1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 111 

BIOASSAY OF 

l-AMINO-2-METHYLANTHRAQUINONE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 82-28-0 

NCI-CG-TR-111 

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

Public Health Service 

National Institutes of Health 

BIDASSAY OF
 

l-AMINO-2-METHYLANTHRAQUINONE
 

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) 78-1366 

REPORT ON THE BIOASSAY OF l-AMINO-2-METHYLANTHRAQUINONE
 

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
 

l-amino-2-methylanthraquinone 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 l-amino-2-methylanthraquinone 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 analysis was performed by 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), and Dr. A. S. Krishna Murthy (3) at the
 

Mason Research Institute, 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).
 

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), using methods
 

selected for the Carcinogenesis Testing Program by Dr. J. J. Gart (9).
 

iii
 
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. 1
 

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

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.
 

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.
 

iv
 
10.	 No wit w h 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.
 

v
 

SUMMARY
 

A bioassay for possible carcinogenicity of technical-grade
 

l-amino-2-methylanthraquinone was conducted using Fischer 344 rats
 

and B6C3F1 mice. l-Amino-2-methylanthraquinone was administered in
 

the feed, at either of two concentrations, to groups of 45 to 50
 

males and females of each species. The high and low time-weighted
 

average concentrations of l-amino-2-methylanthraquinone were 0.20
 

and 0.10 percent, respectively, for male and female rats. For mice,
 

two dosage regimens (designated A and B) were used, but the time-


weighted average concentrations were the same, 0.06 percent. For
 

each species, 50 animals of each sex were placed on test as controls.
 

The period of compound administration was 78 weeks for rats followed
 

by 26 to 28 additional weeks of observation, and 73 weeks for mice
 

followed by 24 to 25 additional weeks of observation.
 

A statistically significant positive association between compound administration and mortality was established for the male and
 

female dose A mice. Dose A mice did not survive sufficiently long
 

to be at risk from late-developing tumors. Survival in all other
 

groups was adequate.
 

The incidence of hepatocellular carcinomas was statistically
 

significant among dosed rats of both sexes. Kidney neoplasms (the
 

combined incidence of tubular-cell adenomas, tubular-cell adenocarcinomas, and adenocarcinomas NOS) were significantly increased among
 

dosed male rats.
 

Administration of the compound was associated with a significant
 

increase in the combined incidence of hepatocellular carcinomas and
 

neoplastic liver nodules in female mice. No other neoplasms occurred
 

in statistically significant positive incidences in male or female
 

mice. l-Amino-2-methylanthraquinone demonstrated nephrotoxic properties in mice of both sexes.
 

Under the conditions of this bioassay, l-amino-2-methylanthraquinone was carcinogenic in Fischer 344 rats, inducing hepatocellular
 

carcinomas in rats of both sexes, and kidney tumors in male rats.
 

The compound was carcinogenic in female B6C3F1 mice, producing an
 

increased combined incidence of hepatocellular carcinomas and neoplastic nodules.
 

VII
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 4
 

A. Chemicals 4
 

B. Dietary Preparation 5
 

C. Animals 5
 

D. Animal Maintenance 6
 

E. Selection of Initial Concentrations 9
 

F. Experimental Design 10
 

G. Clinical and Histopathologic Examinations 14
 

H. Data Recording and Statistical Analyses 15
 

III. CHRONIC TESTING RESULTS: RATS 21
 

A. Body Weights and Clinical Observations 21
 

B. Survival 21
 

C. Pathology 24
 

D. Statistical Analyses of Results 27
 

IV. CHRONIC TESTING RESULTS: MICE 38
 

A. Body Weights and Clinical Observations 38
 

B. Survival 38
 

C. Pathology 41
 

D. Statistical Analyses of Results 43
 

V. DISCUSSION 49
 

VI. BIBLIOGRAPHY 53
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH l-AMINO-2­

METHYLANTHRAQUINONE C-l
 

ix
 
 D-l 

TABLE OF CONTENTS (Concluded)
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH l-AMINO-2­

METHYLANTHRAQUINONE


x
 
Figure Number
 

1
 

Table Number
 

LIST OF ILLUSTRATIONS 

Page 

CHEMICAL STRUCTURE OF l-AMINO-2-METHYLANTHRAQUINONE 2 

GROWTH CURVES FOR l-AMINO-2-METHYLANTHRAQUINONE CHRONIC STUDY RATS 22 

SURVIVAL COMPARISONS OF 1-AMINO-2-METHYLANTHRAQUINONE CHRONIC STUDY RATS 23 

GROWTH CURVES FOR l-AMINO-2-METHYLANTHRAQUINONE CHRONIC STUDY MICE 39 

SURVIVAL COMPARISONS OF 1-AMINO-2-METHYLANTHRAQUINONE CHRONIC STUDY MICE 40 

LIST OF TABLES 

DESIGN SUMMARY FOR FISCHER 344 RATS— 

l-AMINO-2-METHYLANTHRAQUINONE FEEDING 

EXPERIMENT 11 

DESIGN SUMMARY FOR B6C3F1 MICE—l-AMINO2-METHYLANTHRAQUINONE FEEDING EXPERIMENT 12 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

l-AMINO-2-METHYLANTHRAQUINONE 28 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH l-AMINO-2-METHYLANTHRAQUINONE 33 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN MALE 

MICE TREATED WITH 1-AM1NO-2-METHYLANTHRAQUINONE 44 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN FEMALE 

MICE TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 46 

xi
 
LIST OF TABLES (Concluded)
 

Table Number 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH 1-AMINO-2-METHYLANTHRAQUINONE A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH 1-AMINO-2-METHYLANTHRAQUINONE A-7 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH 1-AMINO-2-METHYLANTHRAQUINONE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH 1-AMINO-2-METHYLANTHRAQUINONE B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH l-AMINO2-METHYLANTHRAQUINONE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 

l-AMINO-2-METHYLANTHRAQUINONE C-10 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH l-AMINO2-METHYLANTHRAQUINONE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 

l-AMINO-2-METHYLANTHRAQUINONE D-7 

XI1
 
I. INTRODUCTION
 

l-Amino-2-methylanthraquinone (Figure 1) (NCI No. C01901), an
 

intermediate in the synthesis of anthraquinone dyes and a dye itself,
 

was selected for bioassay by the National Cancer Institute in an
 

attempt to elucidate those chemicals which 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 Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is l-amino-2-methyl-9,10-anthracene­

*
 dione. It is also known as 2-methyl-l-anthraquinonylamine and as
 

Disperse Orange 11 (C.I. [Colour Index] No. 60700).
 

l-Amino-2-methylanthraquinone is used as a dye for a variety of
 

synthetic fibers as well as wool, sheepskins and furs, and additionally, for the surface dyeing of thermoplastics (Society of Dyers and
 

Colourists, 1971a). It may also be used as an intermediate for the
 

production of a variety of dyes including Acid Blue 47, Acid Blue 49,
 

and Solvent Blue 13 (Urso, 1977; Society of Dyers and Colourists,
 

1971b); however, none of these is currently produced commercially in
 

the United States (U.S. International Trade Commission, 1977).
 

*
 

The CAS registry number is 82-28-0.
 
FIGURE 1

 

CHEMICAL STRUCTURE OF 1-AMIIMO-2-METHYLAIMTHRAQUINONE 
Although l-amino-2-methylanthraquinone has not been produced in
 

this country in commercial quantities since 1970, significant quantities of the chemical are imported annually (Bouchard, 1977). In this
 

country the greatest potential for exposure to l-amino-2-methylanthraquinone would be among workers engaged in the dying of textiles. An
 

increased incidence of bladder cancer has been observed among textile
 

workers in Leeds, England (Anthony and Thomas, 1970).
 
II. MATERIALS AND METHODS
 

A. Chemicals
 

Technical-grade l-amino-2-methylanthraquinone was purchased from
 

Carroll Products, Wood River Junction, Rhode Island. Analysis was
 

performed by Midwest Research Institute, Kansas City, Missouri. The
 

melting point (192° to 208°C) suggested the presence of impurities
 

due to its wide range. The value reported in the literature was 205°C
 

(Pollock and Stevens, 1965). Thin-layer chromatography (TLC) showed
 

the presence of at least two impurities. The infrared spectrum was
 

consistent with that reported in the literature (Pouchert, 1975). The
 

nuclear magnetic resonance spectrum was consistent with the structure
 

except for an extra peak in the aromatic region that matched the chemical shift of benzene. The extra peak indicated a possible impurity.
 

Spectra in the ultraviolet and visible range showed X at 245, 280
 

max
 

(shoulder), 305 and 475 nm for a methanol solution of the chemical.
 

The reference spectra showed \ at 246.5 and 305.0 for l-amino-2­ max
 

methylanthraquinone in methanol with molar extinction coefficients (O
 

3 3


of 36.9 x 10 and 6.4 x 10 , respectively (Sadtler Standard Spectra).
 

The shoulder at 280 was present in the literature spectra but no extinction coefficient was reported. The extraneous peak at 475 nm suggested the presence of impurities. The observed e values for the two
 

3 3
 peaks (246.5 and 305 nm) were, respectively, 35.2 x 10 and 4.2 x 10 .
 

Although the two e values for the 246.5 nm peak were comparable, the
 

e's for the 305 nm peak suggested a maximum purity of approximately
 

4
 
68 percent. Since the same solvent (methanol) was utilized in obtaining spectra of the reference and test compound, and linearity of
 

the Beer Lambert Law would be expected at the concentrations tested,
 

the molar extinction coefficients should give a reasonable estimate
 

of purity of this compound. The wide melting point range, the extraneous spots revealed by TLC, and the extraneous peaks in the ultraviolet spectrum and the nuclear magnetic resonance spectrum, all
 

indicated the presence of impurities.
 

Throughout this report the term l-amino-2-methylanthraquinone is
 

used to represent this technical-grade material.
 

B.	 Dietary Preparation
 

The basal laboratory diet for both dosed and control animals
 

(R)
 consisted of Wayne Lab-Blox (Allied Mills, Inc., Chicago, Illinois).
 

l-Amino-2-methylanthraquinone 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 2 weeks.
 

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 the rats and the mice assigned to
 
the dosed groups in the chronic bioassay were supplied by Charles
 

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

mice assigned to the control groups in the chronic bioassay were
 

supplied by ARS/Sprague-Dawley, Madison, Wisconsin.
 

Upon arrival, a sample of animals was examined for parasites
 

and other signs of disease. The remaining animals were quarantined
 

by species for 2 weeks prior to initiation of test. Animals were
 

assigned to groups and distributed among cages so that the 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
 

/s\
 

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. For the first 7 months
 

of the bioassay containment was in stainless- and galvanized-steel
 

wire-mesh cages suspended above newspapers. During this period newspapers were replaced daily and cages and racks were washed weekly.
 

For the remainder of the bioassay, suspended polycarbonate cages
 

equipped with disposable nonwoven fiber filter sheets were used for
 

rats. Fresh corncob bedding (SAN-I-CEL , Paxton Processing Company,
 
Paxton, Illinois) and clean cages were provided twice weekly during
 

this period. Once every 2 weeks the disposable filters were replaced
 

and the stainless steel cage racks (Fenco Cage Products, Boston,
 

Massachusetts) were cleaned.
 

Mice were housed by sex, ten per cage for the first 11 months
 

of the bioassay and five per cage thereafter. Containment was in
 

polycarbonate cages fitted during periods of compound administration
 

with perforated stainless steel lids, and with stainless steel wire
 

bar lids during the final observation period. Both types of lids
 

were supplied by Lab Products, Inc., and nonwoven fiber filter
 

bonnets were secured over all. Clean cages, lids, filters, and
 

bedding were provided three times weekly when cage populations were
 

ten and twice weekly when the cage populations were reduced to five.
 

Reusable filter bonnets and pipe racks were sanitized once every 2
 

®
 weeks throughout the study. Ab-sorb-dri hardwood chip bedding
 

(Wilner Wood Products Company, Norway, Maine) was provided for the
 

®
 first 6 months of the bioassay, corncob bedding (SAN-I-CEL, ) for the
 

(R)
 next 12 months, and another corncob bedding (Bed-o-Cobs , The Andersons Cob Division, Maumee, Ohio) was used for the remainder of the
 

study.
 

Water was available ad libitum for both species from 250 ml
 

water bottles equipped with rubber stoppers and stainless steel sip-


per tubes. Glass water bottles were used for the first 7 months and
 

polycarbonate bottles were used thereafter. Bottles were replaced
 
twice weekly and, in the case of rats because of their greater water
 

consumption, refilled as needed between changes. Wayne Lab-Blox®
 

meal was used throughout the period of chemical administration. The
 

treated or untreated food, replenished daily, was available ad
 

libitum to the appropriate groups of both rats and mice. Alpine^
 

aluminum feed cups (Curtin Matheson Scientific, Inc., Woburn,
 

Massachusetts) equipped with stainless steel baffles were used to
 

dispense food and were replaced weekly. During the final observation
 

period, rats were provided food pellets on the cage floor while mice
 

obtained food pellets from a food hopper incorporated into the cage
 

lid.
 

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

*
 

containing 3-amino-4-ethoxyacetanilide (17026-81-2); 4-nitroanthranilic acid (619-17-0); 5-nitroacenaphthene (602-87-9); and 5-nitro-otoluidine (99-55-8). Control rats were housed in a room with other
 

rats receiving diets containing 3-nitro-p-acetophenetide (1777-84-0);
 

2-methyl-l-nitroanthraquinone (129-15-7); and amitrole (61-82-5).
 

Dosed mice were housed in a room with other mice receiving
 

diets containing 3-amino-4-ethoxyacetanilide (17026-81-2); 4-nitroanthranilic acid (619-17-0); 5-nitro-o-anisidine (99-59-2); 2,4-dinitrotoluene (121-14-2); N,N-dimethyl-p-nitrosoaniline (138-89-6);
 

2,5-toluenediamine sulfate (6369-59-1); 2,4-diaminoanisole sulfate
 

(615-05-4); 2-aminoanthraquinone (117-79-3); 3-nitro-p-acetophenetide
 

*
 

CAS registry numbers are given in parentheses.
 
(1777-84-0); 1-nitronaphthalene (86-57-7); 5-nitroacenaphthene (602­

87-9); APC (8003-03-0); and amitrole (61-82-5). Control mice shared
 

a room with other mice receiving diets containing p-cresidine (120­

71-8); fenaminosulf (140-56-7); 4-chloro-m-phenylenediamine (5131-60­

2); and cinnamyl anthranilate (87-29-6).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

l-amino-2-methylanthraquinone 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
 

ten groups, each consisting of five males and five females. The
 

chemical was incorporated into the basal laboratory diet and supplied
 

ad libitum to nine of the ten groups of each species in concentrations
 

of 0.03, 0.06, 0.12, 0.24, 0.50, 1.50, 2.50, 3.50, and 4.50 percent.
 

The tenth 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 7 weeks, followed by a 1-week observation
 

period during which all animals were fed the basal diet.
 

The highest dosage causing no deaths, no compound-related gross
 

abnormalities, and no mean 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.
 

Compound-related mean body weight depression, mortality, and
 

gross lesions were observed in both species during the subchronic
 
test. Mean body weight gain, expressed as a percentage of the weight
 

gained by the controls, was calculated and recorded at the end of the
 

observation period.
 

All rats receiving doses of 1.50 percent or higher, all male
 

mice receiving doses of 0.24 percent or higher, and all female mice
 

receiving doses of 0.50 percent or higher died during the 8-week sub-


chronic study. Two male rats receiving 0.5 percent, two male mice
 

receiving 0.12 percent, and four female mice receiving 0.24 percent
 

l-amino-2-methylanthraquinone in their diet died. Compound-related
 

gross lesions encountered at dosages above 0.24 percent in rats and
 

0.06 percent in mice included pitted, enlarged, discolored kidneys;
 

enlarged lymph nodes; and reddened adrenals.
 

In rats receiving 0.24 percent l-amino-2-methylanthraquinone,
 

the mean male weight gain was 94 percent and the mean female weight
 

gain was 78 percent of the weight gained by the respective controls.
 

