1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 49 

BIOASSAY OF 

ACRONYCINE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 7008-42-6 

NCI-CG-TR-49 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

ACRONYCINE
 

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) 78-849
 

BIOASSAY OF
 

ACRONYCINE
 

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
 

acronycine 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 greater incidence of cancer than control animals,
 

do not necessarily mean that the test chemical is not a carcinogen, inasmuch as the experiments are conducted under a limited
 

set of circumstances. Positive results demonstrate that the test
 

chemical is carcinogenic for animals under the conditions of the
 

test and indicate that exposure to the chemical is a potential
 

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

animal carcinogens requires a wider analysis.
 

CONTRIBUTORS; This bioassay of acronycine was conducted by
 

Southern Research Institute, Birmingham, Alabama, 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 and doses were determined by Drs. D. P.
 

Griswold*-, J. D. Prejean^, E. K. Weisburger^, and J. H.
 

Weisburger2>3. Ms. J. Belzer1 and Mr. I. Brown* were responsible
 

for the care of the laboratory animals and the administration of
 

the test chemical. Data management and retrieval were performed
 

by Ms. C. A. Dominick*. Histopathologic examinations were
 

performed by Drs. S. D. Kosanke* and J. C. Peckhanr, and the
 

diagnoses included in this report represent their interpretation.
 

The reported neoplasms and chemical-related lesions were reviewed
 

iii
 
by Dr. J. F. Hardisty^, who prepared the interpretive pathology
 

summary included in this report.
 

Animal pathology tables and survival tables were compiled at EG&G
 

Mason Research Institute^. The statistical analyses were
 

performed by Dr. J. R. Joiner", using methods selected for the
 

bioassay program by Dr. J. J. Gart^. Chemicals used in this
 

bioassay were obtained through Mr. C. A. Hewitt^. Chemical
 

analyses were performed by Drs. J. Stewart" and R. H. Iwamoto",
 

and the analytical results were reviewed by Dr. S. S. Olin°. The
 

structural formula was supplied by NCI^.
 

This report was prepared at Tracer Jitco" under the direction of
 

NCI. Those responsible for the report at Tracer Jitco were Dr.
 

Marshall Steinberg, Director of the Bioassay Program; Dr. L. A.
 

Campbell, Deputy Director for Science; Drs. J. F. Robens and C.
 

H. Williams, toxicologists; Dr. R. L. Schueler, pathologist; Dr.
 

G. L. Miller, Ms. M. S. King and Mr. W. D. Reichardt, bioscience
 

writers; and Dr. E. W. Gunberg, technical editor, assisted by Ms.
 

Y. E. Presley.
 

The statistical analysis was reviewed by members of the
 

Mathematical Statistics and Applied Mathematics Section of NCI':
 

Dr. John J. Gart, Mr. Jun-mo Nam, Dr. Hugh M. Pettigrew, and
 

Dr. Robert E. Tarone.
 

The following other scientists at NCI^ were responsible for
 

evaluating the bioassay experiment, interpreting the results, and
 

reporting the findings: Dr. Kenneth C. Chu, Dr. Cipriano Cueto,
 

Jr., Dr. J. Fielding Douglas, Dr. Dawn G. Goodman, Dr. Richard A.
 

Griesemer, Dr. Harry A. Milman, Dr. Thomas W. Orme, Dr. Robert A.
 

, and Dr. Jerrold M. Ward.
 

Southern Research Institute, 2000 Ninth Avenue South,
 

Birmingham, Alabama.
 

iv
 
^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

%ow with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

^Experimental Pathology Laboratories, P.O. Box 474, Herndon,
 

Virginia.
 

5

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

Rockville, Maryland.
 

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

Maryland.
 

^Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute, National
 

Institutes of Health, Bethesda, Maryland.
 

"Stanford Research Institute, Life Sciences Division, Menlo Park,
 

California.
 

^Developmental Therapeutics Program, Division of Cancer
 

Treatment, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

V 

SUMMARY
 

A bioassay of acronycine for possible carcinogenicity was
 

conducted by administering the test chemical by intraperitoneal
 

injection to Sprague-Dawley rats and B6C3F1 mice.
 

Initially, groups of 35 rats of each sex were administered
 

acronycine at one of two doses, either 7.5 or 15 mg/kg body
 

weight, in a vehicle composed of 0.05% polysorbate 80 in
 

phosphate-buffered saline. Control groups of each sex consisted
 

of 10 untreated rats (untreated controls) and 10 rats injected
 

with the vehicle (vehicle controls). Because of high mortality
 

rates in the dosed animals, new dosed groups of 35 rats of each
 

sex were started later at a dose of 3.75 mg/kg. Additional
 

groups of 10 untreated and 10 vehicle controls of each sex were
 

also started. The rats were administered the acronycine or the
 

vehicle for 51 or 52 weeks, then observed for an additional 28-30
 

weeks. All surviving rats were killed at 80-82 weeks.
 

Initially, groups of 35 mice of each sex were administered
 

acronycine at one of two doses, either 12.5 or 25 mg/kg body
 

weight, in a vehicle composed of 0.05% polysorbate 80 in
 

phosphate-buffered saline. Control groups of each sex consisted
 

of 10 untreated mice (untreated controls) and 10 mice injected
 

with the vehicle (vehicle controls). Because of high mortality
 

rates in the dosed animals, two additional dosed groups were
 

started later: 35 mice of each sex at 6 mg/kg and 40 mice of
 

each sex at 2 mg/kg, together with 10 untreated controls and 10
 

vehicle controls of each sex for the groups dosed at 6 mg/kg, and
 

20 untreated controls and 20 vehicle controls for the groups
 

dosed at 2 mg/kg. Periods of administration of the chemical to
 

the mice varied from 25 weeks to 92 weeks, depending on toxicity
 

or length of time of survival. Surviving control animals were
 

killed at 78-105 weeks.
 

Acronycine was toxic to rats and mice of each sex at the doses
 

used in this bioassay, as shown by the high mortality rates in
 

all but the low-dose groups and by the lower mean body weights in
 

dosed rats and mice at all doses throughout most of the bioassay.
 

vii
 
Because of this high number of deaths, time-adjusted statistics
 

are used for the analyses of all incidences of tumors.
 

In male rats, the dose-related trend in the mid- and high-dose
 

groups for the incidence of osteosarcoma at all sites was
 

significant (P = 0.002) using the respective vehicle-control
 

group (vehicle controls 0/8, mid-dose 13/30, high-dose 12/18).
 

Comparisons of the individual groups with respective controlgroups were also significant for the mid-dose (P = 0.022) and
 

high-dose (P = 0.002) groups, but not for the low-dose group.
 

In female rats, osteosarcoma was observed only in 1/8 high-dose
 

animals.
 

Sarcomas and other related tumors of the peritoneum were observed
 

in all three dosed groups of both male and female rats, but in
 

none of the control groups (males: low-dose 5/30, mid-dose 3/26,
 

high-dose 7/16; females: low-dose 1/35, mid-dose 5/30, high-dose
 

13/28). In both sexes, the dose-related trends were significant
 

(males, P = 0.006; females, P = 0.002), and the comparison of the
 

incidences in the high-dose females with the vehicle-control
 

group was significant (P = 0.016). None of the incidences in the
 

individual dosed groups of males were significant when compared
 

with vehicle controls. However, since the tumors were observed
 

in all dosed groups but did not occur in historical-control
 

animals at this laboratory, they are considered to be related to
 

the administration of the chemical.
 

In female rats, the incidence of all tumors of epithelial origin
 

of the mammary gland was significant only at the low dose
 

(low-dose vehicle controls 1/10, low-dose 22/35, P = 0.004).
 

Adenocarcinomas of the mammary gland were observed in seven lowdose, five mid-dose, and two high-dose female rats, but in no
 

control females. The reverse dose relationship of both benign
 

and malignant tumors was probably due to the higher number of
 

early deaths which occurred in the high-dose group.
 

In mice, the low survival in all dosed groups except the lowdose animals precluded an evaluation of the significance of the
 

incidences of tumors. Lymphomas occurred in low-dose groups of
 

viii
 
both males and females; however, the incidence of lymphoma in
 

different control groups was highly variable. The high incidence
 

in the low-dose vehicle controls may have been due to a
 

procedural problem associated with the possibility of transfer of
 

tumor cells or oncogenic viruses during the intraperitoneal
 

injection of the test chemcial.
 

It is concluded that under the conditions of this bioassay, the
 

low survival of the dosed and control mice and the possible
 

procedural problems associated with the intraperitoneal injection
 

of the chemical did not allow a determination to be made of the
 

carcinogenicity of acronycine in this species. In Sprague-Dawley
 

rats, acronycine in the vehicle of 0.05% polysorbate 80 in
 

phosphate-buffered saline was carcinogenic, producing tumors of
 

the mammary gland in females, osteosarcomas in males, and
 

sarcomas and other related tumors of the peritoneum in both males
 

and females.
 

ix
 

TABLE OF CONTENTS
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dosage Preparation. 3
 

C. Animals 4
 

D. Animal Maintenance 5
 

E. Subchronic Studies 9
 

F. Designs of Chronic Studies 10
 

G. Clinical and Pathologic Examinations 17
 

H. Data Recording and Statistical Analyses 18
 

III. Results -Rats 25
 

A. Body Weights and Clinical Signs (Rats) 25
 

B. Survival (Rats) 25
 

C. Pathology (Rats) 31
 

D. Statistical Analyses of Results (Rats) 35
 

IV. Results - Mice 41
 

A. Body Weights and Clinical Signs (Mice) 41
 

B. Survival (Mice) 41
 

C. Pathology (Mice) 47
 

D. Statistical Analyses of Results (Mice) 49
 

V. Discussion 51
 

VI. Bibliography 57
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in
 

Rats Given Intraperitoneal Injections
 

of Acronycine 59
 

Table Al Summary of the Incidence of Neoplasms in
 

Male Rats Given Intraperitoneal Injections of
 

Acronycine (Control Groups) 61
 

xi
 
Table A2 Summary of the Incidence of Neoplasms in 

Male Rats Given Intraperitoneal Injections 

of Acronycine (Treated Groups) 64 

Table A3 Summary of the Incidence of Neoplasms in 

Female Rats Given Intraperitoneal Injections 

of Acronycine (Control Groups) 68 

Table A4 Summary of the Incidence of Neoplasms in 

Female Rats Given Intraperitoneal Injections 

of Acronycine (Treated Groups) 71 

Appendix B Summary of the Incidence of Neoplasms in 

Mice Given Intraperitoneal Injections of 

Acronycine 75 

Table Bl Summary of the Incidence of Neoplasms in 

Male Mice Given Intraperitoneal Injections 

of Acronycine (Control Groups) 77 

Table B2 Summary of the Incidence of Neoplasms in 

Male Mice Given Intraperitoneal Injections 

of Acronycine (Control and Treated Groups) 80 

Table B3 Summary of the Incidence of Neoplasms in 

Female Mice Given Intraperitoneal Injections 

of Acronycine (Control Groups) 83 

Table B4 Summary of the Incidence of Neoplasms in 

Female Mice Given Intraperitoneal Injections 

of Acronycine (Control and Treated Groups) 86 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Given Intraperitoneal 

Injections of Acronycine 89 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Given Intraperitoneal 

Injections of Acronycine (Control Groups) 91 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Given Intraperitoneal 

Injections of Acronycine (Treated Groups). 94 

xii
 
Table C3 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Given Intraperitoneal 

Injections of Acronycine (Control Groups) 98 

Table C4 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Given Intraperitoneal 

Injections of Acronycine (Treated Groups) 101 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Given Intraperitoneal 

Injections of Acronycine 105 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Given Intraperitoneal 

Injections of Acronycine (Control Groups) 107 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Given Intraperitoneal 

Injections of Acronycine (Control and 

Treated Groups) 110 

Table D3 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Given Intraperitoneal 

Injections of Acronycine (Control Groups) 115 

Table D4 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Given Intraperitoneal 

Injections of Acronycine (Control and 

Treated Groups) 118 

Appendix E Analyses of the Incidence of Primary Tumors 

in Rats Given Intraperitoneal Injections 

of Acronycine. .....*. 123 

Table El Time-adjusted Analyses of the Incidence of 

Primary Tumors in Male Rats Given Intraperitoneal 

Injections of Acronycine, Using Vehicle Controls... 125 

Table E2 Time-adjusted Analyses of the Incidence of 

Primary Tumors in Female Rats Given Intraperitoneal 

Injections of Acronycine, Using Vehicle Controls... 133 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Given Intraperitoneal Injections of 

Acronycine. 145 

xiii
 
Table Fl Time-adjusted Analyses of the Incidence of 

Primary Tumors in Male Mice Given Intraperitoneal 

Injections of Acronycine 147 

Table F2 Time-adjusted Analyses of the Incidence of 

Primary Tumors in Female Mice Given 

Intraperitoneal Injections of Acronycine 149 

TABLES 

Table 1 Design of Chronic Studies of Acronycine 

in Rats 11 

Table 2 Design of Chronic Studies of Acronycine 

in Male Mice 13 

Table 3 Design of Chronic Studies of Acronycine 

in Female Mice. 15 

FIGURES 

Figure 1 Growth Curves for Male Rats Treated with 

Acronycine 26 

Figure 2 Growth Curves for Female Rats Treated with 

Acronycine 27 

Figure 3 Survival Curves for Male Rats Treated with 

Acronycine 28 

Figure 4 Survival Curves for Female Rats Treated with 

Acronycine. 29 

Figure 5 Life Table for Male Rats Treated with 

Acronycine: Osteosarcoma of the 

Musculoskeletal System 37 

Figure 6 Growth Curves for Male Mice Treated with 

Acronycine 42 

Figure 7 Growth Curves for Female Mice Treated with 

Acronycine 43 

xiv
 
Figure 8 Survival Curves for Male Mice Treated with 

Acronycine 44 

Figure 9 Survival Curves for Female Mice Treated with 

Acronycine 45 

xv
 

I. INTRODUCTION
 

Acronycine (CAS 7008-42-6, NSC 403169, NCI C01536), an alkaloid
 

derived from the bark of the Australian scrub ash (Lahey and
 

Thomas, 1949), has been investigated as an experimental anticancer drug. In preclinical screening tests in mice, broadspectrum antitumor activity of acronycine was demonstrated
 

(Svoboda et al., 1966). Phase I clinical trials were conducted
 

but have not been reported (Carter, 1971). Acronycine inhibits
 

cellular uptake of two extracellular nucleosides (uridine and
 

thymidine) necessary for DNA and RNA synthesis, apparently by
 

interfering with their transport across cell membranes (Dunn et
 

al., 1973).
 

Acronycine was selected for screening in the carcinogenesis
 

program in an attempt to evaluate the carcinogenicity of certain
 

drugs that may be used for prolonged periods in humans.
 

1 

II. MATERIALS AND METHODS
 

A. Chemical
 

ACRONYCINE
 

OCH,
 

Acronycine, which is the name used most commonly for 3,12-dihydro-6-methoxy-3,3,12-trimethyl-7H-pyrano(2,3-c)acridin-7-one, was
 

obtained through the Developmental Therapeutics Program, Division
 

of Cancer Treatment, National Cancer Institute, in two batches
 

that were manufactured by the Commonwealth Scientific and
 

Industrial Research Organization, East Melbourne, Australia. The
 

identity of the chemical was confirmed in analyses performed by
 

Stanford Research Institute for the Developmental Therapeutics
 

Program. These analyses included melting point of the chemical
 

and its picrate salt; elemental analyses (C, H, N) for C2()Hi9N03;
 

and infrared, ultraviolet, and nuclear magnetic resonance
 

spectra. Thin-layer chromatography showed only trace impurities.
 

3 
No attempt was made to identify or quantitate these impurities.
 

All data indicate that these batches of acronycine were nearly
 

100% pure.
 

The bulk chemical was stored in a brown glass bottle. This
 

bottle was enclosed in a plastic bag containing Drierite and was
 

refrigerated at 5°C.
 

B. Dosage Preparation
 

Test solutions were prepared daily by adding a specified amount
 

of the drug to a vehicle composed of 0.05% polysorbate 80 in
 

phosphate-buffered saline. This mixture was emulsified in a
 

10-ml Potter-Elvehjem tissue grinder with a Teflon pestle for 20
 

seconds. Each concentration (0.02, 0.06, 0.125, 0.15, 0.25, 0.3,
 

or 0.6%) was administered on the day of preparation.
 

C. Animals
 

Sprague-Dawley rats obtained from Charles River Breeding
 

Laboratories, Inc., Wilmington, Massachusetts, were used for all
 

groups of this species. B6C3F1 mice obtained from A. R. Schmidt,
 

Madison, Wisconsin, were used for the upper mid- and high-dose
 

groups and respective controls; B6C3F1 mice from Charles River
 

Laboratories, for the lower mid-dose groups and controls; and
 

B6C3F1 mice from Litton Bionetics, Frederick, Maryland, for the
 

4 
low-dose groups and controls. The rats used in the chronic
 

studies were 30-42 days old on arrival at the laboratory; the
 

mice were 30-32 days old. All animals were quarantined (rats: 5
 

days in the original study, 12 days in the rerun; mice: 5 days
 

in the original study, 10 days in the first rerun, 13 days in the
 

second rerun). Animals having no visible signs of disease were
 

assigned to control and treated groups and earmarked for
 

individual identification.
 

D. Animal Maintenance
 

Animals were placed on study at different intervals during a
 

4-year period. Some techniques for animal care were improved
 

during this time, and as a result, the animal groups placed on
 

study at the beginning of the bioassay (high- and mid-dose rats,
 

high- and upper mid-dose mice) were exposed to somewhat different
 

environmental conditions than the groups started later (low-dose
 

rats; lower mid- and low-dose mice).
 

During all of the studied, animals were housed in temperatureand humidity-controlled rooms. The temperature range was
 

20-24°C, and the relative humidity was maintained at 40-60%. The
 

room air was changed 15 times per hour and passed through both
 

intake and exhaust fiberglass roughing filters. In addition to
 

natural light, illumination was provided by fluorescent light for
 

5 
9 hours per day. Wayne® Lab Blox animal meal (Allied Mills,
 

Inc., Chicago, 111.) and water were supplied daily and were
 

available ad libitum.
 

All animals were housed in solid-bottom stainless steel cages
 

(Hahn Roofing and Sheet Metal Co., Birmingham, Ala.). Rats were
 

housed five per cage, and mice in the original groups (high- and
 

upper mid-dose) were housed seven per cage; mice in later groups
 

(lower mid- and low-dose) were housed five per cage, due to a
 

reduction in cage size. The bottoms of the rat cages were lined
 

with Iso-Dri hardwood chips (Carworth, Edison, N. J.), and cage
 

tops were covered with disposable filter bonnets beginning at
 

week 34 for the high- and mid-dose groups of rats and at week 1
 

for the low-dose group of rats.
 

Mouse cages were provided with Sterolit® clay bedding (Englehard
 

Mineral and Chemical Co., New York, N. Y.), except for the cages
 

of the low-dose mice, which were provided with Betta-Chip®
 

hardwood bedding (Northeastern Products Corp., Warrensburg, N.Y.)
 

from week 84 until termination of the study. Filter bonnets were
 

installed on cages of the low-dose mice in week 32.
 

Bedding was replaced once per week; cages, water bottles, and
 

feeders were sanitized at 82°C once per week; and racks were
 

cleaned once per week, except during the later studies with
 
low-dose rats and low-dose mice, when clean cages and fresh
 

bedding were provided twice per week.
 

Rats and mice were housed in separate rooms. Control animals
 

were housed with respective treated animals. Animals treated
 

with acronycine were maintained in the same rooms as animals of
 

the same species being treated with the following chemicals:
 

RATS
 

Gavage Studies
 

cholesterol (p-(bis(2-chloroethyl)amino)phenyl)acetate
 

(phenesterin) (CAS 3546-10-9)
 

estradiol bis( (p-(bis(2-chloroethyl)amino)phenyl)acetat:e)
 

(estradiol mustard) (CAS 22966-79-6)
 

Intraperitoneal Injection Studies
 

4'-(9-acridinylamino)methansulfon-m-aniside monohydrochloride
 

(MAAM) (NSC 141549)
 

5-azacytidine (CAS 320-67-2)
 

beta-2'-deoxy-6-thioguanosine monohydrate (beta-TGdR)
 

(CAS 789-61-7)
 

1,4-butanediol dimethanesulfonate (busulfan) (CAS 55-98-1)
 

emetine dihydrochloride tetrahydrate (CAS 316-42-7)
 

3,3'-iminobis-l-propanol dimethanesulfonate (ester)
 

hydrochloride [IPD] (CAS 3458-22-8)
 

(+)-4,4'-(l-methyl-l,2-ethanediyl)bis-2,6-piperazinedione
 

(ICRF-159) (CAS 21416-87-5)
 

N,3-bis(2-chloroethyl)tetrahydro-2H-l,3,2-oxazaphosphorin-2­

amine-2-oxide (isophosphamide) (CAS 3778-73-2)
 

N-(2-chloroethyl)-N-(1-methyl-2-phenoxyethyl)benzylamine
 

hydrochloride (phenoxybenzamine hydrochloride) (CAS 63-92-3)
 

N-(l-methylethyl)-4-((2-methylhydrazino)methyl)benzamide
 

monohydrochloride (procarbazine) (CAS 366-70-1)
 

tris(l-aziridinyl)phosphine sulfide (thio-TEPA) (CAS 52-24-4)
 

2,4,6-tris(dimethylamino)-s-triazine (CAS 645-05-6)
 

7 
MICE
 

Feed Studies
 

4-acetyl-N-((cyclohexylamino)carbonyl)benzenesulfonamide
 

(acetohexamide) (CAS 968-81-0)
 

anthranilic acid (CAS 118-92-3)
 

l-butyl-3-(p-tolylsulfonyl)urea (tolbutamide) (CAS 64-77-7)
 

4-chloro-N-((propylamino)carbonyl)benzenesulfonamide
 

(chlorpropamide) (CAS 94-20-2)
 

5-(4-chlorophenyl)-6-ethyl-2,4-pyrimidinediamine
 

(pyrimethamine) (CAS 58-14-0)
 

2,6-diamino-3-(phenylazo)pyridine hydrochloride (phenazopyridine
 

hydrochloride) (CAS 136-40-3)
 

L-tryptophan (CAS 73-22-3)
 

N-9H-fluoren-2-ylacetamide (CAS 53-96-3)
 

N-(p-toluenesulfonyl)-N'-hexamethyleniminourea
 

(tolazamide) (CAS 1156-19-0)
 

1-phenethylbiguanide hydrochloride (phenformin) (CAS 114-86-3)
 

pyrazinecarboxamide (pyrazinamide) (CAS 98-96-4)
 

4,4'-sulfonyldianiline (dapsone) (CAS 80-08-0)
 

4,4'-thiodianiline (CAS 139-65-1)
 

ethionamide (CAS 536-33-4)
 

reserpine (CAS 50-55-5)
 

Gavage Studies
 

cholesterol (p-(bis(2-chloroethyl)amino)phenyl)acetate
 

(phenesterin) (CAS 3546-10-9)
 

estradiol bis((p-(bis(2-chloroethyl)amino)phenyl)acetate)
 

(estradiol mustard) (CAS 22966-79-6)
 

Intraperitoneal Injection Studies
 

4'-(9-acridinylamino)methansulfon-m-aniside monohydrochloride
 

(MAAM) (NSC 141549)
 

5-azacytidine (CAS 320-67-2)
 

beta-2'-deoxy-6-thioguanosine monohydrate (beta-TGdR)
 

(CAS 789-61-7)
 

1,4-butanediol dimethanesulfonate (busulfan) (CAS 55-98-1)
 

emetine dihydrochloride tetrahydrate (CAS 316-42-7)
 

3,3'-iminobis-l-propanol dimethanesulfonate (ester)
 

hydrochloride [IPD] (CAS 3458-22-8)
 

(+)-4,4'-(l-methyl-l,2-ethanediyl)bis-2,6-piperazinedione
 

(ICRF-159) (CAS 21416-87-5)
 

N,3-bis(2-chloroethyl)tetrahydro-2H-l,3,2-oxazaphosphorin-2­

amine-2-oxide (isophosphamide) (CAS 3778-73-2)
 

8 
N-(2-chloroethyl)-N-(l-methyl-2-phenoxyethyl)benzylamine 

hydrochloride (phenoxybenzamine hydrochloride) (CAS 63-92-3)
 

N-(l-methylethyl)-4-((2-methylhydrazino)methyl)benzamide
 

monohydrochloride (procatbazine) (CAS 366-70-1)
 

tris(l-aziridinyl)phosphine sulfide (thio-TEPA) (CAS 52-24-4)
 

2,4,6-tris(dimethylamino)-s-triazine (CAS 645-05-6)
 

hexamethylmelamine (CAS 14fe-82-3)
 

E. Subchronic Studies
 

Subchronic studies were conducted to estimate the maximum
 

tolerated doses of acronycine, on the basis of which low and high
 

concentrations (hereinafter referred to as "low doses" and "high
 

doses") were determined for administration in the chronic studies
 

using one or both sexes of each species. In subchronic studies
 

of acronycine, male and female Sprague-Dawley rats and male Swiss
 

mice were administered the test chemical by intraperitoneal
 

injection three times pet week for 45 days at one of five
 

different doses. Following administration of the chemical, all
 

surviving animals were observed for an additional 45 days.
 

