National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 67 

1978 

BIOASSAY OF 

A MIXTURE OF ASPIRIN, 

PHENACETIN, AND CAFFEINE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 8003-03-0 

NCI-CG-TR-67 

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

National Institutes of Health 

Public Health Service 

BIOASSAY OF
 

A MIXTURE OF ASPIRIN, PHENACETIN,
 

AND CAFFEINE
 

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) 78-1317 

REPORT ON THE BIOASSAY OF APC
 

FOR POSSIBLE CARCINOGEN1CITY
 

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 APC
 

conducted for the Carcinogenesis Testing Program, Division of Cancer
 

Cause and Prevention, National Cancer Institute (NCI), National Institutes of Health, Bethesda, Maryland. This is one of a series of
 

experiments designed to determine whether selected chemicals have
 

the capacity to produce cancer in animals. Negative results, in
 

which the test animals do not have a significantly greater incidence
 

of cancer than control animals, do not necessarily mean the test
 

chemical is not a carcinogen because the experiments are conducted
 

under a limited set of circumstances. Positive results demonstrate
 

that the test chemical is carcinogenic for animals under the conditions of the test and indicate a potential risk to man. The actual
 

determination of the risk to man from animal carcinogens requires a
 

wider analysis.
 

CONTRIBUTORS: This bioassay of APC was conducted by Mason Research
 

Institute, Worcester, Massachusetts, initially under direct contract
 

to the NCI and currently under a subcontract to Tracor Jitco, Inc.,
 

prime contractor for the NCI Carcinogenesis Testing Program.
 

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

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

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

by Mr. G. Wade (3) and Ms. E. Zepp (3). Chemical analysis was performed by Midwest Research Institute (4) and the analytical results
 

were reviewed by Dr. N. Zimmerman (5).
 

Histopathologic examinations were performed by Dr. D. W. Hayden
 

(3) and Dr. A. S. Krishna Murthy (3) at the Mason Research Institute,
 

and the diagnoses included in this report represent the interpretation of these pathologists. Histopathology findings and reports were
 

reviewed by Dr. R. L. Schueler (6).
 

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

statistical analysis was performed by Mr. W. W. Belew (5) and Dr. J.
 

R. Joiner (6), using methods selected for the Bioassay Program by Dr.
 

J. J. Gart (8).
 

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

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

(5), the task leader, Dr. M. R. Kornreich (5), the senior biologist,
 

Ms. P. Walker (5) and the technical editor, Ms. P. A. Miller (5).
 

The final report was reviewed by members of the participating organizations .
 

The statistical analysis was reviewed by members of the Mathematical Statistics and Applied Mathematics Section of the NCI: Dr.
 

J. J. Gart (8), Mr. J. Nam (8), Dr. H. M. Pettigrew (8), and Dr. R.
 

E. Tarone (8).
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

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

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

A. Griesemer (1), Dr. H. A. Milman (1), Dr. T. W. Orme (1), Dr. R. A.
 

Squire (1,9), Dr. J. M. Ward (l), and Dr. C. E. Whitmire (1).
 

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

2.	 Now with the Naylor Dana Institute for Disease Prevention, American Health Foundation, Hammon House Road, Valhalla, New York.
 

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

4.	 Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

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

Boulevard, McLean, Virginia.
 

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

Maryland.
 

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

Rockville, Maryland.
 

8.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

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

IV
 
9.	 No wit w h the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

V 

SUMMARY
 

A bioassay of a mixture of aspirin, phenacetin, and caffeine
 

(APC) for possible carcinogenicity was conducted using Fischer 344
 

rats and B6C3F1 mice. APC was administered in the feed, at either
 

of two concentrations, to groups of 50 male and 49 or 50 female animals of each species. For each species, 50 animals of each sex were
 

placed on test as controls. The high dose used in the chronic study
 

for the male and female rats and mice was 1.4 percent. The low dose
 

administered to the male and female rats and mice was 0.7 percent.
 

After a 78-week period of compound administration, observation of
 

the rats continued for up to an additional 35 weeks and observation
 

of the mice continued for an additional 16 weeks.
 

No significant association was established between administration
 

of APC and mortality in rats or female mice; however, there was a significant positive association between treatment and mortality in male
 

mice. For both species the survival in all groups was adequate for
 

statistical analysis of tumor incidence.
 

In rats, a variety of endocrine tumors were observed with greater
 

frequency in the male rats treated with APC than in the male control
 

rats. These same tumors were not observed with similar frequencies
 

in the female rats or in the mice of either sex. The endocrine tumors observed most frequently in the treated male rats were adenomas
 

and carcinomas of the pituitary gland. The incidences of these tumors
 

proved to be statistically inconclusive.
 

In rats, a transitional-cell carcinoma of the bladder was observed in one low dose and one high dose females. A tubular-cell
 

adenocarcinoma of the kidney was seen in one low dose female and one
 

low dose male. A fifth neoplasm, a transitional-cell papilloma of
 

the kidney, was seen in one high dose female. The occurrence of these
 

urinary tumors, although considered important, was not statistically
 

significant.
 

In mice, there was no statistically significant positive association between APC administration and the incidence of tumors in either
 

sex.
 

Under the conditions of this bioassay evidence was not sufficient
 

for the carcinogenicity of APC in Fischer 344 rats or in B6C3F1 mice.
 

Vll
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 5
 

A. Chemicals 5
 

B. Dietary Preparation 5
 

C. Animals 7
 

D. Animal Maintenance 7
 

E. Selection of Initial Concentrations 9
 

F. Experimental Design 10
 

G. Clinical and Histopathologic Examinations 13
 

H. Data Recording and Statistical Analyses 15
 

III. CHRONIC TESTING RESULTS: RATS 20
 

A. Body Weights and Clinical Observations 20
 

B. Survival 20
 

C. Pathology 23
 

D. Statistical Analyses of Results 26
 

IV. CHRONIC TESTING RESULTS: MICE 37
 

A. Body Weights and Clinical Observations 37
 

B. Survival 37
 

C. Pathology 40
 

D. Statistical Analyses of Results 40
 

V. DISCUSSION 46
 

VI. BIBLIOGRAPHY 49
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH APC A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH APC B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH APC C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH APC D-l
 

IX
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF APC 6 

2 GROWTH CURVES FOR APC CHRONIC STUDY 

RATS 21 

3 SURVIVAL COMPARISONS OF APC CHRONIC 

STUDY RATS 22 

4 GROWTH CURVES FOR APC CHRONIC STUDY 

MICE 38 

5 SURVIVAL COMPARISONS OF APC CHRONIC 

STUDY MICE 39 

LIST OF TABLES 

Table Number Page 

1 DESIGN SUMMARY FOR FISCHER 344 RATS— 

APC FEEDING EXPERIMENT 11 

2 DESIGN SUMMARY FOR B6C3F1 MICE--APC 

FEEDING EXPERIMENT 12 

3 ANALYSES OF THE INCIDENCE OF PRIMARY 

TUMORS AT SPECIFIC SITES IN MALE RATS 

TREATED WITH APC 27 

4 ANALYSES OF THE INCIDENCE OF PRIMARY 

TUMORS AT SPECIFIC SITES IN FEMALE 

RATS TREATED WITH APC 31 

5 APC-TREATED MALE RATS WITH MORE THAN 

ONE ENDOCRINE TUMOR 36 

6 ANALYSES OF THE INCIDENCE OF PRIMARY 

TUMORS AT SPECIFIC SITES IN MALE MICE 

TREATED WITH APC 41 

7 ANALYSES OF THE INCIDENCE OF PRIMARY 

TUMORS AT SPECIFIC SITES IN FEMALE 

MICE TREATED WITH APC 43 

x
 
LIST OF TABLES (Concluded) 

Table Number Page 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH APC A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH APC A-8 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH APC B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH APC B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH APC C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH APC C-8 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH APC D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH APC D-9 

XI
 

I. INTRODUCTION
 

APC (NCI No. C02697), an abbreviation often used for mixtures
 

of aspirin, phenacetin and caffeine, is a nonprescription analgesic
 

preparation sold for relief of headache, muscular aches and pains,
 

arthritis, and other common afflictions. The active constituents of
 

the drug mixture were selected for bioassay by the National Cancer
 

Institute because they have been implicated in the induction of human
 

renal carcinomas through clinical studies performed in Switzerland
 

and Scandinavia (Weisburger, 1975; Juusela, 1973).
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this preparation is 2-(acetyloxy)-benzoic acid, mixture with N-(4-ethoxy-phenzl) acetamide and 3,7-dihydro-l,3,7-trime­

*
 thyl-lH-purine-2,6-dione. Other names for the various components
 

are acetylsalicylic acid and acetol for aspirin; acetophenetidine and
 

p-acetophenetidide for phenacetin; and theine and 1,3,7-trimethylxanthine for caffeine.
 

Aside from its well-known analgesic properties, APC is antipyretic and anti-inflammatory and also acts as a central stimulant
 

(Windholz, 1976). Thus, for many conditions (such as colds, osteoarthritis, rheumatoid arthritis, and so forth) the drug mixture
 

serves a multifold purpose.
 

