 

1 (2*) 

 (48) 

 (46) 

 (48) 

18 (38X) 

11 (23X) 

HIG H DOSE 

06-0112 

50 

49 

49 

(49) 

(48) 

(48) 

(48) 

7 (15)!) 

1 (2)!) 

(48) 

(42) 

2 (5X) 

1 (2%) 

(46) 

9 (201!) 

7 (15*) 

7 (15*) 1

 J2J1 

D-Q
 
HEBATCTCIE£I S 

#HEBCLYBP H NOTE S 

INFLAMMATION , NOS 

HYPERPLASIA , NOS 

IIYBFH NCC E 

HEMORRHAG E 

INFLAMMATION , NOS 

NECROSIS, NOS 

HYPERPLASIS , NOS 

RETICDLOCYTOSIS 

PLASMACYTOSIS 

HYPERPLASIA , tEM ATOPOIET 1C 

HYPERPLASIA , LYMPHOI D 

• PANCREATI C L.NOt E 

HYPERPLASIA , BHTICULU M CELL 

CIBCUIATCF Y SYSTE M 

*CABDIOVJSCULA R SYSTE 

FERIVfSCULITI S 

#MYOCARDIU M 

INFLAF f ATICN, FOCAL 

FIBROSIS, FOCAL 

DIGESTIV E SYSTEM 

ISALIVAE Y GLAN t 

PERIVASCOLA E CUFFING 

ILIVER 

NECSCSIS, FOCAL 

NECROSIS, COAGULATIVE 

METAMORPHOSI S FATTY 

HYPERPLASIS , NODULA R 

HYPERPLASTI C NODUL E 

_ -HJPERPLASIA^.FQCA L __ 

t NUMBER CI ABIBAL S WITH TISSUE

» NUHBE R CE ANIMAL S NECROPSIED 

TABLE D2 (CONTINUED) 

HIGH DOSE LOW DOSE CONTROL (OKIE) CONTROL (U NTR) 

06-0118 06-0030 

2 (IX) 

(U9) (15) 

2 (IX) 

1 (2X) 

(1") (27) 

9 (20X) 

3 (7X) 

1 <2X ) 

1 <2X ) 

« (9X) 

(11) (27) 

1 (IX) 

(50) (17) 

(50) (16) 

1 (2X) 

(18) (29) 

3 (6X) 1 (3X> 

(50) (16)

7 (11X) 1 (2X) 

1 (2X) 

2 (HX) 

 EXJMINE D MICHOSCOPICSLLY 

LCW DOSE 

06-0(132 

(18) 

(39) 

1 (3X) 

18 (16X) 

1 (3X) 

1 (3X) 

(39) 

(18) 

(17) 

1 (2X) 

(18) 

(1*7) 

11 (23X) 

2 (IX) 

1 <2X) 

1 J2SL 

HIGH EOSE 

06-0112 

(1*6) 

(36) 

7 (19X) 

2 <6X) 

(36) 

(19) 

1 <2X) 

(1*9) 

(11) 

(17) 

5 (11X) 

5 (11X) 

D-IO 
TABLE D2 (CONTINUED) 

ASGIEC1ASIS 

HEMSTOPOIESIS 

mVER/PERIPCBTAL 

HYPERPLASIA , LYMPHOI D 

•GALLBLADDER 

INFLAPPATICN , NO S 

tEANCREA S 

INFLAHPATICN , NO S 

IPANCREAII C EUCT 

LYMPHOCYTIC INFLAMMATORY

&STOMAC H 

IKFLABfATICN , NOS 

INFLAMMATION, FOCAL 

HYPERKEBATOSI S 

ACANTHOSI S 

• GASTRIC MUCOSA 

HYPERPLASIA , EOCAL 

IPEYERS EATCH 

HYPERPLASIA , NOS

 

lEUOCEKU H 

ECTCPIA 

*COLON 

PARASITIS M 

URINAR Y SYSTE M 

* KID KEY 

GLOMEBULONBPHBITIS , NOS 

INFLAMMATION , NOS 

GLONERULONEPHRTTIS, FOCAL 

INFLAMMATION , INTERSTITIAL 

• KIDNEY/TUBUL E 

MINERALIZATION 

* NUMBE R CE ANIHAL S WITH TISSU E

» KDMBE R CI ANIMAL S NECROPSIED 

C^NWUNTB, 

06-0118 

(50) 