Treated groups each consisted of five animals, untreated-control
 

groups consisted of 10 animals, and vehicle control (0.05%
 

polysorbate in buffered saline) groups consisted of 10 animals.
 

All animals were observed daily and weighed once per week.
 

The first subchronic study was on female rats using 60, 150, 300,
 

600, or 1,200 mg/kg body weight for each injection. Weight
 

depression and deaths occurred at all doses. Thus, a second
 

9 
study was performed, using 1.5, 3.75, 7.5, 15, or 30 mg/kg. Male
 

Sprague-Dawley rats which were available at the time were used.
 

Four animals receiving 30 mg/kg died, but no deaths occurred in
 

the remaining four groups. Mean body weights were depressed
 

during the period of chemical administration, but the animals
 

recovered and no weight depression greater than the 15% limit was
 

present at day 90. No gross abnormalities were observed. Low
 

and high doses for chronic studies using rats were set at 7.5 and
 

15 mg/kg.
 

All mice treated at the doses originally selected (100, 250, 500,
 

1,000, or 2,000 mg/kg) died by week 6. A second study was
 

performed using doses of 2.5, 6.25, 12.5, 25, or 50 mg/kg. By^
 

day 90, only one animal treated at a dose of 50 mg/kg had survived. No deaths occurred in the groups receiving 2.5, 6.25, or
 

25 mg/kg, although one animal treated at 12.5 mg/kg died during
 

week 8. Weight gains were not affected in these latter four
 

groups, and no gross abnormalities were observed at necropsy.
 

Low and high doses for chronic studies using mice were set at
 

12.5 and 25 mg/kg.
 

F. Designs of Chronic Studies
 

The designs of the chronic studies are shown in tables 1, 2, and
 

3.
 

10
 
Table 1. Design of Chronic Studies of Acronycine in Rats
 

Sex and 

Test 

Group 

Initial 

No. of 

Animals3 

Acronycine 

Doseb 

(mg/kg) 

Time 

Treated 

(weeks ) 

on Study 

Untreated 

(weeks ) 

Male 

Low-Dose 

Untreated-Control0 

Low-Dose 

Vehicle-Control0 

Low-Dosec 

10 

10 

35 

0 

Qd 

3.75 

52 

52 

81 

28 

28 

Mid- and High-Dose 

Untreated-Control 

Mid- and High-Dose 

Vehicle-Control 

Mid-Dose 

High-Dose 

10 

10 

35 

35 

0 

Qd 

7.5 

15 

52 

52 

51e 

82 

30 

29 

Female 

Low-Dose 

Untreated-Control0 

Low-Dose 

Vehicle-Control0 

Low-Dose0 

10 

10 

35 

0 

Qd 

3.75 

52 

52 

80 

28 

28 

Mid- and High-Dose 

Untreated Control 

Mid- and High-Dose 

Vehicle Control 

Mid-Dose 

High-Dose 

10 

10 

35 

35 

0 

Od 

7.5 

15 

52 

52 

52 

82 

30 

29-30 

28-29 

a

Ages of rats when placed on study: mid- and high-dose males, 40 days;
 

mid- and high-dose females, 47 days; low-dose males and females, 42
 

days.
 

^Acronycine was administered intraperitoneally in a vehicle consisting
 

of polysorbate 80 in phosphite-buffered saline at a volume of 0.25
 

ml/100 g body weight three times per week; doses were based on
 

individual weights. The saflie needle for injection was used for each
 

group of five animals within a cage.
 

11
 
Table 1. Design of Chronic Studies of Acronycine in Rats
 

(continued)
 

G

Because of high mortality in treated groups, new treated and control
 

groups were started 77 weeks after the original start of the study.
 

^Vehicle controls received only polysorbate 80 in phosphate-buffered
 

saline at the same volume as the treated animals. The same bottle of
 

vehicle solution was used for all vehicle-control animals on study at
 

any given time.
 

e

All high-dose males died or were killed by week 51.
 

12
 
Table 2. Design of Chronic Studies of Acronycine in Male Mice
 

Sex and Initial Acronycine Time on Study
 

Test No. of Doseb Treated Untreated
 

Group Animals3 (mg/kg) (weeks ) (weeks)
 

Low-Dose
 

Untreated-Control0 20 0 105
 

Low-Dose
 

Vehicle-Control0 20 0^ 71e
 

Low-Dosec 40 2 92e
 

Lower Mid-Dose
 

Untreated-Control^ 10 0 79
 

Lower Mid-Dose
 

Vehicle-Control^ 10 0<* 52 26
 

Lower Mid-Dose^ 35 6 49e
 

Upper Mid-Dose and
 

High-Dose Untreated
 

Control 10 0 78
 

Upper Mid-Dose and
 

High-Dose Vehicle
 

Control 10 Qd 3i 47
 

Upper Mid-Dose 35 12.5 31 14
 

High-Dose 35 25 25e
 

a

Ages of mice when placed on study: upper mid-dose and high-dose 35
 

days; lower mid-dose, 42 days; low-dose, 43 days.
 

^Acronycine was administered intraperitoneally in a vehicle
 

consisting of polysorbate 80 in phosphate-buffered saline at a
 

volume of 1 ml/100 g body weight three times per week; closes were
 

based on the mean-weight of the animals in each cage. The same
 

needle for injection was used for each group of five animals
 

(restarted groups) or seven animals (original groups) within a cage.
 

°Because of high mortality in the treated animals, treated and control
 

groups, designated "low-dose," were started 97 weeks after the
 

original start of the study.
 

^Vehicle controls received only polysorbate 80 in phosphate-buffered
 

saline at the same volume as the treated animals. The same bottle of
 

vehicle solution was used for all vehicle-control animals on study at
 

any given time.
 

13
 
Table 2. Design of Chronic Studies of Acronycine in Male Mice
 

(continued)
 

e

All animals died or were killed by the times indicated.
 

fBecause of high mortality in the treated animals, treated and control
 

groups, designated "lower mid-dose," were started 46 weeks after the
 

original start of the study.
 

14
 
Table 3. Design of Chronic Studies of Acronycine in Female Mice
 

Sex and 

Test 

Initial 

No. of 

Acronycine 

Dose*5 

Time 

Treated 

on Study 

Untreated 

Group Animals3 (mg/kg) (weeks ) (weeks ) 

Low-Dose 

Untreated-Control0 20 0 105 

Low-Dose 

Vehicle-Controlc 20 Qd 56e 

Low-Dosec 40 2 87e 

Lower Mid-Dose 

Untreated-Controlf 10 0 79 

Lower Mid-Dose 

Vehicle-Control 10 Qd 52 26 

Lower Mid-Dose^ 35 6 49e 

Upper Mid-Dose and 

High-Dose Untreated 

Control 10 0 79 

Upper Mid-Dose and 

High-Dose Vehicle 

Control 10 0^ 31 47 

Upper Mid-Dose 35 12.5 31 10e 

High-Dose 35 25 25e 

a

Ages of mice when placed on study: upper mid-dose and high-dose, 35
 

days; lower mid-dose, 42 days; low-dose, 43 days.
 

°Acronycine was administered intraperitoneally in a vehicle
 

consisting of polysorbate 80 in phosphate-buffered saline at a
 

volume of 1 ml/100 g body weight three times per week; doses were
 

based on mean weight of the animals in each cage. The same needle
 

for injection was used for each group of five animals (restarted
 

groups) or seven animals (original groups) within a cage.
 

c

Because of high mortality in the treated animals, treated and control
 

groups, designated "low-dose," were started 97 weeks after the
 

original start of the study.
 

d

Vehicle controls received only polysorbate 80 in phosphate-buffered
 

saline at the same volume as the treated animals. The same bottle of
 

vehicle solution was used for all vehicle-control animals on study at
 

any given time.
 

15
 
Table 3. Design of Chronic Studies of Acronycine in Female Mice
 

(continued)
 

e

All animals died or were killed by the times indicated.
 

fBecause of high mortality in the treated animals, treated and control
 

groups, designated "lower mid-dose," were started 46 weeks after the
 

original start of the study.
 

16
 
G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity, and
 

animals that were moribund were killed and necropsied. The
 

animals were weighed individually each week, every 2 weeks, or
 

once per month, depending on the schedule in use at the time the
 

animals were weighed. Palpation for masses was carried out at
 

each weighing.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of major tissues, major organs, and all gross lesions
 

from killed animals and from animals found dead. The following
 

tissues were examined microscopically: skin, muscle, lungs and
 

bronchi, trachea, bone and bone marrow, spleen, lymph nodes,
 

thymus, heart, salivary gland, liver, gallbladder and bile duct
 

(mice), pancreas, esophagus, stomach, small intestine, large
 

intestine, kidney, urinary bladder, pituitary, adrenal, thyroid,
 

parathyroid, 'mammary gland, prostate or uterus, testis or ovary,
 

brain, and sensory organs. Peripheral blood smears were prepared
 

from each animal killed. Occasionally, additional tissues were
 

also examined microscopically. The different tissues were
 

preserved in 10% buffered formalin, embedded in paraffin,
 

sectioned, and stained with hematoxylin and eosin. Special
 

staining techniques were utilized when indicated for more
 

definitive diagnosis.
 

17
 
A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals were
 

missing, cannibalized, or judged to be in such an advanced state
 

of autolysis as to preclude histopathologic evaluation. Thus,
 

the number of animals from which particular organs or tissues
 

were examined microscopically varies, and does not necessarily
 

represent the number of animals that were placed on study in each
 

group.
 

H. Data Recording and Statistical Analyses
 

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

System (Linhart et al., 1974). The data elements include
 

descriptive information on the chemicals, animals, experimental
 

design, clinical observations, survival, body weight, and individual pathologic results, as recommended by the International Union
 

Against Cancer (Berenblum, 1969). Data tables were generated for
 

verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear on the possibility of carcinogenicity are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

18
 
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) for testing two groups for equality and Tarone's (1975)
 

extensions of Cox's methods for testing for a dose-related trend.
 

One-tailed P values have been reported for all tests except the
 

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

at a specific anatomic site (numerator) to the number of animals
 

in which that site is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic examination was required to detect lesions prior to histologic sampling
 

(e.g., skin or mammary tumors), or when lesions could have
 

appeared at multiple sites (e.g., lymphomas), the denominators
 

consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

19
 
significantly higher proportion of tumors than did the control
 

animals. As a part of these analyses, the one-tailed Fisher
 

exact test (Cox, 1970) was used to compare the tumor incidence of
 

a control group with 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) 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), was also used. Under the
 

assumption of a linear trend, this test determines if the slope
 

of the dose-response curve is different from zero at the onetailed 0.05 level of significance. Unless otherwise noted, the
 

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

20
 
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 an animal died naturally or was sacrificed
 

was entered as the time point of tumor observation. Cox's
 

methods of comparing these curves were used for two groups;
 

Tarone's extension to testing for linear trend was used for three
 

groups. The statistical tests for the incidence of tumors which
 

used life-table methods were one-tailed and, unless otherwise
 

noted, in the direction of a positive dose relationship.
 

Significant departures from linearity (P < 0.05, two-tailed test)
 

were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each treated group compared to its control was calculated
 

21
 
from the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as pt/pc where pt is the true binomial
 

probability of the incidence of a specific type of tumor in a
 

treated group of animals and pc is the true probability of the
 

spontaneous incidence of the 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% 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 (P < 0.025 one-tailed test when the control
 

incidence is not zero, P < 0.050 when the control incidence is
 

zero) has occurred. When the lower limit is less than unity, but
 

the upper limit is greater than unity, the lower limit indicates
 

the absence of a significant result while the upper limit
 

22
 
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.
 

23
 

III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of both male and female rats at all doses were
 

lower than those of the vehicle and untreated controls during
 

most of the study (figures 1 and 2). Fluctuations in the growth
 

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

diminishes, the mean body weight may be subject to wide
 

variation.
 

Rales were noted in a few animals of both treated and control
 

groups. No other clinical signs were recorded that could be
 

related to toxicity or early deaths. To control respiratory
 

disease, the mid- and high-dose groups and respective controls
 

received oxytetracycline in the drinking water at 0.6 mg/ml
 

during weeks 34 to 40 and at 0.3 mg/ml during weeks 40 to 44.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered acronycine by
 

injection at the doses of this experiment, together with the
 

untreated and vehicle controls, are shown in figures 3 and 4.
 

25
 
Figure 1. Growth Curves for Male Rats Treated With Acronycine 

26 
10 20 30 40 50 60 70 80 90 

TIME ON STUDY (WEEKS) 

Figure 2. Growth Curves for Female Rats Treated with Acronycine 

27 
Figure 3. Survival Curves for Male flats Treated with Acronycine 
Figure 4. Survival Curves for Female Rats Treated with Acronycine 

29 
In male rats, the results of the Tarone test for positive doserelated trend in mortality over the period of the bioassay are
 

significant (P < 0.001), using either set of controls. Also,
 

each of the treated groups has a significantly lower survival
 

than either control group. In the high-dose group, only 2/35
 

(6%) animals survived to week 47 of the study, and the median
 

time on study was 31 weeks; however, the first observed tumors
 

occurred as early as week 32. In the mid-dose group, 1/35 (3%)
 

animals survived to the end of the study, 9/35 (31%) survived to
 

week 52, the median time on study was 42 weeks, and the first
 

observed tumor occurred at week 35. In the low-dose group, 9/35
 

(26%) animals survived to termination of the study, 20/35 (57%)
 

survived to week 52, the median time on study was 56 weeks, and
 

the first observed tumor occurred at week 48. At least 70% of
 

the animals in the control groups (10/10 in either set of lowdose controls and 7/10 in either set of mid- and high-dose
 

controls) lived to the end of the study. The early deaths of the
 

male treated rats may have suppressed the incidences of lateappearing tumors.
 

In female rats, the results of the Tarone test are significant
 

(P < 0.001), using the high-dose, the mid-dose, and either set of
 

control groups, and an indicated departure from linear trend is
 

observed (P = 0.008), due to the steep increase in deaths in the
 

30
 
female high-dose rats. The survival of the low-dose group did
 

not differ significantly from that of either of its control
 

groups. In the high-dose group, only 4/35 (11%) animals survived
 

to week 80, 14/35 (40%) survived to week 52, the median time on
 

study was 46 weeks, and the first observed tumor occurred at week
 

39. In the mid-dose group 19/35 (54%), in the low-dose group
 

23/35 (66%), and in the controls at least 80% of the animals
 

(9/10 of the low-dose vehicle controls, 8/10 of the mid- and
 

high-dose vehicle controls, 8/10 of the low-dose untreated
 

controls, and 8/10 of the mid- and high-dose untreated controls)
 

survived to termination of the study. The early deaths of the
 

high-dose female rats may have suppressed the incidences of lateappearing tumors.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

Appendix A, tables A1-A4; findings on nonneoplastic lesions are
 

summarized in Appendix C, tables C1-C4.
 

A variety of neoplasms were observed in the control and treated
 

rats. There was a high incidence of neoplasms observed in the
 

treated rats when compared with the untreated- or vehiclecontrol rats. The treated female rats had a higher incidence of
 

adenocarcinoma of the mammary gland than the control female rats.
 

31
 
In the treated male and female rats, there were malignant
 

neoplasms of mesenchymal tissue, especially of the peritoneal
 

cavity; these included poorly differentiated sarcomas,
 

fibrosarcomas, hemangiosarcomas, malignant mesotheliomas, and
 

osteosarcomas. Similar neoplasms were not observed in any of the
 

control rats.
 

Malignant neoplasms of the mammary gland were observed in seven
 

low-dose, five mid-dose, and two high-dose female rats. No
 

malignant mammary neoplasms were observed in the control females.
 

The reverse dose relationship of these neoplasms was likely due
 

to the higher number of early deaths and killed moribund animals
 

which occurred in the mid- and high-dose groups. Although the
 

malignant mammary neoplasms varied in histologic appearance, they
 

were classified as adenocarcinomas. These neoplasms were highly
 

cellular and were characterized as focal proliferations of hyperchromatic glandular epithelium. The proliferating epithelium
 

formed small nests and acini which were supported by a fibrous
 

stroma. Papillary proliferation of the mammary epithelium was
 

observed in one of the adenocarcinomas, and large cystic areas
 

were present in a second adenocarcinoma.
 

A high incidence of osteosarcoma (low-dose 3/31, mid-dose 13/32,
 

high-dose 12/34) was observed in the treated male rats. Most of
 

these neoplasms were observed grossly as enlargements involving
 

32
 
the long bones of the limbs. Two osteosarcomas involved
 

vertebrae. Occasionally, the neoplasm appeared to involve only
 

soft tissues, and primary bone involvement was not observed. The
 

osteosarcomas were characterized as anaplastic spindle-cell
 

neoplasms which were forming varying amounts of osteoid. Several
 

of the osteosarcomas had metastasized to other organs, most
 

frequently, to the lung and the liver.
 

Other types of malignant mesenchymal neoplasms, especially of
 

tissues of the peritoneal cavity, were observed frequently in
 

treated male and female rats. Although all of these neoplasms
 

were poorly differentiated spindle-cell tumors, they were
 

variable in histologic appearance.
 

Some of the neoplasms were undif ferentiated and composed of very
 

pleomorphic spindle cells. These neoplasms were highly cellular
 

and contained undifferentiated mesenchymal cells, poorly
 

differentiated spindle cells, and multinucleated giant cells.
 

The neoplasms were rapidly proliferating, and contained numerous
 

mitotic figures. These undifferentiated sarcomas were classified
 

as sarcomas, NOS (not otherwise specified).
 

Other poorly differentiated neoplasms appeared to be composed of
 

malignant fibroblasts which were producing varying amounts of
 

collagen. These neoplasms were classified as fibrosarcomas.
 

33
 
A third group of malignant mesenchymal neoplasms found in the
 

treated rats were forming clefts and blood-filled spaces lined by
 

pleomorphic, hyperchromatic endothelial cells. These neoplasms
 

were classified as hemangiosarcomas.
 

A fourth type of neoplasm observed in the treated rats was
 

classified as mesothelial sarcoma (malignant mesothelioma).
 

These neoplasms were nodular growths arising from the serous
 

membranes lining the peritoneal cavity. They were characterized
 

as papillary projections consisting of a fibrous core covered by
 

large mesothelial cells.
 

The mesenchymal neoplasms described above appeared to be highly
 

malignant, as evidenced by a high incidence of invasion into
 

adjacent organs and soft tissues and/or metastasis to other
 

sites. Many of these neoplasms were generalized and involved the
 

serosal surfaces of the abdominal viscera.
 

A variety of nonneoplastic lesions were present in both treated
 

and control animals. The only lesion which appeared to be
 

related to the injection was chronic inflammation in the
 

peritoneal cavity, involving the serosal surfaces of the
 

mesentery and visceral organs. There were also focal areas of
 

coagulative necrosis observed in the liver. These lesions
 

occurred in one vehicle-control rat and several treated rats.
 

34
 
In the judgment of the pathologists, acronycine, at the doses
 

used in this bioassay, induced malignant neoplasms in both male
 

and female rats. Adenocarcinoma of the mammary gland in female
 

rats and malignant neoplasms of mesenchymal tissues in both male
 

and female rats were observed only in the treated groups.
 

D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the time-adjusted
 

statistical analyses of the incidences of those primary tumors
 

that were observed in at least two animals in one group and with
 

an incidence of at least 5% in one or more than one group.
 

Time-adjusted analyses eliminate animals that died before week 52
 

on study unless a tumor was found at the specific site before
 

this time; in the latter instance, the analysis is based on
 

animals that survived at least as long as the animal in which the
 

first tumor was found. The untreated controls are not included
 

in the tables and the analyses, since the test conditions of the
 

vehicle controls more closely resembled those of the treated
 

animals.
 

In male rats, the result of the Cochran-Armitage test for
 

positive dose-related trend in the incidence of osteosarcoma of
 

the musculoskeletal system is significant (P = 0.019), using the
 

mid- and high-dose vehicle-control group, the mid-dose group, and
 

35
 
the high-dose group, and the results of the Fisher exact test
 

show that the incidences in the mid- and high-dose groups are
 

significantly higher than that in the vehicle-control group (P =
 

0.027 and P = 0.013, respectively); however, the probability
 

level in the mid-dose group is above the 0.025 level for
 

significance required by the multiple comparison criterion. The
 

life table of the incidence of this tumor in the male rats is
 

shown in figure 5. The result of the Tarone test is significant
 

(P < 0.001) when the mid- and high-dose groups are used with
 

their designated control group; however, the result of the Cox
 

test comparing the lowTdose group and its vehicle-control group
 

.is	 not significant. The statistical conclusion is that the
 

incidence of osteosarcoma of the musculoskeletal system in male
 

rats is associated with the administration of acronycine. No
 

such tumor was observed in female rats.
 

Two osteosarcomas of the liver were found in the high-dose male
 

rats. The result of the Cochran-Armitage test on the incidence
 

of this tumor is significant (P = 0.048), using the mid- and
 

high-dose vehicle-control group, the mid-dose group, and the
 

high-dose group; however, the results of the Fisher exact test
 

are not significant. Results of statistical tests on the
 

incidences of this tumor in female rats are not significant.
 

When osteosarcomas of all sites are considered together, the
 

36
 
Figure 5. Life Table for Male Rats Treated with Acronycine:

 

Osteosarcoma of the Musculoskeletal System 

37 
result of tbe Cochran-Armitage test is significant (P = 0.002) in
 

the male rats, using the mid- and high-dose vehicle-control
 

group, the mid-dose group, and the high-dose group. The results
 

of the Fisher exact test indicate that the incidences in both the
 

mid- and high-dose groups are significantly higher than that in
 

the control group (P =» 0.022 and P = 0.002, respectively). The
 

statistical conclusion is that the incidence of osteosarcomas at
 

all sites in male rats is dose associated. Results of statistical tests on the incidences of these tumors in female rats are
 

not significant.
 

The result of the Cochran-Armitage test on the incidence of
 

cortical adenoma of the adrenal in male rats is significant (P =
 

0.045), using the mid- and high-dose vehicle-control group, the
 

mid-dose group, and the high-dose group, but the results of the
 

Fisher exact test are not significant. Results of statistical
 

tests on the incidences of this tumor in female rats are not
 

significant.
 

The results of the Fisher exact test show that the incidence of
 

fibroadenoma of the mammary gland in low-dose female rats is
 

significantly higher (P = 0.007) than that in the low-dose
 

vehicle controls; however, the incidences of the tumor in the
 

mid- and high-dose groups are not significant. When all tumors
 

of the mammary gland, except fibroma, are combined for analysis,
 

38
 
the results of the Fisher exact test show that the incidence in
 

the low-dose group is significantly higher (P = 0.004) than that
 

in the low-dose vehicle controls; however, the result of the
 

Cochran-Armitage test using the mid- and high—dose groups and the
 

appropriate control indicates a significant trend (P = 0.034) in
 

the negative direction. This significant negative trend is due,
 

principally, to the lower incidence observed in the high-dose
 

group. The life-table analysis made using the times of observations of this tumor also yielded a significant negative trend (P
 

= 0.017). As shown in the section concerning survival of the
 

female rats, the high-dose group evidences a steep decrease in
 

survival compared with the other groups.
 

In female rats, five sarcomas, NOS, of the peritoneum were found
 

in the high-dose group, but none were observed in the other
 

groups studied. The result of the Cochran-Armitage test on the
 

incidence of this tumor is significant (P = 0.010), using the
 

mid- and high-dose vehicle-control group, the mid-dose group, and
 

the high-dose group, but the results of the Fisher exact test are
 

not significant. Results of statistical tests on the incidences
 

of this tumor in male rats are not significant.
 