Precise production figures for APC preparations are not available; however, the U.S. International Trade Commission (1976)
 

*
 

The CAS registry number is 8003-03-0.
 

1 
3 3
 

reported a production of 25,434 x 10 pounds and 9835 x 10 pounds of
 

aspirin and acetanilide derivatives, respectively, in 1975.
 

According to the Handbook of Non-Prescription Drugs (1977),
 

seven products that contain aspirin, phenacetin, and caffeine are
 

presently on the market; two of these products contain acetaminophen
 

(an acetanilide derivative related to phenacetin) in addition to the
 

other ingredients. The preparations are in tablet, capsule, or powder
 

form and usually contain approximately 227 mg of aspirin, 160 mg of
 

phenacetin, and 32 mg of caffeine per unit dose. One preparation
 

contains 455 mg of aspirin and 325 mg of phenacetin while another
 

contains only 15 mg of caffeine.
 

Exposure to APC is both widespread and extensive because of its
 

ei'ficacy and ease of availability. Use patterns range from an occasional tablet to authorized ingestion of as many as 30 tablets per
 

day in rheumatoid or other conditions requiring large doses of anti-


inflammatory and analgesic agents (Barr and Perro, 1973). In the
 

latter instances, as much as 4.6 kg of APC may be ingested annually.
 

Consumption of large quantities of APC also occurs among those who
 

abuse these drugs for relatively minor ailments.
 

Adverse health effects attributable to aspirin, phenacetin, and
 

caffeine may be expected to occur following excessive exposure to
 

these compounds in APC mixtures (Stecher, 1968). Aspirin has been
 

classified as moderately toxic following acute ingestion and slightly
 

toxic as an allergen, while phenacetin and caffeine are considered
 

2 
moderately toxic following both acute and chronic ingestion (Sax,
 

1975).
 

A variety of gastrointestinal disturbances and/or central nervous
 

system (CNS) effects have been reported following ingestion of average
 

to large doses of all three compounds by humans (Sax, 1975; Stecher,
 

1968). Excessive ingestion of aspirin may result in gastric ulcer,
 

and abuse of aspirin in the form of APC preparations has reportedly
 

led to a significant increase in the incidence of gastric ulcer among
 

populations where abuse of the drug is rife (Medical Journal of
 

Australia, 1976). Hematological disorders including hemolytic anemias
 

and leucocytosis, among others, are associated with prolonged ingestion of phenacetin (Stecher, 1968). Convulsions, coma, circulatory
 

collapse, respiratory failure, and death may result from overdoses of
 

either aspirin or phenacetin (Stecher, 1968).
 

Analgesic nephropathy, a form of renal disease characterized by
 

renal papillary necrosis and subsequent renal failure, is part of a
 

wider clinical syndrome associated with the abuse of APC mixtures by
 

humans (Nanra, 1976). Aside from the gastrointestinal, CNS, and hematological disorders mentioned previously, neuropsychiatric disorders,
 

ischemic heart disease, toxemia of pregnancy, and premature aging are
 

also observed as part of this syndrome (Nanra, 1976). Although controversy continues about the relative roles of aspirin and phenacetin
 

in the genesis of this disease, evidence suggests that phenacetin is
 

necessary for the major nephrotoxic effect of APC with aspirin having
 

3 
an additive effect in the induction of renal damage (Caughay et al.,
 

1974; Macklon et al., 1974; Gault, 1975; Nanra, 1976).
 

Evidence suggests that APC may be carcinogenic in humans
 

(Juusela, 1973). Three out of seventeen patients with renal pelvic
 

carcinoma were definite abusers of APC preparations while two other
 

patients had also possibly abused the drug. In addition, long-term
 

feeding of phenacetin to female Sprague-Dawley rats (0.535 percent
 

of the diet for 110 weeks) was associated with increased incidences
 

of mammary carcinoma and carcinoma of the ear duct (Johansson and
 

Angervall, 1976).
 

4 
II. MATERIALS AND METHODS 

A. Chemicals
 

Aspirin, phenacetin, and caffeine (Figure 1) were purchased from
 

J. T. Baker Chemical Company. Analysis was performed by Midwest
 

Research Institute. Melting points were determined and comparisons
 

with the literature values are given below:
 

Melting Point Melting Point 

Chemical Literature (°C) Tested Compound (°C) 

Aspirin 56.7 56.7 - 57.2 

Phenacetin 56.7 ­ 57.2 56.1 - 57.2 

Caffeine 80.6 ­ 81.1s 80.6 ­ 81.1s 

(s = Sublimes) 

Thin-layer chromatography with ultraviolet visualization showed only
 

one spot for each compound. Results of both melting point determinations and thin-layer chromatography suggested that all compounds were
 

of high purity.
 

Aspirin, phenacetin and caffeine were combined in the ratio of
 

50:46:4 percent by weight. For the remainder of this report, this
 

mixture is referred to as APC.
 

B.	 Dietary Preparation
 

The basal laboratory diet for both treated and control animals
 

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

APC was administered to the treated animals as a component of the
 

diet. The chemical was mixed in the diet using a 6 kg capacity
 

5 
ASPIRIN

 

PHENACETIN 

CAFFEINE 

FIGURE 1

 

CHEMICAL STRUCTURES OF ASPIRIN, PHENACETIN, CAFFEINE 

6 
Patterson-Kelly stainless steel twin-shell V-blender. The treated
 

diets were prepared once weekly and stored in double plastic bags in
 

the dark at 4°C.
 

C. Animals
 

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

through contracts of the Division of Cancer Treatment, National Cancer
 

Institute. Animals of both species were supplied by Charles River
 

Breeding Laboratories, Inc., Wilmington, Massachusetts. Mice were
 

obtained in one shipment, while rats were received in two (one for
 

treated groups and one for controls).
 

Upon arrival, a sample of animals was examined for nematode
 

infestation and other signs of disease. The remaining animals were
 

quarantined for 2 weeks prior to initiation of test. Animals were
 

assigned to groups and distributed among cages so that average body
 

weight per cage was approximately equal for a given sex and species.
 

D. Animal Maintenance
 

All animals were housed by species in rooms having a temperature
 

®
 

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

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

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

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

Rats were housed five per cage by sex in stainless- and
 

galvanized-steel wire-mesh cages suspended above newspapers for the
 

7 
first 11 months of the bioassay. During this time the newspapers
 

were replaced daily and the racks and cages were washed weekly. For
 

the remainder of the bioassay, rats were housed in suspended polycarbonate cages equipped with disposable nonwoven fiber filter sheets.
 

Corncob bedding, clean cages, and clean stainless steel cage racks
 

were provided twice weekly. Disposable filters were replaced with
 

the same frequency.
 

Mice were housed by sex, ten per cage for the first 18 months
 

and five per cage thereafter, in shoe box type polycarbonate cages
 

fitted with stainless steel lids and nonwoven fiber filter bonnets.
 

®
 Hardwood chips (Ab-sorb-dri , Wilner Wood Products Company, Norway,
 

®
 

Maine) were supplied as bedding for the first 8 months. SAN-I-CEL
 

(Paxton Processing Company, Paxton, Illinois) corncob bedding was
 

utilized during months 9 through 20 and was replaced, due to the
 

increasing dust content of SAN-I-CEL , by Bed-o-Cobs (The Andersons
 

Cob Division, Maumee, Ohio) for the remainder of the bioassay. Clean
 

cages, lids, and bedding were provided three times per week for the
 

first 18 months and twice weekly thereafter. Reusable filters and
 

pipe racks were sanitized once every 2 weeks throughout the study.
 

Water was available from 250 ml water bottles fitted with rubber
 

stoppers and stainless steel sipper tubes. Glass water bottles were
 

utilized for the first 4 months for rats and the first 9 months for
 

mice. These were then replaced by polycarbonate bottles for the remainder of the bioassay. Bottles were replaced twice weekly. Wayne
 

8 
Lab-Blox®was used throughout the bioassay. Water and the treated or
 

untreated food were available ad libitum to both rats and mice.
 

All rats used in this study were housed in a room with other
 

*
 rats receiving diets containing 4-nitroanthranilic acid (619-17-0);
 

5-nitro-o-toluidine (99-55-8); hydrazobenzene (530-50-7); 3-amino-9­

ethylcarbazole hydrochloride; 2,4-diaminoanisole sulfate (615-05-4);
 

2-aminoanthraquinone (117-79-3); 3-amino-4-ethoxyacetanilide (17026­

81-2); 6-nitrobenzimidazole (94-52-0); and 1-nitronaphthalene (86-57­

7).
 

All mice in this bioassay were housed with other mice receiving
 

diets containing the following compounds: acetylaminofluorene (53­

96-3); amitrole (61-82-5); 2-methyl-l-nitroanthraquinone (129-15-7);
 

N,N-dimethyl-p-nitrosoaniline (138-89-6); 2,4-dinitrotoluene (121-14­

2); 3-amino-4-ethoxyacetanilide (17026-81-2); 3-nitro-p-acetophenetide (1777-84-0); 4-nitroanthranilic acid (619-17-0); urea (57-13-6);
 

N-butylurea (592-31-4); sodium nitrite (7632-00-0); dulcin (150-69­

6); acetamide (60-35-5); p-tolylurea (622-51-5); and hexanamide
 

(628-02-4).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

APC for administration to treated animals in the chronic studies,
 

subchronic toxicity tests were conducted with both rats and mice.
 