Body weight gain was 100 and 80 percent in the males and 84 and 75
 

percent in the females receiving 0.06 and 0.12 percent, respectively,
 

as compared to their respective controls.
 

The high concentration selected for administration in the
 

chronic study was 0.06 percent for both species.
 

F. Experimental Design
 

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

treated and untreated observation periods, and the time-weighted
 

average concentrations) are summarized in Tables 1 and 2.
 

10
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

l-AMINO-2-METHYLANTHRAQUINONE FEEDING EXPERIMENT
 

INITIAL
 

GROUP
 

SIZE
 

MALE
 

CONTROL 50
 

LOW DOSE 50
 

HIGH DOSE 50
 

FEMALE
 

CONTROL 50
 

LOW DOSE 45
 

HIGH DOSE 48
 

a
 

Concentrations are
 

1-AMINO-2­

METHYLANTHRAQUINONE
 

CONCENTRATION 3


0
 

0.03
 

0.12
 

0
 

0.06
 

0.24
 

0
 

0
 

0.03
 

0.12
 

0
 

0.06
 

0.24
 

0
 

OBSERVATION PERIOD


TREATED UNTREATED


 (WEEKS) (WEEKS)


0 108


16


6?
 

26
 

16


62
 

28
 

0 108


16


62
 

27
 

16


62
 

28
 

percentages in feed.
 

 TIME -WEIGHTED
 

 AVERAGE
 

 CONCENTRATION
 

0
 

 0.10
 

 0.20
 

0
 

 0.10
 

 0.20
 

b 2 (concentration X weeks received)
 Ti mp— we i crh fe>H avertIOP rnnr pn traf i n
 receiving chemical)
 

11
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

l-AMINO-2-METHYLANTHRAQUINONE FEEDING EXPERIMENT
 

l-AMINO-2­

INITIAL METHYLANTHRA­ OBSERVATION PERIOD TIME -WEIGHTED 

GROUP QUINONE TREATED UNTREATED AVERAGE 

SIZE CONCENTRATION3

 (WEEKS) (WEEKS) CONCENTRATION 

MALE
 

CONTROL 50	 0 0 98 0
 

DOSE A 50	 0.03 16 0.06
 

0.12 26
 

0.03 31
 

0 24
 

DOSE B 50	 0.06 73 0.06
 

0 24
 

FEMALE
 

CONTROL 50	 0 0 98 0
 

DOSE A 50	 0.03 16 0.06
 

0.12 26
 

0.03 31
 

0 24
 

DOSE B 49	 0.06 73 0.06
 

0 25
 

a
 Concentrations are percentages in feed.
 

2 (concentration X weeks received)
 T i mp— wp i ah t~ pH avprv ic*P r nnr pn t~ Tfl t" i nn — : : ;
 2(weeks receiving chemical)
 

12
 
At initiation of the study all rats were approximately 6 weeks
 

old. Dosed rats received initial dietary concentrations of 0.06
 

and 0.03 percent. Throughout this report those rats initially
 

receiving the former concentration are referred to as the high dose
 

groups, while those initially receiving the latter concentration are
 

referred to as the low dose groups. In week 17 high and low concentrations were increased to 0.24 and 0.12 percent, respectively, for
 

the remaining 62 weeks since no compound-related mean weight depression had been observed. After the 78-week dosing period the animals
 

were observed for up to 28 additional weeks.
 

At initiation of the study all mice were approximately 6 weeks
 

old. The group of mice initially receiving 0.03 percent of the test
 

compound in the diet is referred to as the dose A group throughout
 

this report due to the fact that for 26 weeks these mice were
 

receiving l-amino-2-methylanthraquinone at twice the concentration
 

being fed to the mice started on test at a concentration of 0.06 percent, referred to as the dose B group throughout this report. Dose B
 

mice received a dietary concentration of 0.06 percent for the entire
 

period of compound administration. Dose A mice received an initial
 

concentration of 0.03 percent. In week 17, the concentration for
 

dose A mice was increased to 0.12 percent, as no compound-related
 

mean weight depression had been observed. After 42 weeks on test,
 

the low concentration was decreased because of animal deaths from
 

toxicity to the original level of 0.03 percent, and this level was
 

13
 
maintained for the remaining 31 weeks of the dosing period. As the
 

result of variations in dietary concentrations fed to dose A mice
 

during this bioassay, the time-weighted average concentration of
 

l-amino-2-methylanthraquinone fed to all dosed groups of mice was
 

0.06 percent of the diet. After the 73-week dosing period the mice
 

were observed for up to 25 additional weeks.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the experiment. Body weights were recorded twice weekly for the first 12
 

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

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

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

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.
 

14
 
Tissues were preserved in 10 percent buffered formalin, embedded
 

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

to microscopic examination. An occasional section was subjected to
 

special staining techniques for more definitive diagnosis.
 

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

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

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

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

A few tissues were not examined for some animals, particularly
 

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

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,
 

15
 
as recommended by the International Union Against Cancer (Berenblum,
 

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

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

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect
 

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

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
 

16
 
to detect lesions prior to histologic sampling (e.g., skin or mammary
 

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

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

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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
 

17
 
relationship. This method also provides a two-tailed test of departure from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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.
 

18
 
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
 

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

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

c
 

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

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

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

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

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

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

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
 

19
 
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.
 

20
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Relatively consistent dose-related mean body weight depression
 

was observed in both male and female rats (Figure 2) and was readily
 

apparent after week 16 in males and week 22 in females.
 

During the course of the bioassay, palpable subcutaneous masses
 

were the clinical sign most commonly reported. They occurred in six
 

female controls, four male controls, two low dose females, one low
 

dose male, one high dose female, and one high dose male. Three high
 

dose males and three control females were observed to have white
 

discoloration of the eyes. Isolated clinical observations included
 

one high dose male blinded in one eye, one high dose male suffering
 

from severe posterior ataxia, and emaciation of one low dose male.
 

B. Survival
 

The estimated probabilities of survival for male and female
 

rats in the control and l-amino-2-methylanthraquinone-dose groups
 

are shown in Figure 3. For both males and females there was no
 

statistically significant association between dosage and mortality.
 

A sufficient number of males were at risk from late-developing
 

tumors as 62 percent (31/50) of the high dose, 90 percent (45/50)
 

of the low dose, and 68 percent (34/50) of the control rats survived
 

on test until the end of the study. Five high dose males were sacrificed in week 79; five control males were sacrificed in week 80.
 

21
 
750- -750 

UNTREATED CONTROL 

MALE RATS LOW DOSE 600- - 600 

HIGH DOSE 

t­ 450 — -450 

X 

<3 

ui 

8 30°­ — 300 

co 

< 

LU 

150­ -150 

I 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

750- •750 

UNTREATED CONTROL 

55 

600­ FEMALE RATS 

LOW DOSE -600 

cc 

HIGH DOSE 

H 450- -450 

8 30°­ -300 

CD 

< 

LU 

150- - 150 

I \ \ 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR 1-AMINO-2-METHYLANTHRAQUINONE CHRONIC STUDY RATS 

22 
PROBABILITY OF SURVIVAL PROBABILITY OF SU

o 

o o o 

•«» 01 

I I I I 

co 

33 

O 

o 

en

O 

CO 

O 

2 

O in

 

O 

> co 

7C 

CO 

3D 

> 

D 

O 

m 

O 

30 

O 

O 

en 

c 

•D 
For females the survival was also adequate as 56 percent (28/50)
 

of the high dose, 78 percent (39/50) of the low dose, and 70 percent
 

(35/50) of the control rats survived on test until the end of the
 

study. Five high dose females were sacrificed in week 79; five control females were sacrificed in week 80.
 

C.	 Pathology
 

Histopathologic findings on neoplasms in rats are tabulated in
 

Appendix A (Tables Al and A2); findings on nonneoplastic lesions are
 

tabulated in Appendix C (Tables Cl and C2).
 

Hepatocellular carcinomas occurred in 2/48 (4 percent), 7/50 (14
 

percent), and 10/48 (21 percent) control, low dose, and high dose
 

male rats, respectively, and in 1/49 (2 percent), 3/45 (7 percent),
 

and 10/44 (23 percent) control, low dose, and high dose female rats,
 

respectively. In addition, neoplastic nodules of the liver were
 

found in 1/48 (2 percent), 18/50 (36 percent), and 14/48 (29 percent)
 

control, low dose, and high dose male rats, respectively, and 2/49 (4
 

percent), 8/45 (18 percent), and 1/44 (2 percent) control, low dose,
 

and high dose female rats, respectively. Morphology of the neoplastic
 

nodules and hepatocellular carcinomas was similar to that described
 

by Squire and Levitt (1975). Neoplastic nodules were small and
 

compressed the adjacent parenchyma in areas. Cells were large and
 

cytoplasm was acidophilic. Nuclei were hyperchromatic and a few
 

mitotic figures were present. Hepatocellular carcinoma involved a
 

part or an entire lobe of the liver. Lobular architecture was
 

24
 
distorted and liver plates were several cells thick. Pleomorphism in
 

size of neoplastic hepatocytes was noted. Cytoplasm of the cells was
 

acidophilic or vacuolated. Nuclei were large and nucleoli were
 

prominent. Mitotic figures were not numerous.
 

Neither renal tubular-cell neoplasms nor renal tubular-cell
 

hyperplasia was seen in the control rats of either sex. A dose-


related spectrum of changes ranging from hyperplasia to adenoma to
 

adenocarclnoma observed in the kidneys of dosed rats is summarized
 

in the following table: 

Control

MALES 

Low

 Dose

 High 

 Dose Control

FEMALES 

Low

 Dose

 High 

 Dose 

Number of Animals with 

Kidneys Examined 

Histopathologically (48) (50) (48) (49) (45) (43) 

Renal Tubular-Cell 

Hyperplasia 11 13 

Renal Tubular-Cell 

Adenoma 

Renal Tubular-Cell 

Adenocarcinoma 0 0 4 0 0 0 

Adenocarcinoma NOS 0 1 0 0 0 0 

Carcinoma NOS 0 0 0 1 0 0 

Renal Pelvis Transitional-Cell Carcinoma 0 1 1 0 1 0 

A focal increase of tubular cells with a basophilic cytoplasm
 

and large vesicular nuclei was considered renal tubular-cell hyperplasia. Renal tubular-cell adenomas were nodular and were demarcated
 

25
 
from the rest of the renal parenchyma. Cells were arranged in a
 

tubular pattern or occurred as a solid mass. Cytoplasm of cells was
 

basophilic and nuclei were vesicular. There were a few mitotic
 

figures. Two rats had multiple tumors of this nature.
 

Renal tubular-cell adenocarcinomas were large and circumscribed.
 

They had replaced much of the normal renal tissue. In areas, the
 

neoplasms had compressed adjacent tubules or glomeruli. Cells were
 

arranged in a trabecular pattern. Thin strands of fibrovascular
 

tissue dissected the tumor parenchyma into nodules of varying sizes
 

and shapes. In areas, tumor cells attempted to form tubules.
 

Cytoplasm of cells was either vacuolated or acidophilic. Nuclear
 

pleomorphism was not evident and mitotic figures were not numerous.
 

Clusters of lymphocytes, varying degrees of hemorrhage, and areas of
 

necrosis were present in the tumor mass.
 

A transitional-cell carcinoma of the kidney was diagnosed in
 

1/45 low dose female rats. Transitional-cell carcinomas of the renal
 

pelvis occurred in 1/50 low dose male rats and 1/48 high dose male
 

rats. In the low dose male rat, the carcinoma metastasized to the
 

lung. Because of the small number of rats with this type of neoplasm, no clear-cut effect of l-amino-2-methylanthraquinone on the
 

transitional-cell epithelium could be demonstrated.
 

This histopathologic examination provided evidence for the carcinogenicity of l-amino-2-methylanthraquinone in Fischer 344 rats
 

for the following reasons:
 

26
 
(1)	 there was an increase in the incidence of neoplastic
 

nodules of the liver and hepatocellular carcinomas
 

in dosed rats; and
 

(2)	 hyperplastic and neoplastic lesions of renal tubules
 

occurred in dosed rats in a dose-related fashion,
 

predominantly in males.
 

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 tumor in either sex where at least two such tumors were
 

observed in any of the control or l-amino-2-methylanthraquinone-dosed
 

groups and where such tumors were observed in at least 5 percent of
 

the group.
 

High numbers of liver tumors were observed in both male and
 

female rats. In males the Cochran-Armitage test showed a significant
 

(P = 0.012) positive association between dosage and the incidence of
 

hepatocellular carcinomas. The Fisher exact test results supported
 

these findings by a significant (P = 0.014) comparison of high dose
 

to control. In females again the Cochran-Armitage test showed a significant (P = 0.001) positive association between dose and the
 

incidence of hepatocellular carcinomas. The Fisher exact test
 

comparing high dose to control was also significant (P = 0.002).
 

When incidences were combined so that the numerator represented a rat
 

with either a hepatocellular carcinoma or a neoplastic nodule, for
 

both sexes the Cochran-Armitage test (P _£ 0.005) and both the high
 

dose and the low dose Fisher exact test comparisons (P _<_ 0.004) were
 

27
 
S3
 

00
 

TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONEc
 

TOPOGRAPHY: MORPHOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

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
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

2/48(0.04)
 

P = 0.012
 

106
 

3/48(0.06)
 

P < 0.001
 

P = 0.009
 

_—
 

99
 

6/48(0.13)
 

N.S.
 

98
 

LOW
 

DOSE
 

7/50(0.14)
 

N.S.
 

3.360
 

0.681
 

31.860
 

104
 

25/50(0.50)
 

P < 0.001
 

8.000
 

2.694
 

38.152
 

104
 

1/49(0.02)
 

N.S.
 

0.163
 

0.004
 

1.274
 

104
 

HIGH
 

DOSE
 

10/48(0.21)
 

P = 0.014
 

5.000
 

1.143
 

44.920
 

100
 

24/48(0.50)
 

P < 0.001
 

8.000
 

2.686
 

38.147
 

76
 

2/49(0.04)
 

N.S.
 

0.327
 

0.034
 

1.720
 

99
 
TABLE 3 (CONTINUED) 

TOPOGRAPHY: MORPHOLOGY CONTROL 

Lung: Alveolar/Bronchiolar Adenoma or 

Alveolar /Bronchiolar Carcinomab

P Values0

 0/48(0.00) 

 N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

l-o 

Kidney: Tubular-Cell Adenomab

P Values0

 0/48(0.00) 

P = 0.017 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Kidney: Tubular-Cell Ad eno carcinoma 

or Adenocarcinoma NOS^

P Values0

 0/48(0.00) 

P = 0.025 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

LOW
 

DOSE
 

3/49(0.06)
 

N.S.
 

Infinite
 

0.590
 

Infinite
 

104
 

5/50(0.10)
 

P = 0.031
 

Infinite
 

1.212
 

Infinite
 

97
 

1/50(0.02)
 

N.S.
 

Infinite
 

0.052
 

Infinite
 

104
 

HIGH
 

DOSE
 

2/48(0.04)
 

N.S.
 

Infinite
 

0.296
 

Infinite
 

105
 

6/48(0.13)
 

P = 0.013
 

Infinite
 

1.602
 

Infinite
 

97
 

4/48(0.08)
 

N.S.
 

Infinite
 

0.928
 

Infinite
 

89
 
TABLE

TOPOGRAPHY: MORPHOLOGY
 

Kidney: Tubular-Cell Adenoma, Tubular-


Cell Adenocarcinoma or Adenocarcinoma
 

NOSb
 

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
 

Adrenal: Pheochromocytoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

3 (CONTINUED) 

CONTROL
 

0/48(0.00)
 

P = 0.001
 

1/41(0.02)
 

P = 0.017
 

108
 

10/47(0.21)
 

N.S.
 

99
 

LOW
 

DOSE
 

6/50(0.12)
 

P = 0.015
 

Infinite
 

1.537
 

Infinite
 

97
 

10/46(0.22)
 

P = 0.006
 

8.913
 

1.361
 

376.318
 

70
 

10/49(0.20)
 

N.S.
 