(50) 

(48) 

2 (4X) 

(48) INFILTR 

(49) 

1 <2X) 

1 (2X) 

2 (4X) 

(49) 

(48) 

7 (15X) 

(48) 

(38) 

(50) 

4 (8X) 

1 (2X) 

12 (24X) 

(50) 

1 (2X) 

 EXAMINE D HICROSCOPICALLY 

LOW DOSE CONTROL (UHTR ) 

06-0030 

1 (2X) 

(46) 

1 (2X) 

(47) 

(39) 

(39) 

1 (3X) 

(«5) 

3 (7X) 

(45) 

(45) 

1 (2X) 

(45) 

1 (2X) 

(43) 

(45) 

2 (4JJ) 

1 (2X) 

9 (2 OX) 

(45) 

LCW DCSE 

06-0032 

(47) 

(48) 

1 (2X) 

(47) 

1 (2X) 

(47) 

(48) 

7 (15X) 

5 (10X) 

5 (10X) 

5 (10X) 

(48) 

4 (8X) 

(48) 

2 (4X) 

(48) 

(46) 

(47) 

9 (19X) 

(47) 

HIGH EOSE 

06-0112 

2 (4X) 

(47) 

(49) 

(40) 

1 <3X) 

(40) 

(46) 

2 (4X) 

2 (4X) 

(46) 

(39) 

4 (10X) 

(39) 

(33) 

2 (6X) 

(46) 

7 (15X) 

1 <2X ) 

8 (17*) 

(46) 

1 12il 

0-11 
• KIDNEY/PELVI S 

ISFLABtATICN , ACUTE/CHRONIC 

IURINAR Y £LADDEH 

I KFLAKATICN , NOS 

HYPERPLASIA , EPITHELIAL 

ENEOCRIN E SYSTE M 

*EITUI1ARY 

HYPOPLASIA, FOCAL 

tAERENAL/CAPSULE 

HYPERPLASIA , NCS 

tAERENA L CCRTEX 

NCDUL E 

HYPERPLASIA , NOS 

•THYROID 

INFLAfiCATICN , FOCAL 

NECRCSIS, FOCAI 

HYPERPLASIA , PAPILLAR Y 

HYPERPLASIA , ADENOMATOU S 

REPRCEOCT1VE SYSTE K 

*MAMBAH Y GLAN I 

HYPEHELASIA , NO S 

tUTERD S 

HYDRCBETH A 

ABSCESS, NOS 

HYPERPLASIA , ADENOBATOU S 

METAPLASIA , SQUABOUS 

#UTEBUS/HKi:OBETHIU B 

INFLAff.ATICN , SOS 

INFLAMMATION, ACUTE/CHRONIC 

HYPERPLASIA , NOS 

HYPERPLASIA , CYSTIC 

BETAPLASIA. SODAMOUS 

* NUMBE R CE ANIMAL S WITH TISSUE

* KUMBEB CE ANIMALS NECROPSIED 

TABLE D2 (CONTINUED) 

HIGH DOSE CONTROL(UNTB) 

06-0118 

LOW DOSE CONTROL (UNTR ) 

06-0030 

LCD DCSE 

06-0032 

HIGH DOSE 

06-0112 

(50) («5) 

1 (2X) 

(47) (16) 

(US ) 

1 (2X) 

(42) (17) 

2 (4*) 

7 (15S) 

(11) 

1 <2X ) 

(U2) (37) (42) (36) 

2 <6X) 

(48) 

5 (10X) 

(411) (46) 

8 (17%) 

(44) 

3 (7X) 

C*8) 

1 (2X ) 

1 (2«) 

(44) 

1 (2%) 

2 (5S) 

(16) (44) 

(tt)

1 (2X) 

2 (5S) 