*
 

CAS registry numbers are given in parentheses,
 

9 
Animals of each species were distributed among six groups, each consisting of five males and five females. APC was incorporated into
 

the basal laboratory diet and fed ad libitum to five of the six rat
 

groups and five of the six mouse groups in concentrations of 0.09,
 

0.18, 0.36, 0.71, and 1.42 percent. The sixth group of each species
 

served as a control group, receiving only the basal laboratory diet.
 

The dosed dietary preparations were administered for a period of 4
 

weeks, followed by a 2-week observation period during which all
 

animals were fed the basal diet.
 

A dosage inducing no mortality or mean body weight depression in
 

either sex was to be selected as the initial high dose in the chronic
 

bioassay. No mortality, compound-related gross abnormalities, or
 

mean body weight depression were reported for any of the treated animals. The initial high concentration selected for use in the chronic
 

bioassay for both rats and mice was 1.4 percent, the highest concentration administered in the subchronic test.
 

F. Experimental Design
 

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

treated and untreated observation periods) are summarized in Tables
 

1 and 2.
 

The low dose, high dose, and control rats were all approximately
 

6 weeks old at the time they were placed on test. The high and low
 

10
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

APC FEEDING EXPERIMENT
 

INITIAL APC OBSERVATION PERIOD 

GROUP CONCENTRATION TREATED UNTREATED 

SIZE (PERCENT) (WEEKS) (WEEKS) 

MALE 

CONTROL 50 0 0 109 

LOW DOSE 50 0.7 78 

0 34 

HIGH DOSE 50 1.4 78 

0 34 

FEMALE 

CONTROL 50 0 0 109 

LOW DOSE 50 0.7 78 

0 34 

HIGH DOSE 49 1.4 78 

0 35 

11
 
GROUP 

SIZE 

MALE 

CONTROL 50 

LOW DOSE 50 

HIGH DOSE 50 

FEMALE 

CONTROL 50 

LOW DOSE 50 

HIGH DOSE 50 

TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

APC FEEDING EXPERIMENT
 

INITIAL APC OBSERVATION PERIOD
 

CONCENTRATION
 

(PERCENT)
 

0
 

0.7
 

0
 

1.4
 

0
 

0
 

0.7
 

0
 

1.4
 

0
 

12
 

TREATED
 

(WEEKS)
 

0
 

78
 

78
 

0
 

78
 

78
 

UNTREATED
 

(WEEKS)
 

96
 

16
 

16
 

96
 

16
 

16
 
concentrations of APC administered to males and females were 1.4 and
 

0.7 percent, respectively. These concentrations were administered
 

in the diet for a total of 78 weeks followed by 34 or 35 weeks of observation during which the untreated diet was available.
 

The low dose, high dose, and control mice were all approximately
 

6 weeks old at the time they were placed on test. The high and low
 

concentrations of APC administered to males and females were 1.4 and
 

0.7 percent, respectively. These concentrations were administered in
 

the diet for a period of 78 weeks followed by an observation period
 

of 16 weeks during which the untreated diet was available.
 

Both rat and mouse control groups received the untreated basal
 

diet and were maintained and observed in a manner similar to the
 

treated groups.
 

G. Clinical and Histopathologic Examinations
 

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

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

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

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

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

the bioassay and for three consecutive days each month thereafter.
 

The presence of tissue masses and lesions was determined by monthly
 

observation and palpation of each animal.
 

13
 
A necropsy was performed on each animal regardless of whether it
 

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

bioassay. The animals were euthanized by carbon dioxide inhalation,
 

and were immediately necropsied. The histopathologic examination
 

consisted of gross and microscopic examination of major tissues,
 

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

possible, from animals found dead.
 

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

nodes, thymus, heart, salivary gland, liver, gallbladder (mice),
 

pancreas, esophagus, stomach, small intestine, large intestine,
 

kidney, urinary bladder, pituitary, adrenal, thyroid, parathyroid,
 

testis, prostate, seminal vesicle, brain, muscle, ear, uterus,
 

mammary gland, and ovary.
 

Tissues for which slides were prepared were preserved in 10 percent buffered formalin, embedded in paraffin, sectioned,, and stained
 

with hematoxylin and eosin prior to microscopic examination. An
 

occasional section was subjected to special staining techniques for
 

more definitive diagnosis.
 

A few tissues were not examined for some animals, particularly
 

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

preclude histopathologic interpretation. Thus, the number of animals
 

14
 
for which particular organs, tissues, or lesions were examined microscopically varies and does not necessarily represent the number of
 

animals that were placed on experiment in each group.
 

H. Data Recording and Statistical Analyses
 

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

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

recommended by the International Union Against Cancer (Berenblum,
 

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

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental results
 

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect on
 

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

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

15
 
testing a dose-related trend. One-tailed P-values have been reported
 

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

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

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

histologically. However, when macroscopic examination was required
 

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

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

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

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

16
 
used, it is discussed in the narrative section. It is not, however,
 

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

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

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

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

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

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

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

tumors. In this analysis, deaths that occurred before the first tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

17
 
during which animals died naturally or were sacrificed was entered as
 

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

these curves were used for two groups; Tarone's extension to testing
 

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

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

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

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


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

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

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

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

t c t
 

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

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

c
 

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

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

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

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

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

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

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

18
 
would be within the interval calculated from the experiment. When
 

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

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

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

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

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

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

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

detected under the conditions of this test.
 

19
 
III. CHRONIC TESTING RESULTS: RATS
 

A.	 Body Weights and Clinical Observations
 

The mean body weights for both high and low dose male and female
 

rats were consistently lower than those for controls (Figure 2). The
 

only clinical abnormalities observed in the rat groups were a small
 

crusted cutaneous lesion in one control male and three palpable subcutaneous nodules in low dose males.
 

B.	 Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and APC-treated groups are shown in Figure 3.
 

For both male and female rats the Tarone test did not indicate
 

a significant association between dosage and mortality. For males,
 

despite the sacrifice of five rats from each group in week 78, 78
 

percent (39/50) of the high dose and 66 percent (33/50) of the controls actually survived until the end of the study. Eighty percent
 

(40/50) of the low dose males survived until the end of the study.
 

For females, despite the sacrifice of five rats from each group
 

in week 78, 78 percent (39/49) of the high dose and 70 percent (35/50)
 

of the controls survived until the end of the study. Seventy-four
 

percent (37/50) of the low dose females survived until the end of the
 

study.
 

Thus, the actual survival in both sexes was adequate for meaningful statistical analyses of tumor incidence.
 

20
 
FIGURE 2

 

GROWTH CURVES FOR ARC CHRONIC STUDY RATS 

21 
FIGURES

 

SURVIVAL COMPARISONS OF APC CHRONIC STUDY RATS 

22 
C.	 Pathology
 

Histopathologic findings on neoplasms in rats are tabulated in
 

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

tabulated in Appendix C (Tables Cl and C2).
 

The only tumors which were observed more frequently in APC-


treated rats than in the controls occurred in the endocrine system.
 

Endocrine tumors were found in more low dose males (31/50 or 62 percent) and high dose males (26/49 or 53 percent) than were found in
 

control males (16/47 or 34 percent). Many of these male rats had
 

more than one type of endocrine tumor. In female rats, total endocrine tumor incidence was not affected by APC feeding.
 

The most common endocrine tumors in both sexes were those of
 

the pituitary gland. These consisted of adenomas defined as well-


circumscribed nodules in the anterior lobe composed of a single cell
 

type which compressed adjacent normal tissue and of carcinomas which
 

exhibited nuclear pleomorphism and atypicality and some degree of
 

local invasion. Pituitary tumors were found in 8/47 (17 percent)
 

control males, 18/47 (38 percent) low dose males, and in 12/44 (27
 

percent) high dose males. In female rats, there were fewer pituitary
 

tumors in the animals that were fed APC (15/43 or 35 percent high
 

dose, 14/44 or 32 percent low dose) than in the controls (20/46 or 43
 

percent).
 

Among male rats adrenal tumors were observed more frequently in
 

rats fed APC in their diets than in controls. The tumor found most
 

23
 
frequently in the adrenal was a pheochromocytoma consisting of a
 

rounded nodule of atypical medullary cells which enlarged the
 

medulla, compressed adjacent normal pheochromocytes and frequently
 

distorted the surrounding cortex. Adrenal pheochromocytomas were
 

observed in 7/47 (15 percent) control males, 17/49 (35 percent) low
 

dose males, and 9/48 (19 percent) high dose males. In addition,
 

well-circumscribed adrenal cortical adenomas were observed in 0/47
 

male controls, 3/49 (6 percent) low dose males, and 1/48 (2 percent)
 

high dose males.
 

Both follicular-cell and C-cell tumors were found in the thyroid,
 

Treatment with APC did increase the incidence of C-cell tumors in
 

thyroids of male and female rats.
 