When sarcoma and other related tumors of the peritoneum are
 

considered together, the results of the statistical tests are
 

significant in each sex. The results of the Cochran-Armitage
 

39
 
test, using the mid- and high-dose vehicle-control group, the
 

mid-dose group, and the high-dose group indicate probability
 

levels of P = 0.006 in males and P = 0.002 in females, and the
 

Fisher exact comparisons of the incidences in the high-dose
 

groups with those in the control groups are P = 0.033 in males
 

and P = 0.016 in females; however, the P value for the males is
 

above the 0.025 level required for significance by the multiple
 

comparison criterion. The statistical conclusion is that the
 

incidence of these tumors is dose associated in female rats.
 

In summary, the statistical tests indicate dose association in
 

the incidence of osteosarcoma of the musculoskeletal system and
 

in osteosarcoma at all sites in male rats, and also in sarcoma
 

and other related tumors of the peritoneum in female rats.
 

40
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of the high-, upper mid-, and lower mid-dose
 

male mice and of all treated female groups were generally lower
 

than those of the untreated- and vehicle-control groups (figures
 

6 and 7), while the weights of the low-dose males were more
 

comparable to those of the control groups. Fluctuations in the
 

growth curve may be due to mortality; as the size of a group
 

diminishes, the mean body weight may be subject to wide
 

variation.
 

Abdominal distention was the only consistent clinical sign
 

reported in the treated animals; it occurred in all but the
 

high-dose group, in which the time of survival was very short.
 

To control respiratory disease, propylene glycol vapor was used
 

during weeks 11 to 22 in the room housing the low-dose mice.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered acronycine by
 

injection at the doses of this experiment, together with the
 

untreated and vehicle controls, are shown in figures 8 and 9.
 

41
 
TIME ON STUDY (WEEKS) 

Figure 6. Growth Curves for Male Mice Treated with Acronycine 

42 
Figure 7. Growth Curves for Female Mice Treated with Acronycine 

43 
Figure 8. Survival Curves for Male Mice Treated with Acronycine 

44 
Figure 9. Survival Curves for Female Mice Treated with Acronycine 

45 
In each sex, the result of the Tarone test for positive doserelated trend in mortality over the period of the bioassay is
 

significant (P < 0.001), using the high-dose group, the upper
 

mid-dose group, and the vehicle-control groups; all animals in
 

the treated groups died before the end of the study. The median
 

number of weeks on study of male mice was 18 for the high-dose
 

group, 30 for the upper mid-dose, 48 for the lower mid-dose, and
 

89 for the low-dose. In the low-dose group of male mice, 33/40
 

(82%) animals were alive after week 52 on study, and no tumor was
 

observed before this time. In the lower mid-, upper mid-, and
 

high-dose groups, all 35 male mice in each group died before week
 

52. No tumor was observed in the lower mid- and high-dose
 

groups, but in the upper mid-dose group, a carcinoma of the bile
 

duct was observed at week 30 on study.
 

In females, the median number of weeks on study was 17 for the
 

high-dose, 31 for the upper mid-dose, 48 for the lower mid-dose
 

and 74 for the low-dose groups. In the low-dose group, 31/40
 

(78%) animals lived to week 52 on study, and no tumor was
 

observed before week 52. All 35 female mice in each of the three
 

other treated groups (lower mid-, upper mid-, and high-dose
 

groups) died before week 52. No tumor was observed in the lower
 

mid- and high-dose groups, while in the upper mid-dose group, two
 

tumors were observed, one at week 29 (adenocarcinoma, NOS, of the
 

46
 
bile duct) and the other at week 32 (granulocytic leukemia of the
 

bone marrow). The survival rates of the control groups within
 

each sex are not comparable, since, in male mice, the percentage
 

survivals to 78 weeks among the upper mid- and high-dose, lower
 

mid-dose, and low-dose vehicle-control groups are 5/10 (50%),
 

7/10 (70%), and 0/20 (0%), respectively; among the corresponding
 

untreated-control groups, they are 10/10 (100%), 9/10 (90%), and
 

16/20 (80%). In females, the percentage survivals to 78 weeks
 

among the three vehicle-control groups are 9/10 (90%), 8/10
 

(80%), and 0/20 (0%); among the untreated-control groups, they
 

are 9/10 (90%), 10/10 (100%), and 19/20 (95%). The early deaths
 

of the treated mice of both sexes may have suppressed the
 

incidences of late-appearing tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

Appendix B, tables B1-B4; findings on nonneoplastic lesions are
 

summarized in Appendix D, tables D1-D4.
 

A variety of neoplasms were observed at approximately the same
 

incidence in the control mice as in the low-dose mice. No
 

neoplasms were observed in any of the high-dose mice, and very
 

few neoplasms were observed in upper and lower mid-dose mice.
 

There was a high incidence of early deaths and killed moribund
 

47
 
animals in these three treated groups of animals during the
 

exposure period, which may be related to the unusually low
 

incidence of neoplasia observed in these groups.
 

There were cases in this study in which some types of neoplasms
 

occurred only in treated mice. These have been observed as
 

spontaneously occurring neoplasms in this strain of mouse. The
 

nature and low incidence of these neoplasms in this study provide
 

no evidence that they are related to the administration of
 

acronycine.
 

A variety of nonneopla.stic lesions were observed in both control
 

and treated mice. The only apparent acronycine-induced lesions
 

observed in this study were acute and chronic inflammatory
 

lesions involving the thoracic and abdominal viscera, renal
 

medullary necrosis, and bile duct hyperplasia in several mice.
 

In the judgment of the pathologists, the results of this microscopic examination of mice receiving acronycine at any of the
 

four doses are inconclusive. Although there were no obvious
 

acronycine-induced neoplasms observed in the treated animals when
 

compared with control animals, the high incidence of early deaths
 

and killed animals in the treated groups precludes a definitive
 

conclusion on the effect of acronycine in mice in this study.
 

48
 
D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the time-adjusted statistical analyses of the incidences of those primary tumors that
 

were observed in at least two animals in one group and with an
 

incidence of at least 5% in one or more than one group. The
 

untreated controls are not included in the tables and the
 

analyses, since the test conditions of the vehicle controls more
 

closely resembled those of the treated animals. This bioassay
 

originally started with 25 mg/kg as the high dose. In both
 

sexes, survival was low, and no tumors were observed in the
 

high-dose groups. In the groups of male and female mice
 

receiving 12.5 mg/kg (upper mid-dose groups) survival was also
 

low, and only one tumor, a carcinoma of the bile duct, was
 

observed among the male mice. In the upper mid-dose females, one
 

animal had leukemia and another had adenocarcinoma of the bile
 

duct. Subsequently, two other groups were started at doses of 6
 

mg/kg (lower mid-dose group) and 2 mg/kg (low-dose group). No
 

tumors were observed in the lower mid-dose group. Since the
 

survival and numbers of tumors observed in all groups except for
 

the low-dose group and its control group were so low that
 

meaningful analysis was precluded, only the low-dose group and
 

its control group were subjected to statistical analysis. A
 

49
 
summary of all tumors in all treated groups is given in tables
 

B1-B4 of Appendix B.
 

No significant increase in incidences of tumors in the treated
 

groups was observed when compared with their control groups,
 

although statistical analysis of the incidence of tumors in the
 

mice was performed using all mice evaluated histopathologically
 

and also using only those animals that lived beyond week 52 or
 

beyond the week of the first observation of a specific tumor,
 

whichever number of weeks was smaller. In each sex, the
 

incidences of lymphoma in the low-dose groups were lower than
 

those observed in the respective controls. When the incidences
 

of lymphoma in the untreated-control groups are compared with
 

those of the corresponding vehicle-control groups, no significant
 

difference Is observed between the lower mid-dose vehicle
 

controls (0/10 in each sex) and the lower mid-dose untreated
 

controls (0/9 in males and 0/10 in females); however, a
 

significant difference is observed between the low-dose vehicle
 

controls (13/17 in males and 19/19 in females) and the low-dose
 

untreated controls (3/18 in males and 6/19 in females). These
 

extremely high incidences in the vehicle-control groups compared
 

with the untreated groups may indicate procedural difficulties.
 

Overall, the shortened life spans of the treated and vehiclecontrol groups of mice precluded meaningful evaluation.
 

50
 
V. DISCUSSION
 

Acronycine was toxic to both sexes of rats and mice when administered by intraperitoneal injection at the doses used in this
 

bioassay. This is shown by the high mortality rates in all but
 

the low-dose groups, and by the lower mean body weights in dosed
 

rats and mice at all doses throughout most of the study. Because
 

of this high number of deaths, time-adjusted statistics were used
 

for the analyses of all incidences of tumors.
 

In male rats, the dose-related trend in the mid- and high-dose
 

groups for the incidence of osteosarcoma of all sites was
 

significant (P = 0.002) using the respective vehicle-control
 

group (vehicle controls 0/8, mid-dose 13/30, high-dose 12/18).
 

Comparisons of the individual groups with respective control
 

groups were also significant for the mid-dose (P = 0.022) and
 

high-dose (P = 0.002) groups, but not for the low-dose group.
 

Most of these neoplasms were observed grossly as enlargements of
 

the long bones of the limbs, but occasionally, the tumors
 

appeared to involve only soft tissues, and primary bone
 

involvement was not observed. In female rats, osteosarcoma was
 

observed only in 1/8 high-dose animals.
 

Sarcomas and other related tumors of the peritoneum (listed in
 

the appendixes as sarcoma, NOS; mesothelioma, NOS; malignant
 

51
 
mesothelioma; and fibrosarcoma of the peritoneum or multiple
 

organs) were observed in all three dosed groups of both male and
 

female rats, but in none of the control groups (males: low-dose
 

5/30, mid-dose 3/26, high-dose 7/16; females: low-dose 1/35,
 

mid-dose 5/30, high-dose 13/28). In both sexes, the doserelated trends were significant (males, P = 0.006; females, P =
 

0.002), and the comparison of the incidences in the high-dose
 

females with the vehicle-control group was significant (P =
 

0.016). None of the incidences in the individual dosed groups of
 

males were significant when compared with vehicle controls.
 

However, since the tumors occurred in all dosed groups but did
 

not occur in any of the historical-control animals at this
 

laboratory, they are considered to be related to administration
 

of the chemical.
 

In female rats, the incidence of all tumors of epithelial origin
 

of the mammary gland was significant only at the low dose
 

(low-dose vehicle controls 1/10, low-dose 22/35, P = 0.004).
 

Adenocarcinomas of the mammary gland were observed in seven
 

low-dose, five mid-dose, and two high-dose female rats, but in no
 

control females. The reverse dose relationship of both benign
 

and malignant tumors was probably due to the higher number of
 

early deaths which occurred in the high-dose group.
 

All mice of each sex of the three upper dosed groups had died by
 

52
 
week 52. Among the low-dose mice, 33/40 males and 31/40 females
 

lived to week 52 on study; however, only 5/20 male and 1/20
 

female low-dose vehicle controls lived beyond 1 year. Among the
 

high-, upper, and lower mid-dose groups, only one tumor was
 

observed in males and two in females in the upper mid dose. Even
 

among the low-dose groups, no tumor was observed in a statistically significant incidence.
 

Lymphomas were observed at lower incidences in the low-dose male
 

mice (10/37) and low-dose females (6/37) than in the
 

corresponding male (13/17) and female (19/19) low-dose vehicle
 

controls. However, the incidences in the upper mid-dose and
 

high-dose vehicle-control and the lower mid-dose vehicle-control
 

groups were not increased. When the incidences of lymphoma in
 

the untreated- and vehicle-control groups were compared, no
 

significant differences were observed between the lower mid-dose
 

vehicle controls (0/10 in both sexes) and the lower mid-dose
 

untreated controls (0/9 in males and 0/10 in females); however, a
 

significant difference was observed between the low-dose vehicle
 

controls (13/17 in males and 19/19 in females) and the low-dose
 

untreated controls (3/18 in males and 6/19 in females).
 

This high incidence in the low-dose vehicle controls may have
 

been due to a procedural problem. The same needle for injection
 

was used for each group of five animals within a cage, and
 

53
 
furthermore, the same bottle of vehicle solution was used for all
 

vehicle-control animals. Thus, the possibility of transfer of
 

tumor cells or oncogenic viruses cannot be excluded.
 

Nonneoplastic lesions of the peritoneal cavity, i.e., inflammation and fibrosis, were found in rats and mice from each of the
 

dosed groups,, but not in any control animals.
 

Since 1966, acronycine has been tested as an antineoplastic agent
 

in humans; however, no long-term studies in animals or humans
 

have been reported. In a 6-month study for pulmonary tumor
 

response in strain A mice, Stoner et al. (1973) found that
 

intraperitoneal injection of total doses of 0.53 to 2.60 rag/kg of
 

acronycine did not elicit a carcinogenic response.
 

The vehicle used for the acronycine for all groups in this
 

bioassay contained polysorbate 80, which in itself has been
 

implicated as a carcinogen, but only in the production of local
 

sarcomas following subcutaneous injections (Grasso et al., 1971).
 

However, in these bioassays no local sarcomas were observed in
 

the vehicle-control animals administered polysorbate 80 by
 

intraperitoneal injection.
 

It is concluded that under the conditions of this bioassay, the
 

low survival of the dosed and control mice and the possible
 

procedural problems associated with intraperitoneal injection of
 

54
 
the chemical do not allow a determination to be made of the
 

carcinogenicity of acronycine in this species. In Sprague-Dawley
 

rats, acronycine in the vehicle of 0.05% polysorbate 80 in
 

phosphate-buffered saline was carcinogenic, producing tumors of
 

the mammary gland in females, osteosarcomas in males, and
 

sarcomas and other related tumors of the peritoneum in both males
 

and females.
 

55
 

VI. BIBLIOGRAPHY
 

Armitage, P., Statistical Methods in Medical Research, John Wiley
 

& Sons, Inc., New York, 1971, pp. 362-365.
 

Berenblum, I., ed. , Carcinogenicity Testing; A Report of the
 

Panel on Carcinogenicity of the Cancer Research Commission
 

of the UICC, Vol. JZ. International Union Against Cancer,
 

Geneva, 1969.
 

Carter, S. K., Clinical trials and combination chemotherapy.
 

Cancer Chemotherp. Reports Part III 2(1);81-97, 1971.
 

Cox, D. R. , Regression models and life tables. J. R. Statist.
 

Soc. B 34;187-220, 1972.
 

Cox, D. R., Analysis of Binary Oaf a, Methuen & Co., Ltd. London,
 

1970, pp. 48-52.
 

Dunn, B. P., Gout, P. W., and Beer, C. T., Effects of the
 

antineoplastic alkaloid acronycine on nucleoside uptake and
 

incorporation into nucleic acids by cultured L5178Y cells.
 

Cancer Res. 33;2310-2319. 1973.
 

Gart, J. J., The comparison of proportions: a review of
 

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Stat. Inst. 39;148-169. 1971.
 

Grasso, P., Gangolli, S. D., Goldberg, L., and Hooson, J.,
 

Physicochemical and other factors determining local sarcoma
 

production by food additives. Fd. Cosmet. Toxicol.
 

1:463-478, 1971.
 

Kaplan, E. L. and Meier, P., Nonparametric estimation from
 

incomplete observations. J. Am. Statist. Assoc. 53;457-481,
 

1958.
 

Lahey, F. N. and Thomas, W. C., Alkaloids of the Australian
 

Rutaceae: Acronychia Baueri. Australian J. Sci. Res.
 

2A::423, 1949.
 

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

Peters, J. A., Carcinogenesis bioassay data system. Comp.
 

and Biomed. Res. _7:230-248, 1974.
 

57
 
Miller, R. G., Jr., Simultaneous Statistical Inference,
 

McGraw-Hill Book Co., New York, 1966, pp. 6-10.
 

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

Kaufman, D. G., Respiratory tract carcinogenesis in hamsters
 

induced by different numbers of administrations of benzo(a)
 

pyrene and ferric oxide. Cancer Res. 3.2; 1073-1081, 1972.
 

Stoner, G. D. , Shimkin, M. B., Kniazeff, A. J., Weisburger, J. H.
 Weisburger, E. K., and Gori, G. B., Test for carcinogenicity
 

of food additives and chemotherapeutic agents by the
 

pulmonary tumor response in strain A mice. Cancer Res.
 

.33:3069-3085, 1973.
 

Svoboda, G. H., Poore, G. A., Simpson, P. J., and Boder, G. B.,
 

Alkaloids of Acronychia Baueri Schott I. J. Pharm. Sci.
 

.55: 758-768, 1966.
 

Tarone, R. E. Tests for trend in life table analysis.
 

Biometirika 62:679-682. 1975.
 

58
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN RATS
 

GIVEN INTRAPERITONEAL INJECTIONS OF ACRONYCINE
 

59
 

TABLE A1 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS GIVEN INTRAPERITONEAL 

INJECTIONS OF ACRONYCINE (CONTROL GROUPS) 

LOW DOSE MID AND HIGH LOW DOSE 

UNTREATED DOSE UNTREAT- VEHICLE 

CONTROL ED CONTROL CONTROL 

»»IH»LS INITIMIY IN STODY 10 10 10 

ARIBkLS BECBOJSIED 10 9 10 

»«IBHLS KXAHINED HISTOPHTHOIOGICM.LY 10 9 10 

KTEGOHENTUBY 5YE1EH 

HONE 

BISPIBHTOBY SI£1EH 

•LONG (10) (9) (10) 

JUVEOL»F!/BRCNCHIOIJU HDINOHA 

HEH^TOPOIETIC EYSTPH 

NONE 

CIRCOLHTORY SYSTB'n 

NONE 

CIGESTIVF S'ISIEn 

NONE 

OflNHR Y SYStEn 

HONE 

tBDOCBINI SYSTEB 

IPITHITAEY (5) (8) (1) CHBOnOPHQBf ADENOn t 

aU52J12IB2EI._UB£I£oai_ J-JJJS1 . 

I NUHBEP OF XKIHAtS WITH TISSUE EXAMINE D nlCBOSCCPICHLIY 

* NUMBER OF MUflALS NICBOPSIID 

61 

MID AND HIGH DOSE VEHICLE

 

CONTROL 

1C 

e 

8 

(8) 

1 (13») 

CM 

2 (29*) 
TABLE A1 CONTROL MALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE MID AND HIGH UNTREATED DOSE UNTREATCONTROL ED CONTROL 

ACIDOPHIl ADENOH A 

HDREHAL 

CORTICAL KEHCHA 

(10) (9) 

UNPRODUCTIVE SYSTFB 

•BAflBAR Y GLAlit (10) (9) ADENOCARCINOKA , NOS 

JERVOUS STSTTfl 

NONE 

SPECIAL SENS? CBGAN S 

NONE 

PDSCDIOSKELFTAI SYSTEB 

NONE 

ECtI CAVITIES 

»OXE 

All OTHER STSTIBS 

NONE 

JNIHAL DISPOSITICS SBHKAR Y 

AMIBALS INITIALLY IN STODI 11 10 NATURA L CEATR9 3 HORIBDHE SACBITICE 

SCHl!tOl!C SACRIFICE 

ACCIDENTAIIY KILLED 

TEBSIHAL SUCBIFICE 10 7 ANIHAI. BISSING 

3 INCLDDfS AtJIOtTJBD ANIMALS 

I HUBBfR OF ANIBALS SITH TISSUE BXAflHED BICROSCCPICAltT 

* RUBBER Of ARIBAIS NECROPSIED 

LOW DOSE 

VEHICLE 

CONTROL 

(1C) 

(10) 

10 

1C 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

1 (1«X) 

(8) 

1 (13») 

(8)

1 (13t) 

10 

2 

1 

7 

62 
TABLE A1 CONTROL MALE RATS: NEOPLASMS (CONTINUED) 

u 

LOW DOSE 

NTREATED CONTROL 

MID AND HIGH 

DOSE UNTREATED CONTROL 

LOW DOSE 

VEHICLE 

CONTROL 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

10HOE SlIBHAB Y 

TOTAL ANIMAL S KITH PRIMAR Y

TOTAL PRIMAR Y TIIHORS 

 TUHOBS« 1 

1 

U 

6 

TOTAL ANIHAL S WIT H BFNIG I TOTA L EFNIG N TUMORS  TOMOR S II 

5 

TOTAL ANIMAL S WITH MALIGNAN T TOBORS 

TOTAL MALIGNAN T TUMOR S 

1 

1 

1 

1 

TOTAL ANIBAL 5 WITH SECONDAR Y

TOTAL SiECCK'CAR Y TDBORS TOBOBS* 

TOTAL ANIBAL 5 KITH TnflORS

BEBIRN OR flALIGNAHT 

TOTAL UNCEPTAI N TBBORS 

 BNCEBTAINTOTA L ANIfiM S «ITH TUMOR S

PRIMAR Y O B MT1ASTATIC 

TOTAL UHCEFTAIN TDMORS 

 DHCEBTAIN­

* FRIHAP ? TOHCRS : ALL TUHOR S EXCEPT SKCOKDABT TOnORS 

I SECONDAR Y TIJPCRS: HFTHSTATIC TOHOPS OB TUBOBS INVASIV E INTO AN ADJACEN T OBGAN 

63 
TABLE A2 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS GIVEN INTRAPERITONEAL 

INJECTIONS OF ACRONYCINE (TREATED GROUPS) 

LOW DOSE MID DOSE HIGH DOSE 

~ c. ANIMAL S JHtTIAII Y I S STUDY 35 35 

ANIHAL S NECEOESIE D 31 32 3U 

ANIMAL S EXAMINF E HISTOPATH OLOGICALL Y 30 31 JU 

INTEGUB'NTA F V EYSTF H 

'SOBCIIT TTSSD E {3D (32) (3») 5ARCCHA , NOS 1 (3*) FIBBCH A 1 («) 1 (3*) 

"IBHCSUFCCtlA 1 (3X) OSTSCSAFCCP A 1 |3 H) 1 <3«) 

RESPIRATOR Y SYSTE d 

«HING 

ALVEOLAR/EBCNCHI01A R ADENOH A 

HEBANGIOSAFCOIIA , HETASTATI C 

nSTEOSARCCRA , BSTASTATIC 

(30 ) 

1

1

2

 (3*) 

 (3») 

(If) 

(31) 

9 (29*) 

I 31) 

1C (29%) 

fFfllTOPOIETIC SYSTE1 

HtYBP H BOD E (30) (20) mi OSTEOS ^RCC^A , fIFTASTATIC 1 (5*) 1 (SKI 

• Mf'SENTE R 1C I, NCD E (30) (2") <:i) nESOTHEUCBA , H"TASTATIC 1 (3? ) 

CIRCOLATCR T SYSTEP 

NON E 

CIGtSTIV * SYSTI P 

tllVE R (29 ) (31) (3«) HEPATOCFLIBIA S APENOB A 1 (3*) 

!lE.£ATOCFLLi'I.flR CAR£ISaa« 1 JJJS1 

• NUUBP R OP ANIHAL " KITH TISSU E EXASIIBE D BICBOSCCPICA ttY 

» NUBBFH OF AMMAt S NECROPSIFi; 

64 
TABLE A2 TREATED MALE RATS: NEOPLASMS (CONTINUED)

 

FIBROSABCCHA 

HEHANGICSAfCCflA 

OSTSOSARCCHA 

OSTECSABCCBA, HETASTATIC 

• BILE DCCT 

BILE DOCT CAFCINOBA 

•PANCREAS 

riBROSAKCCFA 

OSTECSARCCBA, BETASTATIC 

•STOHACH 

OSTEOSABCCTA. B'TASTATTC 

•LARGE INTESTINE 

SQUABOUS CEIL CARCINOMA 

URINABY SYSTEr 

•KIDNFY 

OSTEOSABCCPA, BET ASTATIC 

ENDOCRINE SIST?n 

IADRENAL 

CORTICAL (TFHOBA 

CORTICAL CJRCINOBA 

OSTFOSAECCBA, BTTASTATIC 

REPRODUCTIVE SYSTE M 

»BAMMAR Y GLAK C 

"IBROADENCMA 

•1ESTIS 

INTFRSTITIJL-CELL TDMOR 

KEBVOUS SYSTFR 

NOUE 

SPECIAL SFNSt C8GAN S 

&ZS.I 

« NOBBTR 0" ANIMAL S WITH TISSUE

* NDMBTR OF ASIBAI S NTCROPSIIt: 

LOW DOSE MID DOSE 

t (1«») 

3 (10X) 1 (13*) 

(3D (32)

 

2 <6K) 

(2K) (26) 

1 («*) 

1 («*> 

(29) (30) 

1 (3*1 

(2«) (29) 