0.959
 

0.396
 

2.330
 

70
 

HIGH
 

DOSE
 

10/48(0.21)
 

P = 0.001
 

Infinite
 

2.980
 

Infinite
 

89
 

8/39(0.21)
 

P = 0.012
 

8.410
 

1.211
 

361.434
 

79
 

6/48(0.13)
 

N.S.
 

0.587
 

0.191
 

1.634
 

105
 
TOPOGRAPHY : MORPHOLOGY
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Testis: Interstitial-Cell Tumor
 

c
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Body cavities: Mesothelioma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

CONTROL
 

0/39(0.00)
 

P = 0.032
 

45/47(0.96)
 

N.S.
 

——
 

80
 

0/48(0.00)
 

P = 0.027
 

LOW
 

DOSE
 

3/47(0.06)
 

N.S.
 

Infinite
 

0.503
 

Infinite
 

104
 

48/50(0.96)
 

N.S.
 

1.003
 

0.931
 

1.080
 

70
 

1/49(0.02)
 

N.S.
 

Infinite
 

0.053
 

Infinite
 

104
 

HIGH
 

DOSE
 

5/46(0.11)
 

P = 0.042
 

Infinite
 

1.078
 

Infinite
 

79
 

43/48(0.90)
 

N.S.
 

0.936
 

0.865
 

1.067
 

76
 

4/49(0.08)
 

N.S.
 

Infinite
 

0.909
 

Infinite
 

79
 
TABLE 3 (CONCLUDED) 

o
 

Treated groups received time-weighted average doses of 0.10 or 0.20 percent in feed.
 

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

£

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

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

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

control group is given beneath the incidence of tumors in the treated group when P < 0.05;
 

otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact
 

tests a negative indication (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.
 

u>

6 The probability level of the test for departure from linear trend is given beneath the con-
 M

 trol group when P < 0.05.
 
___ 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE*
 

TOPOGRAPHY: MORPHOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

U)
 

Liver: Hepatocellular Carcinoma or
 

Neoplastic Nodule"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

1/49(0.02)
 

P = 0.001
 

108
 

2/49(0.04)
 

P = 0.005
 

92
 

7/49(0.14)
 

P = 0.020(N)
 

106
 

LOW
 

DOSE
 

3/45(0.07)
 

N.S.
 

3.267
 

0.274
 

167.567
 

105
 

11/45(0.24)
 

P = 0.004
 

5.989
 

1.402
 

52.986
 

104
 

2/45(0.04)
 

N.S.
 

0.311
 

0.033
 

1.529
 

104
 

HIGH
 

DOSE
 

10/44(0.23)
 

P = 0.002
 

11.140
 

1.690
 

469.425
 

105
 

11/44(0.25)
 

P = 0.004
 

6.125
 

1.444
 

54.128
 

105
 

1/44(0.02)
 

P = 0.042(N)
 

0.159
 

0.004
 

1.165
 

98
 
TOPOGRAPHY : MORPHOLOGY
 

Pituitary: Adenoma NOS or Chromophobe
 

Adenoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenoma, Fibroadenoma
 

or Adenocarcinomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polyp
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED)
 

CONTROL
 

18/44(0.41)
 

N.S.
 

90
 

18/49(0.37)
 

P < O.OOl(N)
 

80
 

12/49(0.24)
 

P = 0.012(N)
 

80
 

LOW
 

DOSE
 

14/40(0.35)
 

N.S.
 

0.856
 

0.458
 

1.562
 

76
 

6/45(0.13)
 

P = 0.009(N)
 

0.363
 

0.130
 

0.858
 

76
 

10/44(0.23)
 

N.S.
 

0.928
 

0.399
 

2.099
 

98
 

HIGH
 

DOSE
 

20/39(0.51)
 

N.S.
 

1.254
 

0.745
 

2.090
 

58
 

3/44(0.07)
 

P < O.OOl(N)
 

0.186
 

0.038
 

0.579
 

106
 

2/42(0.05)
 

P = 0.009(N)
 

0.194
 

0.022
 

0.807
 

106
 
TABLE 4 (CONCLUDED)
 

TOPOGRAPHY: MORPHOLOGY CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Thyroid: C-Cell Adenoma or C-Cell 

Carcinoma" 3/40(0.07) 2/43(0.05) 1/38(0.03) 

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

Relative Risk (Control) 

Lower Limit 

Upper Limit 

0.620 

0.054 

5.138 

0.351 

0.007 

4.140 

Weeks to First Observed Tumor 108 104 105 

Treated groups received time-weighted average doses of 0.10 or 0.20 percent in feed.
 

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

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

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

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

control group is given beneath the incidence of tumors in the treated group when P < 0.05;
 

otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact
 

tests a negative designation (N) indicates a lower incidence in the treated group(s) than
 

in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
significant. Based upon these results the administration of 1-amino2-methylanthraquinone was associated with an increased incidence of
 

hepatocellular carcinomas in both male and female rats.
 

In male rats significant numbers of kidney neoplasms were also
 

noted. When incidences were combined so that the numerator represented male rats with either a tubular-cell adenoma, a tubular-cell
 

adenocarcinoma, or an adenocarcinoma NOS, then the Cochran-Armitage
 

test (P = 0.001) and the Fisher exact tests comparing both high dose
 

to control (P = 0.001) and low dose to control (P = 0.015) were
 

significant. Based upon these results, administration of l-amino-2­

methylanthraquinone was associated with an increased incidence of
 

tubular-cell neoplasms of the kidney in male rats.
 

For male rats the Cochran-Armitage test also indicated a significant (P = 0.017) positive association between dose and the combined
 

incidence of adenomas NOS or chromophobe adenomas of the pituitary.
 

The Fisher exact tests confirmed this finding for the comparison of
 

both low dose (P = 0.006) and high dose (P = 0.012) to control. In
 

historical control data collected by this laboratory for the NCI
 

Carcinogenesis Testing Program, however, 37/334 (11 percent) of the
 

untreated males had a pituitary adenoma, compared to the 1/41 (2
 

percent) observed in the control group for this bioassay. Additionally,
 

the incidences in several historical control groups were above those
 

incidence rates observed in these dosed groups.
 

36
 
For males the Cochran-Armitage test indicated significant associations between dose and both the incidence of mesotheliomas of the
 

tunica vaginalis and the incidence of C-cell thyroid neoplasms. In
 

both cases, however, the Fisher exact tests were not significant under the Bonferroni criterion.
 

For females the possibility of significant negative associations
 

between dose and incidence were observed for mammary tumors and for
 

endometrial stromal polyps. For the mammary tumors, however, historical control data showed 125/589 (21 percent) of the untreated
 

Fischer 344 female rats with either an adenoma, a fibroadenoma, or
 

an adenocarcinoma of the mammary gland—compared to the 18/49 (37
 

percent), 6/45 (13 percent), and 3/44 (7 percent) observed in the
 

control, low dose, and high dose groups, respectively, in this
 

bioassay. The Cochran-Armitage test showed a significant negative
 

association for leukemia or malignant lymphoma, but the Fisher exact
 

tests were not significant under the Bonferroni criterion.
 

Summarizing these results, the statistical conclusions were that
 

the incidences of hepatocellular carcinomas in both male and female
 

rats and of kidney tumors in male rats were associated with the
 

administration of l-amino-2-methylanthraquinone.
 

37
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Relatively consistent and severe dose-related mean body weight
 

depression was observed in the female mice and, to a lesser extent,
 

in the male mice (Figure 4). The inconsistency observed in the
 

weight pattern of the dose A males after week 48 and until week 76
 

may have been indirectly due to a reduction of the concentration of
 

the chemical in the food beginning in week 43. This reduction was
 

initiated because of the numerous deaths experienced by the dose A
 

group. The net result may have been increased food consumption and
 

subsequent weight gain by the remaining, perhaps most healthy,
 

animals.
 

No clinical abnormalities were recorded for mice of either sex.
 

B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and l-amino-2-methylanthraquinone-dosed groups are
 

shown in Figure 5. For both males and females the Cox tests indicated
 

the survival of the dose A groups was significantly (P < 0.001) lower
 

than that of the dose B groups or the control groups. This appears
 

to have been associated with an increase in dosage for the dose A
 

groups in week 17 from 0.03 to 0.12 percent l-amino-2-methylanthraquinone in their feed. In week 43 the dosage for the dose A groups was
 

changed back to 0.03 percent. Dose B groups received the chemical at
 

a dietary concentration of 0.06 percent. As a result of these dosage
 

38
 
T

 

60 75 

TIME ON TEST (WEEKS) 

T 

45 60 75 105 120

 

TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR 1-AMINO-2-METHYLANTHRAQUINONE CHRONIC STUDY MICE 

39 

15 
PROBABILITY OF SURVIVAL PROBABILITY OF SURVIV

<D O ° O O o c3 ro -^ 01 CO cD o 

o — 1 1 1 1 1

 i

l l 1 1 1 1 1 1 1 

(A 

C 

30 | 

| 1 1 

> _ 1 r~ cji 1 

|

n 

O C D " 0 0 ° o p o Oi Cn ^ m m H

^ oX m > g

 -D m m H 

pa r 

~ CO _ 8 <= 

!••• 

en 

o 1 

!••• 

-n 

1 r-' 

j....1

 - _l r- ^"" ^ —l •&• 

2 IJ 01 — — ^ 

z ^ 

O m m 

KJ 0 r* 0 

z

z s H r m H^ TM o,

 

"^ m • — ,../ J

 rs 

s >«" m ./ m 

m H A " •" I w CO " i • 

> ^ — :"" 1 

I

i j 1 •'"• 

O • 

m j 

o g- 8­ 3D 

O ! ..: 

; i :

:

0 

CO Tl 

m 

c °— ^ D " > m m

 

S 

g 

0 O O m m m

 ro —

 o 1 | I | 1 i | i | i | i 

1

 | 

1 c3 o O O O - Zl o O O 
changes, the time-weighted average concentrations of l-amino-2-methylanthraquinone received by the dose A groups and the dose B groups
 

were approximately the same (0.062 and 0.06 percent, respectively).
 

By the end of week 43, 56 percent (28/50) of the dose A group
 

males and 46 percent (23/50) of the dose A group females had died.
 

As such, there were not adequate numbers of dose A group mice at risk
 

from late-developing tumors.
 

For males, however, there were adequate numbers of dose B group
 

and control group mice at risk from late-developing tumors, as 74 percent (37/50) of both the dose B group and the control group survived
 

on test until the end of the study. Five dose B group males were
 

sacrificed in week 79; five control males were sacrificed in week 78.
 

The survival of female dose B and control mice was also adequate
 

as 74 percent (37/50) of the dose B group and 70 percent (35/50) of
 

the control group survived on test until the end of the study. Five
 

control mice were sacrificed in week 78; five dose B group mice were
 

sacrificed in weeks 79 and 80.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are tabulated in
 

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

tabulated in Appendix D (Tables Dl and D2).
 

Hepatocellular carcinomas occurred in both control and dosed
 

mice and did not appear to be compound-related. In male mice, this
 

tumor was found in 10/45 (22 percent) control, 1/36 (3 percent)
 

41
 
dose A, and 8/45 (18 percent) dose B mice. In female mice, hepatic
 

tumors were seen in 4/45 (9 percent) control, 2/34 (6 percent) dose
 

A, and in 12/44 (27 percent) dose B mice.
 

Adenocarcinoma of the kidney, morphologically similar to that in
 

rats, was found in two dose B male mice. The occurrence of these
 

tumors is of interest in view of the occurrence of renal tumors in
 

the rats.
 

Compound-related nonneoplastic lesions involved only the kidney.
 

The incidence of glomerulonephritis (glomerulosclerosis) and interstitial (diffuse) fibrosis in these mice is shown in the following
 

table:
 

MALES FEMALES
 

Control Dose A Dose B Control Dose A Dose B
 

Number of Animals with 

Kidneys Examined 

His topatho logically (45) (37) (45) (43) (37) (42) 

Glomerulonephritis NOS 0 24 42 0 23 31 

Interstitial Fibrosis 0 9 32 0 12 13 

Degenerative changes in renal tubules ranged from loss of cytoplasmic basophilia to necrosis. Large tubular cells with basophilic
 

cytoplasm and vesicular nuclei suggested regeneration. Clusters of
 

inflammatory cells were present in the cortex.
 

In areas, some of the renal tubules were cystic, the glomeruli
 

were atrophic, and the Bowman's space distended. Both the basement
 

42
 
membrane and mesangium were thickened in a few glomeruli. Interstitial fibrosis was present in many mice.
 

The results of this histopathologic examination provided evidence
 

for the carcinogenicity of l-amino-2-methylanthraquinone in B6C3F1
 

mice, as administration of the compound was associated with increased
 

numbers of liver tumors in female mice. l-Amino-2-methylanthraquinone was also nephrotoxic at the doses used to both sexes of B6C3F1
 

mice as shown by the occurrence of glomerulonephritis and interstitial
 

fibrosis.
 

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 tumor in either sex where at least two such tumors were
 

observed in any of the control or l-amino-2-methylanthraquinone-dosed
 

groups and where such tumors were observed in at least 5 percent of
 

the group. Because the time-weighted average dose received by the
 

dose A group was approximately the same as that received by the dose
 

B group, it was inappropriate to use the Cochran-Armitage test with
 

these data. Because the manner in which the dosages were changed
 

resulted in poor survival in the dose A group, the following analyses
 

are based solely upon those mice surviving at least 52 weeks.
 

In female mice a number of liver neoplasms were observed. When
 

incidences were combined so that the numerator represented female
 

mice with either hepatocellular carcinomas or neoplastic nodules,
 

43
 
___ 

TABLE 5
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

a,e
 SPECIFIC SITES IN MALE MICE TREATED WITH 1-AMINO-2-METHYLANTHRAQUINONE


TOPOGRAPHY :MORPHOLOGY
 

Lung: Alveolar /Bronchiolar Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 •P­ •P-


Lung:	 Alveolar /Bronchiolar Carcinoma
 

or Alveolar /Bronchiolar Adenoma*3
 

P	 Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Circulatory System: Hemangiosarcoma
 

P	 Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

4/45(0.09)
 

97
 

11/45(0.24)
 

78
 

0/46(0.00)
 

DOSE A
 

0/10(0.00)
 

N.S.
 

0.000
 

0.000
 

4.357
 

0/10(0.00)
 

N.S.
 

0.000
 

0.000
 

1.212
 

0/12(0.00)
 

N.S.
 

DOSE B
 

1/43(0.02)
 

N.S.
 

0.262
 

0.005
 

2.505
 

79
 

5/43(0.12)
 

N.S.
 

0.476
 

0.141
 

1.350
 

79
 

3/45(0.07)
 

N.S.
 

Infinite
 

0.617
 

Infinite
 

97
 
TABLE 5 (CONCLUDED)
 

TOPOGRAPHY : MORPHOLOGY CONTROL DOSE A DOSE B 

Liver: Hepatocellular Carcinoma 

P Values0 

10/45(0.22) 1/11(0.09) 

N.S. 

8/44(0.18) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

0.409 

0.010 

2.336 

0.818 

0.310 

2.081 

Weeks to First Observed Tumor 93 76 79 

Treated groups received time-weighted average doses of approximately 0.06 percent in feed.
 

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

"The probability level for the Fisher exact test for the comparison of a treated group with
 

the control group is given beneath the incidence of tumors in that 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.
 a
 

"These analyses were based solely upon animals surviving at least 52 weeks.
 
TABLE 6
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH 1-AMINO- 2-METHYLANTHRAQU INONE3'
 

TOPOGRAPHY : MORPHOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma or
 

Neoplastic Nodule^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma°
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

4/44(0.09)
 

78
 

4/44(0.09)
 

78
 

12/45(0.27)
 

95
 

DOSE A
 

2/16(0.13)
 

N.S.
 

1.375
 

0.132
 

8.336
 

97
 

2/16(0.13)
 

N.S.
 

1.375
 

0.132
 

8.336
 

97
 

1/18(0.06)
 

N.S.
 

0.208
 

0.005
 

1.366
 

97
 

DOSE B
 

9/43(0.21)
 

N.S.
 