Other endocrine tumors occurred in males only. They consisted
 

of one adenoma of the parathyroid in a control animal and rare islet-


cell adenomas (2/47 or 4 percent high dose, 3/47 or 6 percent low
 

dose, 3/45 or 7 percent control) and carcinomas (1/47 or 2 percent
 

high dose, 1/45 or 2 percent control).
 

Because phenacetin had been linked with the occurrence of renal
 

papillary necrosis and tumors of the urinary tract in man (Johansson
 

et al., 1974), special attention was paid to the urinary system. The
 

chronic renal degenerative disease characteristic of old rats was
 

seen in all groups of animals, being more frequent in males than in
 

females. Its incidence was not affected by APC feeding.
 

24
 
Renal necrosis was not diagnosed in any male rat or in any
 

female of the control and low dose groups. Among the high dose
 

females, there were four rats with renal medullary necrosis, in two
 

cases associated with hyperplasia of the transitional-cell epithelium
 

of the pelvis. One instance of epithelial hyperplasia was noted in
 

the urinary bladder of a control male, another in the bladder of a
 

high dose male. The only nonneoplastic lesion of the bladder observed in the females was a calculus in a high dose female. This was
 

associated with hydronephrosis.
 

A transitional-cell carcinoma of the bladder was observed in one
 

low dose and one high dose females. A kidney tubular-cell adenocarcinoma was seen in one low dose female and in one low dose male. A
 

transitional-cell papilloma of the kidney was also seen in one high
 

dose female. However, it is concluded that this experiment produced
 

no conclusive evidence that AFC induced transitional-cell carcinomas
 

in any portion of the urinary tract. Two of the three high dose
 

female rats exhibiting renal medullary necrosis also showed benign
 

hyperplasia of the renal pelvic epithelium, a much lower incidence of
 

this lesion than that reported by Johansson and Angervall (1976) in
 

Sprague-Dawley females fed phenacetin alone.
 

The gastrointestinal tract was examined carefully for possible
 

toxic lesions which might be associated with aspirin. No nonneoplastic gastric lesions were noted in either the male or female controls. One instance of acute gastric inflammation was found in a
 

25
 
low dose male rat, and one case of focal gastric inflammation in a
 

high dose female. There were two instances of gastric epithelial
 

hyperplasia in high dose males.
 

Tumors of the stomach and intestine were only infrequently
 

observed. One mucinous adenocarcinoma of the colon was found in a
 

male control. This had metastasized to the stomach and pancreas.
 

One papillary adenocarcinoma of the ileum was found in a high dose
 

male. No gastric or intestinal tumors were found in any females.
 

APC was nephrotoxic in less than 10 percent of the high dose
 

female rats. Nephrotoxicity was not apparent in male rats.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

rats are summarized in Tables 3 and 4. The analysis for every type
 

of tumor that was observed in more than 5 percent of any of the APC-


dosed groups of either sex is included.
 

For male rats both the incidence of pituitary neoplasms and the
 

incidence of pheochromocytomas of the adrenal gland were high in the
 

low dose group. The Fisher exact test was significant both for the
 

pituitary tumors (P = 0.018) and the pheochromocytomas (P = 0.022) in
 

comparing low dose to control. The Fisher exact tests comparing high
 

dose rats to controls and the Cochran-Armitage tests, however, did
 

not indicate statistically significant results for either the pituitary tumors or the pheochromocytomas. For females no tests showed a
 

positive association between APC administration and tumor incidence.
 

26
 
___ 

TABLE 3 

ANALYSES OF THE INCIDENCE OF PRIMARY

SPECIFIC SITES IN MALE RATS TREATED

TOPOGRAPHY : MORPHOLOGY CONTROL 

Skin: Fibroma*5 0/47(0.00) 

P Values0 N.S . 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Hematopoietic System: Malignant Lymphoma 

or Leukemia" 5/47(0.11) 

P Values0 N.S . 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 85 

Liver: Neoplastic Nodule or Hepatocellular 

Carcinoma" 5/47(0.11) 

P Values0 N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 108 

 TUMORS AT 

 WITH APCa 

LOW 

DOSE 

3/50(0.06) 

N.S . 

Infinite 

0.569 

Infinite 

105 

4/50(0.08) 

N.S . 

0.752 

0.159 

3.285 

100 

4/50(0.08) 

N.S. 

0.752 

0.159 

3.285 

98 

HIGH 

DOSE 

0/49(0.00) 

N.S . 

2/49(0.04) 

N.S. 

0.389 

0.038 

2.214 

112 

1/49(0.02) 

N.S. 

0.192 

0.004 

1.627 

112 
TOPOGRAPHY : MORPHOLOGY 

Pituitary: Adenoma or Carcinoma NOS 

P Values0 

Departure from Linear Trend 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Adrenal: Cortical Adenoma 

P Values 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Adrenal: Pheochromocytoma 

P Values0 

Departure from Linear Trend 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

TABLE 3 (Continued) 

CONTROL 

8/47(0.17) 

N.S. 

P = 0.044 

108 

0/47(0.00) 

N.S. 

7/47(0.15) 

N.S. 

P = 0.016 

108 

LOW 

DOSE 

18/47(0.38) 

P = 0.018 

2.250 

1.045 

5.337 

98 

3/49(0.06) 

N.S. 

Infinite 

0.576 

Infinite 

112 

17/49(0.35) 

P = 0.022 

2.329 

1.021 

6.006 

102 

HIGH 

DOSE 

12/44(0.27) 

N.S. 

1.602 

0.668 

4.074 

1/48(0.02) 

N.S. 

Infinite 

0.053 

Infinite 

112 

9/48(0.19) 

N.S. 

1.259 

0.457 

3.655 

78 

112 
TABLE 3

TOPOGRAPHY : MORPHOLOGY
 

Thyroid: C-Cell Carcinomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma or C-Cell Carcinoma
 

P Values0
 

Relative Risk ( Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pancreatic Islets: Islet-Cell Adenoma
 

or Islet-Cell Carcinomab
 

P Values0
 

Relative Risk ( Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 (Continued) 

CONTROL
 

2/46(0.04)
 

N.S.
 

108
 

4/46(0.09)
 

N.S.
 

108
 

4/45(0.09)
 

N.S.
 

85
 

LOW
 

DOSE
 

5/48(0.10)
 

N.S.
 

2.396
 

0.414
 

24.185
 

98
 

9/48(0.19)
 

N.S.
 

2.156
 

0.652
 

8.969
 

98
 

3/47(0.06)
 

N.S.
 

0.718
 

0.112
 

4.010
 

112
 

HIGH
 

DOSE
 

5/46(0.11)
 

N.S.
 

2.500
 

0.436
 

25.197
 

95
 

8/46(0.17)
 

N.S.
 

2.000
 

0.580
 

8.488
 

95
 

3/47(0.06)
 

N.S.
 

0.718
 

0.112
 

4.010
 

112
 
TABLE 3 (Concluded) 

TOPOGRAPHY : MORPHOLOGY CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Testis: Interstitial-Cell Tumorb 

P Values0 

44/47(0.94) 

P < O.OOl(N) 

46/49(0.94) 

N.S. 

27/49(0.55) 

P< O.OOl(N) 

Departure from Linear Trend6 P = 0.005 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

1.003 

0.907 

1.109 

0.589 

0.511 

0.756 

Weeks to First Observed Tumor 78 100 104 

a

Treated groups received concentrations of 0.7 or 1.4 percent in feed.
 

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

c

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

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

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

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

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

e

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

group when P < 0.05.
 
_ __ 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH APCa
 

TOPOGRAPHY : MORPHOLOGY 

Hematopoietic System: Leukemia or 

Malignant Lymphoma'5 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Liver: Neoplastic Nodule or Hepato cellular 

Carcinoma" 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Pituitary: Adenoma or Carcinoma NOS 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

CONTROL 

5/48(0.13) 

N.S. 

_ — 

96 

0/47(0.00) 

N.S. 

20/46(0.43) 

N.S. 

94 

LOW 

DOSE 

8/50(0.16) 

N.S. 

1.536 

0.479 

5.571 

82 

4/47(0.09) 

N.S. 

Infinite 

0.929 

Infinite 

88 

14/44(0.32) 

N.S. 

0.732 

0.396 

1.319 

96 

HIGH 

DOSE 

1/49(0.02) 

N.S. 

0.196 

0.004 

1.662 

112 

3/45(0.07) 

N.S. 

Infinite 

0.631 

Infinite 

112 

15/43(0.35) 

N.S. 

0.802 

0.444 

1.420 

112 
TOPOGRAPHY : MORPHOLOGY
 

Adrenal: Cortical Adenoma or Cortical
 

Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Adenoma or
 

Follicular-Cell Carcinomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (Continued)
 

CONTROL
 

4/47(0.09)
 

N.S.
 

96
 

1/47(0.02)
 

N.S.
 

109
 

0/46(0.00)
 

N.S.
 

LOW
 

DOSE
 

4/48(0.08)
 

N.S.
 

0.979
 

0.193
 

4.964
 

112
 

2/48(0.04)
 

N.S.
 