1 (3K) 

(29) (31) 

1 (3^) 

(28) (3D 

1 <!>*) 2 (6*) 

1 (3*) 

1 (UK) 

(31) (32) 

1 (31) 

(2B) (30)

 

1 (»*) 

 EXUBIBE D MICSOSCCPICALLY 

65 

HIGH DOSE 

1 (3*) 

1 (3*) 

2 (6*) 

5 (15X) 

(31) 

(30) 

1 (3*) 

(30) 

(28 ) 

(33) 

1 (3*) 

(33 ) 

1 (12K) 

3 (9*) 

<3<4) 

(32) 
TABLE A2 TREATED MALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE 

HOSCOLOSKELITH SYSTEB 

• BOHE 

OSTEOSAECCPA 

(31) 

3 (10*) 

(32) 10 (31*) 

(31) 

8 (2««) 

•VERTEBRA 

OSTEOSARCCPA (31) (32) 

1 <3«) 

(31) 

1 (3*) 

• SKF.IZTAI BUSCtE (31) (32) 43d) OSTECSAFCCFA. BETASTATIC 1 <3*> 

JODI CAVITIES 

• ABTIOBIHAI CAVIT Y 

FIBEOSABCOKA (31) (32) r<o 

~1 (3*) 

• PERITONEUM 

SARCCBA , SCS (31) 

2 (6*) 

(32) (31) 1 (3*) 

FIBROSABCOBA 

BESOTHEIICPA, DOS 1 <3K ) 1 (3*) 

2 (6%) 

HESOTHELIOHA, BALISHAST 

HEBANGIOSA5COBA, BETASTATIC 

OSTEOSARCOBA 

2

1

 (6*) 

 (3*) 

1 (31) OSTECSAECCBA, BETASTATIC 1 (3») 

•PFRITONIAL CAVIT Y 

FIBRCSABCCBA (31) (32) (3d) 

1 (3X) 

•BESENTERY 

OSTECSAECCPA (31) (32) 

1 (3*) 

(3d) 

Alt OTHER SYSTIBS 

•SUlflPlE ORGAN S 

SAECOHA, HCS, BETASTATIC (31) 

2 (6*) 

(32) (3d) 

FIBROSARCOBA 

OSTEOSARCCHA, BETASTATIC 1 (3*) 

2

2

 (6*) 

 (6«) 

2 (6X) 

CIAPHSAU M j 

« HUHBFR OF ABIBA1S WITH TISSUE EIABIKED HICROSCOPICA1LY 

» BOBBER OF ABIBAL S NECBOPSIED 

66 
TABLE A2 TREATED MALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE 

AIIBU L DISICSITICB SOHHABY 

• NIHALS INITIALL Y III STUDY 35 35 1C

 

HATOBAL DEMH8 17 17 "9 HORIBUN C SACRIFICE 9 17 2f

 SCHECUIIC SACRIFICE

 

ACCIDENTAIIY KILLED 

TERMINA L SKRIPICE 9 1 

ANIBB L BISEI1IG 

3 INCLODIS AD1CIYZE D ABIHAL S 

•IOIIOR sunnAR H 

TOTAL ANinA U IIITH PRIflARI THHORS* 17 16 15 TOTAL PRIBAR Y THMORS 21 23 28

 

TOTAL AKIBAL ; »ITH BENIRH TDIIORS 5 (1 ll TOTAL BEMG N TOBORS 5 U H 

TOTAL JlNIBAIE BITH KALIGMANT TOHOBS 12 16 in TOTAI BMIGNABT TOBOBS 15 18 2U

 

TOTAL ASIBAI S BITH SKONnARY TOBORS* 8 12 10 

TOTAL SECOSEAR Y TOBORS 13 19 22

 

TOTAL AHIBAL ; BITH TonoRs OHCSRTAIH- BEKIGll OR BAI1GHAN T 1 1

 

TOTAL UNCERTAI N TOBORS 1 1

 

TOTAL AVIHAI S BITH TOHOPS ONCIRTAIH-

FRIHAR Y OP BETASTATIC 

TOTAL OHCEFTMN TOBORS 

» PRIMAR Y TOHOPS: ALL TDBORS EXCEPT SECONDARY TDBOBS 

« SECONDARY TIlfOBS: BETASTAIIC TDHORS OR TBHORS IHVUSIVE I»TO AS ADJACIM ORGAS 

6* 
TABLE A3

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL GROUPS) 

LOW DOSE MID & HIGH DOSE IJNTREATED UNTREATED 

CONTROL CONTROL 

JNIHAL S INITIJIII Y I N STUDY 10 10 

ANIMAL S HFCFOISIE D 10 9 

ANIBAL S EXAB1NII ) HISTOPATHOLOGICAILY 10 8 

INTEGUBSNTAB Y JYS1EB 

NON E 

BESPIRATCBY SYETf F 

HTFACBE A (10) (8)

 

CARCINCBA , BOS, BETASTATIC 

ILUN G (10) (7)

 ALVECIAF/EFCNCHIOLA R CAECINOH A

 

FEBATOPOKTIC SYSTEH 

HONE 

CTKCOLATCBr STSTEf 

NON E 

CJGESTITF SYSTfR 

• ESOPHAGU S (10) (8)

 

CARCIHOMA , NOS, BFTASTATIC 

DBIHAE Y SYSTEB 

NOHE 

fSEOCRIBI SYSTFB 

IPITOITA8Y (7) (7)

QH?99QEBO^j APJNOHA 3 O3I) j_aai. t HDBBEH OF KRIHM.S WITH TISSUE EXAIUHED HICROSCCPICAltT 

« NUMBER OF ASIHUI S NECROPSIID 

LOW DOSE 

VEHICLE 

CONTROL 

1C 

10 

10 

(10) 

(10) 

<10) 

<6)

2 (331 I)

MID & HIGH DOSE 

VEHICLE 

CONTROL 

10 

9 

9 

(9)

1 (11*) 

(9)

1 (11 «) 

(6) 

1 (1711) 

m1 (11*1 

68 
TABLE A3 CONTROL FEMALE RATS: NEOPLASMS (CONTINUED) 

CHRCHOPHCEE ORCINOHA 

IADRENAL 

CORTICAL AtENCflA 

•THYHOIt 

CABCINCMA.KOS 

REPBODOCTIVE SYSTEn 

'BAHHARY GLA»r 

PIBRCADENCHA 

•CERVIX DTERI 

SQDABOOS CtLl PUPILIOBA 

KERVOUS SYSTB H 

NONE 

SPECIAL SJSSE CEGANS 

NONE 

HOSCOLOSKEIITBI SYSTEM 

NONE 

tocr CAVITIES 

NONE 

All OTHER SYSTEM S 

I NOHBER OF ANIHAL S WITH TISSDE

* NDHBER OF AKIHAI S NEC80PSIED 

LOW DOSE 

VEHICLE 

CONTROL 

(1C) 

OC) 

(10) 

1 (10*) 

(10), 

1 (10*) 

MID & HIGH DOSE 

VEHICLE 

CONTROL 

(9) 

1 (11*) 

(I) 

1 (1»*) 

(9) 

3 (33X) 

(9) 

LOW DOSE 

UNTREATED 

CONTROL 

(10) 

(10) 

(10) 

II (40*) 

(10) 

MID & HIGH DOSE 

UNTREATED 

CONTROL 

1 (1M) 

(8) 

(7) 

(9) 

1 (11*) 

(8) 

 EXAMINE D BICROSCCPICAllt 

69 
TABLE A3 CONTROL FEMALE RATS: NEOPLASMS (CONTINUED)

 

MID & HIGH DOSE VEHICLE CONTROL 

10 

2 

8 

3 

7 

3 

•; 

2 

2 

1 

2 

LOWDOSE MID & HIGH DOSE 

Ul ^TREATED UNTREATED 

(2ONTROL CONTROL 

AKIHA L DISFCEI1ION SOBBARY 

AHIHALS IKITIALIY IN STDDY 10 10

 NATURA L tFMHJ 1 1

 

BORIBOND SACRIFICE 1 1

 

SCHItOLJT! SACRIFICE 

ACCIDENTALLY KILLED 

TERBINA l SACRIFICE a 8

 

ANIHA L HISJIH G 

s iNCLtins AnTdYZ^p AHIMAI.S 

1DROG SUBMA8 1 

TOTAL ANIBAI S »ITH PBIHAR Y TOIIOES* 5 H

 

TOTAL PBIBAH Y TUBORS 7 5 

TOTAL AHIHAL S SITH BENIGK TOHORS 5 3 

TOTAL BINIG1I TOHORS 7 11

 

TOTAL AHIHAL S SITH nALIGHAMT TDBORS 1

 

TOTAI BAIIGNAS T TOBORS 1

 

TOTAL ANIBAI S BITH SECONDARY TOHORS* 

TOTAL SECCHtASl TDHORS

 

TOTAL ANIBAL S WITH TOBOSS ONC1RTAI11-

BENIGN OR BAIIGNAN T 

TOTAL ONCEETAIN TDBOSS

 

TOTAL ANIBAL S SITH TOHORS DNCIRTAIK-

PFIHAR Y OF HETASTATIC 

TOTAL UNCIFTAIN TUBORS 

LOW DOSE VEHICLE

 CONTROL

 

10 

1 

9 

l| 

D 

4 

U 

* FRIHABY TOHORS: All TOHORS EXCEPT SECONDARY TOHOHS
 

« SECONDARY TUBORS: HETASTATIC TOHORS OB TOMOHS INVASI7H INTO AH ADJACIKT ORGAN
 

70 
TABLE A4 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (TREATED GROUPS) 

fNIBALS INITIAIIY IN STDDT 

ANIBAL S NECFOEEIE D 

UNIBAI S EXABINTl : HISTOPATHOLOGICALLY 

INTEGUHENTAS Y SYSTEM 

• SKIN 

FIBROSARCCf A 

*SDBCUT TISSUI 

CARCINOMA , NO S 

SARCCBA, NCS 

FIBHCBA 

FIBROSAFCCB A 

LIPOBA 

BHABDOHYOSAECOB A 

BISPIHATOR Y SYETE K 

• LONG 

CARCISOBA . SOS. BETASTATIC 

ALVSOLAR/EfOBCHIOLA S ADESOBA 

AlVECLAR/EECNCHIOLA B CARCINOB A 

SARCCBA. SCS 

SARCCBA. KCS, BETASTATIC 

HEBANGIOSAFCOtA , BFTASTATIC 

HEBATOPOIETIC SYSTEB 

IIYBPH SOBH 

CAECINOHA , NOS, BETASTATIC 

ITHYBOS 

CARCINOBA , NOS, BFTASTATIC

 

CIRCtUATCBY SYSTEP 

NONE 

LOW DOSE MID DOSE HIGH DOSE 

35 35 zc 

35 

35 

32 

32 

3U 

-. i 

(35) (32) 

1 (3*) 

(3») 

(35) 

It

2

1

 (11*) 

 (6)1) 

 (3*) 

(32) 

2 (6K) 

|3«) 

1

1

 (3*) 

 (3*) 

1 O*> 1 (3%) 

(35) (32) 

2 (6*) 

(33)

1 (12*) 

1

1

 (3*) 

 (31!) 

1

1

1

 (3*) 

 (3*) 

 (3%) 

(35) (12) I2B) 

1 («) 

(1«) (16) (6)

1 <n«) 

' 

I STJBBER OF ANIBAtS ilTH TISSDE EXABINBD BICROSCCPICAL1T * NUBBEH OF ANIBAI S NECROPSIID 

71 
TABLE A4 TREATED FEMALE RATS: NEOPLASMS (CONTINUED)
 

tIGESTIVE SYSTIB 

•LIVER 

CARCIHOBA, BOS, BRTASTATIC 

HEPATOCEltULAR ADENOBA 

HSPATOCEILUIAF CARCISOHA 

HEBANGICSAFCOB A 

OSTEOSABCCBA. BETASTATIC 

• PANCREAS 

CARCINOHA.ROS 

SARCCBA , HOS 

OSTECSABCCTA. BETASTATIC 

ISTOBACH 

SARCCHA , NCS 

OSTEOSABCCBA. BETASTATIC 

IIIPDH 

FIBROSAHCCB A 

ILARGE INTESTIH E 

SARCCBA. NCS 

OflNAH I STSTSP 

•KIDNEY 

FIBROMA 

inRINAS l BLItCEB 

FAPIILOBA, SOS 

OSTEOSABCOBA, HETASTATIC 

ISDOCBISI SISTIB 

•PITUITARY 

CHROBOPHOEI »EENOBA 

1ADBENAL 

CORTICAL >E!NOBA 

CORTICAL CABCIMOB A

 

IADBENAL COBTIX 

£QBII£ii_U!Ia£Bi 

t HDHBES OF ANIHALS HITH TISSUE

* NDHBER OF AIINAt S HECROPSItt 

LOW DOSE MID DOSE 

(35) (32) 

it (13*) 

1 (3*) 5 (1IU) 

(3t) (30) 

(35) (30) 

(35) (29) 

(35) (29) 

(35) (31) 

(32) (26) 

1 (It*) 

(28) (30) 

4 (1VX ) 1 (3*) 

(32) (30) 

9 (30*) 

(32) (30) 

J J3.SL­

 EXABIHED BICBOSCCPICA1LI 

72 

HIGH DOSE 

(33) 

1 (3*) 

1 (3*) 1 (3*) 

(31)

1 (3*) 1 (3*) 

1 (3*) 

(32) 

1 (3*1 

1 (3*) 

(3C) 

1 (3*) 

(31) 

2 (6*1 

(32) 

1 (3*) 

(31) 

1 (3*) 

(30) 

1 (3X) 

(3D

•7 (23*) 

1 (3*) 

131) 
TABLE A4 TREATED FEMALE RATS: NEOPLASMS (CONTINUED) 

HWBODUCtlVE SJSTFfl 

»BA!!HARY GLAN t 

ADENCCARCIKBA . SOS 

PAPILLAE! ADENOCARCINOB A 

CYSTADENOHA, SOS 

CYSTADKHOCARCINOBA, DOS 

FIBRCBA 

FIBRCADENCB A 

tCTSROS. 

SARCCBA. SCS 

LEIOBYOSAECCB A 

ENDOBFTFIA L STBOBAL POLYP 

IOVAHY 

LEIOBYOBA 

SERVOUS SYSTIf 

NONE 

SPECIAL S?NS! CEGAN S 

NONE 

BUSOTLOSKILETAI SYSTEB 

•SKELETAL BHECLt 

SARCOBA, NCS 

ECEY CAVITIES 

•ABBOBINAL CAVIT Y 

PIBBOSAECCP A 

•PERITOBEUB 

SARCCBA, KCS 

FIBRCSAECCf A 

BESOTHELICBA, BALIGNAN T 

t NUBBER OF ANIBAL S BITH TISSOF

» NOBBER OF ASIBAI S NECHOPSirD 

LOW DOSE MID DOSE 

(35) (32) 

6 (17*) » (13*) 

1 (IK) 

1 (3*) 

1 <3X) 

2 (6*) 

2fl (571!) 13 (01*) 

(31) (32) 

1 (3K) 

5 (15*) 

(31) (31) 

1 (3«) 

(35) (32) 

(35) (32) 

1 (3*) 

(35) (32) 

2 (6*) 

1 <3H ) 

 EXABINE D BICROSCCPICALLY 

HIGH DOSE 

(31) 

2 (6*) 

1 (3*) 

3 (9*) 

(35) 

1 (3*) 

1 (3*) 

(31) 

(31)

1 (3*) 

(31) 

1 <«) 

|3«)

5 (15*) 

2 (6*) 

73 
TABLE A4 TREATED FEMALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE MID DOSE HIGH DOSE 

«BKENTIRY 

SARCCBA, HCS 

HI OTH'R SYSTIHS 

*BDLTIPtF CRGAB S

 

SARCCBA, SCS 

FIBRCSABCCBA 

»NIBA L DISEOSITIOS S3BBARY 

ANIHALS INITIALLY III STUDY 

NATDRA L DEATH S 

BOHIBONt SACRIFICE 

SCHTtDLID SACRIFICE 

ACCIDENTAIIY KIILED

 

TERHIKAI J^CEiriCE 

ANIRA L HISSING 

8 ISCL17DES ADICLY7ED ANIHAI.S 

TOBOB SOB1IABY 

TOTAL AMIHAt E »ITH PRIMAR Y TDHORS* 

TOTAI PRItHR Y TOHORS 

TOTAL ANIBAL S BITH BEMIGH TDBORS 

TOTAI BEK1G H TDHORS 

TOTAL A»IHAL: UITH n ALIGN* NT TORORS TOTAI BAIIG»AST TOBORS 

TOTAL AHIBAL S KITH SECONDARY TOBORSt 

TOTAL SECONDAR Y TOBORS 

TOTAL ANIBAL S KITH TUBORS DMCIRTAINEENIGN OR HAIIGNAN T 

TOTAL DNCEETAIN TOMOPS 

TOTAL A1IBALS WITH TOBOBS DNCERTAINPRIHARJ 08 HJTASTATIC 

TOTAI OSCIfTAIN TOBORS 

(35) (32) 

(35) (32) 

2 (6t) 

2 <6») 

35 35 5 7 

8 9 

22 19 

31 20

 

51 52

 

21 18

 

32 35

 

16 12

 

19 17

 

1 

1 

(31) 

1 (3*) 

<3"> 

3 (9%) 

1 (3*) 

35 

9 

21 

f 

25 

« Q 

1C 

15 

;o

29 

e 

13 

* TBIHABY TCHORS: ALL TUBOHS EXCEPT SBC ODD ART TOUORS 

I SFCONDABI TUBORS: DETASTATIC TUflORS OR TDHORS IKTASIVE IliTO AH ADJACIDI ORGAB 

74 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MICE
 

GIVEN INTRAPERITONEAL INJECTIONS OF ACRONYCINE
 

75
 

TABLE 81

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL GROUPS) 

LOWER UPPER 

MID DOSE MID AND HIGH 

uNTREATED DOSE UNTREATCONTROL ED CONTROL 

»»IBALS IHITIAIIY IB STUDY 10 10 »BIBALS NFCBOFEIED 9 10 

AIIBALS EXABIHID HISTOPATBOLOGICALIY 9 10 

IBIEGUBF.IITABY SYS1FH 

• SKIN (9) (10) 

PAPIllCBA, HOS 

•SOBCOT TISSUI m (10) 

SABCCHA, NCS 

RESPIRATORY SYSTEB 

«IOHG m (10) 

HEPATOCEIIUIAF CA8CIBOBA, BETAST 1 (10X) ALVEOLAS/EEONCHIOLAR ADENOBA 1 (10X) 

ALVECIAS/BFCSCHIOIAB CAPCIBOHA 

BIBATOPOIFTIC SYSTEM 

*BOI.riPIF OEGAMS (9) (10) BALIG.IYBPBOBA, DHDIPFER-TYPE 

LYBPHOCYTIC ItUKEBIA 

IIHGUINAL LYBIH NODE (9) (9) 

SARCCBA, «CS, BETASTATIC 

CIRCOLATCBI SYETIB 

NONE 

tIGESTITI SYSTII! 

ItlVEH (9) (10) 

HSPATOCEIIOIA8 ADEROBA 2 (20») 

BEPATOCEHyiAB CABCIHOI|A 1 MU» 2 J1SSL 

I ROBBEB OF ANIHAL S »ITH TISSDE EXANIHED BICBOSCCPICAILY 

* HDHBER OF ANIBAt S NECROPSIED 

77 

LOW DOSE 

UNTREATED 

CONTROL 

20 

20 

20 

(20) 

1 (5%) 

(20) 

2 (10X) 

(20) 

1 (5X) 

2 (10X) 

(20) 

3 (15X) 

(20) 

1 (5«) 

(20) 

« (20X) 

a_nm 

LOWER 

MID DOSE 

VEHICLE 

CONTROL 

10 

10 

in 

(10) 

(10) 

(10) 

(10) 

3 (30*) 

(10) 

(10) 

1 (10*) 

1 (10*1 

UPPER 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

10 

10 

10 

(10) 

(10) 

(10) 

(10) 

(9) 

(10) 

. 
TABLE B1 CONTROL MALE MICE: NEOPLASMS (CONTINUED) 

LOWER UPPER 

MID DOSE MID AND HIGH 

UNTREATED DOSE UNTREATCONTROL ED CONTROL 

CRINAR Y SKSTTH 

NONE 

fNCOCRINF SJSTl-fl 

HONE 

RfPRODDCTIVI S5STIII 

NCNE 

BIRVOUS SKSTf f 

NCNE 

SPBCTAl SFNSF. ORGANS 

HONE 

CCSCDLOSKELETM. EYSTEfl 

•fEHUB (9) (10)

 

CSTECCHCSi:fCBA 

EOD? CAVITISS 

*(IESEllTtRY (9) (10)

 

LIPOBA 

AII rams sys'tins 

_-HQl!£_; „ _„„ ^^ 

» WHBFR OF AHIHAt S HTTH TISSUE FXAHI1IED HICROSCCPICAH.T 

* BOBBER OF AISIBAI S HECROPSIEC 

'78 

LOW DOSE 

UNTREATED 

CONTROL 

(2C) 

1 C5*> 

(20) 

LOWER 

MID DOSE 

VEHICLE 

CONTROL 

HO) 

(10) 

UPPER 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

(10) 

(10) 

1 (10*) 

COTIHOI5 ON 
TABLE B1 CONTROL MALE MICE: NEOPLASMS (CONTINUED) 

1 

Ml 

UN' 

CO 

.OWER 

ID DOSE 

fREATED iNTROL 

UPPER 

MID AND HIGH 

DOSE UNTREATED CONTROL 

LOW DOSE 

UNTREATED 

CONTROL 

LOWER 

MID DOSE 

VEHICLE 

CONTROL 

UPPER 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

AURAL DISPOSITION SOHNABY 

AHIHALS INITIALLY IH STUDY 

HATOBAL DEATH S 

BORIBUBD SACBIWCB 

SCHEtOlID SHCRIFICE 

ACCIDENTALLY KILLED 

TERBINAL SACRIFICE 

AHIBM. HISSING 

1P 

1 

9 

10 

10 

;o 

5 

e 

9 

10 

2 

1 

7 

10 

5 

5 

8 INCLUDIS AOTCLY7ID ANIBALS 

1UBOE SUBBAB Y 

TOTAL ANIKAI S KITH PBIBA8Y TUBOBS* 

TOTAt PBIBJR Y THBORS 

1 

1 

It 5 

13 

17 

5 

5 

1 

1 

TOTAL »»IflAL £ KIT H BFHIGH TDBORS 

TOTAl BFH3C.» TDBOBS 

3 

3 

6 

7 

1 

1 

1 

1 

TOTAL A»Ifl»l £ 1IITH BMIGBAKT TOBORS 

TOTAI (IA11ENAST TDBORS 

1 

1 2 

2 

9 

10 II 

II 

TOTAL ABIBA1 5 KITH SECONDARY

TOTAI SICCKCUB Y THIIORS 

 TUBOBSI 1 

1 

1 

1 

TOTAL ANIBAIJ BITH TDBOBS

EEIIIGN OB BAIIGNAN T 

TOTAL OHCEtTAIN TIMORS 

 ONCtRTAINTOTAL A1IIBMS »ITB TUBOBS OHCIRTAINPBIBAS Y OB BETASTATIC 

TOTAL HBCFFTAI N TOBORS 

* FRTBAR Y TDHCBS: All TnBORS EXCEPT SICOTOA8T TDBORS 

« SECONDARY TOBOBS: HETXSTATIC TUBORS OR TOBOFS I»VASIV! I»TO AN ADJACUI ORGAN 

79 
TABLE 62

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE GIVEN INTRAPERITONEAL 

INJECTIONS OF ACRONYCINE (CONTROL AND TREATED GROUPS) 

LOW DOSE 

VEHICLE LOW DOSE 

CONTROL 

HilHULS JNITI»ilJ III STODY 20 00 KIBALS HISSING 

HilBALS NECJOfEIf C 20 no ANIBUL S EXABTNI D HISTOPATHOLOGICALLY 20 no 

IKTIGUHFNIA M JYEIFB 

NCNE 

ffSPIBATOR Y E1J1EB 

• LUNG (18) (10) ALVEOLAR/EFCHCHIOtAH ADHNOB A 1 <3*> 

HEBATOPOIETIC EtSlEU 

»TOLTIPIE OBG»H S <2P) (It?) BAIIGKANT 1YBPHOBA". HOS 2 (10*) RALTG.LYHPfcOBA , DMDIFFE8-TYP? 5 (251) 6 (15*) 

HALIG.tYdPHOIIA , IYBPHOCYTIC TYPE 6 (SOU) 3 (8<) 

ISPLEEH (19) (3P) 

HSnANGIOSVGCOn A 1 (3*1 

ITHYBUS (5) (7) BALIG.LYBPHCBA , LYHPHOCYTIC TYPF 1 (1HX) 

CIRCDLATCRY SYSTEf 

NONE 

tTGESTIVt SYSlfB 

«BILE COCT (20) <«0) BILE DUCT CARCTNOBA 

CRINA8 Y SYSlt P 

(ONE 

I NHBBER OF ANIKAL S KITH TISSUE EIAIIINED BICHOSCCPICAIIT 

• NUBBER OF ANIMAL S HFCROPSIEC 

LOWER 

MID DOSE 

35 

35 

35 

(35) 

(35) 

(35) 

(15) 

(35) 

UPPER 

MID DOSE 

35 

1 

3« 

33 

(33) 

(31) 

(30) 

(2) 

(31) 

1 <3X) 

~~ ^! 