2.302
 

0.700
 

9.502
 

97
 

12/43(0.28)
 

P = 0.022
 

3.070
 

1.021
 

12.053
 

97
 

5/43(0.12)
 

N.S.
 

0.436
 

0.141
 

1.350
 

90 
TABLE 6 (CONCLUDED)
 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE A DOSE B 

Pituitary:

P Values0 

 Adenoma NOS 6/37(0.16) 0/11(0.00) 

N.S. 

3/34(0.09) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

0.000 

0.000 

1.903 

0.544 

0.095 

2.328 

Weeks to First Observed Tumor 98 97 

treated groups received time-weighted average doses of approximately 0.06 percent in feed.
 

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

"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 that 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.
 

a

These analyses were based solely upon animals surviving at least 52 weeks.
 
the Fisher exact text indicated that the incidence of liver tumors
 

was significantly (P = 0.022) greater in the dose B group than in the
 

control. In historical data collected by this laboratory for the NCI
 

Carcinogenesis Testing Program, 13/350 (4 percent) untreated female
 

B6C3F1 mice had one of these tumors, compared to the 4/44 (9 percent),
 

2/16 (13 percent), and 12/43 (28 percent) observed in the control,
 

dose A, and dose B groups, respectively, in this bioassay. Based
 

upon these results, the administration of l-amino-2-methylanthraquinone was associated with an increased incidence of liver neoplasms in
 

female mice.
 

No other test at any other site in either sex was statistically
 

significant.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

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

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

Tables 5 and 6, the value one is included; this indicates the absence
 

of statistically significant results. It should also be noted that
 

many of the confidence intervals have an upper limit greater than
 

one, indicating the theoretical possibility of tumor induction in
 

mice by l-amino-2-methylanthraquinone that could not be established
 

under the conditions of this test.
 

48
 
V. DISCUSSION
 

Under the conditions of this bioassay adequate numbers of animals
 

survived sufficiently long to be at risk from late-developing tumors
 

in all except the male and female dose A mouse groups. The poor survival may be attributable to the concentration of l-amino-2-methylanthraquinone administered to these groups from weeks 17 through 42
 

(0.12 percent). Although dose A mouse groups were started as the low
 

dose groups, the concentration given of the test chemical in feed
 

from weeks 17 through 42 was twice the highest concentration received by the dose B groups.
 

Hepatocellular carcinomas were observed, respectively, in 2/48
 

(4 percent), 7/50 (14 percent), and 10/48 (21 percent) of the control,
 

low dose, and high dose male rats and in 1/49 (2 percent), 3/45 (7
 

percent), and 10/44 (23 percent) of the control, low dose, and high
 

dose female rats. The Cochran-Armitage tests indicated a significant
 

positive association between dosage and the incidences of these neoplasms in both sexes and the Fisher exact comparison of the high dose
 

to the control group for each sex supported these findings. Neoplastic
 

liver nodules were detected in 1/48 (2 percent), 18/50 (36 percent),
 

and 14/48 (29 percent) of the control, low dose, and high dose male
 

rats and in 2/49 (4 percent), 8/45 (18 percent), and 1/44 (2 percent)
 

of the control, low dose, and high dose female rats, respectively.
 

For each sex the Cochran-Armitage test revealed a significant positive
 

association between compound administration and the incidence of these
 

49
 
nodules. In the males, the high dose to control Fisher exact comparison supported this finding but in females only the low dose to control Fisher exact comparison supported the association. When all the
 

female rats in each group having either hepatocellular carcinomas or
 

neoplastic liver nodules were combined and the resulting incidences
 

of females with these tumors were statistically analyzed, both the
 

high dose to control and the low dose to control Fisher exact tests
 

indicated significant positive associations between compound administration and the occurrence of these neoplasms.
 

A spectrum of compound-related renal changes was noted, ranging
 

from hyperplasias to adenomas and adenocarcinomas, particularly among
 

the male rats. Statistical analyses of these kidney tumors, using
 

the Cochran-Armitage test, revealed significant associations between
 

dosage and the incidence of tubular-cell adenomas and the combined
 

incidence of tubular-cell adenomas, tubular-cell adenocarcinomas, and
 

adenocarcinomas NOS. The Fisher exact comparisons of high dose to
 

control supported both of these associations in male rats.
 

The only other statistically significant positive association
 

between chemical administration and increased tumor incidence in rats
 

was demonstrated for males with pituitary adenomas. The Cochran-


Armitage test indicated the positive association and it was supported
 

by both the high and low dose Fisher exact comparisons. The incidence
 

of pi-tuitary adenomas in male rat controls (1/41 or 2 percent) was unusually low compared to historical controls (37/334 or 11 percent).
 

50
 
In addition, the incidences of these neoplasms in some of the historical control groups from this laboratory have closely approximated
 

the incidences observed in the dosed male rats in this bioassay.
 

For this reason, the statistical results based on observed tumor
 

incidences are not considered sufficient proof that the compound
 

induced pituitary adenomas in male rats.
 

When those female mice having hepatocellular carcinomas were
 

combined with those having neoplastic liver nodules and the resulting
 

incidence of dose B females having these tumors was compared to the
 

incidence in control females, a significant positive association
 

between compound administration and tumor incidence was demonstrated.
 

No other neoplasms occurred in statistically significant positive
 

incidences in male or female mice.
 

The detection of adenocarcinomas of the kidney in two dose B
 

male mice was of interest, considering the renal abnormalities reported in rats. The only compound-related nonneoplastic lesions in
 

the mice were glomerulonephritis and interstitial (diffuse) fibrosis,
 

both of which occurred only in dosed animals. As a result, the
 

compound was determined to be nephrotoxic in mice at the concentrations administered in the feed.
 

Under the conditions of this bioassay, l-amino-2-methylanthraquinone was carcinogenic in male and female Fischer 344 rats, inducing hepatocellular carcinomas in rats of both sexes. It also induced
 

renal neoplasms in male rats. The compound was carcinogenic in
 

51
 
female B6C3F1 mice, producing an increased incidence of liver tumors
 

(i.e., the combined incidence of neoplastic nodules and hepatocellular
 

carcinomas).
 

52
 
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.
 

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

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

Union Against Cancer, Geneva, 1969.
 

Bouchard, D.R., Product Manager, Organic Intermediates, BASF Wyandotte
 

Corporation, Porsippcency, N.J. Personal communication, 1977.
 

Chemical Abstracts Service. The Chemical Abstracts Service (CAS)
 

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

Chemical Society, Washington, B.C., 1977.
 

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

Co., Ltd., London, 1970.
 

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

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

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

Tests, Confidence Limits, and Adjustments for Stratification."
 

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

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

Observations." Journal of the American Statistical Association
 

23_:457-481, 1958.
 

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

"Carcinogenesis Bioassay Data System." Computers and Biomedical
 

Research 7:230-248, 1974.
 

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

Co., New York, 1966
 

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

York, p. 154, 1965.
 

53
 
Pouchert, C., The Aldrich Library of Infrared Spectra, 2nd edition.
 

Spectra No. 788H. Aldrich Chemical Company, Milwaukee, Wisconsin,
 

1975.
 

Sadtler Standard Spectra. Sadtler Research Laboratories, Philadelphia, Pennsylvania. UV No. 3788.
 

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 2.
 

Yorkshire, England, 1971a.
 

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

Yorkshire, England, 1971b.
 

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 communication, 1977.
 

U.S.	 International Trade Commission, Synthetic Organic Chemicals,
 

United States Production and Sales, 1975. USITC Publication
 

804, U.S. Government Printing Office, Washington, D.C. , 1977.
 

Wynder, E.L., J. Onderdonk, and N. Mantel, "An Epidemiological
 

Investigation of Cancer of the Bladder." Cancer 16:1388-1407,
 

1963.
 

54
 
Review of the Bioassay of l-Amino-2-Methylanthraquinone*
 

for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DREW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

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

academia, industry, organized labor, public interest groups,
 

State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 

biostatistics, toxicology, pathology, and epidemiology.
 

Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

Subgroup of the Clearinghouse is charged with the responsibility 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 l-Amino-2-Methylanthraquinone for
 

carcinogenicity.
 

The reviewer said that the compound induced liver
 

tumors in both sexes of treated rats and in female mice.
 It also induced kidney tumors in male rats and was nephro­ toxic in mice. The reviewer opined that the hepatic effect
 

in male mice may have been masked by the high spontaneous
 

incidence of liver tumors in this sex. He noted the
 

negative trend for mammary tumors in treated female rats.
 The reviewer considered the experimental design acceptable
 

67
 
and he moved that the report on the bioassay of l-Amino-2­ Methylanthraquinone 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.
 

<1U.S. GOVERNMEN T PRINTIN G OFFICE : 1978-260-899/3195
 

68
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE
 

TABLE A1 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 

CONTROL (ONTO) 

01-0070 

LOU DOSE 

01-0066 

HIGH D3SE 

01-0067 

ANIMALS INITIALLY IN STUDY 50 

ANIMALS HISSING 1 

ANIMALS NECROPSIED U8 

AMIHALS EXAMINED HI STOPATHOL3GICALLY *» 148 

50 

19 

«8 

50 

KTPGUHINTART SYSTEM 

•SKIN 

SQUAMOUS CELL PAPILLOMA 

SQUA10US CELL CARCINOMA 

BASAL-CELL CARCINOMA 

("8)

1 (2*)

1 (2*) 

1 (2*> 

 (1»9) 

1 (2%) 

(U 91 

•SOBCUT TISSUE 

FIBROMA 

'IBR3SARCOBA 

(U8)

2 («*) 

 (U9) f U 9) 

2 (UX) 

1 (28) 

RESPIRATORY SYSTEM 

• LUNG 

CARCINOMA, NOS, HETASTATIC 

TRANSITIONAL-CELL CARCINOMA, MET 

ALVEOLAR/BRONCHIOLAR ADENOMA 

ALVF3LAB/BROHCHIOLAR CARCINOMA 

OSTBOSARCOMA, MFTASTATIC 

(t8) 

1 (2*) 

1 (2*) 

(1)9) 

1 (2X) 

3 (6%) 

(US) 

2 (i*!5) 

HSMATOPOIETIC SYSTEM 

•MULTIPLE ORGANS 

MALIGNANT LYMPHOMA, SOS 

LEUK'MIA.NOS 

MYELOHONOCYTIC LEUKEMIA 

(U8) 

1 (2%) 

5 (10X| 

(U9 ) (19) 

1

1

 (2*) 

(2%) 

•SPLBEN 

OST^OSAPCOMA, HE^ASTATIC 

BTELOBOHOCYTIC LEUKEMIA 

(U9 ) 

1 (2*) 

(U8 ) 

CIRCULATOR Y SYSTEM 

» NUMBE R O F ANIMAL S IITH TISSUE EXAMINE D

» NUMBE R OF ANIMAL S NECHOPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

 MICROSCOPICALL Y 

A-3 
TABLE


DIGESTIV SYSTEM
 

*LIVER
 

NEOPLASTIC NODULE
 

HEPATOCELLULAR CARCINOMA
 

•PANCREAS
 

ACINAR-CEL L ADENOM A 

tSTOMACH 

SQUABOUS CELL PAPILLOMA
 

SQOABOnS CELL CASCINOMA
 

URINARY SYSTEM
 

tKIDNEY
 

ADENOCARCINOMA, NOS
 

TUBULAR-CELL ADENOMA
 TUBULAR-CFLL ADENOCARCINOMA
 

IKIDNEY/PELVIS
 

TRANSITIONAL-CELL CARCINOMA
 

tURT NARY BLADDER
 

TRANSITIONAL-CELL PAPILLOMA
 

ENDOCRIN" SYSTEM
 

ADENOMA, NOS
 

CHROMOPHOBF, ADENOMA
 

tADRENAL
 

CORTICAL ADFNOMA
 

PHEOCHROMOCYTOHA
 

SANSLIONEtlBOMA
 

t^HYROTD
 

FOLLICULAR-C^LL CARCINOMA
 

C-CELL ADENOMA
 

C-CELL CARCINOMA
 

^PANCREATIC ISLETS
 

ISLET;CELL__ADENOMA
 

 Al (CONTINUED)
 

CONTROL (UNTB) 

01-0070 

(18)
 

1 (2X)
 

2 (IX)
 

(45)
 

(U8)
 

(US)
 

(45)
 

1 (2X)
 

(41)
 

1 (2X)
 

(47)
 

1 (2X)
 

10 (21X)
 

1 (2X)
 

(39)
 

(15)
 

LOW DOSE 

01-0066 

(50)
 

18 (36X)
 

7 (14X)
 

(48)
 

(17)
 

1 <2X)
 

1 (2X)
 

(50)
 

1 (2%)
 

5 (10X)
 

(50)
 

1 (2X)
 

(18)
 

(46)
 

2 (IX)
 

8 (17X)
 

(49)
 

10 (20X)
 

(47)
 

1 (2X)
 

2 <4X)
 

1 (2X)
 

(18)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-4
 

HIGH DOSE
 

01-0067
 

(48)
 

14 (29X)
 10 (21X)
 

(47)
 

1 (2X)
 

(47)
 

1 (2X)
 

(48)
 

6 (13X)
 

4 (8X)
 

(48)
 

1(2%)
 

(15)
 

(39)
 

1 <3J)
 

7 (18X)
 

(48)
 

6 (13X)
 

(16)
 

3 (7X)
 

2 (4X)
 

(17)
 

—1-J2X1-.
 
TABLE Al (CONTINUED) 

CONTROL ((JNTH) L0« DOSE HIGH DOSE 

01-0070 01-0066 01-0067 

REPRODUCTIVE SYSTEM 

*MAMMA8Y GLAND (148) (149) (19) 

PAPILLARY ADENOCARCINOMA 1 (2«) 

FIBROADENOMA 1 (2%) 

*PBEPUTTAL GLAND (148) (149) (149) 

CARCINOMA,NOS 2 (U%) 

SQUA10US CELL CARCINOMA 1 (2X) 

ADENOMA. NOS 1 (2X) 

iTESTIS (17) (50) (148) 

INTERSTITIAL-CELL TOMOR 145 (96*) U8 (96X) 43 (90*) 

NERVOUS SYSTEM 

NONE 

SPECIAL SENS'! ORGANS 

NONE 

M'JSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

• BODY CAVITIES (148) (149) (<49) 

MESOTHELIOMA, NOS 1 (2%) It (3*) 

•PERITONEUM (148) (149) ((49) 

MESOTHELIOMA, NOS 1 (2%) 

ALL OTHER SYSTEHS 

NONE 

* NUMBE R O P ANIMAL S WITF TISSUE EXAMINE D MICROSC3PICALL Y 

* NUP1BFP OF ANIMAL S NECFOPSIED 

A-5 
TABLE Al (CONCLUDED)
 

CONTROL (DNTR) LOW DOSE HIGH DOSE 

01-0070 01-0066 01-0067 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

NATURAL DEATHS 5 10
 

MORIBUND SACRIFICE 5 (I
 

SCHEDULED SACRIFICE 5 5
 

ACCIDFNTALLY KILLED
 

TERMINAL SACRIFICE 3U 31
 

ANIMAL MISSINS 1
 

3 INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOmAL ANIMALS WITH PRIMARY TUMORS* 1)5 U8 U5
 

TOTAL PRIMARY TUMORS 81 115 111
 

TOTAL ANIMALS WITH BENIGN TUMORS U 5 U8 1*5
 

TOTAL BENIGN TUMORS 66 79 73
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 10 17 17
 

TOTAL MALIGNAN T TUMORS 13 17 20
 

TOTAL ANIMALS WITH SECONDARY TUMORSI 2 1
 •"DIAL SECONDARY TUMORS 3 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OH MALIGNANT 2 19 17
 

TOTAL UNCERTAIN TUMORS 2 19 18
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

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

A-6
 
TABLE A2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE

 

CONTROL (UNTR) LOW DOSE HIGH DDSE
 

02-0070 02-0066 02-0067
 

ANIMALS INITIALLY IN STTIDY 50 459 488
 

ANIMALS NECROPSIED 49 45 44
 

ANIMALS EXAMINED HI STOPATHOLOGICALLY ** 19 45 42
 

INTEGUMENTARY SYSTFM 

*SKTN

SQUAMOUS CELL CARCINOMA 

 (l»9) (45) 

1 (2%) 

(44) 

*SOBCUT TISSUE

FIBROMA 

 (1)9) (45) 

1 (2%) 

(44) 

1 <2X) 

RESPIRATORY SYSTFM
 

ILUNG (49) (44) (43)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 2 (5X)
 

HEMATOPOIF.TIC SYSTEM
 

*MULTIPLE ORGANS (49) (45) (44)
 

MALIGNANT LYMPHOMA, NOS 2 (4X)
 

MYELOMONOCYTIC LEUKEMIA 1 (8%) 1 (2*)
 

LYMPHOCYTIC LEUKEMIA 1 (2%)
 

*UPPER TRUNK (49) (45) (44)
 

MYELOMONOCYTIC LEUKEMIA 1 (2%)
 

tlYMPH NODE (42) (44) (40)
 

MALI-5. LYMPHOMA, HISTIOCYTIC TYPE 1 (2X)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

*LTVER (49) (45) (44)
 ____ N.EOPLASTIC_NODULE
 

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

* NUMBE R OF ANIMAL S NECROPSIKD **EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

a <50 ANIMAL S WER E INITIALL Y IN "HE STUDY, BUT 5 I N THF LOW-DOS E GR3U P AND 2 IN THE 

HIGH-DOSE GROUP WERE FOUND TO BE MALE ANIMALS IN FEMALE GROUPS. 