1.958
 

0.105
 

113.119
 

108
 

3/46(0.07)
 

N.S.
 

Infinite
 

0.602
 

Infinite
 

108
 

HIGH
 

DOSE
 

2/44(0.05)
 

N.S.
 

0.534
 

0.050
 

3.523
 

113
 

2/44(0.05)
 

N.S.
 

2.136
 

0.115
 

123.141
 

112
 

2/43(0.05)
 

N.S.
 

Infinite
 

0.316
 

Infinite
 

112
 
TABLE

TOPOGRAPHY : MORPHOLOGY
 

Thyroid: C-Cell Adenoma or C-Cell Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polyp
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

4 (Continued) 

CONTROL
 

3/46(0.07)
 

N.S.
 

109
 

14/48(0.29)
 

P = O.OOl(N)
 

76
 

15/47(0.32)
 

P = 0.043(N)
 

78
 

LOW
 

DOSE
 

8/46(0.17)
 

N.S.
 

2.667
 

0.690
 

14. 739
 

101
 

7/50(0.14)
 

N.S.
 

0.480
 

0.180
 

1.156
 

105
 

14/48(0.29)
 

N.S.
 

0.914
 

0.463
 

1.796
 

96
 

HIGH
 

DOSE
 

4/43(0.09)
 

N.S.
 

1.426
 

0.255
 

9.221
 

112
 

2/49(0.04)
 

P = O.OOl(N)
 

0.140
 

0.016
 

0.563
 

113
 

7/46(0.15)
 

N.S.
 

0.477
 

0.182
 

1.119
 

78
 
TABLE 4 (Concluded) 

o
 

Treated groups received concentrations of 0.7 or 1.4 percent in feed.
 

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

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

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

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

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

The 95% confidence interval on the relative risk of the treated group to the control group.
 
Based upon these results the statistical conclusion is that
 

there was not adequate evidence to demonstrate that APC had a carcinogenic effect on Fischer 344 rats.
 

The possibility of a negative association between treatment
 

and incidence was noted for mammary fibroadenomas in females.
 

Interstitial-cell tumors of the testis were significant in males;
 

these tumors, however, are well known to have a high and variable
 

incidence (Cockrell and Garner, 1976).
 

Because many of the male rats were observed to have more than
 

one type of endocrine tumor, additional analyses were conducted based
 

upon the combinations of different endocrine tumors shown in Table 5.
 

No significant positive Fisher exact test results were observed when
 

the Bonferroni criterion was applied.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

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

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

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

absence of statistically significant results. It should also be
 

noted that many of the confidence intervals have an upper limit
 

greater than one, indicating the theoretical possibility of tumor
 

induction in rats by APC that could not be established under the
 

conditions of this test.
 

35
 
TABLE 5
 

APC-TREATED MALE RATS WITH MORE THAN ONE ENDOCRINE TUMOR
 

LOW HIGH
 

TYPE OF TUMOR CONTROL3 DOSE3 DOSE3
 

Pituitary Adenoma NOS and Adrenal
 

Pheochromocytoma 1/47 5/46 1/43
 

Pituitary Adenoma NOS and Adrenal
 

Cortical Adenoma 0/47 1/46 0/43
 

Pituitary Adenoma NOS and Thyroid
 

C-Cell Neoplasmb 2/46 1/45 1/41
 

Pituitary Adenoma NOS and Pancreatic
 

Islets: Islet-Cell Neoplasm0 1/45 1/44 0/42
 

Adrenal Fheochromocytoma and Thyroid
 

C-Cell Neoplasmb 1/46 4/47 1/45
 

Adrenal Pheochromocytoma and Parathyroid Adenoma NOS 1/24 0/26 0/25
 

Adrenal Pheochromocytoma and Adrenal
 

Cortical Adenoma 0/47 1/49 0/48
 

Adrenal Cortical Adenoma and Thyroid
 

C-Cell Neoplasmb 0/46 0/47 1/45
 

Thyroid C-Cell Neoplasmb and Thyroid
 

Follicular-Cell Adenoma 0/46 0/48 1/46
 

Pituitary Adenoma NOS and Adrenal
 

Pheochromocytoma and Pancreatic
 

Islet: Islet-Cell Neoplasm0 1/45 0/43 1/41
 

Pituitary Adenoma NOS and Adrenal
 

Pheochromocytoma and Thyroid
 

C-Cell Neoplasm 0/46 1/44 0/40
 

Pituitary Adenoma NOS and Adrenal
 

Pheochromocytoma and Thyroid C-Cell
 

Neoplasm and Adrenal Cortical
 

Adenoma 0/46 1/44 0/40
 

TJumber of animals with these tumors/number of animals with tissues
 

examined from all sites listed.
 

Thyroid: C-cell adenoma or C-cell carcinoma.
 p
 

Pancreatic Islets: Islet-cell adenoma or Islet-cell carcinoma.
 

36
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

The mean body weights in treated mice of both sexes were generally depressed in comparison with controls (Figure 4).
 

No clinical abnormalities were recorded for any treated or control mice.
 

B.	 Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and APC-treated groups are shown in Figure 5.
 

For male mice the Tarone test indicated a significant (P = 0.013)
 

positive association between dosage and mortality. The survival was
 

high, however, with 72 percent (36/50) of the high dose group surviving until the end of the study. Five high dose mice were sacrificed
 

in week 78. Eighty-eight percent (44/50) of the low dose and 84
 

percent (42/50) of the control group survived until the end of the
 

study. Five control group males were sacrificed in week 78.
 

For female mice there was no significant association between
 

dose and mortality. Eighty-two percent (41/50) of the high dose and
 

74 percent (37/50) of the control group females survived until the
 

end of the study, despite the sacrifice of five mice from each group
 

in week 78. Eighty-six percent (43/50) of the low dose group survived
 

until the end of the study.
 

In both sexes survival was adequate for meaningful statistical
 

analysis of tumor incidence.
 

37
 
FIGURE 4

 

GROWTH CURVES FOR ARC CHRONIC STUDY MICE 

38 
FIGURES

 

SURVIVAL COMPARISONS OF ARC CHRONIC STUDY MICE 

39 
C. Pathology
 

Histopathologic findings on neoplasms in mice are tabulated in
 

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

tabulated in Appendix D (Tables Dl and D2).
 

Results of this histopatologic examination did not indicate any
 

difference in tumor incidence among mice fed APC in the diet as
 

compared to control animals. In contrast to rats, tumors of the
 

endocrine system were infrequently observed in mice and were not
 

associated with APC administration. No tumors of the kidney or
 

urinary bladder were found in treated or control mice of either sex.
 

The results of the histopathologic examination for nonneoplastic
 

lesions (Appendix D) indicate that APC was not toxic to the kidneys
 

or gastrointestinal tracts of treated mice.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 6 and 7. The analysis for every type
 

of tumor that was observed in more than 5 percent of any of the
 

APC-dosed groups of either sex is included.
 

For both male and female mice there were no statistically significant positive associations between dosage and tumor incidence.
 

Based upon these results there was no conclusive evidence of the
 

carcinogenicity of APC in mice at the dose levels used in this
 

experiment.
 

40
 
__ _ 

_ __ 

TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH APCa
 

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar /Eronchiolar Adenoma or
 

Alveolar/Bronchiolar Carcinoma*5
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or Malignant
 

Lymphoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

6/48(0.13)
 

N.S.
 

—_
 

96
 

4/48(0.08)
 

N.S.
 

96
 

7/48(0.15)
 

N.S.
 

78
 

LOW
 

DOSE
 

9/46(0.20)
 

N.S.
 

1.565
 

0.542
 

4.924
 

94
 

7/48(0.15)
 

N.S.
 

1.750
 

0.477
 

7.660
 

93
 

11/46(0.24)
 

N.S.
 

1.640
 

0.637
 

4.551
 

94
 

HIGH
 

DOSE
 

12/47(0.26)
 

N.S.
 

2.043
 

0.778
 

6.082
 

94
 

4/48(0.08)
 

N.S.
 

1.000
 

0.196
 

5.071
 

94
 

6/47(0.13)
 

N.S.
 

0.875
 

0.261
 

2.813
 

94
 
TABLE 6 (Concluded) 

a 

Treated groups received concentrations of 0.7 or 1.4 percent in feed. 

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

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

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

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

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

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

TABLE 7
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH APCa
 

TOPOGRAPHY : MORPHOLOGY 

Lung: Alveolar /Bronchiolar Adenoma or 

Alveolar/Bronchiolar Carcinoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Hematopoietic System: Leukemia or 

Malignant Lymphoma'3 

P Values 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Liver: Hepatocellular Carcinoma 

P Values 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

CONTROL 

4/46(0.09) 

N.S. 

96 

5/48(0.10) 

N.S. 

96 

1/47(0.02) 

N.S. 

96 

LOW 

DOSE 

7/45(0.16) 

N.S. 

1.789 

0.491 

7.797 

94 

6/45(0.13) 

N.S. 

1.280 

0.350 

4.939 

94 

2/45(0.04) 

N.S. 

2.089 

0.113 

120.488 

94 

HIGH 

DOSE 

4/47(0.09) 

N.S. 