HIGH DOSE 

35 

2<» 

12 

(12) 

(2")) 

(12) 

m 

(29) 

80 
TABLE B2 CONTROL & TREATED MALE MICE: NEOPLASMS (CONTINUED) 

LOW DOSE 

VEHICLE 

CONTROL 

LOW DOSE LOWER 

MID DOSE 

UPPER 

MID DOSE HIGH DOSE 

IHCOCHINE SJSTin 

tUDREBA L 

PHEOCHRCHCCYTCB A 

ITHYBOIC 

FOLlICBLAB-CIlt CABCINOPA 

(20) 

(18) 

(37) 

1

(38) 

1

 (3*) 

 (3*) 

(32 ) 

(If ) 

(29) 

(17) 

(11) 

(7) 

REPRODUCTIVE SYSTFB 

ITESTIS 

HBBANGIOSJBCCB A 

(20) (38) 

1 (3*) <3t) (31) (12) 

KEHVOU S SYSTIR 

NONE 

SPECIAL SENS! CFGAN S 

NONE 

(OSCULOSKEIETAI SYSTEB 

•KNEE JCINT 

OSTEOCHCHCfOB A 

(20) (10) 

1 (3*) 

(35) (31) (2S) 

BODY CAVITIES 

NONE 

« NBBBER OP ANIBAL S HITH TISSUE EXABINE D BICRCSCCPICALLY 

« NDBBER OF A1IIBAIS MKROPSIE D 

All OTHm SYSTIBS 

•BUITIPIE ORGAN S 

._ SJFCCBi^ NCJ, 

(20) (10) 

2-lSil 

(35) (31) (29) 

CCNIINOID 

81 
TABLE B2 CONTROL & TREATED MALE MICE: NEOPLASMS (CONTINUED) 

* PBIHAE Y T1JHOBS: »H IDHOBS EICEPT SECONDARY TUMORS 

I SECOHD/lBY TOtOBS: METHSTATIC TDMORS OB TDBOFS I»V»SIVE INTO UN »DJ»Cf»T ORG»S 

82 
TABLE B3

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE GIVEN INTRAPERITONEAL 

INJECTIONS OF ACRONYCINE (CONTROL GROUPS) 

LO WER MID DOSE UPPER MID AND 

UNTREATED HIGH DOSE 

CONTROL UNTREATED CONTROL 

miBULS INITIAItY IN STUDY in 10 >1!1 1! JUS NECBOfSISD 10 10 HIIHULS EIABINID HISTOPATKOIOGICALLY 10 10 

III1EGUBEHTABY SYS1EB 

• SDBCDT TISSOI (10) (10) 

SARCOBA, NCS 

HEBANGIOB H 

BISPMATCRY SYSTIH 

NO KB 

FIBATOPOIETIC SYSTEB 

•BOITIPI* OBGAHS (10) (10) BALIG.LYBPtOHA. DSDIPPER-TYFF 

BAIIG. IYBIBOBA, IYBPBOCYTIC TYPI 

BALI6. IYBPHCBA, HISTIOCYTIC TYPI 

IYHPHOCYTIC IEOKEBIA 3 (10») 

ICDODKHOH (10> (10) 

HALIG.LYHPBOBA, ONDIPFEF-TYPE 

CISCOIATCRY SYSTP.F 

NONE 

tIGESTIVI SYSTIB 

ILIVEB (10) (10) HBPHTOCf IIDLAR ADEROBK 

ORINAB Y SYSTEK 

BONE 

t millBEB OF ANIBALS SITH TISSUE EXAHINED HICBOSCCPICAILY 

* NURBER OF AIIIHAIS BFCBOPSIEC 

LOW DOSE 

UNTREATED 

CONTROL 

;o 

19 

19 

(19) 

1 <5«) 2 (111) 

|19) 

3 [16*) 1 (5«) 

1 «5«) 

<19> 

1 (51) 

(19) 

1 (5X) 

LOWER MID 

DOSE VEHICLE 

CONTROL 

10 

10 

10 

(10) 

(10) 

1 (10*) 

(10) 

(10) 

UPPER MID AND 

HIGH DOSE 

VEHICLE CONTROL 

10 

10 

10 

(10) 

(10) 

1 (10*) 

(10) 

(10) 

83 
TABLE B3 CONTROL FEMALE MICE: NEOPLASMS (CONTINUED) 

LOWER MID DOSE UPPER MID AND 

UNTREATED ! HIGH DOSE 

CONTROL UNTREATED CONTROL 

INCOCBINE SYSTEB 

• PITUITARY (7) (9)

 

CHPOBOPHOEE JCEBOHA 

UNPRODUCTIVE SYSTE M 

HCTEBUS (10)

 

HEBANGIOSABCCBA m1 pit) 

BERVOUS SYSTFt 

NONE 

SfECIAl SENSE CEGANS 

*HARDERIA!< GIANC (10) (10) 

PAPILLARY CYSTADENOBA, DOS 

HISCOLOSKILETAI SYSTEn 

NONE 

EOTY CAVITIES 

NONE 

All OTHES SYSTEBS 

*BHITIP1E OBGANE (10) (10) 

. LEElSSifSCBi- - — 

* NUBBEB OF ANIBAL S »ITH TISSOE EIABINED BICSOSCCPICAltY 

* N0BBER OF AKIBAI S NECROPSIIE 

LOW DOSE 

UNTREATED 

CONTROL 

1")

3 (ia«) 

(19) 

(19) 

1 (5X) 

(19) 

J_J5Si_. 

LOWER MID , 

DOSE VEHICLE 

CONTROL 

(8) 

(11) 

UPPER MID AND 

HIGH DOSE 

VEHICLE CONTROL 

(9) 

(9) 

(10) (10) 

(10) 

_

(10) 

 ,»__ ....^-..^ 

84 
TABLE B3 CONTROL FEMALE MICE: NEOPLASMS (CONTINUED) 

LOW 

ur 

c 

ER MID DOSE 

JTREATED 

JONTROL 

UPPER MID AND 

HIGH DOSE 

UNTREATED CONTROL 

ANIHAL DISECSniON SDBMARY 

ANIMALS IHITIALtY IS STUDY 

NATURAL CIJTB? 

nORIBUND SACRlFICF 

SCHEtOLIC SACEIFICE 

ACCIDENTALLY KILLED 

TEEHIHAL SACRIFICE 

ANIHAL HIEEING 

10 

10 

10 

1 

9 

1 INCLUDES AOTCLY!ED ANIBALS 

1011OE SDBBAE1 

TOTAL ANIBALS KITH PFIflAfiT TOBOES* 

TOTAL PBIBAEY TUBOBS 

3 

3 

1 

1 

TOTAL ANIBAL: BITH BENIGS TDBOKS 

TOTAl BINI6N TOBOBS 

TOTAL ANIBALE KITH HALIGNAMT THBOBS 

TOTAI BAUGNAST TOBOBS 

3 

3 

1 

1 

TOTAL ANIMAIS NITH SECONDABY TOBOBS* 

TOTAL SFCOKDABY TnHORS 

TOTAL ANIBALS KITH TOBOBS ONCEBTAINEENIGN OB BAIIGNANT 

TOTAL ONCEBTAIN TOBOBS 

TOTAL ANIKAIS »ITH TOHORS ONCERTAINEBIBABY OP HETASTATIC 

LOW DOSE 

UNTREATED 

CONTROL 

;o 

6 

a 

10 

13 

15 

7 

7 

e 

3 

LOWER MID 

DOSE VEHICLE 

CONTROL 

10 

1 

1 

8 

1 

1 

1 

1 

UPPER MID AND 

HIGH DOSE 

VEHICLE CONTROL 

10 

1 

9 

1 

1 

1 

1 

EBIBAB Y TDBCBS: ALL TnBOES EXCEPT SECONDARY TUBOSS 

SECONDARY TUBOBS : HETASTATIC TUHOfiS OB TOBOBS INVASIV E INTO AN ADJACIM ORGA N 

85
 
TABLE B4

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE GIVEN INTRAPERITONEAL 

INJECTIONS OF ACRONYCINE (CONTROL AND TREATED GROUPS) 

LOW DOSE LOW DOSE 

VEHICLE ' 

CONTROL 

»SIBALS 1NIITIII1 IH STUDY 20 <l(! 

JKIMAL S NFCBCfSTFB 20 39 

HIIHALS FXABINI D HISTOPATHOLOGICA 1LY 20 39 

JNTTGUBENTAf Y SYSTE M 

KONE 

65SPIRATCR Y SYSTtr 

«tON G (17) (38) 

ALVSOLAR/EBCIICHTOLA B CABCINCB A 1 <3t) 

tEHATOPCIETK: SYSTE H 

•BOLTIPIl! CBGAN S (20) (39) 

MALIGNAM T lYrFHOBA . NO S 1 (20*) 

BALIG.LYBFHCB A, U!JDIFF?P-TYPF 13 (65«) 1 (3*) BALI!5.LYflf HOBA , LYBPHOCYTIC TYPE 2 (K*) <l (1C*) 

BALIG.LYHEHOBA , HISTIOCYTIC TYPE 1 (3*) 

IBONE BABBO S (18) (39) 

GBANDLO C YT1C IEOKTBIA 

• THYBO S m ( I D ALVEOLAB/tBCIICHIOLA B CA, BETASTA 1 (9?) 

CIBCOLATCBY SYSTIB 

NON E 

I1GFSTIVE SYSTJ B 

IlIVEP (20) (39) 

HEPATOC1IIDIA B CABCISOBA 1 (3*) 

•EILE COCT (2P) (39) 

^DEfUqABCJUIIA . DOS 

« NDBBEB OF ANIMAL S WITH TISSUE EXAMINE D BICBCSCCPICAILY 

* NUMBER OP ABIBAI S NFCROPSIIt 

LOWER 

MID DOSE 

3K 

33

i -3 

(33) 

(33) 

(-'I) 

(5;> 

CD 

(33) 

UPPER 

MID DOSE 

35 

33 

33 

(33) 

(33) 

(32) 

1 (3«) 

(7) 

(32) 

(33) 

i_U21— 

HIGH DOSE 

35 

32 

15 

('*) 

(32) 

(11) 

(1) 

(IS) 

(32) _ 

86 
TABLE 84 CONTROL & TREATED FEMALE MICE: NEOPLASMS (CONTINUED) 

"TM~ ~ ' T ' ' ' " " TM TM~ ' Tin - - r -i 

tCDODEIQH 

ADEICHATOBS POLIP. (OS 

OB1NABY STSTEB 

»O»B 

IICOCBINI SIS1IH 

DO HE 

EIPBODOCTITt SYSTIB 

*H»flfl»RY GH«t 

»DE»OCAEClNCa» . NOS 

IISVODS STSTIP 

NONE 

SPICIAI SfKSf CEGAK S 

HOVE 

BDSCD10SKEIITAI SISTEB 

NONE 

ECEY CAVITIIS

 

NONE 

HL OTHIF SYSTIBS 

*B01TIPtE OEGHN S 

SABCSHA, K r 

I HUBBEB OF ANIBALS HITH TISSDB

» NOBBEB OP AHIBA1S NECBOPSIID 

LOW DOSE LOW DOSE LOWER UPPER HIGH DOSE 

VEHICLE MID DOSE MID DOSE 

CONTROL 

(12) (39) 

1 (3K) 

C6) (32) («) 

(20) (39) (33) (33) (32) 1 (3*1 

(20) (39) 133) (33) (32) _______ _ .. 

„ . ,.„,.., , i_J551__. ... . i 

 EXAMINED BICBOSCCPICALLT 

87 
TABLE B4 CONTROL & TREATED FEMALE MICE: NEOPLASMS (CONTINUED) 

LOW DOSE 

VEHICLE 

CONTROL 

LOW DOSE LOWER 

MID DOSE 

UPPER 

MID DOSE 

HIGH DOSE 

AHHA L DISfCEniCN SOBBAR Y 

ANIMAL S INITIALL Y IN STUDY 

NATUFA L CUTHa 

BORTEUN D SACRIFICE 

SCHECULf C SACRIFICE 

ACCIDENTALL Y KILLED 

TERMINA L SACRIFICE 

ANIBA L BISETN G 

20 

1U 

6 

140 

7 

31 

2 

35 

14 

21 

35 

12 

23 

35 

29 

6 

8 INCLUDES AUTCLYZE D ANIHAL S 

10BOE SDHHAR 1 

TOTAL ANIBAI S »ITH PRIBAR Y

TOTAL PHII1JR Y TO nOBS 

 TUBORS* 19 

19 

9 

12 

2 

2 

TOTAL AHinAL S WIT H BFNIGN

TOTAL BENIG B TIJHOHS 

 TUHOBS 1 

1 

TOTAL AHIIIAL S HITH BALIGHAN T TOBORS 

TOTAI BAIIGNAH T TUKORS 

19 

19 

9 

11 

2 

2 

TOTAL ANIBAL E HITH SECONDAR Y TOBORS' 

TOTAL STCCHEAB Y THBORS 

tt 1 

1 

TOTAL ANIBAL S WIT H TUBORS UNCERTAI N 

BENIGN OR HAIIGNAN T 

TOTAL UNCEFTAI N TIMORS 

_ 

TOTAL ANIRAI S HITH TOBORS UNCERTAI N 

ERIBAR Y OB BETASTATIC 

TOTAL UNCEETAI H TUBOPS 

* ERIIHRY TUMCRS : ALL TOBOES EXCEPT SECONDART TUHOHS 

I SECONDftBY TUnORS : HETftSTATIC TOHORS OR TOHOBS INVASIV E INTO AN ADJACIK1 ORGA 

88 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS GIVEN INTRAPERITONEAL INJECTIONS
 

OF ACRONYCINE
 

89
 

TABLE C1 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL GROUPS)

 

LOW DOSE 

UNTREATED 

CONTROL 

MID AND HIGH DOSE 

UNTREATED CONTROL 

LOW DOSE 

VEHICLE 

CONTROL 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

ANIBAIS INITIAUY IN STDDT 

AKIBAL S NEC60ISIED 

AHIBAL S ZJABINJ D HISTOPATHOLOGICALLY 

10 

10 

10 

10 

9 

9 

10 

10 

10 

10 

8 

S 

INTEGUBENTAB Y SISTEfl 

HONE 

FISPIRATOBY SYS1EB 

*TRACHVA INPLABBATICII,

INPLABBATICN,

 CHRONIC 

 CHBOHIC SBPFURATIV 

«9) (9)

2 (22*) 

(8) (8) 

1

1

 (13*) 

 (13*) 

tlDNG/BBONCHIOL! 

HYPEBPLASIA , LYBPHOID 

(10) (9) 

3 (33*) 

(1C) (8) 

1 (13X) 

• LONG 

BBOKCaOPNIUNCKIA, NOS 

BBOHCHOENKaBORIA SUPPORATIVf 

PHEnnONIA , CHRONIC BOBIHB 

(10) 

5 (50*1 I 

(9) 

1 (11*) 

(10) 

7 (70*) 

(8) 

1

2

 (13X) 

 (25X) 

KEBATOPOIETIC SYSTfH 

•BONE BABBO H 

ATBOPBY, »CS 

HYPEFPIASIA, NOS 

(9)

1 (11%) i (9) 4 (41*) 

(10) 

2 (20*) 

(8) 

2

1

 (25X) 

 (13*) 

IBARDIBOLA B L. NODE 

HYPEBPLASIA. PLASBA CELI (7) (9) («) 

1 (17*) 

(7) 

CIBCOLATCBY SYSIEB 

IIMDOCABOIDB 

PI BROS IS, FOCAL 

(9) (9) (10) 

1 (10*) 

(6) 

• CEI.IAC A8TIB1 

— QGSEIEBillCB. K95 

(10) (9)

1 M1X.1 <10) (8) 

i NUBBBR

* RDKBER

 OF ARIBALS HITB TISSUE EIAHINED

 OF ANIHAIS lECROPSIID 

 MICROSCOPICALLY 

91 
TABLE C1 CONTROL MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

VEHICLE 

CONTROL 

(10) 

(10) 

7 (70*) 

(10) 

(10) 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

(6) 

(8) 

1 (13*) 

1 (13*) 

2 (25*) 

(8) 

(8) 

LOW DOSE 

UNTREATED 

CONTROL 

EIGESIIVt SJISTIB 

ISBALI. INTIiSTINF (8)

 

PEHHHT1IRITIS 

UPIHARY SYSTEfl 

• KIDNE Y (9)

 CALCOLIJS , SOS

 

IHFLAHIUTION, INTERSTITIAL 

I»FL»HH»TICH, CHBOKIC 3 (33 * ) 3 (33*) 

MID AND 

HIGH DOSE UNTREATED CONTROL 

m 

1 (11*) 

(»)

2 (22X) 

IKIDNEY/TUIIOI! 

BIIIER»L15r»TIOII (9) 

ESDOC8IHE

NODE 

 SYSTIB 

RtPRODDCTIV! SYSTIH 

tPBOSTUTE 

HFLABHJTICS , SDPPU8ATIVS (9) 

SEFVOUS

KOKE 

 SYSTEfl 

SPICIAL

NODE 

 SEHSF CEGAH S 

BOSCOIOSKItlTAI SYSTEB 

HONE 

ECDY CAVITItS 

sons . _ . . 

(9)

1 (11*) 

(7) 

2 (29*) 

« MOHBER OP AHIBALS KITH TISSOE EKAHISED BICROSCCPICA ttY 

* HUBBER OP AKIBAIS NECROPSIID 

92 
TABLE C1 CONTROL MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

• 

LOW DOSE MID AND LOW DOSE MID AND HIGH 

UNTREATED HIGH DOSE VEHICLE DOSE VEHICLE 

CONTROL UNTREATED CONTROL CONTROL CONTROL 

DLL OTH8B STSTIHS 

HOSE 

SP1CIAL HOBEHCIOGY SBHHUHY 

»0 IISION ItrOFTID 3 1 1 1 

JUTOLYSIS/HO SECROPSY 1 2 

I NOHBER OF AHIBAI S KITH TISSUE EXAHIKSD HICSOSCCPICAI.LY 

* ROBBER OF »N1B»I3 NECBOPSIED 

93 
TABLE C2 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (TREATED GROUPS) 

•- 1 

LOW DOSE MIDDOS.E HIGH DOSE 

ABIHAL S ISniULY IN STODT 35 35 35 ASTHALS «ECfOfSIIC 31 32 34 AHIBAL S IXABIHI D HISTOPATHCLOGICA IIY 30 31 3« 

INTfGUBSHTABY SYSTEH 

*SOBCOT TISSOI (31) (32) |3«) EPIDIRBA L INCIUSION CYST 1 O*> HEBORRHAG F 1 (3X) IHFtABBATICH , BECROTIZING 1 <3») IHFIABHATICH, POCAI GR AHDIOBATOO i n») 

RESPTBATCRY SYSTEB 

•TBACHEA (29) (31) (31) IBF1AMBATICN, SOPPDRATIVE 2 (7») 1 (3t) 2 (6*) INFIABBATICH , ACOTF/CHROHIC 2 (6«) » (13%) 

IN?IABMATICI(, CH80SIC SUPPORATIV 1 (3%) 

tlDNG/BROBCHOS (30) (31) (31) IHFIABBATICH , »OS 1 (3*) 

•LUNG (30) (31) O«) HEBOBDIIACE

 2 (6*) BBONCHCFKEDPCBH , DOS

 1 <3X) 

INFtABBATICll, INTERSTITIAL 1 (31) BHONCHCESEOBCNH SDPPDRATIVE 1 (3») 3 (10*) 1 (3X) PHEOBOHIA, CHBOMIC BBRIHE 8 (27*) ^ > 

HYP^FPIASIA, ALVTOLAR EPITHEtlOB 1 (3t) 

HEBATOPOIETIC SYSTEM 

IBON E BARRC H (28) (29) (32) ATROPHY , NCS 6 (21*) 8 (28*) 19 (59X) 

ISPLEEH (27) (31) (33) FIBROSIS

 1 (3*) 

PE|)ATOFCIE£IS 3 (11*1 w » __ —— — -.^i — nr» 1 

• HUHBER OF ANIBALS WITH TISSUE EXAHIIED BICROSCCPICAltY 

« SUBBER OF ABIBAI S NECROPSIIC 

94 
TABLE C2 TREATED MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID DOSE 

• BtDIASTINAL I. SODS (30) (20) 

HEBORRHAG I 

• PANCREATIC I. NOD E (30) (20) RYPF.BPLASIJ, LYBPHOID 1 (3*)

 

IHESENTSRIC I. NODE (30) (20) CONGESTION, NCS 1 (5K) HEBOEBHAG K 1 (5*) HYPFEPLASIA , LYNPHOI D 1 (3») 

CIRCOLATCHY SYSTIP 

IflYOCAREIOB (29) (30)

 BEBORRHAG t 1 (3X)

 INFtABBATICN , INTERSTITIAL 1 (3*)

 INFLABBATICN, CHRONIC POCAt 1 (3»)

 OFLABHATION , CHRONIC SDPPORATIV 1 <3X )

 

• PnLBOKAR Y ARTER Y (31) (32)

 AHTERIOSCLIROSIS, NOS 1 (3K)

 

DIGESTIVE 3YSTFB 

niVEH (29) (31) HEBORRHAG F 1 (3X) 2 (6X) INFLABHATIOH, NECROTIZING 1 <3K) INPIABHATION , CHRONIC 

IBFIABBATICN, CHRONIC NEC80TIZIN 1 (3*) FIB80SIS 1 (3%) SECRCSIS, KOS 

HEC80SIS, COAGOLATIJE 1 (UK) CYTOtOGIC CEGINERATIOH 1 (3*) HYPEBPLASIA , NODDLAR 5 (17«) HYPERPIASIA, LYBPHOID 1 (3K) 

•LIVER/PERIPOPTAI, (29) (31)

 

FIBROSIS, IIFTOSE 

*EILE DUCT (31) (32)

 FIBRCSTS, IOCAI 1 (3*)

 

HYPERPLASIA , SOS 2 («*)

 

(PANCREAS (21) (26)

 

IMfLftinATKh , INTEBSTITIAI, 

« BOBBER OF ANIBALS WITH TISSUE EXABINED BICHOSCCPICALtY 

» NUBBF.R OF .ABIBA1S SECHOPSIID 

HIGH DOSE 

CD 

1 (5*1 

(21) 

(21) 

(32) 

<|3t) 

|3«) 

1 (3t) 

1 (3*) 

1 (3X) 

1 (3%) 

(31) 

1 (3X) 

(31) 

<3C> 

1 (JS1 

95 
TABLE C2 TREATED MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID DOSE 

FIBRCSI;-

HYPEFPLflSTft , BO:; 

HETAPLASIH , OSSEOUS 1 (IK) 

• COLON <2U ) (29)

 

INPLAHMATICN , HEBORRHAGI C 

•CECUB (21) (29)

 

HEFinRRHAG f 

URINAR Y SYSTPB 

IKIDNFY (29) (31)

 MFLAMHATICN , CHRONIC 7 (214*)

 

IURINAR Y BLATTE R (21) (29) 

FIBHOSI 5 

IBDOC8INE STSTf M 

lADREHA t CUtTI X (28) (31) 

HY:PEBPL»SIJ, noon us 

ITHYBOTC (26) (26)

 

CYSTIC I'OItlCtFS 1 (UK ) 

REPRODDCTIVl: SYSTF M 

IPROSTATI: (26) (31) 