A-7
 
TABLE A2 (CONTINUED) 

CONTROL (DNTR) 

02-0070 

HEPATOCHLLULAR CARCINOMA 1 (2%) 

•PANCREAS 

ADENOCARCINOMA, NOS, METASTATIC 

(147) 

•STOMACH 

ADENOCARCINOMA, NOS, METASTATIC 

(« 9) 

HISH DOSE
 

02-0067
 

10 (23X)
 

(41)
 

(42)
 

(43)
 

1 (2*)
 

(39)
 

13	 (33X)
 7 (18X)
 

(41)
 

(38)
 

1 (3*)
 

1 (3*)
 

1 (3X)
 

(41)
 

(««)
 

3 (7X)
 

(44)
 

URINAR Y SYSTEM
 

•KIDNEY
 

CARCINOMA, NOS
 

TRANSITIONAL-CELL CARCINOMA
 

TUBDLAB-CELL ADENOMA
 

ENDOCRINE: SYSTEM
 

•PITUITARY
 

ADENOMA, NOS
 

CHROMOPHOBE ADFNOMA
 

• ADRENAL
 

CORTICAL ADENOMA
 

CORTICAL CARCINOMA
 

PHEOCHROMOCYTOMA
 

••"HY30ID
 

ADENOMA, NOS
 

FOLLIC'ILAR-C"LL CARCINOMA
 

C-CELL ADENOMA
 

C-CELI. CARCINOMA
 

PAPILLARY CYSTADENOKA, NOS
 

•PANCREATIC	 ISLETS
 

ISLET-CELL ADENOMA
 

REPRODUCTIVE SYSTEM
 

*MAMMAR Y GLAND
 

ADENOMA, NOS
 

ADENOCARCINOMA, NOS
 

"IBROADENOMA
 

*CLTTORAL 1LAND
 

ADENOMA,. NOS
 

» NUMBER OF ANIMAL S BITH TISSUE 

* NUM3E R OF ANIMAL S NECROPSIE D 

(19)
 

1 (2X)
 

(ltd)
 

18 (11*1
 

(49)
 

1 (2%)
 

2 (I**)
 

(40)
 

1 (3X)
 

2 <5X»
 

1 (3X)
 

(47)
 

1 (2X)
 

(49)
 

2 (4%)
 

16 (33*|
 

. .

(49) j mi
 

EXAMINE D MICROSCOPICALLY 

IOW DOSE
 

02-0066
 

3 (7X)
 

(45)
 

1 (2X)
 

(45)
 

1 (2*)
 

(45)
 

1 (2X)
 

(40)
 

5 (13X)
 

9 (23X)
 

(45)
 

1 (2X)
 

1 (2X>
 

(43)
 

2 (5X)
 

(45)
 

(45)
 

1 (2X)
 

2 (4X)
 U (9X)
 

(45)
 

A-8 
TABEL A2 (CONTINUED)
 

• UTERUS


ADENOCARCINOMA, NOS


LEIONYOSARCOMA


ENDOMETRIAL SIROMAL POLYP


ENDONETRIAL STROMAL SARCOMA


*UTERUS/-ENDOMETRIOM


ADENOCARCINOMA, NOS


tOVARY


ADENOCARCINOHA, NOS, HETASTATIC


NERVOUS SYSTEM
 

IBPAIN


SQUANODS CELL CARCINOMA, INVASIV


OLIGODENDPOGLIOMA


SPECIAL SENSE ORGANS
 

NONE
 

MUSCIILOSKELETA L SYSTEM 

NON E 

BOCY CAVITI'S
 

*PFRITONEUM


MESOTHELTOMA, NOS


ALL OTHSR SYSTEMS
 

SITE UNKNOWN
 

SQUAMOUS CELL CARCINOMA


OMFNTUM
 

A DENO C AR CINQMAj., NOSt_MET ASTATIC


CONTROL(ONTR)


02-0070


 (49)


 1 (2X)
 

 12 (24*|


 (49)


 2 (4U)
 

(17)


 (49)


 1 (2X)
 

(49)


 1(2%)
 

 LOW DOSE HIGH D3SE
 

 02-0066 02-0067
 

 (44)	 (12)
 

1 (2X)
 

 10 (23X) 2 (5X)
 

1 (2X)
 

 (44) (42)
 

 (44)	 (42)
 

1 (2X)
 

 (45)	 (42)
 

1 (2)1)
 

 (45) (44)
 

1
 

1
 

*	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NBMBSR OF ANIMALS NECROPSIED
 

A-9
 
TABLE A2 (CONCLUDED) 

CONTBOL(UKTR ) LOU DOSE HIGH D3SE 

02-0070 02-0066 02-OOS7 

ANIMAL DISPOSITION SUMMAR Y
 

ANIMALS INITIALLY IH STUDY 50 «3
 

NATURAL DEATHS 3 13
 

MORIBUND SACRIFICF 7 2
 

SCHEDULED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 35 28
 

ANIMA L MISSING
 

9 INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 45 31 28
 

TOTAL PRIMARY TUMORS 73 55 13
 

TOTAL ANIMAL S WITH BENTGN TDMORS 37 23 23
 

TOTAL BENIGN TUMORS 5« 3« 30
 

TOTAL ANIMAL S WITH MALIGNANT TUMORS 13 11 11
 

TOTAL MALIGNAN T TUMORS 16 13 12
 

•"OTAL ANIMAL S WITH SECONDARY TUMORS* 2
 

TOTAL SECONDARY TUMORS 5
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BFNIGN OB MALIGNAN T 3 8 1
 

"•OTAL UNCERTAIN TUMORS 3 8 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PPTMARY OR METASTATC
 

TOTAL UNCERTAIN TUMORS
 

« PRIMAR Y TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 * SECONDARY TUMORS: METASTATT TUMORS OB TUMORS INVASIVE INTO AN ADJACEST OE3AN
 

A-IO
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE
 

TABLE B1 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 

CONTROL (UNTR)

05-0077

 DOSE A

 05-0066

 DOSE B 

 05-0067 

ANIMAL S

ANIMAL S

ANIMAL S

 INITIALL Y IN STUD Y

 NECROPSIED

 EXAMINE D HISTOPATH3L03ICALLY* *

 50

 46

 45

 50

 35

 31

 50 

 46 

 45 

INTEGUMENTAR Y SYSTEM 

NON E 

RESPIRATORY SYSTEM
 

tLIJNS


HEPATOCELLULAR CARCINOMA, METAST


ALVEOLAR/BRONCHIOLAR ADENOMA


ALVEOLAR/BRONCHIOLAR CARCINOMA


HEMATOPOIETIC SYSTEM
 

*MHLTIPLE ORGANS


MALIGNANT LYMPHONA, NOS


ISPLEEN


HEMANGIOSARCOMA


MALIGNANT LYMPHOMA, NOS


MALIG.LYMPHOMA, HISTIOCYTIC TYPE


*MANDIBULAR L. NODE


MALI3.LYMPHOMA, HISTIOCYTIC TYPE


CIRCULATORY SYSTEM
 

*HE»PT


HEMANGIOSARCOMA


DIGESTIVE SYSTEM
 

(SALIVARY 3LAND


HEMANGIOSARCOMA


 (1*5)

 1 (2X) 

7 (16X)

 4 (9X)

 (21) (U3) 

 1» (9S) 

1 (2?) 

 (46) (35) (46) 

1 (2X) 

 (45)

 1 (2X) 

 (22) (43) 

2 (5X) 

1 (2X) 

 (35)

 1 (3*) 

 (13) (36) 

 (44) (21) (42) 

1 (2X) 

 (43) (21) (40) 

J_J3%1_ 

* NUMBER OF ANIMALS HI^H TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE R OF ANIMAL S NECROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
*1IVEP


HEPATOCELLULAR CARCINONA


*STOMACH


SQUAMOUS CELL PAPTLLOMA


UBTNAB Y SYSTEM
 

*KIDN"Y


ADENOCARCINOMA, NOS


TQBOLAR-CELL ADFNOCARCINOMA


SNDOCHINS SYSTF1
 

NOUE
 

REPRODUCTIVE SYSTEM
 

NONE
 

NEPVOHS SYSIKM
 

NONE
 

SPECIAL SENSE ORGAN S
 

*EAR CANAL


SQUAMODS CELL CARCINOMA


BOSCULOSKELETAL SYSTEM
 

NON?
 

BODY CAVITIES
 

NONE
 

ALL OTHSH SYSTEMS
 

NONE
 

TABLE Bl (CONTINUED) 

CONTRO L (DNTR )

05-0077

 (U5)


 10 (22*)


 (1*2)


 1 (2%)
 

 (1*5)


 (16)


 1 (2%)
 

 DOSE A DOSE B 

 05-0066 05-0067 

 (36)	 (1*5)
 

1 (3X) 8 (18X)
 

 (22) (43)
 

 (37) (15)
 

1	 (2X)
 

1	 (2X)
 

 (35) (1*6)
 

*	 NUMBER OF ANI1ALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUHBE9 OF ANIMAL S NECROPSIED
 

B-4
 
TABLE Bl (CONCLUDED) 

CONTROL (UNTR ) DOSE A DOSE B 

05-0077 05-0066 05-0067 

ANIMAL DISPOSITION SUMMAR Y
 

ANIMAL S INITIALLY IN STUDY SO 50 50
 

NATURA L DFATH5) 7 36 7
 

MORIBUND SACRIFICE 1 1 1 1
 

SCHEDULED SACRTFICF 5 5
 ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 37 37
 A"JIMAL MISSING
 

5) INCLUDES AUTOLYZED ANIMALS
 

TONOH SUMMAR Y
 

TOTAL ANIMAL S KITH PRIMAR Y TUMORS' 21 18
 

TOTAL PRIMARY TUMORS 25 21
 

TOTAL ANIMAL S WITH BFNTGN TUMORS 8 4
 

TOTAL BENIGN TUMORS 8 u
 

TOTMAL ANIMAL S WITH MALIGNAN T IUMORS 15 1 16
 

TO~AL MALIGNAN T TUMORS 17 1 17
 

TO"AL ANIMAL S WITH SECONDARY TUMORS* 1
 

TOTAL SECONDARY TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 TOTAL UNCFRTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPPTMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMAR Y TUM03S: ALL TUMORS EXCEPT SECONDARY TUMORS
 

« SECONDARY TUMORS: M'TASTATIC TUMORS OH TUMORS INVASIVE INTO AN ADJACENT OR3AN
 

B-5
 
FABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE

 

CONTROL(UNTR) DOSE A DOSE B 

06-0077 06-0066 06-0067 

ANIMAL S INITIALL Y IN STUDY 5 0 50 49S 

ANIMAL S

ANIMALS

 NECPOPSIE D

 EXAMINE D HISTOPATHOL03ICALLY*¥ 4 6  15 

38 

34 

1)5 

44 

INTGUMENTAR Y SYSTE M 

(46) (38) (15) 

FIBR3SARCOM A 2 (4X) 

*SUBCU T TISSUE (46) (39) (« 5) 

I EIOPIYOSARCOM A 1 (2%) 

RESPIRATOR Y SYSTE M 

»LUN T (15) (27) (43) 

ALVEOLAR/BRONCHIOLA R ADENOM A 1 (2%) 

ALVOLAR/BRONCHIOLA R CARCINOM A 

HEMATOPOIETIC SYSTEM
 

• MULTIPLE ORGANS (46) (38) (45 ) 

MAIIGNANT LYBPHOMA, NOS 3 (7%) 1 (2*) 

MALI3.LYMPHOMA, UNDIFFER-TYPE 1 (2%} 

MALIG.LYMPHOKA, HISTIOCYTIC TYPE 6 (13*| 1 (3X) 4 (9X) 

LYMPHOCYTIC LEUKEMIA 1 (2%) 

tPEYERS PATCH (13) (23 ) (44 ) 

MALI3.LYMPHOHA, HISTIOCYTIC TYPE 1 (2%) 

CIRCULATOR Y SYSTE M 

NON E 

DIGESTIVE SYSTE M 

ULIVfl H (1(5) (34 ) (44 ) 

___ _ NEOPLASTIC_NODUL E 

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

* NUMBE R OF ANIMAL S NECSOPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

3> 5 0 ANIMAL S WPR E INITIALL Y IN THF STUDY, BUT ON E WAS FOUN D TO BE A MAL E ANIMA L 

IN A FEMALE GROUP. 