0.979 

0.193 

4.955 

94 

5/47(0:11) 

N.S . 

1.021 

0.251 

4.153 

94 

3/47(0.06) 

N.S . 

3.000 

0.251 

154.014 

93 
TABLE 7 (Concluded) 

TOPOGRAPHY : MORPHOLOGY CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Pituitary: Adenoma NOS 

P Values0 

2/42(0.05) 

N.S . 

4/38(0.11) 

N.S . 

2/41(0.05) 

N.S . 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

2.211 

0.336 

23.279 

1.024 

0.078 

13.545 

Weeks to First Observed Tumor 96 94 94 

Treated groups received concentrations of 0.7 or 1.4 percent in feed. 

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

Q 

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

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

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

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

The 95% confidence interval on the relative risk of the treated group to the control group. 
To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

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

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

Tables 6 and 7, the value one is included; this indicates the
 

absence of statistically significant results. It should also be
 

noted that all of the confidence intervals have an upper limit
 

greater than one, indicating the theoretical possibility of tumor
 

induction in mice by APC that could not be established under the
 

conditions of this test.
 

45
 
V. DISCUSSION
 

In both sexes of rats and in female mice there were no significant associations between administration of APC and mortality; however, there was a significant positive association between compound
 

administration and mortality in male mice. In both species, adequate
 

numbers of APC-treated animals survived sufficiently long for meaningful statistical analysis of tumor incidence.
 

In rats tumors of the endocrine system were observed almost twice
 

as often in treated males as in controls (i.e., 16/47 [34 percent],
 

31/50 [62 percent], and 26/49 [53 percent] in the control, low dose,
 

and high dose groups, respectively). The endocrine tumors most frequently observed were adenomas and carcinomas of the pituitary gland
 

(8/47 [17 percent], 18/47 [38 percent], and 12/44 [27 percent] in the
 

control, low dose, and high dose groups, respectively). Only one of
 

the Fisher exact tests, the one comparing low dose and controls, provided a statistically significant result and it was not supported by
 

significance from either the other male rat Fisher exact test or the
 

Cochran-Armitage test.
 

Pheochromocytomas of the adrenal were also a major contributor
 

to the frequency with which endocrine tumors were observed in male
 

rats. In the control, low dose, and high dose groups the incidences
 

of this tumor were 7/47 (15 percent), 17/49 (35 percent), and 9/48
 

(19 percent), respectively. The Fisher exact test for the comparison
 

of the incidence of adrenal pheochromocytomas in the low dose to that
 

46
 
in the control group was significant. The high dose to control Fisher
 

exact comparison, however, was not.
 

Johansson et al. (1974) indicated a relationship between phenacetin exposure and papillary necrosis and tumors of the urinary tract
 

in man. Renal medullary necrosis was observed in only four high dose
 

females. The presence of five neoplasms in the urinary system of rats
 

receiving APC was not considered sufficient evidence to incriminate
 

APC, since similar tumors occasionally appear in control animals in
 

this program.
 

The gastrointestinal tract was examined for the toxic lesions
 

which might have been expected to be associated with ingestion of
 

aspirin. One case of acute gastric inflammation was observed in a
 

low dose male rat and one case of focal gastric inflammation in a
 

high dose female rat.
 

Although adequate numbers of animals survived sufficiently long
 

for meaningful statistical analysis of tumor incidence, no effect of
 

APC administration on tumor incidence was observed and no conclusive
 

evidence was provided for APC-induced toxic lesions in the kidney,
 

stomach, or intestine.
 

Johansson and Angervall (1976) reported that long-term feeding
 

of phenacetin (0.535 percent of the diet for 110 weeks) to female
 

Sprague-Dawley rats produced mammary carcinoma in 5/30 test animals
 

(1/30 controls) and carcinoma of the ear duct in 4/30 test animals
 

(0/30 controls). Johansson and Angervall concluded that phenacetin
 

47
 
had weak carcinogenic activity in Sprague-Dawley rats. In the chronic
 

bioassay of APC performed at Mason Research Institute, Fischer 344
 

rats were fed phenacetin for 78 weeks, at levels of 0.644 percent of
 

the diet for the high dose groups and 0.322 percent of the diet for
 

low dose groups. No mammary gland carcinomas or carcinomas of the
 

ear duct were reported in rats fed APC in this bioassay, although one
 

mammary adenocarcinoma was observed in a female control rat. Johansson and Angervall also observed urothelial hyperplasia of the renal
 

papillae in 26/30 rats and dilation of the vasa recta in 28/30 rats.
 

In this APC bioassay, renal pelvic epithelial hyperplasias were
 

reported in 2/46 high dose female rats but no renal pelvic epithelial
 

hyperplasias were observed in other female rat groups or in male rat
 

groups. No vascular dilation was reported among rats in this APC
 

bioassay. Differences between histopathologic results of Johansson
 

and Angervall's phenacetin bioassay and results of this APC bioassay
 

could have been due to the different strains of rats used, to the
 

presence of aspirin and caffeine, or to expected variability in
 

spontaneous tumor incidence.
 

Under the conditions of this bioassay sufficient evidence was
 

not provided for the carcinogenicity of APC in Fischer 344 rats or
 

in B6C3F1 mice.
 

48
 
VI. BIBLIOGRAPHY
 

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

J. Wiley & Sons, New York, 1971.
 

Barr, W.H. and R.P. Perro, "Internal Analysis." Handbook of Non-


Prescription Drugs. The American Pharmaceutical Association,
 

Washington, B.C., pp. 36-50, 1973.
 

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

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

Union Against Cancer, Geneva, 1969.
 

Caughay, D.E., I.C. Isdale, J.M. Tweed, B.L.J. Tresdwell, J.K. Laing,
 

and J. Kirk, "Aspirin and the Kidney." British Medical Journal
 

_I:593-596, 1974.
 

Chemical Abstracts Service, The Chemical Abstracts Service (CAS)
 

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

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

Cockrell, B. and F.M. Garner, "Interstitial-Cell Tumor of the Testis
 

in Rats." Comparative Pathology Bulletin 8(2):2-4, 1976.
 

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

Co., Ltd., London, 1970.
 

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

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

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

Tests, Confidence Limits, and Adjustments for Stratification."
 

International Statistical Institute Review 3_9_: 148-169, 1971.
 

Gault, M.H., "The Clinical Course of Patients with Analgesic Nephropathy." Canadian Medical Association Journal 1:204-207, 1975.
 

Handbook of Non-Prescription Drugs, Fifth edition. "Products 10
 

Internal Analgesic." The American Pharmaceutical Association,
 

Washington, D.C., pp. 129-133, 1977.
 

Johansson, S. and L. Angervall, "Urothelial Changes in the Renal
 

Papillae in Sprague-Dawley Rats Induced by Long Term Feeding of
 

Phenacetin." Astor Pathologico et Microbiologico Scandinavia
 

Section A 84:375-383, 1976.
 

49
 
Johansson, S., L. Angervall, U. Bengtsson, and L. Wahlqvist, "Urothelial Tumors of the Renal Pelvis Associated with Abuse of
 

Phenacetin-containing Analgesics." Cancer 33:743-753, 1974.
 

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

Analgesic Abuse." Annales Chirurgiae et Gynaecologiae Fenmiae
 

£3:386-390, 1973.
 

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

Observations." Journal of the American Statistical Association
 

21:457-481, 1958.
 

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

"Carcinogenesis Bioassay Data System." Computers and Biomedical
 

Research 7:230-248, 1974.
 

Macklon, A.F., A.W. Craft, M. Thomson, and D.N. Kerr, "Aspirin and
 

Analgesic Nephropathy." British Medical Journal 1:597-600, 1974.
 

Medical Journal of Australia, "Aspirin and Chronic Gastric Ulcer."
 

Vol. I, pp. 104-105, 1976.
 

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

Co., New York, 1966.
 

Nanra, R.S., "Analgesic Nephropathy." Medical Journal of Australia
 

_!: 745-748, 1976.
 

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

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

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

Sax, N.I., Dangerous Properties of Industrial Materials. Van Nostrand
 

Reinhold Company, New York, 1975.
 

Stecher, P.G., editor, The Merck Index: An Encyclopedia of Chemicals
 

and Drugs, Eighth edition. Merck and Company, Inc., Rahway,
 

New Jersey, 1968.
 

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

j>2_:679-682, 1975.
 

U.S International Trade Commission, Synthetic Organic Chemicals.
 

United States Production and Sales, 1975. USITC Publication
 

804, Washington, D.C., 1977.
 

50
 
Weisburger, E., Chief, Carcinogen and Toxicology Branch, National
 

Cancer Institute, Bethesda, Maryland. Personal communication,
 

1975.
 

Windholz, M., editor, The Merck Index: An Encyclopedia of Chemicals
 

and Drugs, Ninth edition. Merck and Company, Inc., Rahway, New
 

Jersey, 1976.
 