IKPLAdmiTICB. SHPPURATIT E 1 (3*) 

tERTOU S STSTEf 

NONE 

SPECIA L SEN S E CPGAH S 

NONE 

POSCOLOSKEIITAI SYSTEM 

'BONE (3') (32)

 

OSTKOPF1POEI3 

-Jl£Ef£lJS51J!*_JIS!S,. „ _„

 

» NUflB^R OF ANIHJU. S KITH TISSOE ESUHINED HICROSCCPICALtT 

» NDBBFR OF AKIBAI S NECBOPSIEC 

HIGH DOSE 

1 (3*) 

1 (3«) 

lie)

1 (1*1 

(28) 

1 («*) 

(33) 

(33) 

1 (3*) 

(33) 

2 (6X) 

<;:) 

(32) 

(31) 

1 (3X > 

-. 1 .(3.S1 

96 
TABLE C2 TREATED MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE MID DOSE 

tODT CATTMIS 

•PEFITOHEOri (31) (32) IHFLAIIflimOII, SOPPURATIVE 

ABSCISS. ICS 

INFLAIVBATICII. CHBOIIIC 1 («) 

INPLAHHATI01I. CHRONIC

FIBROSIS 

 DISPOSE 1 <3X) 

1 (3X) 

ADHBSIOII, SOS 1 (3X) HETAPLASIK, OSSEOUS 1 OX) 1 (3») 

HI OTHER SrSTIBS 

*BUITIPIE OFGANS (31) (32) FIBROSIE 1 O«) 

SPECIAL CORfHCIOGT SUMHABT 

HO LFSIOII FEfORTED l| 

NO Hf CROPS T P18PORBED 1 

KOTC/RKCRCESI/HO HISTO 1 1 

AUTOltSIS/JC HK8OPST 3 3 

I SDHBEB CF HHIUIS KITH TISSOI M*BINED B ICFOSCOPICIlt,LT 

» NOBBEP Of XIRAIS KKCROPSIFD 

HIGH DOSE 

(3») 

1 (3X) 

1 (3«) 

S (15«) 

2 (6X) 

131) 

1 

1 

97 
TABLE C3

 

SIUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

GIVEN INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL GROUPS)

 

— —— LOW DOSE MID AND HIGH 

UNTREATED DOSE UNTREATCONTROL ED CONTROL 

nna»LS INITIAII Y IN STITDY 10 10 AlilBALS BECECISIFD 10 9 fNIBAL S EXABTNI D BISTOPATHOlOGICAllY 10 8 

IBTEGUHENTAF Y SYS1H1 

*SHBCOT TISSDt (10) (9)

 imaAKHATTCH, CHROHIC JCCAL

 

RESPTBATCR Y SYS1FP 

•TRACHEA (10) (8) 

IBELABHATICII, HOS 1 <1«) mnAniiATioM, ACUTE/CHROHIC 2 (25*) 

• LtJHG/BFONCHICIH (10) (7)

 HYPIRPLASIA , IYHPHOID

 

• LOSG (10) (7) mrLABHATICll, INTEPSTITIAL 1 (1«*> P»!nBO»IA, CHROHIC HORIN E 1 <10») 

FEBATOPOIETIC SYSTEH 

IBCNE HARF.CW (10) (8) 

ATROPHY . NCS 5 («3*) 

ISPLEEK (10) (8)

 HEnATOtOIISIS 1 (10*)

 

CIRCDLATCR Y SYSTEB 

MORE 

tiGisnvr SYSTIB 

•HIPATIC CAPEOII (10) (8) 

SBCEB5I5. tOASHUTIXB n i ..«.«. i • ( 

I NUMBER OT ANlnAt S IITH TI5SOE EIAKIRED BICROSCCPICALLT 

» BOMBfR OP ASIBAt S HECBOPSIIt 

LOW DOSE VEHICLE CONTROL 

10 

10 

10 

(10) 

1 (10*) 

(10) 

(10) 

(1C) 

1 (10*) 

«») 

(1C) 

(9)

J..U..1SL. 

MID AND HIGH 

DOSE VEHICLE CONTROL

 

10 

9 

9 

P) 

(9) 

(•»)

1 (23*) 

(9) 

m 

H («4t) 

(9) 

(9) 

n r .i* . V_M.I_... .,UI, ,. . 

98 
TABLE C3 CONTROL FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID AND HIGH LOW DOSE 

UNTREATED DOSE UNTREAT- VEHICLE 

CONTROL ED CONTROL CONTROL 

BBINAR Y STSTIB 

IKIDKFT (10) (8) (1C) CALC0105, NOS 3 (38*) I BFIAB NATION, CHBOSIC 2 (20*) 4 (DO*) 

ENDOCRIN? STSTIM 

ROUE 

BIPRODDCTIVI SYSTEB 

*BABHAR T GLAK t (10) <9) (10)

 CYST, HOS 1 (11*)

 

lOTEROS/EHCOBfTRIUn (10) (8) (10) INFLANHATICH, SUPPDRATIVE 3 (30*) 3 (38*) 2 (20«) 

IHrlABHATION. CHRONIC SOPPORHTIV 1 (10X) 

KRVOOS SISTBH 

NONE 

SPICIA1 SENS? CB6ANS 

NONE 

BOSCOLOSPE1ITAI SYSTEM 

HONE 

BODY CATJTIIS 

*PERITOIIEnB (10) (9) (10)

 IHFIAIIBATICS, CHRONIC SDPPORATIV 1 (10X)

 

All OTHEB SISTIBS 

DONE 

I WOBBER Of AHIBA15 WITH TISSOE EIAHIVBD HICROSCCPICALLT 

* NDHBER OF AKIBAIS HECROPSItD 

MID AND HIGH DOSE VEHICLE

 

CONTROL 

W 

1 (11*) 

(9) 

(9) 

2 (22*) 

(9) 

99 
TABLE C 3 CONTROL FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID AND HIGH LOW DOSE MID AND HIGH 

UNTREATED DOSE UNTREAT- VEHICLE DOSE VEHICLE 

CONTROL ED CONTROL CONTROL CONTROL 

SPECIAL MOeiHClOGY SOBflAR Y 

NO LKICN FEFCBTED 2 2 2 

NECROPSY FIPF/NO HISTO PE8FORHED 1 

AUTOIY3IS/KC HKP.OPSY 1 1 

• SUBBEF CF AMKA1 S WITH TISSUE !X»HINED HICBOSCOPICIUU 

* NONBEB OF ANIfUI S NECROPSIED 

100 
TABLE C4 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (TREATED GROUPS) 

LOW DOSE MID DOSE HIGH DOSE 

ANIBAL S INITIUL Y IN STDDT 35 35 35 

ABIBAL S mcsossitt 35 32 31 

AHIBAL S BXANINI D HISTOPATHOIOGICA ILY 35 32 33 

IHTFGUBESTAR Y SYSTEB 

»SKI N (35) (32) (3U) 

OLCER, CHFCNI C 1 (31) 

RESPIRATOR Y SYETE B 

•TRACHEA 

INFLABBATICN ,

ISFLABBATIOH,

INPLABBATICN ,

 NOS 

 ACOTTVCHBOS 1C 

 CHBOHIC 

(35) (30) 

1 <3«> 

2 (7*) 

(32) 

1

1 <3X) 

 (3*) 

(LUNG/BPONCHC5 

BROHCHIICTASIS 

INFLAHBATICB , NO S 

(35) (32) 

1 (3*) 1 (3*) 

(33) 

• LUN G 

HEBOPEHAG f 

INFLABBATICS , INTERSTITIAL 

BRO»CHCENFOBCNI A SHPPURATIV H 

PNEnrCNIA , CHFONIC BURIN E 

INFLABBATICN , CHRONIC 

(35) 

5 <1H» ) 

(32) 

2 (6%) 2 (6*) 

1 (3%) 

(33) 

1

1

 (3*) 

 (3*) 

EEBATOPCIET1C SYSTEB 

«BCNE BARSC V (35) (32) UD 

ATROPHY , NCS 3 (9«) 7 (22t) 10 (32*) 

ISPLE'N (35) (31) (21) HEBATOPOIfSI S 10 (29K) 1 (3K) 

IAXILLAR Y IYBP H NODE (35) (12) (2S) HYPERPLASIA , PLASBA CELL 1 (3T) 

CISCDLATCRY SYSTF P 

E9JI£ _. 

t NUBBER OF ABIBAL S HITH TISSUE F.XABINED BICBOSCCPICA1LY 

* NUMBE R OF AMflMS NFCBOPSirC 

101 
TABLE C4 TREATED FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)

 

EIGFSTIV I SYSTf B 

ULIVEF 

BEBOR?H,»G1C CYST 

IllFLA1fl»TICM, CHROMIC 

"IBRC3IS, ElfFUSF. 

NECRC5IS. KOS 

NECBC3IS, CCAGOLATIV E 

HYPF.BPLASIA , MODULAR 

•LIVEB/CENTRIIOEULA R 

NECROSIS, COKGULATIVF. 

*EILE DUCT 

CYST, NOS 

HYPEBPLASIA , NOS 

HYPFFPLASIA . CYSTIC 

«PAHCEE» S 

?IBR05IS 

•STOHACH 

FIBROSIS 

OBIHAH Y SYSTPB 

IKIDHF Y 

CALCDLOS , »OS 

HYDBCHSPHFOEI 5 

INn.»nBATIOH, SOPPUBATI?B 

IBFIADBATICII, CHBOKIC 

IKIDNEY/TOBOI! 

NEPHROSIS , NOS 

EKDOCHTH11 SYSTIfl 

lADRENA I 

IHPLJBBATIC1I, CHROMIC 

AKGIICTASIS 

lADRENA I COSIfX 

HYPEBPl»SI» . BODOIAR 

I NtlBBEB OF AMIHAL S WITH TISSUE

» NUBBER OF AHIBAI S MECROPSIED 

LOW DOSE MID DOSE 

(35) (32) 

1 (3%) 

1 (3*) 

1 (3X> 

1 (3X) 

1 (3*> 1 (3X) 

05) (32 ) 

1 (3») 

(35) (32) 

1 (3*1 

1 (3X) 

1 (3*1 

(34) (30) 

1 (3%) 

(35) (30) 

1 (3*) 

(35) (31)

1 (3*) 

1 (3*) 

3 (9*1 1 (3%) 

(35) (31) 

(32) (30) 

1 (3«) 

1 (3*1 2 (7K) 

(32) (30) 

 EXHBIM8D BICRCSCCPICALIY 

HIGH DOSE 

(33) 

1 (3K) 

3 (9%) 

(33 ) 

(31) 

1 (3*) 1 <3X ) 

(31) 

(32 ) 

(32 ) 

1 (1*) 

(32 ) 

1 (3K) 

(31) 

(3D 

_ J-J3.H 

102 
TABLE C4 TREATED FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID DOSE 

REPROD0CTTV E SiSTEB 

• BAHtiAR Y GLAB T (35) (32) IHFIASIBATICN, NKCROTIZING 1 (3X) HYPEBPLASIA , CYSTIC

 1 <3X) 

• VAGIN A (35) (32)

 TNFtABBATICN , SUPPURATIV E

 

• DTERUS/ENCOniTRIU B (3H) (32) INFLAHBATICN , SUPPOBATIVF 6 (18*) 1 (3X) HYPEfPLASIA , CYSTIC 2 (6K) 

•OVAHY/OVItUCT (31) (32)

 HEflOBBBAG F 1 (3*)

 

«OVAB Y (31) (31)

 IHFLABnATICN , SOPPIIBATIVE 1 <3X )

 ISFLABBATICN , CHRONIC

 1

 0*) 

^BRVOt;S SYSTEP 

HONE 

SPICIAl SENSE C5GAN S 

HONE 

CDSCOLOSKELETAI SYSTEfi 

NONE 

ECCY CAVITIES 

'PERITONEUM (35) (32) 

INFtAHHATICN . CHRONIC 2 (6K) 

PIBBCSIS 1 (3X) 

Alt OTHEB SYSTIBS 

DIAPHRAG B 

UEt»flflllISH^-£JilQja£ 3 , , .

I HDHBER OF ANIBALS »ITH TISSOE EXABINED HICSOSCOPICA1LY
 

* NOHBER OF AHIHAIS NECROPSTID
 

HIGH DOSE 

«31) 

1 OX) 

|3«)

1 (3X) 

«32) 

«3J) 

(31) 

(31)

2 (6*) 1

 (3«) 

__, 

103
 
TABLE C4 TREATED FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE MID DOSE HIGH HOSE 

ACIPOSI TISSU E 

INFLAflflATICN , CHROHIC FOCAL 1

 

SPICIAl PORPfiOIOGY SOHHAR T 

DO tISICK HECBTFE 1 1 1

 

AOTO/!(?C!10PST/IIO HISTO 1

 

AUT01YSIS/KC BECBOPSY 1 1

 

I BOBBER C* AUfAt S WITH TISSU! FJMIINED HICBOSCOPICA1LY 

* BOBBER OP mn»IS HECHOPSIED 

104 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE GIVEN INTRAPERITONEAL INJECTIONS
 

OF ACRONYCINE
 

105
 

TABLE D1

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL GROUPS) 

I 

LOWER MID DOSE INTREATED CONTROL 

UPPER 

MID AND HIGH DOSE UNTREAT ED CONTROL 

LOW DOSE UNTREATED CONTROL LOWER MID DOSE VEHICLE CONTROL 

UPPER MID AND HIGH 

DOSE VEHICLE CONTROL 

KIBALS INITIAIIY III STUDY 

AKIBAL S NECECSSIED 

HHTBALS IXABIH1 D HISTOPATHOlOGICAItY 

10 

9 

9 

10 

10 

10 

:c 

20 

;o 

10 

10 

10 

10 

10 

10 

JNTEGUBEBTAK Y SYSTEB 

»SKI8 

W.CEP, FOCA L 

FIBROSIS 

FIBRCSIS, IOCAI 

ACARIASI S 

19) (10) (20) 

1

1

2

1

 (5») 

 (S«) 

 (10*) 

 (5*) 

(10) (10) 

*SDBCOT TIESCI 

GRANULATION , TISSUE (9) (10) <2C) 

1 (5«) 

(10) (10) 

BESPIHATOR Y SYSTEB 

HIDNG 

INFLABBATICN . IHTPRSTITIAl 

BHONCHOFNEOBONIA SOPPORATIVE 

HYPERPIASIJ, ALVEOIAR EPITHELIDH 

HYPEfPLASIA , tYHPHOID 

(9) 

1 (11*) 

(10) (20) 

1

2

 (5«) 

 (10*) 

(10) 

1 (10X) 

(10) 

HJBATOPOIETIC SYSTEB 

(SPLEJK 

AISOPHY. SCS 

HYPEEPLA5IJ, 8EH ATOPOIETIC 

HYPSFPLASIA, LYBPHOID 

HEMATOPOIEEI S 

(9) 

1 (11*) 

(10) CO) 

3 (15*) 

m 

1 (11*) 1 (11*) 

(10) 

1 (10*) 

ILYBP H NODE 

ATROPHY , NCS m (9) (2C) (10) (9)

1 (11*) 

IBEDIASTTNA 1 I.80DE 

ATBOEHY. BCS 

(9) (9) i;o> (10) 

1 (10*) 

(9) 

IPASCREATIC I.NOtE __ fllP-EEfLiSlii-llHEHQII! 

• NUBBER OF HNIHAT. S WITH TISSUE

» NUBBEB OF AKIBAI S NECHOPSIIC 

(9) (9) 

 EXABIHED BICRCSCCPICA1LY 

(20) (10) 

— i.nau. (9) 

CdlTIHUID OH 

107 
TABLE D1 CONTROL MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

IMFSENTfRI C I. HCDE 

HEHORRHAG l 

ATROEHY . DCS 

ANGIECTASIS 

• IHYtlUS 

ATROPHY , DCS 

CIRCULATOR Y SYETE f 

fMYOCAEtltl d 

TNFLAHnATICN , INTERSTITIAL 

DIGESTIV E SYStl K 

•LIVER 

NECROSIS , KOS 

HYPEETEOPHY , NO S HYPF,EPLASI» , MODULA R 

HYPSRPLAST1C RODOLE 

HYPSBPLASIA , H'flATOPOIETIC 

HYPEfFLASIA , LYflPHOTD 

tLIVBR/CF.mPILOBULAR 

DEGIHEBATICN, NOS 

»ITVER/HEPATCCYTT S 

HYPFRPLASIJ , SOS 

UBINA8 Y SYSTEC 

• KIDNE Y 

HYDRONEPHRCSI S 

INFLAflBATICB , CHRONIC 

IHFAFCT , KCS

 

• DRIIIARY BLttTE R 

INFLAnnATICN , CHRONIC 

ESDOCHIN E SYSIf B 

JADRENA I 

FIBROSIS 

__ CiI£IFI£iJIC_Hl_l!a5 

« OTHBER OF ANIHAL S 8ITH TISStJT

LOW DOSE 

UNTREATED 

CONTROL 

(20) 

(1) 

(SO) 

1 (5*) 

(20) 

1 (5*) 

<2C ) 

|20) 

CO) 

2 (10*) 1 (5*) 

(2C) 

(18) 

LOWER 

MID DOSE 

VEHICLE 

CONTROL 

(10) 

(1)

1 (IOC* ) 

(10) 

1 (1C*) 

(10) 

1 (1C*) 

2 (20*) 

2 (20*) 

(10) 

1 (10*) 

(10) 

(10) 

(8) 

(10) 

1 (10*) 

1 _[10»1 

UPPER 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

(9) 

(D

1 (100*) 

(10) 

(10) 

(10) 

(10) 

(10) 

1 (10*) 

(10) 

2 (20*) 

(9) 

LOWER 

MID DOSE 

UNTREATED 

CONTROL 

<9)

1 (1U) 1 (111) 1 (11*) 

(3) 

(9) 

(9) 

2 (22*) 

m 

m 

(9) 

(9) 

(8) 

 FXABIHE D HICEOSCCPICALLY 

UPPER 

MID AND HIGH 

DOSE UNTREAT-

ED CONTROL 

(9) 

(10) 

(10) 

1 (1C*) 

(10) 

(10) 

1 (10*) 

(10) 

(10) 

(10) 

108 
TABLE 01 CONTROL MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOWER UPPER LOW DOSE 

MID DOSE MID AND HIGH UNTREATED 

UNTREATED DOSE UNTREAT- CONTROL 

CONTROL ED CONTROL 

BIPHODOCTIVI SJSTIH 

IPBOSTATF (2) (10) (20)

 

mPLAHHATICH, SIJPPU»ATIVF 

IRFLAHflATION, CHROHIC SOPPOHATIV

 

I1ESTIS (9) (10) (20)

 

CAICIPICATICS, SOS 

SIBVOUS SYSTIB 

ROUE 

SPFCIAt S«»SF CBGAHS 

MORE 

BDSCOlOSBEIfTAI SYSTEM 

ROME 

tODT CATITIIS 

HONE 

All OTHER SISTfBS 

RORE 

SPECIAL BOEEHCIOGY SOflHABT 

NO LESION IEPOBTED 5 5 it 

ADTOIISIS/RO IECBOPSI 1 

I HDHBER OP lUlnHtS SIIH TISSOI EMHIHED HICPOSCOPICHLLT 

* HUHBE8 OF ASINAI S HECBOPS1ED 

LOWER 

MID DOSE 

VEHICLE 

CONTROL 

(10) 

1

(10) 

1

2 

 (10*) 

 (10«) 

UPPER 

MID AND HIGH 

DOSE VEHICLE 

CONTROL 

(10) 

1 [10*) 

(10) 

7 

109 
35 

TABLE 02 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL AND TREATED GROUPS) 

LOW DOSE LOW DOSE 

VEHICLE 

CONTROL 

»NIBALS INITIAII! III ST0DY 20 40 

X1IBAI.S BISSIHG 

HIIBALS «P,CEOP£IEC 20 00 

HIBALS EIABINID HISTOPATHOtOGICALLY 20 40 

IBTEGUBENTAF Y SIS1PB 

*SKIH (20) (40) 

HEBATOBA , (OS 

»SUBCUT TISSD1! (20) (10) 

HEBOPRHAG l 

BISPIRATC8Y SYSTffl 

ITS AC HE* (16) (38) 

INFLABBATICN , SOPPDRATIVE 1 (6*) 

HONG (18) (40) 

HEBORRHAGl 

IHPtABHATICN, INTERSTITIAL 2 (11*) 1 (3«) 

BROHCHOPBtBNONIA SOPPURATIVF 1 <6X) 2 (S*) 

HYP5EPLASIA, ITBPHOin 1 (3«) 

BEBATOPOIBTIC SYSTBB 

IBONE BARRC B (20) (38) 

ATROPHY, «CS 

HYPSEPLASI*. HFBATOPOIETIC 

HYPEEPLASIA , GRANOIOCYTIC 1 (3*) 

ISPLEEIf (19) (38) 

ATROPHY. HOS 

IHEDIASTINAL I. NODE (17) (40) 

HEBORRBIGI 

ATROPHY, «CS 

IBESENTEBIC L. NODE (") (40) 

__AI&Q£flIJ.££S . 

t NUBBER OF ARIBALS WITH TISSOF EXABIHED BICROSCCPICALLT
 

* HOBBTR OF AflBAlS HECROPSIIO
 

LOWER MID DOSE 

3« 

35 

35 

(35) 

(35) 

(31) 

(35) 

5 (6*) 

1 (3*) 

(34) 

1 (3*) 

1 (3X) 

(35) 

2 (6X) 

131) 

(3D

1 (3X1 

UPPER

MID DOSE 

35 

1 

34 

33 

(34) 

(34) 

(29) 

(33) 

(33) 

(30) 

1 (3*1 

(31) 

2 (6X) 

(31) 

 HIGH DOSE 

29 

12 

(29) 

1 (3X) 

(29) 

1 (3%) 

(11) 

(12)

1 (8*1 

(12)

1 <8X) 

(12)

1 <8X) 

(10) 

1 (10K) 

7 (70X) 

(10) 

110
 
TABLE D2 CONTROL AND TREATED MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

LOW DOSE 

VEHICLE LOW DOSE 

CONTROL 

BIPEBPlAStJ. ITRPHDID 1 (3«) 

ITHI80S (5) (7) 

AtSOPHT. (OS 

CIRCOIATCBT STSTIi 

ISYOCAStHJH (20) (39) 

IMFLKIIHATICM, SDPPURITI7P 3 (8*) 

IIFLAHHATICB, CHROR 1C DITFUSE 1 (3*) 

IIUDOCABEIOH (20) (39) 

ill Ft ABa AT ic ii . sos 2 (5*) 

IBFLABHATIOH, FOCAL 1 (3«) 

IHFLAHMATICll, SOPPORATIVF, IRFlAHBATICtl, ACOTF/CHBCIIIC 

•AOBTA (20) (»0) 

IIFIAHBATICII. SnPPDRATHF. 

CIGFSTIY1 STSllB 

IlITIS (20) 139) 

THIOHBOSIS, MOS 

IinARHATICI, SOPPORHTITI! 

ABSCISS, ICS 

TVF1AHHATICII, CN1ONIC SDPPOFHtl? 

IKC8CSIS, IOCAL KECBCSIS, COAGOLATITB 3 (8*) 

HIPEEPLASIA, DODDLAt 1 (5») 

AHGIF.CTASIS 1 13*1 

HIPBBPLASIA, HSBATOPOIMIC 

HJPTfPLASIA, 1TBPHOID 1 (3%) 

HKBATOPOIISIS 1 (5») 

•BILE DDCT (20) (HO) 

IHFIAHI1ATIOH. SOPPDRATIVI 1 (3K) 

INFUBBATICH. CHBOHIC SOPPOSATIV 

BYPEFPLASI*. IDS 1 (3*) 

HIPEFPLASIf, FOCAl 

IPANCRFA S (20) (38) 

I«Fl»Bg»TJCII. liTESSTITIAl 

ATBOEHI. ^IOS 

< IIUNBER OF HHIIUIS IITH TISSU! SIAHIRCD BICBOSCCPICHlir * HOBBER OF MllnJUS HFCBOPSIIC 

LOWER 

MID DOSE 

(15) 15

 (100%) 

(35) 

2 <«*> 

(35) 

3 (9X) 

1 (3X) 

(35) 

1 (3*) 

(3«) 

(«)

[3»> (3*) 

(3*> («)

(3*) 

1 (3*) 

(35) 

3 (9*) 1 (3*) 

5 (11*) 

7 (20X) 

(33) 

J (S«) ^ 2 (6S1 _, 

UPPER 

MID DOSE 

(2)

2 (1CCX) 

(32) 

(32) 

(3») 

(33) 

(3«) 

1 (3*) 

(30) 

HIGH DOSE 

r "' 9

 (100X) 

(12) 

(12) 

(29) 

(12) 

(29) 

1 OX) 

(12) 

1 (8X) 

111 
TABLE D2 CONTROL AND TREATED MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

VEHICLE 

CONTROL 

LOW DOSE 

ATBOEHT, fCCAt HKPSFPHSIA, MODOLAB 

IGASTBIC SOEHOCOSA 

HEBORBUHO I (20) (39) 

DBINABY SYS'tEC 

»KIDM>Y (20) (HO) 

PYELONEPHEITIS. FOCAL 2 (5«) 

INFtAHflATICK, IHTEBSTITIAt INPLAHBATIC1I, StJPPUHATIVE INFUBHUTICH, CHBONIC 

IirLANHftTICI, CHBOHIC FCCAL IHFtAHBATICN, CHBONIC SOPPUEATI V 

FIBROSIS. CIFFOS? 