B-6 
TABLE B2 (CONTINUED) 

CONTROL(UNTR )

06-0077

 DOSE A

 06-0066

 DOSE B 

 06-0067 

HEPATOCELLULA R CARCINOM A

tSTOMAC H

SQUAMOU S CELL PAPILLOM A

 1 (9%)

 ('42)

 3 (7%) 

2 (6% )

 (21)

 9 (20%) 

 (13) 

URINAR Y SYSTE M 

NON E 

ENDOCRIN E SYSTEM 

tPITUTTAR Y

ADENOMA , NOS

• ADRENA L

CORTICA L ADENOM A

IPANCREATI C ISLETS

ISLET-CELL ADENOM A

 (37)

 6 (16X )

 (43 )

 1 (21) 

 ('41 )

 1 (2% ) 

 (19 )

 (27)

 (27 )

 . (35) 

3 (9X ) 

 (U2 ) 

 (1(2) 

REPRODUCTIV E SYSTE M 

'MAMMAR Y GLAN D

ADENOCARCINOHA , NOS

tDTFRU S

ENDOMETRTA L STROMA L POLY P

*OVAR Y

LUTEOM A

 (16)

 1 (2X) 

 (13)

 (11)

 1 (2% ) 

 (38)

 (22 )

 (22)

 (15) 

 (11) 

1 (2)6) 

 (11) 

NERVOU S SYSTE M 

NON E 

SPECIA L SENS E ORGAN S 

NON E 

MUSCULOSKELETA L SYSTE M 

NONE 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBF R OF ANIMAL S NECROPSIE D 

B-7 
TABLE B2 (CONCLUDED) 

CONTRO L (DNTR ) DOSE A DOSE B 

06-0077 06-0066 06-0067 

BODY CAVITIES
 

NONE
 

ALL OTHFR SYSTEMS
 

NONE
 

ANIMAL DISPOSTTTON SUMMAR Y
 

ANIMALS INITIALLY IN STUDY 50 50 « 9 

NATURAL DEATH3 32 6 

MORIBUND SACRIFICE 8 1 

SCHEDULED SACRIFICE 5 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 35 10 37 

ANIMA L MISSING
 

3 INCLUDES AHTOLYZFD ANIMALS
 

TUMO P SUMMAR Y 

TOTA L ANIMAL S WIT H PRIMAR Y TUMORS * 2 2 U 1 6 

TOTAL PRIMAP Y TUMORS 32 <4 22 

TOTAL ANIMAL S WIT H B'NIG N TUMOR S 1 2 It 

•"OTAL BENIG N TUMOR S 13 <l 

TOTAL ANIMAL S WIT H MALIGNAN T TUMOR S 1 8 It 1 2 

TOTAL MALIGNAN T TUMOR S 19 It 15 

TOTAL ANIMAL S WITH SECONDARY TUMORSt
 

TO^AL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT 3
 

"OTAL UNCERTAIN TUMORS 3
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OP METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: AIL TUMORS EXCEPT SECONDARY TUM3RS
 

* SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTD AN ADJACENT OB3AN
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN MALE RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 

<rONTRO L [IINTR ] 1 LOW DOSE 

01-0070 01-0066 

ANIMAL S INITIALL Y I N STDDY 50 50 

ANIMAL S MISSING 1 

ANIMAL S NECROPSIE D 18 19 

ANIMAL S EXAMINE D HI STOPATHOLOGICALLY* * 13 18 

INTEGUMENTAR Y SYSTE M 

*SKTN (18) (19)
 

NOCLEAB-SHAPE ALTERATION
 

HYP"RKERATOSIS 1 (2%)
 

*SUBCUT TISSUE (18) (19)
 

FIBPOSIS 1 (2%)
 NECROSIS, NOS 1 (2JS)
 

NECROSIS, FAT
 

RESPIRATOR Y SYSTE M 

tTRACHEA (15) (17)
 

INFLAMMATION, NOS 8 (17X)
 

INFLAMMATION, ACUTE/CHRONIC 18 (40X)
 

*LUN3/BRONCHUS (18) (19)
 

BRONCHIECTASIS 3 (6*) 1 (2X)
 

INFLAMMATION, NOS 3 (6%)
 

INFLAMMATION, FOCAL 1 (8X)
 

INFLAMMATION, SOPPURATIVE
 

ILUNS/BRONCHIOLE (18) (19)
 

INFLAMMATION, NOS
 

tHING (18) (19)
 MINERALIZATION
 

CONGESTION, NOS 1 (2*)
 LOBA R PNEUMONIA, NOS
 

INFLAMMATION, FOCAL 2 (1*) 1 (2X)
 INFLAMMATION, INmER?TITIAL 22 (15X)
 

INFLAMMATION, SUPPURATIVE
 

ABSCESS, NOS 1 (2X)
 

PNEUMONIA,. CHRONIC MURINE - - 1_12%1
 

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

* NUMBE R OF ANIMAL S NECROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

HIG H DOSE 

01-0067 

50 

19 

18 

(19)
 

1 (2*)
 

(19)
 

1 <2X)
 

(17)
 

1 (9Jt)
 

(18)
 

1 (8%)
 

3 (6S)
 

5 (10%)
 

1 (2X)
 

(18)
 

1 (2*)
 

(18)
 

1 (2%)
 

1 (2*)
 

22 (16%)
 

1 (21)
 

C-3 
TABLE Cl (CONTINUED) 

,RANULOKA, NOS


HEMOSIDEPOSIS
 

HYPFBPLASIA, NOS
 

HYPERPLASIA, EPI^HFLIAL
 

HYPERPLASIA, FOCAL


HYPERPLASIA, ALVFOI.AB EPITHELIUM


HEMATOPOIETIC SYSTEM
 

«BONE MABRO H
 

MYFLOFIBROSI S 

MEGAKARYOCYTOSI S 

HYPFRPLASIA , HFPIAT3POIETIC 

HYPERPLASIA , GBANULOCYTI C 

HYP'BPLASIA , MEGAKARiOCY'"IC 

»SPI EEt< 

>iOS 

FOCAL
 

FE10SIDEROSIS
 

pYpt-pnLASIA , HFH ATOP3IETIC 

• 1YPCPPLASIA , FRYTHBOI D 

HYPZPPLASIA , RETICULO M

"RY'HROPOIESI S 

tlYM-'H MODE
 

INFLA1MA"I01, NOS
 

HYPEtPLASIA, NOS
 

PETICULOCYTOSIS
 

I Y1PHOCYTOSIS
 

PLASNACYTOSIS
 

H'P^RPLASIA, LYMPH3ID
 

*MANDIPULA R L. NODE
 

DILATATION, NOS
 

HYP'RPLASIA, NOS
 

-IPCHLATTRY SYSTEM
 

*HEART
 

FIBROSIS, FOCAL
 

FIBBOSIS, DIFFUSE
 

(tHYOCABDIU I 

 CEL L 

CONTROL (UNTR ) 

01-0070 

1 (2%)
 

1 (2%)
 

1 (2X)
 

(U8)
 

1 (2%)
 

1 (2X)
 

1 (2)5)
 

1 (2X)
 

(48)
 

1 (2X)
 

(42)


(42)


1 (2%)
 

1 (2%)
 

(48) 

11 (23%) 

1 (2%) 

(48) 

INFLAM(1A_TION £ ._I_NTFRSTITIAL _ 

*	 NU13FR OF ANIMALS HI^H TISSUE EXAMINED MICROSCOPICALLY
 

* M!BB"R OF ANIMAL S NFCROPSIED
 

C-4
 

L OW DOSE HIGH D3SE 

01-0066 01-0067 

1	 (2%)
 

2 (4%)
 

1 (2%) 5 (10%)
 

(50) (47)
 

1 (2*)
 

(U9)
 

2 (U%)
 

1 (2%)
 

12 (24X) 7 (15X)
 

15 (31%) t (3X)
 

29 (59%) 23 (U3)t)
 

1 (2*)
 

 (47)	 (46)
 

1 (2X)
 8 (17%)
 

1 (2%)
 

1 (2%) 1 (2%)
 

2 (4JS)
 1 (2%) 2 (4%)
 

 (47) (46)
 

(50) (48)
 

(50) (48)
 
TABLE Cl (CONTINUED) 

CONTROL (UNIR ) LOW DOSE HISH D3SE 

01-0070 01-0066 01-0067 

INFLAMMATION, ACUTE/CHRONIC 

FIBROSIS 

FIBROSIS, FOCAL 

DEGENERATION, NOS 

3 (5X) 

2 (IX) 

1 (2X) 

15 (30%) 3

1

 (5%) 

 (2%) 

(ENDOCARDIUM

INFLAMMATION, FOCAL 

(18) (50) (18) 

1 (2%) 

ICARDIA; VALVE 

INFLAMMATION, ACUTE/CHRONIC 

(18) 

1 (2X) 

(50) (18) 

"CORONARY ARTERY 

MINERALIZATION 

PERIVASCULITIS 

(18) 

1 (2X> 

(19) 

1 (2%) 

(19) 

1 (8!5) 

*PULMOSARY ARTERY 

MINERALIZATION 

(18) 

11 (23X) 

(19) 

1 (8X) 

(19) 

9 (18%) 

DIGESTIVE SYSTEM 

(LIVER (18) (50) (18) 

INFLAMMATION, NECROTIZING 1 (2%) 

ABSCESS, NOS 1 (2%) 

NECROSIS, FOCAL 8 (17X) NECROSIS, COAGULATIVE 2 (IX) 

NECROSIS, HEMORPHAGIC 1 (2%) 

METAMORPHOSIS FATTY 1 (8X) 11 (28X) 8 (17%) 

CHOLESTEROL DEPOSIT 3 (6JS) 

CYTOPLASMIC VACUOLIZATION 3 (6%) 

HYPKRPLASIA, NOS 1 (2X) 

HYP^RPLASIA, FOCAL 8 (17*) 13 (26X) 13 (27%) 

ANGIECTASIS 2 (IX) 2 (1%) 1 (2X) 

ERYTHROPOIESIS 1 (2X) 

*LIVER/:ENTRILOBULAR (18) (50) (18) 

DEGENERATION, EOSINOPHILTC 2 (1*) 

*BILE DUCT 

CALCULUS, NOS (18) (19) 

1 (2%) 

(19) 

INFLAMMATION, NOS 7 (11X) 1 (3!6) 

HYPERPLASIA, NOS 6 (13X| 13 (88X) 17 (95%) 

• PANCREAS (15) (18) (17) 

HEMORRHAGF 1 (2*) 

INFLAMMATION, NOS 17 J35X.1 18 _[33%1 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSC3PICALLY 

* NUMBER OF ANIMALS NECROPSIED 

C-5
 
TABLE Cl (CONTINUED) 

CONTROL (UNTR) 

01-0070 

LOW DOSE 

01-0066 

TNFLAMMATION, ACUTF/CHRONIC 

PSBIAPTEBITIS 

ATROPHY, "OCAI 

6 (13)5) 

1 (2%) 

1 (2%) 

*PANCR?:ATIC DUCT HYPEBPLASIA, NOS 

(U5) (18) 

tPANCHEATTC ACINUS 

INFLAMMATION, NOS 

ATROPHY, NOS 

HYP'RPLASIA, "OCAI. 

(K5) (13) 

tSTOMACH 

EPIDERMAL INCLUSION CYST 

INFLAMMATION, NOS 

!!LCEP, NOS 

INFLAMMATION , ACUTE/CHRONIC 

HYP"RPLASIA, NOS 

HYPVRKVB ATOSIS 

ACANTHOSIS 

(18) 

1 (2)1) 

(") 

1 (2%) 

5 (11*) 

6 (13X) 

11 (23%) 

•GASTRIC MtJCOSA 

DEGENERATION, NOS 

(U8) («7) 

•PFYERS PATCH 

HYPERPLASTA, NOS 

HYPERPLASIA, PETICULIIM CELL 

(K5) 

1 (2X) 

(1*S) 

1 (2%) 

*ILFUM 

HYPERPLASIA, LYMPHOID 

(15) 

1 (2%) 

(18) 

•COLON 

NEMATODIASI S 

(Hit) 

1 (9X) 

(16) 

1 (2%) 

UPINAR Y SYSTE M 

(18) (50) 

RLOdERULON'SPHRITTS , NO S 3 (6%) U6 (92% ) 

INFLAMMATION , ACUTE/CHRONI C 1 (2%) 

FIBPOSI S 

FIBP3SIS, "OCAL 

NKPHROSIS , NO S 1*1 (85X)

 

GLOIFRnLOSCLEROSIS , NO S 

riYP"RPLASIA , TITBOLA R CEL L 11 (22%) 

HYPERPLASIA . EPITHELIA L 

* NUMBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE R OF ANIMAL S N=;CPOPSIF,D
 

HIGH D3SE
 

01-0067
 

(47)
 

3 (6X)
 

(U7)
 

1 (2X)
 1 (2*)
 

3 (6*)
 

(1*7)
 

2 <!*X)
 

1 (2X)
 

1 (2*)
 

1 (28)
 7 (15X)
 

8 (17X)
 

(1*7)
 

1 (2X)
 

(17)
 

9 (19%)
 

(17)
 

(11)
 

(U8 ) 

15 

1 (2% ) 

1 (2% ) 

1 (2%)

13 (27%) 

C-6
 
HIGH DOSE
 

01-0067
 

(18)
 

1 <2X)
 

1 <2X)
 

1 (2X)
 

(18)
 

1 (2X>
 2 (IX)
 

(15)
 

1 <2X)
 

(39)
 

1 (3X)
 

3 (3X)
 

(18)
 

(18)
 

1 (2X)
 

(18)
 

9 (19X)
 

1 (2X)
 

1 (2X)
 

(16)
 

1 (7«>
 

(17)
 

2 (IX)
 

(19)
 

1 llil
 

tKIDNEY/TUBULE
 

MINERALIZATION
 

NECROSIS, NOS
 

NECR3SIS. FOCAL
 

IKIDNEY/PELVIS
 

MINERALIZATION
 

HYPERPLASIA, EPITHELIAL
 

iURINAHY BLADDER
 

HYPERPLASIA, EPITHELIAL
 

METAPLASIA, SQUAMOIIS
 

ENDOCRINE SYSTEM
 

IPITBITARY
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

tADRENAL
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

*ADRENAL CORTEX
 

NODULE
 

HYPERPLASIA, FOCAL
 

IADPENAL MEDDLLA
 

NECROSIS, COAGULATIVE
 

HYPERPLASIA, NODULAR
 

HYPERPLASIA, NOS
 

HYPPRPLASIA, FOCAL
 

•THYROID
 

HYPEHPLASIA, NOS
 

HYPERPLASIA, T-CELL
 

fPANCREATIC ISLETS
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

REPRODUCTIVE SYSTEM
 

*MAMMAH Y GLAND
 

- _ GALACTOCELF


* NUMBE R OF ANIMAL S WITH TISSUE

* NUMBE R OF ANIMAL S NECROPSIED 

TABLE Cl (CONTINUED) 

CONTROL(UNTR)
 

01-0070
 

(18)
 

(U8)
 

1 (2X)
 

(16)
 

1 (2*)
 

(11)
 

3 (7)1)
 

(17)
 

1 (2X)
 

3 (6X)
 

(17)
 

1 (2%)
 

(17)
 

(39)
 

1 (3%)
 

(15)
 

1 (2*)
 

(18)
 

. 1.11*1
 

LOW DOSE
 

01-0066
 

(50)
 

1 (2X)
 

(50)
 

3 (6X)
 

(18)
 

(16)
 

1 (2*)
 

1 (2X)
 

(19)
 

(19)
 

1 (2%)
 

(19)
 

1 (2X)
 

1 (8X)
 

(17)
 

1 (2X)
 

(18)
 

1 (2%)
 

(19)
 

2 11%1 . .
 

 EXAMINE D MICROSCOPICALL Y 

C-7 
TABLE Cl (CONTINUED) 

V
 

HYPERPLASIA, NOS
 

iPROSTATE
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

INFLAMMATION, »CDTE
 

INFLAMMATION, ACUTE FOCAL
 

INFLAMMATION, ACBTE/CHBONIC
 

HYPEBPLASIA, NOS
 

HYPEPPLASIA, FOCAL
 

METAPLASIA, SQDAHOUS
 

•SEMINAL	 VESICLE
 

ATROPHY, NOS
 

tTESTIS
 

MINERALIZATION
 

DEGENERATION, NOS
 

ATBOPHY, NOS
 

HYPSRPLASIA, NOS
 

HYPERPLASIA, INTERSTITIAL CELL
 

ITESTIS/TUBOLE
 

MINERALIZATION
 

DESPERATION, NOS
 

NERVOUS SYSTFM
 

•PRAIN
 

INFLAMMATION, FOCAL SRANULOM4TOO
 

SPECIAL SENSE ORGANS
 

• FYE
 

CATARACT
 

• EY1VCORNKA
 

INFLAMMATION, NOS
 

*FYF,/REriNA
 

ATROPHY, NOS
 

MOSCDL3SKELETA L SYSTEM 

• BONE 

CONTROL (UNTR)
 

01-0070
 

3 (6*>
 

("3)
 

1 (2%)
 

6 (141)
 

8 (19«)
 

2 (5»)
 

(48)
 

2 (4*)
 

(47)
 

39	 (83K)
 

1 (2*)
 

(47)
 

(»7)
 

(48)
 

(48)
 

(48)
 

(48) 

HIGH D3SE
 

01-0067
 

13 (27X)
 

(46)
 

25 (54X)
 

1 (2X)
 

(49)
 

1 (2X)
 

(48)
 

2 <4K)
 

8 (17*)
 

3 <6X)
 

5 (10*)
 

(48)
 

5 (104)
 

1 (2X)
 

(47)
 

1 (2*)
 

(49)
 

3 (S»)
 

(49)
 

(49)
 

2 <4X)
 

(49) 

LOW DOSE
 

01-0066
 

20	 (41*)
 

(48)
 

22 (461)
 

1 (2«)
 

3 (6X>
 

(49)
 

(50)
 

2 (41)
 

5 (10*)
 

1 <2»)
 

(50)
 

(50)
 

(49)
 

2 (4»)
 

(49)
 

1 (2X)
 

(49)
 

1 (2*1
 

(49 ) 