51
 

Review of the Bioassay of a Mixture of
 

Aspirin, Phenacetin, and Caffeine (APC)* 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 carcinogenesis or related fields
 

and, collectively, provide expertise in chemistry, biochemistry,
 

biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse is charged with the responsibility of
 

providing a peer review of reports prepared on NCI-sponsored
 

bioassays of chemicals studied for carcinogenicity. It is in
 

this context that the below critique is given on the bioassay
 

of a mixture of Aspirin, Phenacetin, and Caffeine (APC) for
 

carcinogenicity.
 

The primary reviewer noted the higher incidence of
 

endocrine tumors in treated male rats than in the associated
 

controls, although the incidence was not statistically
 

significant. After briefly describing the experimental design
 

and conditions of test, he raised the question as to whether
 

the increased incidence of endocrine tumors was sufficient
 

cause to consider a retest of APC.
 

The secondary reviewer said that APCs are still commonly
 

used. He questioned the significance of the five urinary
 

tract transitional-cell neoplasms in the treated rats.
 

In reviewing the historical controls, a Program staff member
 

said that the incidence of renal papillomas in male rats
 

was 0 out of 255 and in females 1 out of 235. It was noted
 

that a statement in the report summary is needed in regard
 

to the urinary tract tumors. It was suggested that the
 

53
 
report summary conclusion be rewritten to indicate that the
 

evidence was insufficient to assess the carcinogenicity of
 

APC.
 

It was moved that the report on the bioassay of APC
 

be accepted with the appropriate modifications in the report
 

summary section. It was further moved that the APC be considered for retest. 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, University of Wisconsin Medical Center
 

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.
 

ftU.S. GOVERNMENT PRINTING OFFICE:1978 260-899/3109 1-3 5 4 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH APC
 

TABLE A1

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH APC 

c:ONTROL (UNTR) LOW DOSE HIGH DOSE
 

01-0055 01-0058 01-0059
 

ANIMALS INITIALLY IN STUDY 50 50 50
 ANIMALS MISSING 2
 

ANIMALS NF.CROPSIED «7 50 «9
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** «7 50 «9
 

INTEGUMENTARY SYSTEM
 

*SKIN (H7) (50) (<t9)
 

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

SQOAMOOS CELL CARCINOMA 1 (2%)
 

BASAL-CELL CARCINOMA 1 (2X)
 SARCOMA, NOS 1 (2X)
 FIBROMA 3 (6X)
 FIBROSARCOMA 1 <2X)
 LIPOMA 1 (2X)
 

LEIOMYOSARCOHA 1 <2X)
 

GRANULAR-CELL TUMOR, BENIGN 1 (2X)
 

*SUBCUT TISSUE (K7) (50) (49)
 

BASAL-CELL CARCINOMA 1 (2X)
 

FIBROMA 7 (15X)
 

RFSPIfATORY SYSTEM
 

tTRACHEA (16) (U8) (t8)
 SQOAMOUS CELL CARCINOMA 1 (2X)
 

tLUNG («7) (50) (t9)
 HEPATOCELLULAR CARCINOMA, METAST 1 (2%)
 ALVEOLAR/BRONCHIOLAR ADENOMA 1 (2X)
 ALVEOLAR/BRONCHIOLAR CARCINOMA 2 (IX)
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE ORGANS («7) (50) (U9)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

LEUKEMIA, NOS 1 <2X>
 

... MPIFFERENTIATED LEUKEMIA ..,1.J2,SL
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-3 
TABLK A I (CONTINUED) 

HIG H DOSE 

01-0059 

1 (2«) 

(19) 

(12) 

(17) 

1 (2*) 

(19) 

(49 ) 

1 (2*) 

(17) 

(17) 

(17) 

1 (2%) 

(13) 

(19) 

KYEU'MONOCYTIC LFtT[f FMIA 

I.YMPKOCYTI C LEUKEMI A 

»SPLEFP 

M'JCIKOtI S ADENOCABCINOMA , NETAST A 

MALIC- . LYBPHOMA , LYMPHOCYTI C TYP E 

LFUKrKiA,No s 

IflEPIASHNA L L. NOD E 

MUCIKCII S ADiNOCARCINOMA ,

• PEYEP S PATC H 

MALIGNAN T LYMPHOHA , NO S 

CIPCU1ATOPY SYSTEM 

Ni'N E 

DIGFSTIV r SYSTF K 

*nEA L CAVIT Y 

FIBROSAKCOIU 

*LIVE F 

NEnpl.ASTr c NODUL E 

HEPATOCFLLULA R CARCINOM A 

*PANCPFAS 

•IBCINOU S HDENOCAPCINOHA ,

*STOKACH 

HUCINOQ S ADEHOCABCINOMA ,

HILFII H 

PAPILLAF Y ADENOCAPrlHOM A 

#C010N 

MUCINOTJ S AD3NOCARCHJOH A 

I1PINAP Y SYSTE N 

*KII'N" Y 

 METAST A 

 METAST A 

 fl^TAST A 

TUBULAK-CSL L ADENOCABCINOB A 

CONTPO L (UNTF ) 

01-0055 

1 (2%) 

(It?) 

1 (2%) 

1 (2X ) 

(12) 

1 (7%) 

(17) 

(17) 

1 (2%) 

(17) 

5 (11K) 

(15) 

1 (2% ) 

(17) 

1 (2% ) 

(17) 

(16) 

1 (2« ) 

(17) 

LOW DOSE 

01-0058 

3 ((,%) 

(ST) 

1 (2%) 

(11) 

(13) 

(50) 

(53) 

3 (6%) 

t (2X ) 

(17) 

(19) 

(19) 

(16) 

(50) 

!_.USX_.._ 

» NIIKBEJ1 . O F AKI1AL 5 WIT H TISSU E EXAMINF D MICROSCOPIC ^ L L 1 

* NUI13FR O F ANIMAL S NFCROPSTS D 

A-4 
TABLE A1 (CONTINUED)
 

CONTRO L (UNTR ) LOW DOSE 

01-0055 01-5058 

LIPOM A 1 (2%) 

NEPHFOBLASTOM A 1 (2%) 

ENDCC5IN E SYSTE M 

((PITUITAR Y («7) (17) 

CARCINOMA , NOS 2 (4%) 

ADENOMA , NO S 8 (17X ) 16 (3UJ) 

KADRENA l (U7 ) (H9 ) 

CORTICAL ADENOM A 3 (6%) 

PHEOCHHOMOCYTOMA 7 (15%) 17 (35*) 

((THYROID («6) (U3) 

FOLLICULAB-CEL L ADENOM A 

C-CELL ADENOM A 2 (It*) it (8%) 

C-CELL CARCINOM A 2 (HX ) 5 (10X) 

»PAHATHY80I D (2U ) (27)

 

ADENOMA , NO S 1 (UX )

 

*PANCREATI C ISLETS (t5) (147) 

ISLET-CELL ADENOMA 3 (7*) 3 (6%) 

ISLET-CELL CARCINOM A 1 (2S) 

REPECDUCTIV E SYSTE M 

*PKEPUTIA L GLAN D (47) (50) 

PnENOHA , NOS 1 (2%) 1 (2S) 

*SEMINAL VESICLE (U7) (50)

 

HllCISOrJS ADENOCARCINOMA , NETAST A 1 (2S)

 

KTESTIS («7) (149) 

INTEESTiriAL-CEL L TUBO S UU (911%) 146 (9U% ) 

NFEVOOS SYSTE M 

*BEAIN/KENINGES («5) (50) 

SQUAMO'J S CFL L CARCINOMA , K'TAST A 1 (2*) 

*EFAI N (15) (50) 

ASTROCXTOH A , , 1.12SL 

* NUKEEF OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-5
 

HIGH DOSE 

01-0059 

1 (2*) 

(1414) 

1 (2%) 

11 (25%) 

(148) 

1 (2%) 

9 (19X) 

(146) 

1 (2% ) 

3 (7%) 

5 (11% ) 

(25) 

(147) 

2 (14%) 

1 (2%) 

(149) 

(149) 

(149) 

27 (55%) 

(148) 

(148) 
TABLE A1 (CONTINUED) 

CONTROL (UNTR) 

01-0055 

LOW DOSE 

01-0058 

HIGH DOSE 

01-0059 

SPECIAL SENS! ORGANS 

*EAF CANAL 

SQUAKOUS CELL CARCINOMA 

CERUKISOUS CARCINOMA 

(17) 

1 (2X) 

(50) 

1 (2*) 

1 (2X) 

(«9) 

MUSCULOSKEIETAL SYSTEM 

•STERNUM 

MUCINOUS ADENOCARCINOMA, METASTA 

(«7) 

1 (2X) 

(50) (1*9) 

BODY CAVITIES 

•BODY CAVITIES 

flSSOTHELIOKA, MALIGNANT 

(«7) 

2 <i»») 

(51) (49) 

•MEDIASTINUM 

ALVEOLAP./BFONCHIOLAR CA, METASTA 

(t7) (50) (49) 

1 (2*) 

•ABDOMINAL CAVITY 

I 7.1 Of: YO SARCOM A 

(47) (50) (49) 

1 (2X) 