1 (2%) 

(44) (42) (12) 

1 (2*) 

2 (5X) 

(50) (47) (18) (49) 

1 (2X) 3 (6)1) 

(17) 

13 (28%) 

(43| 

4 (9%) 

(48) 

8 (17X) 

(46) 13 (28*) 

1 (2X) 

2 <4X) 

1 <2S) 

(1*7) 

8 (17X) 

8 (17X) 

6 (13X) 

(U3) 

1 (2X) 

33 (77X) 

(48) 

1 (8JS) 

1 (2X) 

8 (17X) 

28 (58X) 

1 <2X) 

(16) 

4

9

6

 (9X) 

 (20X ) 

 (13X) 

 EXAMINE D MICROSCOPICALLY 

D-I2 
TABLE D2 (CONTINUED) 

LOW DOSE CONTROL (UflTR) LCif DOSE HIGH DOSE 

06-0118 06-0030 06-0032 06-0112 

•OVARY/OVIDUCT (47) (43) (<*8) (U6) 

IhFLAfKATICN , NO S 4 (9X) 1 <2X ) 

SESCESS, NOS 

DEGENERATION , NOS 

1 (2X) 

1 (2X) 

fCVAR Y (48) (1*8) (45) 

CYST, NCS 

INFLAMMATION , NOS 

10 (21X) 

1 (8S) 

7 (15X) 2

2

 (4X) 

 (4X) 

INFLAMMATION , NECROTIZING 1 (2X) 

AESCESS, NOS 1 (2X) 

PERIAR1ERITIS 

DEGENERATION , CYSIIC 

1 (2X) 

3 (6%) 4 (9X) 

HYPERPLASIA , NOS (2X) 

#CVARY/FCLLICL f (48) (44) (48) (45) 

HFBCREHAG E 2 (4X) 

NERVOU S SYSTE M 

NON E 

SPECIAL SENSE ORGAN S 

t.CNF 

fltlSCtUCSKELETA L SYSTE M 

NON E 

BOI Y CAVITIES 

*PLEUR S (50) (47) (48) (49) 

HYPERPLASIA , LYMPHOI D 1 (2X) 

All CTHER SYSTEMS 

COEkTD B JICBOSIS^FAJI \ 

* NUMBE R CF ANIMAL S WITH TISSUE EXAMINE D SICEOSCOPICA LLY 

* kUMBER CE ANIMAL S NECROPSIED 

D-I3 
TABLE D2 (CONCLUDED) 

(ONTR) COKTBOLfUNTB ) 10M DOSE HIGH COSE 

06-0118 06-0030 06-0032 06-0112 

SPECIA L RCEPHOICG Y SUHHAR Y 

NC IESICN EEEOBTED 3 1 1 

JUTO/NECROPSY/EISTO PERF 1 1 2 

AUTOLYSIS/N O SBCBOPSt 3 2 1 

f NUdEE B CF ililBALS WITH TISSUE EXSKINE D HICSOSCOPICSL1Y * SUHBEB CI ANIMAL S NECROPSIED 

D-14 
Review of the Bioassay of 2-Aminoanthraquinone* 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 arid research
 

organizations. Members have been selected on the basis of
 

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

as ad hoc members. The Data Evaluation/Risk Assessment
 

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

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

on the bioassay of 2-Aminoanthraquinone for carcinogenicity.
 

The reviewer said that the compound induced hepatocellular
 

carcinomas in male rats and in both sexes of mice but that
 

the treated female rats died of nephrotoxicity. After a
 

brief description of the experimental design, a discussion
 

ensued on the nature of the "foreign body granulomatous
 

nephritis" found in the treated female rats. In view of the
 

findings, the reviewer said that the compound must be
 

considered to pose a potential carcinogenic risk to humans.
 He added that nephrotoxicity also may pose a human hazard.
 The reviewer moved that the report on the bioassay of 2­ Aminoanthraquinone 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.
 

49
 

<JU.S. GOVERNMEN T PRINTIN G OFFICE : 1978-260-899/3204
 


DHEW Publication No. (NIH) 78-1399