• NOMBE R OF ANIMAL S «ITH TISSUE EI»III»eD IHCROSC3PIC»tLI 

• NIIMBFR OF ANIMAL S NECROPSIED 

C-8 
TABLh Cl (CONCLUDED) 

CONTROL (UNTR) LOW DOSE HIGH D3SE 

01-0070 01-0066 01-0067 

"IBROSIS 1 (2*) 

poq-IUDTIO N 1 (2% ) 1 (2*) 

BOD Y CAVITI' S 

NON F 

A LI OTrfF P SYSTFM S 

ADIPOSE TTSSUF 

IMFLA^MAT ION, A^OTE/CH^ONIC 2 

OBFNTOB N^CRSSIS, NOS 1 1 

SPFCIAL MOH^HOLTGY 

ANI1A L MISSING/N O NECROPS Y 1 

AI7TO/NEC10PSY/HIST O PFRF 1 

AUTO/NEC»OPSY/NO HIS10 1 1 

A1TTLYSIS/N O NFCROPS Y 1 1 1 

# NUPBF R Of AM1ALS KIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* NUMBE R O F ANIMAI S NECR3PSIE D 

C-9
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE RATS TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 

CONTROL (UNTR) LOU DOSE HIGH D3SE
 

02-0070 02-0066 02-0067
 

ANIMALS INITIALLY IN STUDY 50 1453
 

ANIMALS NFCROPSIED 49 45 44
 ANIMAL S EXAMINED HISTnPATHOLOSIOALLY** 49 U5 42
 

INTEGUMENTARY SYSTEM
 

*SKIN (49) (45) (44)
 

EPIDERMAL INCLUSION CYST 1 (2X)
 

*SURCHT TISSUE (49) (45) (44)
 

ABSCESS, NOS 1 (2S)
 

RESPIRATORY SYSTEM 

*T"ACHEA (49) (43) (42) INFLAMMATION, NOS 3 

INFLAMMATION, ACUTE/CHRONIC 15 (31*) 

H.UNT/BRONCHUS (49) (44) (43) 

BRONCHIECTASTS 1 (2X) 

INFLAMMATION, FOCAL 2 (5S) 3 (7X) 

INFLAMMATION, SUPPURATIVE 1 (2%) 

INFLAMMATION, ACUTE/CHRONIC 1 (2%) 

*LUN", (49) (44) (43) 

INFLAMMATION, FOCAL 

INFLAMMATION, INTERSTITIAL 

2

2

 (4X) 

 (4%) 25 (57%) 1 16

 (2*) (37%) 

INFLAMMATION, SHPPURATIVE 1 (2 X ) 

HYPERPLASIA, EPITHELIAL 3 (7*) 2 (5X) 

HYPEHPLASIA, ALVEOLAR EPITHELIUM 1 (2%) 

HEMATOPOIETIC SYSTEM
 

*BONS MARRO W (46 ) (45 ) (43) 

OSTEOSCLEROSIS 1 (2%) 

*SPLEEN (45 ) (43) 

HEMOSIDEROSIS 

* NUMBE R O F ANIMAL S WIT H TISSU E EXAMINE D MICHOSC3PICALL J 

* NUMBE R O F ANIMAL S NECROPSIE D **EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

») *-0 ANI1AL S WER " INITIALL Y IN THE STUDY , BUT 5 IS THE LOW-DOS E GR3U P AND 2 IN 

THE HIGH-DOSE GROUP WERE FOUND TO BE MALE ANIMALS IN FEMALE GROUPS. 

C-IO
 
HYPERPLASIA, HEMATOPOIETIC
 

HYPERPLASIA, ERYTHROID
 

HYPERPLASIA, RETICULUM CELL
 

•LYMPH	 NODE
 

INFLAMMATION, NOS
 

HYPERPLASIA, NOS
 

RETICULOCYTOSIS
 

LYMPHOCYTOSIS
 

PLASMACYTOSIS
 

HYPERPLASIA, RETICOLDN CELL
 

HYPERPLASIA, LYMPHOID
 

•MFDIASTINAL	 L.NODE
 

HYPERPLASIA, NOS
 

PLASMACYTOSIS
 

CIRCULATORY SYSTEM
 

IHEART
 

FIBROSIS, FOCAL
 

FIBROSIS, DIFFnSE
 

PERIARTERITIS
 

•MYOCARDIUM
 

INFLAMMATION. DOS
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, ACUTE/CHRONIC
 

FIBROSIS
 

FIBHOSIS, FOCAL
 

"IBROSIS, DIFFUSE
 

tCAPDIA: VALV E
 

INFLAMMATION, ACUTF/CHRONIC
 

'PULMONARY ARTERY
 

MINERALIZATION
 

DIGESTIVE SYSTEM
 

• LIVES


DEGENERATION, EOSINOPHILTC


NECROSIS, FOCAL


NECROSIS j,_cgAGULATIVE


* NBMBEP OF ANIMALS WITH TISSUE


* NUMBER OF ANIMAL S NECROPSIED
 

TABLE C2 (CONTINUED) 

CONTRO L (ONTR ) LOW DOSE 

02-0070 02-0066 

1 (2%)	 29
 

39 (87%)
 

1 (2%)
 

(42)
 

1 (2%)
 

2 (5%)
 

1 (2%)
 

2 (5%)
 

2 (5%)
 

(1(2) (44) 

1 (2%) 

1 (2%) 

(«9) (45)
 

1 (2X)
 

1 (2%)
 

1 (21)
 

(U9) (45)
 

1 (2X)
 

2 («) 31 (69%)
 

1 (2X)
 

3 (7«)
 2 (4%)
 

1 (2X)
 

l<*9) (45)
 

1 (2X)
 

(U9) (45)
 

9 (18JS) 2 (4X)
 

 (49)	 (45)


 2 (4X)
 

3 (6X) 7 (16%)


 I_i2%)
 

 EXAMINED MICROSCOPICALLY
 

HIG H DOSE 

02-0067 

22 (51%)
 

27 (63%)
 

(40)
 

2 (5%)
 

5 (13%)
 

1 (3%)
 

1 (3%)
 

4 (10%)
 

1 (3)1) 

(140) 

(43)
 

(43)
 

1 (2%)
 25 (58X)
 

8	 (19X)
 

(43)
 

(44)
 

 (44)
 

4 (9%)
 

C-l
 
TABLE C2 (CONTINUED) 

CONTROHUNTR) 

02-0070 

LOW DOSE 

02-0066 

HIGH D3SE 

02-0067 

METAMORPHOSIS TATTY 

CYTOPLASMIC VACUOLIZATION 

HYPERPLASIA, FOCAL 

HYPSRPLASIA, DIFFUSE 

AN3IECTASIS 

4 (8%) 

29 (59%| 

1 (2X) 

4 (9X) 

1 (2X) 25 (56%) 

1 <2X) 

1 (2X) 

3 (7X) 

2 (5X) 

26 (59X) 

*BTL^ DUG1" 

INFLAMMATION, NOS 

HYPERPLASIA, NOS 

HYPERPLASIA, FOCAL 

(49) 

5 (10X) 

1 (2%) 

(45) 

4 <9X) 

33 (73X) 

(44) 

4 (9X) 

38 (36X) 

• PANCREAS 

INFLAMMATION, NOS 

INFLAMMATION, ACUTE/CHRONIC 

ATPOPHY, NOS 

(47) 

4 (9%) 1 (2X) 

(45) 

17 (38X) 

(41) 

19 (46X) 

tPANCPEATIC ACINUS 

HYPERPLASIA, NOS 

HYPFRPLASIA, FOCAL 

(47) (45) 

1 (2X) 

(41) 

1 (2%) 

1 (2X) 

tSTOIACH 

INFLAMMATION, NOS 

ULCER, NOS 

INFLAMMATION, FOCAL 

ULCER, "OCAL 

HYPSRPLASIA, NOS 

HYP'RPLASIA, FOCAL 

HYPEBKERATOSIS 

ACANTHOSIS 

(49) 

1 (2%) 

(45) 

1 (2X) 

1 (2%) 

4 (9%) 

5 (11X) 

(«2) 

1 (2X) 

1 (2X) 

2 (5*) 

7 (17X) 

10 (24X) 

•PEYEBS PATCH 

HYPERPLASTA, NOS 

(49) (45) 

8 (18X) 

(41) 

10 (2KX) 

•COLON 

NS1ATODIASIS 

(44) 

2 (5?) 

(41) 

1 (2%) 

(37) 

5 (14%) 

URINAR Y SYSTE M 

• KIDNE Y (49) (45) (43) 

MINERALIZATION 1 (2%) 1 (2X) POLYCYSTIC KIDNEY 1 (2X) 

SLOM"BULONEPHRITIS, NOS 43 (96)6) 41 (95X) 

NEPHROSIS, NTT 34 (69X) 

HYPEBPLASIA, riikJLAP CELL 3 (7X) 1 (2%) HYPERPLASIAt _EPITHELIAL 

» NUMBER OF ANIMALS WITH TISSDE EXAMINED MICROSCOPICALLY
 

* NUMBE R OF ANIMAL S NECROPSIED
 

C-12
 
HIGH DOSE
 

02-0067
 

(13)


1 (2X)
 

(1*3)
 

(i*1)
 

1 (2X)
 

(39)
 

1 (3X)
 

(11)
 

(1*1)
 

1 (2X)
 

1 (2X)
 

2 (5X)
 

(H1)
 

2 (5X)
 

(38)
 

1 (3X)
 

(38)
 

(1*1)
 

(11)
 

7 (16X)
 

•KIDNEY/TUBULE
 

MINERALIZATION
 

•KIDNEY/PELVIS
 

HYPE8PI.ASIA, FOCAL
 

•URINARY	 BLADDFR
 

HYPERPLASIA, EPITHELIAL
 

ENDOCRINE SYSTEM
 

IPITUITAPY
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

•ADRENAL
 

METAMORPHOSIS FATTY
 

•ADPENAL	 CORTFX
 

NODULE
 

METAMORPHOSIS FATTY
 

HYPERPLASIA, NOS
 

HYPFRPLASIA, FOCAL
 

•ADRENAL	 MEDULLA
 

HYPERPLASIA, NODULAR
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

I^HYROTD
 

HYPERPLASIA, C-CELL
 

•"HYROID	 FOLLICLE
 

NECROSIS, FOCAL
 

• PANCREATIC ISLFTS
 

HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEM
 

*MAMMAR Y SLAND
 

GALACTOCELE
 

IN"LAMBATIONi ACUTE
 

» NUMBE R OF ANIMAL S WITH TISSUE

* NUMBE R OF ANIMAL S NECFOPSIED 

TABLE C2 (CONTINUED) 

CONTROL (UNTR)
 

02-0070
 

(K9)
 

(K9)
 

(<*9)
 

(11)
 

1 (2*)
 

2 <5X)
 

(1*9)
 

3 (6*)
 

(1*9)
 

3 (6X>
 

1 (2X)
 

(i*9)
 

1 (2X)
 

1 (2X)
 

(1*0)
 

(HO)
 

(!*7)
 

(«9)
 

9 (18X)
 

„ 1_12XL
 

LOW DOSE
 

02-0066
 

(15)
 

(15)
 

1 12%)
 

(11)
 

1 (2X)
 

(10)
 

(15)
 

(15)
 

1 (2X)
 

1 (2X)
 

(15)
 

1 (2X)
 

(13)
 

U (9%)
 

(13)
 

1 (2X)
 

(15)
 

1 (2X)
 

(15)
 

S (18%)
 

 EXAMINE D MICROSCOPICALL Y 

C-I3 
HYPBHPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

*VAGINA
 

INFLAMMATION, ACtJTE/CHRONIC
 

tUTERUS
 

HYDROBETRA
 

ABSCESS, NOS
 

HYPERPLASIA, ADEKOMArOOS
 

ICERVIX UTERI
 

INFLAMMATION, ACUTE/CHRONIC
 

HYPERPLASIA, BASAL CELL
 

ACANTHOSIS
 

tUTERUS/ENDOMETRIUM
 

INFLAMMATION, HOS
 

INFLAMMATION, SDPPDRATIVE
 

INFLAMMATION, ACUTE
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

HYPERPLASIA, CYSTIC
 

HYPERPLASIA, STROMAL
 

tOVARY/OVIDUCT
 

INFLAMMATION, NOS
 

INFLAMMATION, ACUTE
 

tOVABY
 

CYST, NOS
 

INFLAMMATION, NOS
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

*EYE
 

INFLAMMATION, SBPPURATITE
 

SYNECHIA, NOS
 

CATARACT
 

*FYF/CORNEA
 

INFLAMMATION.,. CHRONIC
 

TABLE C2 (CONTINUED)
 

CONTROL (DNTR)
 

02-0070
 

23 (47X)
 

2 (4X)
 

(49)
 

1 (2%)
 

(U9)
 

6 (12X)
 

(19)
 

2 (IX)
 

1 (2X)
 

1 <2X)
 

(49)
 

23 (47X)
 

5 (10X)
 

5 (10X)
 

<U9)
 

1 (2X)
 

(47)
 

2 (1%)
 

(49)
 

1 (2X)
 

1 (2X)
 

(49)
 

1.12SL
 

LOW DOSE
 

02-0066
 

22 (49X)
 

(45)
 

(44)
 

1 (2X)
 

6 (14X)
 

(44)
 

(44)
 

20	 (45X)
 

1 (2X)
 

6 (14X)
 

1 (2X)
 

1 (2X)
 

(44)
 

2 (5X)
 

(44)
 

6 (14X)
 

2 (5X)
 

(45)
 

1 (2X)
 

1 <2X)
 

(45)
 

HIGH D3SE
 

02-0067
 

18 (41X)
 

(44)
 

(42)
 

1 (2X)
 

(42)
 

(42)
 

8 (19X)
 

2 (5X)
 

1 (2X)
 

(42)
 

2 (5X)
 

(42)
 

5 (12X)
 

(44)
 

(44)
 

* NUMBER OF ANIMALS HITH TISSDE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-I4
 
*EYE/RETINA


DEGENERATION, NOS


CATARACT


ATROPHY, NOS


MUSCUL3SKELETAL SYSTEM
 

*STERNHM


OSTEOPETROSIS


BODY CAVITIES
 

*BEDIASTINOM


PERIARTEEITIS


ALL OTHER SYSTEMS
 

ADIPOSE TISSUE
 

INFLAMMATION, ACUTE/CHRONIC


INFLAMMATION, CHRONIC


OMENTUM
 

MINERALIZATION


NECROSIS, NOS


SPECIAL MORPHOLOGY SUMMARY
 

AUTO/NECROPSY/NO HISTO


AUTOLYSIS/NO NECROPSY


TABLE C2 (CONCLUDED)
 

CONTROL (ONTH)


02-0070


 (t9)


 1 (2X)
 

(49)


 1 (2X)
 

 (1*9)


 1 (2X)
 

3
 

2
 

1
 

1


 LOU DOSE HIGH DOSE 

 02-0066 02-0067 

 (15) (It) 

1 (2K) 

1 (2*) 

 (<»5) (It) 

 (15) (I»<1) 

1 

2 

U 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NDMBER OF ANIMALS NECROPSIED
 

C-15
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 1-AMINO-2-METHYLANTHRAQUINONE
 

TABLE Dl 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN MALE MICE TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 

CONTROL (ONTR) DOSE A DOSE B 

05-0077 05-0066 05-0067 

ANIMAL S INITIALLY IN STUDY 50 50 50 ANIMAL S NF.CROPSIED 46 35 46 

ANIMAL S EXAMINE D HISTOPATHO LOGICALLY »» 45 34 U5 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

tTRACHEA (44) (18) (42)
 

INFLAMMATION, NOS 1 (6%)
 

*LUNG (45) (21) (43)
 

INFLAMMATION, FOCAL 1 (5X)
 

INFLAMMATION, INTERSTITIAL 5 (24X) 1 (2X)
 

PERIVASCDLITIS 1 (5X) 3 (7X)
 

ARTERIOSCLEROSIS, NOS 1 (2X)
 

HYPEHPLASIA, ADENOHATOUS 1 (2*)
 

HEMATOPOIETIC SYSTEM
 

*BONE MARRO W (45) (19) (43)
 MYELOFIBROSIS 1 (2X)
 

tSPLEEN (45) (22) (43)
 FIBROSIS 1 (2X)
 HEMOSIDEROSIS 1 <5X)
 HYPERPLASIA, NOS 2 (9X) 4 (9X)
 HYPEPPLASIA, HEMATOPOIETIC 3 (7X)
 HYPERPLASIA, EBYTHP3ID 1 (2X)
 HYP"PPLASIA, RETICULBM CELL 3 (7X) 1 (2X)
 HYPEHPLASIA, LYMPHOID 8 (19*)
 H»MATOPOIESIS 1 (2X)
 

#LYMPH NODE (35) (13) (36)
 

HEMORRHASE 1 <3X)
 INFLAMMATION, NOS 1 I8X) 7(19X1 .
 