KECRC5IS, BECailAIlT 1 (3*) 

IKIDMIT/COBTII (2P) (10) 

ATBOPHT, HCS 

HTBOFHY, FOC»t 

IKIDHEY/TOEDL E (20) (10) 

CAST, HCS 

IKIDNyY/fl!lVI£ (20) (HO) 

ABSCESS, NCS 

IDBIKAB1 EtPCtER (20) (38) 

INFIAHBATICH, SOPPOBAIIVE 

HEBOSIDESOSIJ 

IKCOCHIKJ SYSTIB 

KCHE 

BIPBODOCTUt S1STIB 

*SEBINA1 \IISICLE (20) (110) 

IHFUBf A1ICN, SUPPURATIVE 

«TESIIS (20) (38) 

PIBBOS1S, tlTFOSE 

ATBO^gjj JICS 

I NOHBEB Of ANIHALS WITH TISSUE EIAHINED NICROSCCPICA1LY *

 HtinBER 01' AfilHAIS HECROPSIID 

LOWER UPPER HIGH DOSE MID DOSE MID DOSE 

1 (8«) 

1 (3X) 

(32) (301 (12) 

1 (8«) 

(35) (33) (12) 

2 (6«) 1 (8*) 

2 (6X) 

1 (3«) 2 (17*)

 

1 (3«) 

2 (6«) 

1 (8*) 

6 (17*) 

OS) (33) (12) 

1 (8*) 

1 (8*) 

(35) (33) (12) 

1 (3«) 

«35) (33) (12)

1 (8*) 

(33) (33) (12) 

1 (3*) 

1 (8*) 

(35) (3«) (29) 

1 (3«) 

(3«) (31) (12) 

1 (3*) 

1_J8JJ_ 

112 
TABLE D2 CONTROL AND TREATED MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

VEHICLE LOW DOSE CONTROL 

BEBVOUS SYSTIB 

1ERAIN/BINIHGIS (20) («0)

 

IHFLABBATICN, SHPPURATTVE 

SPECIAL SEHSE CFGAH S 

NCNE 

BtSCOLOSKELSTAL SYSTEB 

NOSE 

BCDY CAVITIES 

•PERITOBEtJH (20) (»0) INFLABBATIC« , 1IOS 

INFLAHHATIOH, SUPPOBATIVE 1 (31() IHFLABBATIC8, FIBRINOOS 

IHFLABHATICH ACUTE AND CHEONIC 1 (3X) INFIAHBATICN , ACUTE/CHHOHIC 1 (3«) IBF1ABHATICN. CHBOHIC 

I8Ft»HHATIC», CHRONIC FOCAl 

IMF1ABBATICI1, CHROHIC DIFFDSF 1 (3«) INFIABBATICH, CHROHIC SOPPBRATIV 

INFLABHATICD , PYOGRANnlCBkTODS 

FIBSOSIS 

FIBRCSIS, FOCAt 

*PLEDBA (20) (HO) 

HFIAHMATICN, SDPP0RATIVF. 

til OTHER STS1IBS 

• BULTIPIE OEGAHS (20) (»0) ATROPHY. DCS 

HYPEEP1ASIA. LYHPHOID 1 (3X) 

SPECIAL BOEEBCIOGY SDHHABY 

HQ LESIOH BEPOBTED 4 13 

f RUBBER OF AIIBALS WITH TISSDI EXARIHED HICIOSCOPICALLI 

* MUHBER OF MUBAIS HECROPSIED 

LOWER 

MID DOSE 

<33) 

1 (3«) 

(35) 

1 (3*) 

2 (6*) 

1 (3*) 

1 <3X ) 21 |60K) 

2 (6X) 

1 (3*) 

(35) 

3 I9X) 

(35) 

3 

UPPER 

MID DOSE 

(28) 

(31) 

2 (6X) 

(31) 

(31) 

1 <3X) 

2? 

HIGH DOSE 

(12) 

(29) 

1 (3X) 

1 OX) 

(29) 

(29) 

.. 1 

113 
TABLE D2 CONTROL AND TREATED MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

VEHICLE 

CONTROL 

LOW DOSE LOWER 

MID DOSE 

UPPER 

MID DOSE HIGH DOSE 

»NIBM HSSIKG/NO NECROPSY 1 

HECROPSY EIRF/NO HISTO PEBFOBflED n MJTC/HECHCISY/HO HISTO 1 

HUTOLYSIE/NC HECHOPSY 6 

I HUHBEB OP KKIBAI S HITH TISSD! ETHHIKED HICBOSCCPICALLI 

» 1IUBBEB OP «ma»lS HECROPSIED 

114 
TABLE D3

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL GROUPS) 

LOWER UPPER LOW DOSE LOWER UPPER 

MID DOSE MID AND HIGH UNTREATED MID DOSE MID AND HIGH 

UNTREATED DOSE UNTREAT- CONTROL VEHICLE DOSE VEHICLE 

CONTROL ED CONTROL CONTROL CONTROL 

MIHALS HMIAIIY 18 STUDY 10 10 ;c 10 10 

HIIBAI.S WCBOISIED 10 10 19 10 10 IIIIIUS IXAHIHf D HISTOPATHOIOGICAILY 10 10 19 10 10 

IHTEGDHEHTA5Y SYS1EB 

»SKIN (10) (10) (19) (10) (10)

 

W.CFR, FOCA L 1 (5X)

 

RESPIRATORY SYSTEB 

«IU«G/BFOHCHDE (10) (10) (19) (10) (9)

 

BRONCHIECTASIS 1 (10*) 

INPtMINATICK. SUPPURATHE 1 (10*)

 

HYPEfPLASIA , IYBPHOID 1 (10X) 

ILONG/BROHCHIOIE (10) (10) (19) (10) (9)

 

HIPfF-PLASIA. IYBPHOID 1 (10X) 

ILUNG (10) (10) 11?) (10) (9)

 

IDEHA, HOS 1 (10X)

 BRONCHOPNECnONIA, NOS 1 (10*)

 INPlAHflATICN, IKTERSTITIAl 6 (60S) 1 <5*) 4 («0()

 

RYPERPIASIA, ALVEOLAR EPITBEHOB 1 1 (10*)

 HYPEEPLASIA. LYBPHOID 2 (2CK) 6 (32*) I1 (10*)

 

EEBATOPOIBTIC SYSTEB 

ISPLEFN (10) (10) (19) (10) (10)

 

HECROSIS. COAGOLATIVE 1 (10*) HYPEFPIAEIA , BFB ATOPOIETIC 3 (30X)

 

HYPEEPLASIA, LYBPHOID 3 (16*; 1

 HEMATOPOIFSIS 5 (26*1 1

 

IBESENTERIC I. NODE (10) (1) (19) (10) (10) 

HYPERPLASIA , LYHPHOID 1 (10X) 

ITHYBDS (1) 46) (1) ATROPHY, »CS i (ioox) HYPEtELlSJJ. LYBPHOID 1 (17*1 I , _,_ 

I WJBBEH OF AHIBAL S HITH TISSUE EXAHIHE D MICROSCCPICAIIY 

« HUHBFB OF ABIflAtS NECROFSItD 

115 
TABLE 03 CONTROL FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOWER UPPER MID DOSE MID AND HIGH

 

UNTREATED DOSE UNTREAT-

CONTROL ED CONTROL 

CIRCDLATORY SSSTF.P 

(IHF.ABT (10) (10) 

PERI ART FRIT IS 2 (20t) 

C1GFSTIVE SYSTE M 

ILIVER (10) (10)

 

NECROSIS, COAGDLATIVF 1 (10*)

 

CYTOFLAS11IC VACDOIIZ ATION 

BASOPBIIIC CY10 CHANGE 1 (10K)

 

HYPFRPLA31IC NODDIF 

HYPFFPIASIA, HEtlATOPOIETIC 1 (10J) 

HYPFRPLA5IA, IYBPHOID 2 (20*)

 

*BILE DCCT (10) (10) 

HYPFRP1A 5IA, HEN ATOPOIETIC 

IPANCRFAS (10) (9)

 

CYSTIC t'JCTS 

INFLAJinATIOH, INTERSTITIAL

 

• PANCREATIC KINGS (10) (9)

 ATROPHY, HCS

 

ATROCHY, ?CCAL 

URINARY SYSTPr 

•KIDNEY (10) (10) 

INFLABBATICN, CHRONIC 

INFAECT, NOS 1 (10*) 

• URINAfY BUttF.R (10) <•») HYPFRPLASIJ, IYBPHOID 

EKDOCSINE SYSTJB 

•THYKOIE (5) <8) HYPERPIA:;I>. CYSTIC 

HlPEl£tA::llJ._igitI£BtAEr£SJ1l. 

t NUBBF R OF MIBALS HITH TISSUE EXAHINE D BICBOSCCPICAILY 

* NUSBER OF HSIBAI S NECROPSIED 

LOW DOSE 

UNTREATED 

CONTROL 

(19) 

(19) 

1 (5K) 

1 (5«) 

119) 

(19) 

1 (5*) 

(19) 

1 15*) 

1 (5*) 

(19) 

1 (5*) 

(1?) 

(19) 

1 15S1. 

LOWER MID DOSE 

VEHICLE 

CONTROL 

(10) 

(10) 

1 (10*) 

2 (20*) 

(10) 

1 (10*) 

(10) 

1 (10*) 

(10) 

(10) 

(8) 

1 (13*) 

(6) 

1 (17*) 

<UPPER 

MID AND HIGH DOSE VEHICLE

 

CONTROL 

(9) 

(10) 

(10) 

(10) 

(10) 

(1C) 

(9) 

(10) 

. 

116 
TABLE 03 CONTROL FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

LOWER UPPER 

MID DOSE MID AND HIGH 

UNTREATED DOSE UNTREATCONTROL ED CONTROL 

REPRODUCTIVE SSSTEB 

IUTEBUS (10) m CYST, SOS 

HEflOERHAGF 

1 (10*) i (in) PYOBETRA 

ANGIECTASIS 1 (ii«) 

HCTEBUS/ENEOBITRIUI1 (10) (9) 

HYPEBPIASIA, CYSTIC 8 (80*) 

•OVUBY (8) (3) 

RINERALIZATION 

POLLICatAB CYST. NOS 

ATRCEHY. KCS 

fERVOUS SYSTEB 

NONE 

SPECIAL SENSf CEGAB S 

NONE 

BUSCOLOSKELITAI SYSTEM 

NONE 

EODY CAVITIES 

NONE 

All OTHER SYS1IBS 

ADIPOSE TISSUE 

INFLAHHAT10N, POCAL 

SPICIAL BOBEHOIOGY SOBHABY 

NO 1ESION EEPORTED 7 

jipTOljsjs/Ng NKSQPS? I

 BDHBEB OF AKIBAL S »ITH TISSUE F.XAHINED HICBOSCOPICAtLY 

* NOBBEF OF ABIMAIS NECROPSIKD 

LOW DOSE 

UNTREATED 

CONTROL 

(19) 

(19) 

15 (79*) 

(18) 

1 (6X> 1 (6*) 

1 

1 

LOWER UPPER 

MID DOSE MID AND HIGH 

VEHICLE DOSE VEHICLE 

CONTROL CONTROL 

(10) (9) 

1 (10*) 

(10) (9)

 

9 (9C») 

<6) (6) 

2 (33*) 

, , 

117 

9 
TABLE D4

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE GIVEN 

INTRAPERITONEAL INJECTIONS OF ACRONYCINE (CONTROL AND TREATED GROUPS)

 

LOW DOSE VEHICLE 

CONTROL 

LOW DOSE 

ASIHALS

ASIMALS

JHIBAL S

 INITIHtl IN STUD!

 NECBCISIfD

 UAHIXID HISTOPATHOLOGICAltT

 20 

 20 

 20 

HO 

39 

39 

ISTEGUireHTABT STS1EM 

*SKI8 (20) (39) 

INPLABBATICH, SUPPURATIVE 

• SOBCOT TISSO E (20) (39) IHFLABBATICS, SUPPORATIVl 

ABSCESS, SCS 

FtSPIRATCHY SKSTJB 

ITBACHEA (18) (38) IHFLAflllATICH, SDPPDRATIVF 3 (8*) 

ItDHG (17) (38) 

EDEHA, MOS 

REHOCBHAG E 

INFLABHATICB, IHTERSTIIIAl 

BBOMCHOPllfOBOSIA SDPPORAtTVI 11 (29X) 

IMFIAHMATICN, ACUTE SUPPDB ATIVE 

HYPEIP1ASIA. ITBPHOID 2 <5t> 

HEBAIOPOIETIC STS1FB 

IBONE BABBC H (18) (39) 

ATROPHY, DCS 

HYPEFPIASIA, HOS 

HIPESPLASIA, HEBATOPOIETIC 

ISPLEER (20) (39) 

HEBORRHAGI 

AIBOEHI, «CS 

BIPIEPIASIA, ROS 

BlPIffl-flSIii, BEHATOP9ISIIC 

I RUBBER OF AHIRALS IITH TISSUE EXAHIHED BICROSCCPICALI.Y 

* ROBBER OF ARIBAIS HECROPSIIB 

LOWER MID 

DOSE 

35 

33 

33 

(33) 

1 (3K) 

(33) 

(30) 

(33) 

1 (3K) 

2 (6%) 

1 <3X) 

1 <3X) 

(31) 

1 OX) 

(33) 

5 (15X) 

UPPER MID DOSE

 

35 

33 

33 

(33) 

(33) 

1 (3K ) 

(32) 

(33) 

1 (3)1) 

(32) 

« (13X) 

1 <3X ) 3 (9X) 

(3D 

1 (3X) 3 (10J) 

2 (6t) 

_ -«_um. 

HIGH DOSE 

35 

32 

15 

(32) 

(32) 

1 (3X) 

1 <3X) 

(1t> 

(15) 

2 (13%) 

(1«) 

(15) 

1 (I*) 

118 
TABLE D4. CONTROL AND TREATED FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

, 

RYPBFPIAEIJ. IYBPHOID HEBATOICIEEIS 

ISSDIASTINAL I. RODE

 ATHOPHY, NCS

 

HYPFFPLASIJ, PLASBA CELL 

ITHYHDS 

ATROPHY. DOS 

HY PEE PL A SI A, PLASBA «LL 

CIRCULATORY SYSTFfl 

fBYOCARriOB 

HFLAHBATICH , INTERSTITIAL IHFLABBATIOH, SUPPURATIVE 

IHPlABHATICIi , ACUTF FOCAL 

INFLABBATICN , ACUTE SUFFORATIV E 

lENDOCAEDIO n I8FL»BBATIC» , NOS

 

INFLABBATION , StlPPDRATIVF I8FLAHBATICS. PIBRIHOOS

 

I«Fl«HBATICN, ACDTE 

•AOBTA 

IHFLABBATICB, SHPPURATIVE 

INFLABBATICN , ACUTE SDPPURATIV E 

tlGESTIVE SYSTIB 

ILITER 

IHFLAHHATICN . SOPPORATIVF INFLAHBATICN , NECROTIIIKG 

IMFtABBATICN , ACUTE SDFPflBATHE BECROSIS, TOCAL 

KECRCSIS. CCAGOLATIVE LEOItOCYTOSIS, ROS 

HYPEfPlASIA , LYBPHOID 

ILIVER/PIRIPORTAI 

HECROSIS, HOS

 

•BILE DOCT 

_EI1M1HQ1*-E2S 

LOWTXDSE 

VEHICLE CONTROL 

(20) 

(9) 

(19) 

(19) 

1 (5K) 

(20) 

(20) 

1 <5*> 

(20) 

(20) 

LOW DOSE 

10 (26X) 

1 (3*1 

(31) 

(11) 

(38) 

3 (8*) 

(38) 

2 (5%) 

(39) 

(39) 

1 (3«) 

(39) 

(39) - ___ _ 

LOWER MID UPPER MID DOSE DOSE 

(58) (31) 

5 (16X) 

3 (10X) 

(22) P)

22 (100X) 6 (86*) 

1 (UK) 

<33) (33)

 

1 (3*) 1 (3*)

 

1 <3X) 

1 <3*> 

(33) (33)

 

1 (3X) 10

 (30%) 

1 (3X) 

2 <6X > 

(33) (33) 

3 (9«) 

1 (3X) 

(3D (32) 

1 OX) 1 (3*) 

2 (6X) 1 (3X) 2 (6X) 

1 (13*) 

2 (6X) 

(21) (32) 

1 (3K) 

(33) (33) 

„ ...1 [3J1_ 

HIGH DOSE 

(12) 

3 (25X) 

(»)

<l (100X) 

(IS) 

<15) 

(32) 

(15) 

(15) 

(32) 

I	 NUBBER OF ANIBALS SITH TISSUE EXABIHED BICROSCCPICALtY
 

*	 NUBBFB OF ABIBHIS NECROPSITD
 

119
 
TABLE D4. CONTROL AND TREATED FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE LOW DOSE LOWER MID UPPER MID HIGH DOSE VEHICLE DOSE DOSE 

CONTROL 

INFLANBATICN , SOPPUHATIVE 1 2 (6X) 1 (3*) 

INFLABBATICN , ACUTE SUPFORATIV E 1 (3*) 

HYPFEPLASIA , SOS 3 <9X) 19 (58X1 

HYPEFPLASIA , FOCAL 12 (36*) 1 (3«) HYPKBPLASIA , EIFFDSE 1 <3X > 

HYPFfPIASIA , CISTIC 2 (6*) 

• PANCREA S (16) (35) <3C) (29) (15) INFLABBATICN , FOCAL 1 (7*) 

INFLAnHATION , INTERSTITIAL 1 (6») 1 (3*) NECROSIS , COAGUtATIVE 1 (3*) 

ATROEHY , KC S 1 (3«) 1 (3«) 

ATROEHY , FCCA I 1 (7X) 

•STOMACH (18) (39) (29) (32) (1U) 

HYPEBPUSIA . EIASH A CELL 1 (3t) 

ILARG E INTESTIN E (13) (36) (28) (32) (11 ) 

NEMATODIASIS 1 (U*) 

OBINAB Y SYSTE M 

•KIDNEY (20) (39) (33) (33) (11) 

INFLABBHTICN , INTERSTITIAL 1 (3*) 

INFLABHATICN . SUPPUBATIVE 3 (9X) 

PYELCNEPHFITI S SUPPORATIVF 1 (3*1 

INFLABBATICN , ACUTE SOPPURATIV E 1 (3*) 

INFLABBATICK, CHBONIC 2 (6*) 1 (8X) 

IN"LABBATICS , CHRONIC FCCAL 1 <3«) 1 <3X) 

INFLABBATICN , CHRONIC DIFFUSE 1 (3K) NECRCSIS, CCAGULATIV E 1 (3X) 

NECROSIS, BEEUILAR Y 3 <8t) 6 (1SK ) 

•KIDNEY/CORTI S (20) (39) (33 ) (33) (13) 

ISFLABBATICN , SUPPURATHE 1 <3«) 

•KIDNEY/BECUII A (20) (39) (33) (33) (13) 

ABSCESS, NCS 1 (3*) 

NECROSIS, COAGOLATIV? 1 (3*) 

«KIDN"Y/TDEUL I (20) (39) (33) (33) (13) 

CAST, NOS 5 (6*) 

CALCIFICATION, NOS 1 (3*) 

•KIDNEY/E^LVI S (20) (39) (33) (33) (13) 

_ »SSC.ISS_, J£S . . l-Jiil.. J-lS'l 

I BOBBER OF ANIBAL S BITH TISSUE EXABINE D BICROSCCPICALLY 

* BOBBER OP AMBAI S NECROPSIIE 

120 
TABLE D4. CONTROL AND TREATED FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE VEHICLE CONTROL LOW DOSE LOWER MID DOSE UPPER MID DOSE HIGH DOSE 

INCOCBIN I SISTffl 

NCHE 

BtPBODUCTIVt SYSTf B 

IDTEBOS 

CYST. NOS 

PYOBETHA 

(20) (39) (31)

: (6*) 2 (6%) 

(32) (15) 

lOTEBOS/ENCOKHBIUa 

HYPEBPLASJA , CYSTIC 

«OVABY/OVIEOCT 

MFLAHBATICN, SOPPGBATIVE 

IOVABY 

CYST, NOS 

FOLLICDLAB CYST. HOS 

INFLABBATICN , SOPPUBATIVE 

ATBOFHY , DCS 

(20) 

2

(20) 

(20) 

 (10*) 

(39) 

21 (54X) 

(39) 

(39) 

1 (3*) 

131) 

7 (23*) 

(3D 

(27) 

1 («*) 

1 (»*> 

(32) 

(32) 

1

(29) 

1

1

 (3*) 

 (3*) 

 (3*) 

(15) 

(15) 

(11) 

HIBTOOS SYSTIB 

NONE 

SPECIAL SIHSI CEGABS 

NONE 

BnSCBLOSKEUTAI SYSTEB 

•SKELETAL BDSCLI 

INFLABBATICN, FOCAL 

(20) (39) (33) 

1 (3*) 

(33) (32) 

EODT CAVITIES 

•ABDOMINAL BAIL 

ABSCESS, NOS 

• PERITOSEDB 

... IN£IiftBRKTICIi..BO? ., 

(20) 

(20) 

(39) 

(39) 

« TOHBER OF ANIBALS BITH TISSUE EXABIHF.D BICBOSCCPICALIY 

* NOBBED OF A Hid A IS NECROPSIIC . 

(33) 

(33) 

2 (6*1 

(33) 

(33) 

(32) 

1

(32) 

 (3*1 

121 
TABLE D4. CONTROL AND TREATED FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

LOW DOSE 

VEHICLE CONTROL 

LOW DOSE LOWER MID 

DOSE 

UPPER MID

DOSE 

 HIGH DOSE 

ISTLABBAIIOH .

IHFLAflPIATICB ,

ABSCESS, SCS 

INFLABBATICN ,

INFLABBATICB,

INF1ABBATICN ,

INFIABBATICN ,

PIBRC3IS 

 SDPPDRATIVB 

 FIBRIKOOS 

 ACOTE/CHBCHIC 

 CHRONIC 

 CHRONIC FOCAl 

 CHRONIC SDSPDBATIV 

2

1

 (5*) 

 (3*) 

e (2«X) 

3 (9%) 9 (27t) 

1 (3%) 

1 (3*) 

1 (3X1 

H (12*1 

1 (2« ) 

7 (21*1

1 (3*1 

1 O») 

2 <6X) 

• PLEUB A 

INFIABBATICH ,

IBFLABBATICB,

IBFIABBATICN ,

 NOS 

 SOPPUHATIVK 

 FIBRIMOBS 

(20) (39) (33) 

1 (3K) 

(33)

1 (3») 

1 <3K ) 

 (32) 

All OTHF.R SYSTIBS 

•HDITIPIE OEGAH S 

HYPEEPL»SI» , 1YBPHOID 

(20) (39) 

1 (3K) <33) (33) (32) 

SPFCTAt flOFEHCIOGY SHBBASt 

»0 LF.SIOH FEFORTED 

1IECROPSI PIRF/80 HISTO

AOTOLYSIS/HO NECROPSY 

 PERFORBED 

1 

1 

1 

2 

e

2

 9 17 

3 

«

«

 HDHBEB OP

 HUBBEB OF

 HHIBHIS HITH TISSDE EIABINED

 AHIHIUS HECROPSISD 

 BICBCSCCPICALLT 

122 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN RATS
 

GIVEN INTRAPERITONEAL INJECTIONS
 

OF ACRONYCINE
 

123
 

Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Given Intraperitoneal Injections of Acronycinea, Using Vehicle Controls
 

Topography: Morphology
 

Liver: Hepatocellular Adenoma
 

or Carcinoma^* (52)
 

P Values0 >d
 

Relative Risk (Vehicle Control)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Osteosarcomab (41)
 

P Values0 »d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Low-Dose 

Vehicle 

Control 

Mid- and 

High -Dose 

Vehicle 

Control 

Low 

Dose 

Mid 

Dose 

High 

Dose 

0/10 (0) 0/8 (0) 2/21 (10) 0/11 (0) 0/0 (-) 

— 

__ 

— 

__ 

N.S. 