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

« NUMBF R OF ANIMAL S NECROPSIED 

**FXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-3 
TABLE Dl (CONTINUED)
 

HYPERPLASIA, NOS 

RETICBLOCYTOSIS 

HYPERPLASIA , HEMATOPOIETIC 

HYPERPLASIA , tYMPHOI D 

CIRCULATOR Y SYSTEM 

NONE 

DIGESTIVE SYSTEM 

*LIVEP 

DEGENERATION , NOS 

NECROSIS, FOCA L 

METAMORPHOSI S FATTY 

HYPE8PLASTIC NODUL E 

HYPERPLASIA , FOCAL 

•LIVER/PERIP08TAL 

INFLAMMATION , NOS 

tLIVER/KUPFFER CELL 

HYPERPLASIA , NOS 

#LIVER/HEPATOCYT"S 

HYPERTROPHY , NOS 

•BILE DUCT 

INFLAMMATION , NOS 

• PANCREA S 

INFLAMMATION , NOS 

NECROSIS , FOCAL 

METAMORPHOSI S FATTY 

• PANCREATIC ACINU S 

HYPERTROPHY , FOCAL 

•STOMACH 

HYPERPLASIA , FOCAL 

HYPERKERATOSI S 

ACANTHOSI S 

•PEYERS PATC H 

HYPERPLASIA t _NO S _„ _ 

CONTROL(UNTR ) DOSE A 

05-0077 05-0066 

1 (8X) 

(45) (36) 

1 (2X) 

1 (3X) 

3 <7X) 

1 (3X) 

(45) (36) 

1 (2X) 

(t5) (36) 

2 (4X) 

(45) (36) 

(46) (35) 

1 (2X ) 

(44) (23) 

1 <4X ) 

(44) (23) 

(42) (22) 

1 (2X > 1 <5X ) 

1 (5X) 

(43) (20) 

*	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-4
 

DOSE B 

05-0067 

2 (6X) 

1 OX) 

1 <3X) 

1 (3X) 

(45) 

1 (2X ) 

2 (4X) 

2 (4X ) 

(45) 

(45) 

(45) 

1 (2X) 

(46) 

(42) 

1 (2*) 

1 (2X) 

(42) 

1 (2X) 

(43) 

1 <2X ) 

1 (2X ) 

1 (2X ) 

(44) 

3.J7X1 
TABLE Dl (CONTINUED) 

CONTROL (HNIR ) DOSE A DOSE B 

05-0077 05-0066 05-0067 

*COLON (38) (17) (13) PARASITISM 1 (2*) 

URINARY SYSTEM
 

•KIDNEY	 (1*5) (37) (15)
 CALCULUS, NOS 20
 

MINERALIZATION 1 (2X)
 SLOMERULONEPHRITIS, NOS 2U (55X) 12 (93X)
 PYELONEPHRITIS, NOS 5 (lit*)
 INFLAMMATION, NOS 1 (3X)
 INFLAMMATION, FOCAL 1 (2X)
 INFLAMMATION, TNTERST ITIA.L 5 (11*) 2 (5X)
 INFLAMMATION, CHRONIC 1 <2X ) 2 <5X)
 GLOMERIILONEPHRITIS, CHRONIC 1 (3X)
 INFLAMMATION WITH FIBROSIS 2 (KX)
 

"IBHOSIS, DIFFUSE 9 (2<4X) 32 (71X)
 PSBIVASC0LITIS 2 (KX)
 DEGENERATION, CYSTIC 16 (36X)
 

ARTERIOSCLEROSIS, NOS 1 (2X1
 NEPHROSIS, NOS 1 (2*)
 GLOMERULOSCLEROSIS, NOS 7 (19X)
 

HYPERPLASIA, TUBOLAP CELL 2 (UX)
 

*KIDNEY/TDBULE ("5) (37) (K 5)
 

MINERALIZATION 1 (2X)
 INFLAMMATION, NOS 2 (5%)
 DEGENERATION, NOS 1 (2X)
 DEGENERATION, CYSTIC H (9X)
 METAMORPHOSIS "ATTY 9 (20X)
 

fURINARY BLADDER (20) (U3)
 HYPERPLASIA, EPITHFLTAL 3 (15X) 1 (2X)
 

HYPERPLASIA, PAPILLARY 1 (2X)
 

ENDOCRINE SYSTEM
 

#ADRENAL COPTEX (K3) (2U) (38)
 

HYPERPLASIA, NOS 1 (11X)
 

#THYROID (40) (18) (37)
 

PERIVASCULITI S 1 (6X)
 

*PANCH?ATI C ISLFTS (23) C*2)
 

HYPE R PLA SIAi _NOS_ 

# NUMBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

» NUMBER OF ANIMALS NECROPSIED
 

D-5
 
TABLE Dl (CONCLUDED) 

CONTROL (ONTH ) DOSE A DOSE B 

05-0077 05-0066 05-0067 

BEPFODDCTIVE SYSTEM
 

#TESTIS (1*5) (22) (44)
 

HYPERPLASIA, INTERSTITIAL CELL 1 (2X)
 

*TESTIS/TUBOLE (45) (22) (44)
 

MINERALIZATION 1 (5X)
 

DEGENERATION, NOS 2 (4*)
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGAN S 

NONE 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*ABDOMINAL CAVITY (46) (35) (46)
 

STEATITIS 1 (2%)
 

*PEPICARDIHM (46) (35) (46)
 

INFLAMMATION, FOCAL 1 (2X)
 

ALL OTHER SYSTEMS
 

'MULTIPLE ORGANS (46) (35) (46)
 

PERIVASCULITIS 1 (28)
 

SPECIAL MORPHOLOGY SDHMARY
 

NO LESION REPORTED 8
 

AUTO/NECROPSY/NO HISTO 1 1 1
 

AOTOLYSIS/NO NECPOPSY 4 15 4
 

* NUMBER OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-6
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH l-AMINO-2-METHYLANTHRAQUINONE 

CONTROL (OHTR) DOSE A DOSE B 

ANIMALS INITIALLY IN STUDY

ANTMAL S NECROPSIED

ANIMAL S EXAMINE D HISTOPATHOLOSICALLY* *

INTEGUMENTAR Y SYSTEM 

*SKIN


"IBROSIS


FIBROSIS, FDCAL


RESPIRATORY SYSTEM
 

*LUNG


MINERALIZATION


INFLAMMATION, FOCAL


INFLAMMATION, INTERSTITIAL


PERIARTERITIS


HEMATOPOIETIC SYSTEM
 

tBONE MARRO W

MYELOFIBROSTS


• SPLEEN


HYPERPLASIA, NOS


HYPEFPLASIA, HEMATOPOIETIC


HYP^RPLASIA, ERYTHHOID


HYP'RPLASIA, RETICULUM CELL


HYPERPLASIA, LYMPHOID


HEMATOPOIESIS


*LYMPH NODE


INFLAMMATION, NOS


HYPERPLASIA, LYKPHOID


tMESENTERIC L. NODE


06-0077 06-0066 06-0067 

 50 50 498 

 46 38 45 

 45 34 44 

 (46) (38) (45) 

1 (2%) 

1 (2%) 

 (45) (27) (43) 

1 (4X) 

1 (4X) 

2 (4X) 1 (2*) 

1 (2X) 

 (44) (25) (43) 

4 (16X) 6 (14*) 

 (43) (26) (44) 

4 (9X) 

4 (9t) 

2 (5X) 

2 (5X) 1 (2X) 

4 (9X) 5 (11)t) 

1 (2X) 

 (41) (15) (39) 

1 (7%) 2 (5JS) 

1 (7%) 1 (3X) 

 (41) (15) (39) 

* NUMBE R OF ANIMAL S HITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIE D "EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

9 50 ANIMAL S WER E INITIALL Y IN THE STUDY, BUT ON E WAS FOUN D TO BE A MAL E ANISA L 

IN A FEMALE GROUP. 

D-7 
CIRCULATORY SYSTEM
 

tMYOCARDIUM
 

CALCIFICATION, FOCAL
 

*PULMONARY ARTERY
 

HYPERPLAKIA, NOS
 

DIGESTIVE SYSTEM
 

ILIVSR
 

NECROSIS, FOCAL
 

HYPERTROPHY, FOCAL
 

HYPFRPLASIA, NODULAR
 

HYP'RPLASTIC NODULE
 

HYPERPLASIA, FOCAI,
 

HYPERPLASIA, DIFFUSE
 

HEMATOPOIESIS
 

fLIVER/PERIPORTAL
 

INFLAMMATION, NOS
 

ILIVER/KUPFFER CELL
 

HYPERPLASTA, NOS
 

*BILE DUCT
 

INFLAMMATION, NOS
 

IPANCREAS
 

INFLAMMATION, NOS
 

*STOMACH
 

ULCER, NOS
 

HYPERPLASIA, FOCAL
 

HYPERK^RATOSIS
 

ACANTHOSIS
 

tPFYSRS PATCH
 

HYPERPLASIA, NOS
 

CCOLON
 

PARHSITISM
 

URINARY SYSTEM
 

#KIDNEY
 

MINERALIZATION
 

TABLE D2 (CONTINUED)
 

CONTROL (UNTR)
 

06-0077
 

(45)
 

1 (2*)
 

(46)
 

1 (2*)
 

(45)
 

1 (2%)
 

1 (2X)
 

(H5)
 

1 (2*)
 

(U5)
 

1 (255)
 

(46)
 

1 (2X)
 

(41)
 

(42)
 

(43)
 

(41)
 

(43)
 

DOSE A DOSE B
 

06-0066 06-0067
 

(27) (43)
 

(38) (45)
 

(34)	 (44)
 

2 (5X)
 

1 (2%)
 

1 (2%}
 

3 (9*) 1 (2%)
 

5 (11X)
 

1	 <2X)
 

(34) (44)
 

(34) (44)
 

(38) (45)
 

(27)	 (42)
 

1 (2*)
 

(24)	 (43)
 

1 <2X)
 

1 (2X)
 

2 (5*)
 

2 (5K)
 

(23)	 (44)
 

2 (5X)
 

(22)	 (37)
 

1 (5X)
 

(37) (42)
 

*	 NUMBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBSR OF ANIMALS NECROPSIED
 

D-8
 
TABLE D2 (CONTINUED)
 

CONTROL (UNTR)

06-0077

3 (7% )

1 (9%) 

(13)

1 (2X ) 

(13)

(13)

1 (2% ) 

(11)

(13)

(30)

(11)

(16)

(13)

1 i9%l_

 DOSE A 

 06-0066 

23 (62X) 

1 (3% ) 

2 (5*) 

3 (8X ) 

3 (SX ) 

12 (32X ) 

3 (8X ) 

 (37) 

 (37) 

 (37 ) 

 (23) 

1 (US) 

 (27 ) 

1 (1%) 

 (18) 

 (27 ) 

 (38) 

 (22 ) 

t J18Jj_ 

DOSE B 

06-0067 

31 (71X) 

1 (2X) 

2 (5J) 

13 (31X) 

5 (12X ) 

1 (2X ) 

(12) 

1 (10X ) 

(12) 

3 (7%) 

(12) 

(12) 

1 (2%) 

4 (10X) 

(12) 

5 (12% ) 

1 (2*) 

(35) 

1 (3*) 

(12) 

1 (2X ) 

(15) 

2 (1JI) 

(11) 

6 J15X). 

CLOMFRULONFPHRITTS , NO S 

PYELONEPHRITIS , NO S 

INFLAMMATION , NOS 

INFLAMMATION , "OCAL 

INFLAMMATION , INTFRSTITIA L 

INFLAMMATION , CHRONI C 

FIBROSIS , DIFFflSE 

PERIVASC'II ITIS 

DEGENEPATION , CYSTIC 

GLOMFRULOSCLEROSIS . NO S 

tKIDNEY/GLOMERULU S 

DEGENERATION ,

AMYLDIDOSI S 

• KIDNEY/TUBUL E 

DEGENERATION ,

KKTDNFV/PFLVI S 

INFLAMMATION ,

tURTNAR ? BLADDFP 

INFLAMMATION ,

HYPCRPLASIA ,

"IPOCRINF SYSTEM 

*ADPEN» L CORTE X 

HYPERPLASIA ,

 CYSTTC 

 CYSTI " 

 ACU^F/^HPO N 1C 

 NO S 

EPI T H rLIA L 

 NO S 

HVPERPLASIA , INTRADD;TA I 

• "•HYROID HYPEPPLASIA , PAPILLAR Y 

• PANCREATI C ISL"TS 

HYPEPPLASTA , NO S 

REPRODUCTIV E SYST^ K 

*MAMMAR Y GLAN D 

HYPFROLASIA , SOS 

tU-'RDS 

HYDHOMETR A 

* NUM3BR OF ANIMAL S WITH TISSUE ^XAKINED MICROSCOPICALLY
 

* NUMBE R OF ANIMALS NFC-ROPSI3D
 

D-9
 
TABLE D2 (CONTINUED) 

CONTROL (UNTR ) DOSE A DOSE B 

06-0077 06-0066 06-0067 

HYP'RPLASIA , ADFNOMATOU S 

HYPERPLASIA , STPOMA L 1 (2X) 

tUTERUS/ENDOMFTRin K 

CYST, NOS 

INFLAMMATION , NOS 

(13) 

2 (5*) 

(22) 

1 (5X) 

(11) 

4 (10*) 

INFLAMMATION , ACUTE 1 (2X) FYPEPPLASIA ,

HYPERPLASIA ,

 NO S 

 CYSTIC 

1 (2X) 35 (81X) 1 (5X) 1 (5X) 1 (10X) 

9 (22X) 

•OVARY/OVIDUC T 

INFLAMMATION , NO S 

(« 3) (22) (11)

2 (5X) HYPERPLASIA , PAPILLAP Y 1 (2X) 

#OVAH Y 

CYST, NOS 

(41) 

1 (2X) 

(22) 

1 (5X) 

(11) 

7 (17X) 

HEMORRHAGE 1 (5%) INFLAMMATION, NOS 3 (7X) 

IOVARY/FOLLICLE (1(1) (22) (111) 

HEMORRHAGE 1 (2X) 

NERVOUS SYSTEM 

NONE 

SPECTAL SFNS" ORGANS 

NONE 

•IHSCULOSKFLETA L SYSTE M 

*BON EP IBROSI S (46) (38)

1 (3%) (45) 1 (2X) PESORPTION 1 (3X) 1 (2X) 

"VERTEBRA (46) (38) (45) 

OS^EOSCLEPOSIS 1 (2X) 

BODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONE 

* NUMBE R 0" ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIF.D 

D-IO 
TABLE D2 (CONCLUDED) 

CONTROL(UNTR | DOSE A DOSE B 

06-0077 06-0066 06-0067 

SPECIAL MORPHOLOGY SUMMAR Y 

NO LESION RFPORTED 1 1
 

NFCROPSY PERF/NO HISTO PERFORMED 1
 

AUTO/NECROPSY/HISTO PERF 1
 

AUTO/NECROPSY/NO HISTO 1 It
 

AUTOLYSTS/NO NECROPSY 4 12 14
 

* NUMBE R 0 " ANIMUL S WITH TISSU E EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NKCHOPSIE D 

D-ll 


i 

p. 

tvr

*

t 

DHEW Publication No. (NIH) 78-1366