Infinite 

0.156 

Infinite 

80 

— 

__ 

— 

_

— _ 

— 

__ 

— 

— 

__ 

0/10 (0) 0/8 (0) 0/25 (0) 0/18 (0) 2/7 (29) 

— P = 0.048 — 

— 

— 

— 

— __ 

— 

— 

N.S. 

Infinite 

0.392 

Infinite 

— — — — 41 
__ 

Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Given Intraperitoneal Injections of Acronycine3, Using Vehicle Controls
 

(continued) 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid High 

Topography: Morphology Control Control Dose Dose Dose 

Musculoskeletal System:
 

Osteosarcomab

 (33) 0/10 (0) 0/8 (0) 3/29 (10) 11/26 (42) 8/15 (53)
 

P Valuesc»d P = 0.019 N.S. P = 0.027 P = 0.013
 —
 

Relative Risk (Vehicle Control)f Infinite Infinite Infinite
 

Lower Limit 0.231 1.192 1.442
 

Upper Limit Infinite Infinite Infinite
 

Weeks to First Observed Tumor 

__ 

50 35 33
 

All Sites:
 

Hemangiosarcoma" (46) 0/10 (0) 0/8 (0) 4/23 (17) 0/13 (0) 1/4 (25)
 

P Valuesc'd N.S. N.S. N.S.
 — —
 

Relative Risk (Vehicle Control)f Infinite Infinite
 

Lower Limit 0.452 0.117
 —
 Upper Limit Infinite Infinite
 —
 

Weeks to First Observed Tumor 48 46
 —
 — —
 
___ 

__ 

Tc.tlc El 1 • Timc~ddDucted An—1"sss of ttie Incidence of. Piri-tnsir^-7 TUIQOTS 3_n ^5^ 1^ ^3.ts 

Given Intraperitoneal Injections of Acronycinea, Using Vehicle Controls 

(continued) 

Mid- and
 

Low-Dose High -Dose
 

Vehicle Vehicle Low Mid High
 

Topography: Morphology Control Control Dose Dose Dose
 

All Sites- Osteosarcomab (25) 0/10 (0) 0/8 (0) 4/31 (13) 13/30 (43) 12/18 (67)
 

P Valuesc>d P = 0.002 N.S. P = 0.022 P = 0.002
 

Relative Risk (Vehicle Control) f Infinite Infinite Infinite
 

Lower Limit 0.334 1.248 1.941
 

Upper Limit Infinite Infinite Infinite
 

Weeks to First Observed Tumor 50 35 25
 —
 

Bile Duct: Bile Duct
 

Carcinomab (52) 0/10 (0) 0/8 (0) 2/20 (10) 0/11 (0) 0/0 (-)
 

P Valuesc»d N.S.
 —
 —
 

Relative Risk (Vehicle Control)^ Infinite
 

Lower Limit 0.164
 — —
 Upper Limit Infinite
 — —
 

Weeks to First Observed Tumor 62
 —
 —
 
Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats 

Given Intraperitoneal Injections of Acronycinea , Using Vehicle Controls 

(continued) 

Topography: Morphology
 

Adrenal: Cortical
 

Adenoma** (41)
 

>d
 P Valuesc


Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Cortical Adenoma
 

or Carcinoma*1 (41)
 

P Values0'd
 

Relative Risk (Vehicle Control) f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid High 

Control Control Dose Dose Dose 

0/10 (0) 1/8 (13) 1/24 (4) 2/18 (11) 4/7 (57) 

P = 0.045 N.S. N.S. N.S. — 

Infinite 0.889 4.571 

0.024 0.058 0.629 

Infinite 49.343 153.053 

.._ 72 80 61 41 

0/10 (0) 1/8 (13) 1/24 (4) 3/18 (17) 4/7 (57) 

N.S. N.S. N.S. N.S. — 

Infinite 1.333 4.571 

0.024 0.138 0.629 

Infinite 65.560 153.053 

72 80 61 41 — 
__ __ 

__ 

__ 

__ 

Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Given Intraperitoneal Injections of Acronycine3

, Using Vehicle Controls
 

(continued)
 

Mid- and
 

Low-Dose High -Dose
 

Vehicle Vehicle Low Mid High
 

Topography: Morphology Control Control Dose Dose Dose
 

—
 

Peritoneum: Sarcoma, NOSb (32) 0/10 (0) 0/8 (0) 2/29 (7) 0/26 (0) 1/16 (6) 

P Valuesc » d N.S. N.S. N.S. — — 

Relative Risk (Vehicle Control)f Infinite Infinite 

Lower Limit 0.112 0.030 

Upper Limit Infinite — Infinite 

Weeks to First Observed Tumor 56 32
 

Peritoneum: Fibrosarcoma'3 (32) 0/10 (0) 0/8 (0) 0/29 (0) 0/26 (0) 2/16 (13)
 

P Valuesc»d N.S. N.S.
 — — —
 

Relative Risk (Vehicle Control)f .._
 Infinite
 

Lower Limit 0.169
 

Upper Limit — —
 Infinite
 — —
 

Weeks to First Observed Tumor 32
 — — —
 ——
 
__ __ __ 

Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Given Intraperitoneal Injections of Acronycinea

, Using Vehicle Controls
 

(continued)
 

Mid- and
 

Low-Dose High -Dose
 

Vehicle Vehicle Low Mid High
 

Topography: Morphology Control Control Dose Dose Dose
 

Peritoneum: Mesothelioma^ (52) 0/10 (0) 0/8 (0) 3/20 (15) 1/11 (9) 0/0 (-)
 

P Values0 >d N.S. N.S. N.S.
 —
 —
 _w
 

Relative Risk (Vehicle Control)f Infinite Infinite
 

Lower Limit 0.336 0.044
 

Upper Limit Infinite Infinite
 —
 

—
 

Weeks to First Observed Tumor 58 55
 

Peritoneum: Sarcoma and
 

Other Related Tumorsb (32)+ 0/10 (0) 0/8 (0) 5/30 (17) 3/26 (12) 7/16 (44)
 

P Valuesc»d P = 0.006 N.S. N.S. P = 0.033
 —
 

Relative Risk (Vehicle Control)f Infinite Infinite Infinite
 

Lower Limit 0.470 0.213 1.142
 

Upper Limit Infinite Infinite Infinite
 

Weeks to First Observed Tumor 56 55 32
 —
 —
 

"'These tumors consist of sarcoma, fibrosarcoma, or mesothelioma.
 
__ 

Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Given Intraperitoneal Injections of Acronycinea, Using Vehicle Controls
 

(continued) 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid High 

Topography: Morphology Control Control Dose Dose Dose 

Multiple Organs:
 

Fibrosarcomab (33) 0/10 (0) 0/8 (0) 0/29 (0) 2/26 (8) 2/15 (13)
 

P Valuesc»d N.S. N.S. N.S.
 —
 —


Relative Risk (Vehicle Control)

f Infinite Infinite
 

Lower Limit 0.104 0.181
 —
 Upper Limit Infinite Infinite
 —
 

Weeks to First Observed Tumor 61
 — — —
 

a

Treated groups received doses of 3.75, 7.5, or 15 mg/kg by intraperitoneal injection.
 

"Number of tumor-bearing animals/number of animals examined at site (percent), based on
 

animals that survived at least as long as the number of weeks on study shown in parentheses
 

after the description of morphology.
 

G

Beneath the incidence of tumors in the mid- and high-dose control group is the.probability
 

level for the Cochran-Armitage test when P < 0.05 using only the mid- and high-dose groups in
 

the trend analysis; otherwise, not significant (N.S.) is indicated. Beneath the incidence
 

of tumors in a treated group is the probability level for the Fisher exact test for the
 

comparison of that treated group with its appropriate control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

33 
Table El. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Rats
 

Given Intraperitoneal Injections of Acronycinea, Using Vehicle Controls
 

(continued)
 

"A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^The 95% confidence interval of the relative risk between each treated group and its appropriate
 

control group.
 
Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycinea

, Using Vehicle Controls
 

Topography: Morphology
 

Subcutaneous Tissue:
 

Fibromab (52)
 

P Values0'd
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Subcutaneous Tissue:
 

Fibrosarcomab (52)
 

P Values0'd
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid High 

Control Control Dose Dose Dose 

0/10 (0) 0/9 (0) 2/34 (6) 0/29 (0) 1/14 (7) 

N.S. N.S. N.S. — — 

Infinite —. Infinite 

0.096 0.037 — Infinite Infinite — __ 

79 80 — — 

0/10 (0) 0/9 (0) 1/34 (3) 2/29 (7) 0/14 (0) 

. N = S = N = S, N.S. — 

__ 

Infinite Infinite 

0.017 0.103 — Infinite Infinite — 

58 56 — — — 
__ 

Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycinea, Using Vehicle Controls
 

(continued) 

Mid- and 

Low-Dose High-Dose 

Vehicle Vehicle Low Mid High 

Topography: Morphology Control Control Dose Dose Dose 

Subcutaneous Tissue: Fibroma
 

or Fibrosarcomab (52) 0/10 (0) 0/9 (0) 3/34 (9) 2/29 (7) 1/14 (7)
 

P Values0 >d N.S. N.S. N.S. N.S.
 —
 

Relative Risk (Vehicle Control)f Infinite Infinite Infinite
 

Lower Limit 0.197 0.103 0.037
 

Upper Limit Infinite Infinite Infinite
 

Weeks to First Observed Tumor 58 56 80
 — —
 

Lung: Alveolar/Bronchiolar
 

Adenomab

 (32) 0/10 (0) 0/8 (0) 0/35 (0) 2/31 (6) 0/28 (0)
 

P Valuesc»d N.S. N.S. N.S.
 — __
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.087
 

Upper Limit Infinite
 —
 

—
 

Weeks to First Observed Tumor
 _—
 —
 32 
Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given IntrapeTitOfpsTL Injections of Acronvcine3. Using Vehicle Controls
 

(continued)
 

Topography: Morphology
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma^ (32)
 

P Valuesc»d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Adenoma
 

or Carcinomab (52)
 

P Values0 »d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle . Low Mid High 

Control Control Dose Dose Dose 

0/10 (0) 1/8 (13) 1/35 (3) 2/31 (6) 0/28 (0) 

N.S. N.S. N.S. N.S. — 

Infinite 0.516 0.000 

0.017 0.034 0.000 

Infinite 29.485 5.278 __ __ 

82 76 32 

0/9 (0) 0/8 (0) 0/34 (0) 4/29 (14) 0/13 (0) 

N.S. . N.S. — — __ __ 

Infinite 

0.296 — Infinite — 

— — 

64 — — — — 
__ 

__ 

Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycine3

, Using Vehicle Controls
 

(continued) 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid 

Topography : Morphology Control Control Dose Dose 

Adrenal: Cortical Adenoma
 

or Carcinoma6 (40) 0/10 (0) 1/8 (13) 0/32 (0) 9/28 (32)
 

P Valuesc»d N.S. N.S.
 — —
 

Relative Risk (Vehicle Control)

f 2.571
 

Lower Limit 0.477
 —
 Upper Limit 107.105
 —
 

Weeks to First Observed Tumor _..
 82 58
 

Mammary Gland:
 

Adenocarcinoma, NOS^ (32) 0/10 (0) 0/8 (0) 6/35 (17) 4/34 (12)
 

P Valuesc>d N.S. N.S. N.S.
 —
 

Relative Risk (Vehicle Contol)f Infinite Infinite
 

Lower Limit 0.512 0.252
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 73 32
 — —
 

+

0ne animal in this group was diagnosed with both adenoma and carcinoma. 

High
 

Dose
 

7/23 (30)+
 

N.S.
 

2.435
 

0.418
 

103.211
 

40
 

2/30 (7)
 

N.S.
 

Infinite
 

0.090
 

Infinite
 

61
 
Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycine3

, Using Vehicle Controls
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Fibromab (52)
 

P Valuesc>d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland:
 

Fibroadenomab

 (51)
 

P Valuesc»d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid- and 

Low-Dose High-Dose 

Vehicle Vehicle Low Mid High 

Control Control Dose Dose Dose 

0/10 (0) 0/8 (0) 0/34 (0) 2/29 (7) 1/14 (7) 

N.S. N.S. N.S. — — __ 

Infinite Infinite 

0.093 0.034 — Infinite Infinite 

__ __ _

— _ 

82 80 

1/10 (10) 3/8 (38) 20/34 (59) 13/29 (45) 3/15 (20) 

N.S. P = 0.007 N.S. N.S. — 

5.882 1.195 0.533 

1.209 0.491 0.104 

227.093 5.422 3.235 

80 82 76 51 61 
__ 

Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycine3, Using Vehicle Controls
 

(continued) 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid 

Topography: Morphology Control Control Dose Dose 

Mammary Gland: All Tumors
 

Except Fibromab (32)+ 1/10 (10) 3/8 (38) 22/35 (63) 16/34 (47)
 

P Valuesc>d P = 0.034(N) P = 0.004 N.S.
 —
 

Relative Risk (Vehicle Control) ^ 6.286 1.255
 

Lower Limit 1.310 0.535
 

Upper Limit 237.797 5.627
 

Weeks to First Observed Tumor 80 82 66 32
 

Uterus: Endometrial
 

Stromal Polypb (45) 0/10 (0) 0/8 (0) 5/34 (15) 0/30 (0)
 

P Valuesc>d N.S. N.S.
 —
 —
 

Relative Risk (Vehicle Control)f Infinite
 

Lower Limit 0.415
 —
 Upper Limit Infinite
 —
 

Weeks to First Observed Tumor 80
 —
 — —


"'"These tumors consist of adenocarcinoma, fibroadenoma, papillary adenocarcinoma,
 

cystadenoma, or cystadenocarcinoma. The fibromas are omitted from this combination.
 

High
 

Dose
 

5/30 (17)
 

N.S.
 

0.444
 

0.128
 

2.471
 

61
 

1/19 (5)
 

N.S.
 

Infinite
 

0.025
 

Infinite
 

45
 
__ __ 

__ 

__ 

__ 

__ 

_

__ 

Table E2. Time-adjusted Analyses of

Given Intraperitoneal Injections

(continued) 

Low-Dose
 

Vehicle
 

Topography: Morphology Control
 

Peritoneum: Sarcoma, NOS^ (42) 0/10 (0)
 

P Valuesc>d
 

—
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Peritoneum: Fibrosarcoma^ (52) 0/10 (0)
 

P Values0 »d
 

—
 

Relative Risk (Vehicle Control)*
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 . —
 

 the Incidence of Primary Tumors in Female Rats 

 of Acronycine3, Using Vehicle Controls 

Mid- and 

High-Dose 

Vehicle Low Mid High 

Control Dose Dose Dose 

0/8 (0) 0/35 (0) 0/30 (0) 5/22 (23)
 

P = 0.010 N.S.
 —
 

Infinite
 

—
 

0.533
 —
 —
 Infinite
 —
 —
 _
 

0/8 (0) 0/34 (0) 2/29 (7) 0/14 (0)
 

N.S. N.S.
 — —
 

Infinite
 

0.093
 — Infinite
 —
 

—
 —
 

58
 — —
 —
 

42 
__ 

__ 

__ 

__ 

__ 

Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats 

Given Intraperitoneal Injections of Acronycine3, Using Vehicle Controls 

—
 

(continued) 

Mid- and 

Low-Dose High-Dose 

Vehicle Vehicle Low Mid High 

Topography: Morphology Control Control Dose Dose Dose 

Peritoneum: Mesothelioma^5

 (43) 0/10 (0) 0/8 (0) 1/35 (3) 0/30 (0) 2/21 (10) 

P Values0 » d N.S. N.S. N.S. — — 

Relative Risk (Vehicle Control)f Infinite Infinite 

Lower Limit 0.017 0.129 

Upper Limit Infinite — Infinite 

Weeks to First Observed Tumor 46 43
 

Peritoneum: Sarcoma and
 

Other Related Tumorsb (34)+ 0/10 (0) 0/8 (0) 1/35 (3) 5/30 (17) 13/28 (46)
 

P Valuesc»d P = 0.002 N.S. N.S. P - 0.016
 —
 

Relative Risk (Matched Control)f Infinite Infinite Infinite
 

Lower Limit 0.017 0.390 1.339
 

Upper Limit Infinite Infinite Infinite
 

Weeks to First Observed Tumor 46 51 34
 —
 

~*"These tumors consist of sarcoma, fibrosarcoma, or mesothelioma.
 
Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycinea

, Using Vehicle Controls
 

(continued)
 

Topography: Morphology
 

Multiple Organs:
 

Sarcoma, NOSb

 (34)
 

P Valuesc»d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Multiple Organs:
 

Fibrosarcomab

 (41)
 

P Valuesc»d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid High 

Control Control Dose Dose Dose 

0/10 (0) 0/8 (0) 0/35 (0) 2/30 (7) 3/29 (10) 

N.S. N.S. N.S. — — 

_« 

Infinite Infinite 

0.090 0.192 — Infinite Infinite — __ __ «... 51 34 

0/10 (0) 0/8 (0) 0/35 (0) 2/30 (7) 1/25 (4) 

N.S. N.S. N.S. — — 

— -— Infinite Infinite • 0.090 0.019 — Infinite Infinite — 

51 41 —— — — 
Table E2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycine3

, Using Vehicle Controls
 

(continued)
 

Topography: Morphology
 

All Sites: Osteosarcomab (52)
 

P Valuesc>d
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

All Sites:
 

Hemangiosarcomab (52)
 

P Valuesc»d
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mid- and 

Low-Dose High -Dose 

Vehicle Vehicle Low Mid High 

Control Control Dose Dose Dose 

0/10 (0) 0/8 (0) 0/32 (0) 0/22 (0) 1/8 (13) 

N.S. N.S. — — — 

Infinite — — 0.059 — — Infinite — — __ 

65 

0/10 (0) 0/8 (0) 5/34 (15) 0/29 (0) 1/14 (7) 

N.S. N.S. N.S. — — 

__ 

Infinite Infinite 

0.415 0.034 — Infinite Infinite — 

70 80 — — — 
Table E2. Time-adjusted analyses of the Incidence of Primary Tumors in Female Rats
 

Given Intraperitoneal Injections of Acronycine3

, Using Vehicle Controls
 

(continued)
 

a

Treated groups received doses of 3.75, 7.5, or 15 mg/kg by intraperitoneal injection.
 

"Number of tumor-bearing animals/number of animals examined at site (percent), based on
 

animals that survived at least as long as the number of weeks on study shown in parentheses
 

after the description of morphology.
 

c

Beneath the incidence of tumors in the mid- and high-dose control group is the probability
 

level for the Cochran-Armitage test when P < 0.05 using only the mid- and high-dose groups in
 

the trend analysis; otherwise, not significant (N.S.) is indicated. Beneath the incidence
 

of tumors in a treated group is the probability level for the Fisher exact test for the
 

comparison of that treated group with its appropriate control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

"A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

^The 95% confidence interval of the relative risk between each treated group and its appropriate
 

control group.
 

APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN MICE
 

GIVEN INTRAPERITONEAL INJECTIONS
 

OF ACRONYCINE
 

145
 

Table Fl. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Mice
 

Given Intraperitoneal Injections of Acronycinea

, Using Vehicle Controls
 

Topography: Morphology 

Low -Dose 

Vehicle 

Control 

Hematopoietic System: Lymphoma^3

 (33) 13/17 (76) 

P Valuesc » d 

Relative Risk (Vehicle Control)6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 33 

Multiple Organs: Sarcoma, NOSb (52) 0/5 (0) 

P Values0 >d 

Relative Risk (Vehicle Control)6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Low
 

Dose
 

10/37 (27)
 

P < O.OOl(N)
 

0.353
 

0.218
 

0.687
 

89
 

2/33 (6)
 

N.S.
 

Infinite
 

0.057
 

Infinite
 

81
 
Table Fl. Time-adjusted Analyses of the Incidence of Primary Tumors in Male Mice
 

Given Intraperitoneal Injections of Acronycine3, Using Vehicle Controls
 

(continued)
 

a

The low-dose group received 2 rag/kg.
 

^Number of tumor-bearing animals/number of animals examined at site (percent), based on
 

animals that survived at least as long as the number of weeks on study shown in parentheses
 

after the description of morphology.
 

c

Beneath the incidence of tumors in a treated group is the probability level for the
 

Fisher exact test for the comparison of the treated group with the vehicle-control
 

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

"A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

e

The 95% confidence interval of the relative risk between the treated group and the control
 

group.
 
Table F2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Mice
 

Given Intraperitoneal Injections of Acronycinea, Using Vehicle Controls
 

Topography: Morphology 

Low-Dose 

Vehicle 

Control 

Hematopoietic System: Lymphomab (28) 19/19 (100) 

P Value sc » d 

Relative Risk (Vehicle Control)6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 28 

Multiple Organs: Sarcoma, NOSb (52) 0/2 (0) 

P Values c » d 

Relative Risk (Vehicle Control)6 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor —_ 

Low
 

Dose
 

6/37 (16)
 

P < 0.001 (N)
 

0.162
 

0.000
 

0.289
 

74
 

2/31 (6)
 

N.S.
 

Infinite
 

0.039
 

Infinite
 

85
 
Table F2. Time-adjusted Analyses of the Incidence of Primary Tumors in Female Mice
 

Given Intraperitoneal Injections of Acronycine3

, Using Vehicle Controls
 

(continued)
 

a

The low-dose group received 2 mg/kg.
 

^Number of tumor-bearing animals/number of animals examined at site (percent), based on
 

animals that survived at least as long as the number of weeks on study shown in parentheses
 

after the description of morphology.
 

c

Beneath the incidence of tumors in a treated group is the probability level for the
 

Fisher exact test for the comparison of the treated group with the vehicle-control
 

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

^A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

e

The 95% confidence interval of the relative risk between the treated group and the control
 

group.
 
Review of the Bioassay of Acronycine* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

March 7, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be
 

exposed,, The members of the Clearinghouse have been drawn
 

from academia, industry, organized labor, public interest
 

groups, State health officials, and quasi-public health and
 

research organizations. Members have been selected on the
 

basis of their experience in carclnogenesis 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 carcinogenicity. It is in
 

this context that the below critique is given on the bioassay
 

of Acroriycine for carcinogenicity.
 

The primary reviewer for the report on. the bioassay of
 

Acronycine described the experimental design and conditions
 

under which Acronycine was tested. A dose-related incidence
 

of osteosarcomas occurred in the high dose male rats; one
 

osteosarcoma was observed among the treated females. Other
 

tumors were reported in the peritoneal cavity in both sexes
 

of treated rats. A statistically significant increase also
 

was found in the incidence of mammary gland tumors in treated
 

female rats. Although survival was inadequate to evaluate
 

the carcinogenicity of Acronycine in mice, increases in
 

lymphomas were observed among the treated animals. The
 

primary criticism of the study was the use of excessively
 

high doEie levels. Since Acronycin Is used as a chemotherapeutic agent, the primary reviewer said that it should
 

receive special consideration in assessing human risk.
 

The secondary reviewer commented that Acronycine was
 

probably carcinogenic, although he said the study was
 

151
 
deficient. Another Subgroup member agreed with the conclusion
 

given in the report. However, he felt that the value of the
 

study was diminished as a result of the excessively high
 

dose levels administered and the fact that animals were housed
 

in the same room in which other chemicals were under study.
 

A motion was made that the report on the bioassay of
 

Acronycine be accepted as written. The motion was seconded
 

and approved unanimously.
 

Members present were:
 

Gerald N. Wogan (Chairman), Massachusetts Institute of
 

Technology
 

Arnold Brown, Mayo Clinic
 

E. Cuyler Hammond, American Cancer Society
 

Joseph Highland, Environmental Defense Fund
 

Henry Pitot, McArdle Laboratory
 

George Roush, Jr., Monsanto Company
 

Michael Shimkin, University of California at San Diego
 

*	 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.
 

152
 

•ftll.S. GOVERMMEN T PRINTIN G OFFICE : 1978-260-899/3087
 

DHEW Publication No. (NIH) 78-849