ALL CTHEF SYSTEMS 

NONE 

ANIMA L DISPOSITION SUMMARY 

ANI.1ALS INITIALLY IN STUDY 

NA^OSAL DEATHS 

MQFIEUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TEFKTNAL SAC8IFICF 

ANTMA L MISSINS 

50 

2 

8 

5 

33 

2 

50 

4 

6 

UO 

50 

3 

3 

c, 

39 

S INCLUDES AUIOLYZED ANIMALS 

» NUH3E R

* NUMBE R

 O F ANIMAL S WIT H TISSUE

 OP ftNIMALS NECROPSIED 

 EXAMINE D MICROSCOPICALL Y 

A-6
 
TABLE Al (CONCLUDED) 

(rONTROL (UNTR) LOW DOSE HIGH DOSE
 

01-0055 01 -0058 01-0059
 

THMOF SUMMAR Y
 

TOTAL ANIMALS KITH PRIMARY TUMORS* 147 50 38
 

TOTAL PRIMARY TUMORS 96 118 72
 

TOTAL ANIMALS WITH BENIGN TUMORS 1*7 U7 36
 

TOTAL BENIGN TUHORS 76 96 55
 

TOTAL ANIKAL S HITH MALIGNANT TUMOPS 11 17 1<4
 TOTA1 MALIGNANT TUMORS 15 19 16
 

TOTM ANIMALS WITH SECONDARY TUMORS* 1 2 1
 

TOTA1 SECONDARY TUMOES 6 2 1
 

TOTAL ANIMALS HITH TUMOPS UtiCERTAIN-


EENIGN OR MALIGNANT 5 3 1
 

TOTAL UNCERTAIN TDMORS 5 3 1
 

TOTAL ANIMALS WITH TUMORS UNCE8TAINPEIMAEY OS METASTATIC
 TOTAL UNCERTAIN TUMORS
 

* FRIflAf.Y TUMOBS: ALL TUMOBS EXCEPT SECONDARY TUMORS
 * SECONDARY TUMORS: NETASTATIC TUMORS CR TUflOES INVASIVE INTO AN ADJACENT ORGAN
 

A-7
 
TABLE A2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH APC 

CONTROL (ONTE) LOW DOSE HIGH DOSE
 

02-0055 02-0058 02-0059
 

ANIMALS INITIALLY IN STUD* 50 50 950
 ANIMALS HISSING 2
 ANIMALS NECROPSIED «8 50 49
 

ANIRALS EXAMINED HISTOPATHOLOGICALLY ** «7 50 49
 

INTEGUMENTARY SYSTEM
 

*SKIN (MB) (50) (49)
 

SQUAHOUS CELL PAPILLOMA 1 (2X)
 

SQUAMOUS CELL CARCINOMA 1 (2S)
 FIBROMA 1 (2X)
 

FIBROSARCOHA 1 (2%)
 

LIPOMA 1 (2*)
 

»SUBCBT TISSUE (t8) (50) (49)
 

SQUAMOUS CELL CARCINOMA 1 (2*)
 

RESPIRATORY SYSTEM
 

ILUNG (1*7) (48) (45)
 

HEPATOCELLOLAH CARCINOMA, METAST 1 <2X)
 TUBULAR-CELL ADENOCARCINOMA , MET 1 <2X)
 

HEBATOPOIETIC SYSTEM
 

*HDLTIPLE ORGANS	 («8) (50) (49)
 

MALIGNANT LYHPHOMA, NOS 2 («X)
 LEUKEMIA, NOS 1 <2X) 1 (2X)
 

HYELOMONOCYTIC LEUKEMIA 3 (6X) 4 (8X)
 

ISPLEFN (47) (48) (45)
 

LEOKFNIA.NOS 1 <2X)
 MYELOMOHOCYTIC LEUKEMIA 1 (2«)
 

tLYMPH NODE (tO) (46) (44)
 

HEPATOCELLOLAR CARCINOMA, BETAS! 1 (2X)
 

tSHALL INTESTINE	 (16) (46) (45)
 

JWiISilJS£BaBlt-flI3aQ5niS_TT.Jjj — ___=._»«__— _—_
 ___^___— ...„«. 1.J2SJ, _.
 

* NUMBER OF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED "EXCLUDES

 PARTIALLY AUTOLYZED ANIMALS 

9 50 ANIMAL WERE INITIALLY IN THE STUDY, BUT 1 ANIMAL HAS FOUND TO BE A MALE ANIMAL 

A-8 
TABLE A2 (CONTINUED) 

tTHYMUS
 

THYMOMA
 

CIRCULATORY SYSTEM
 

* HEART
 

HEPATOCELLULAH CARCINOMA, METAST
 

DIGESTIVF SYSTEM
 

#LIVER
 

NEOPLASTIC NODULE
 

HEPATOCELLULAR CARCINOMA
 

TUBULAR-CELL ADENOCARCINOMA , MET
 

•PANCREAS
 

TUBULAR-CELL ADENOC ASCI NOM A, MET
 

IJFIMARY SYSTEM
 

tKIDNEY
 

TRANSITIONAL-CELL PAPILLOMA
 

TUEULAR-CELL ADENOCARCINOMA
 

tURISARY BLADDER
 

TRANSITIONAL-CELL CARCINOMA
 

FNCOCRINE SYSTEM
 

tFITIIITABY
 

CARCINOMA ,NOS
 

ADENOMA, NOS
 

#ADRENAL
 

CORTICAL ADENOMA
 

CORTICAL CARCINOMA
 

FHECCKROMOCYTOMA
 

LIPO^!^
 

tADFENAL MEDULLA
 

.. ..f-ANGLIOSSUROMA ,_ ,„
 

* NUMBE R O f ANIKAL S WIT H TISSUE

* tiUMBIP OP ANIMAL S NECROPSIED 

CONTROL (UNTR)
 

02-0055
 

(39)
 

1 (3X)
 

(U7)
 

(U7)
 

(t6)
 

(47)
 

(«7)
 

(U6)
 

2 (U*)
 

18 (39X)
 

(17)
 

3 (6X)
 

1 (2*)
 

1 <2X>
 

(K7)
 

 EXAMINE D MICROSCOPICALL Y 

A-9 

LOU DOSE HIGH DOSE
 

02-0058 02-0059
 

(30) (33)
 

(«8) (46)
 

1 (2*)
 

(«7) (45)
 

3 (6*) 3 (7%)
 

1 (2X)
 

1 (2*)
 

(U6) (44)
 

1 (2*)
 

(H 7) (46)
 

1 (2X)
 

1 (2X)
 

(47)	 («6)
 

1 (2*) 1 (2X)
 

(4U) (43)
 

1 (2«)
 

14 (32X) 14 (33X)
 

(4^1) («4)
 

4 (8*) 1 (2*)
 

1 (2X)
 

2 (I»X) 2 (5X)
 

1 <2X)
 

(48)	 (44)
 

1.JJSJL,.
 
TABLE A2 (CONTINUED) 

" CONTRO L (UNTR ) LOW DOSE 

02-0055 02-0058 

• THYROI D (16) <K6 ) 

FOLLICULAR-CEL L ADENOM A 1 (2*) 

FOLLICULAR-CELL CARCINOMA 2 (K«) 

C-CEIL ADENOM A 4 (9X) 

C-CEIL CARCINOMA 3 (7X) 4 (9J) 

REPRODUCTIVE SYSTEM 

*MAMMAB Y GLAN D 

ADSNOCAPCINOMA , NO S 

PAPILLAR Y CYSTADENCMA , NO S 

FIBROADENOM A 

C*8) 

1 (2X) 

1 (2%) 

1H (29X) 

(50) 

7 (1«X) 

*VAGINA 

SARCOMA , NOS 

(«8) 

1 (2%) 

(50) 

• UTOUS ADENOCARCINOI«A , NOS 

ADENOC A I N ADENOMATOU S POLY P 

ENDOMETPIA L STROMAL POLYP 

(HI) 

2 (H%) 

15 (32%) 

(K8) 

3

1

la

 (6*) 

 (2«) 

 (29S) 

ICFPVIX UTPHI 

LSIOhYOH A 

(147) (US) 

1 (2X) 

•OVAR Y 

H3PATOCELLULA R CABCINOMS , SFTAST 

3RANULOSA-C2L L TUMOR 

GRBNULOSA-C5L L CARCINOM A 

(16) 

1 (2%) 

(K8) 

1 (2*) 

NFFVOU S SYSTE M 

• BRAI N (H7 ) (K3 ) 

ASTBOCYTOM A 1 (2«) 

SPECIAL SENSE ORGAN S 

NON= I 

MUSCULOSKELETA L SYSTEM 

NON E 

t NUMBE R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBFR OF ANIMALS NECROPSIEC
 

A-10
 

HIGH DOSE 

02-0059 

(43) 

2 (5«) 

2 (5X) 

2 (5X) 

(19) 

2 («X) 

(49) 

(16) 

7 (15X) 

(16) 

(15) 

1 <2X) 

(16) 
TABLE A2 (CONCLUDED)
 

BODY CAVITIES
 

*«'::DIAS'':INUM
 TUBULAP-CELL ADENOCARCINOMA, MET
 

ML OTHER SYSTEMS
 

NONE
 

ANIMA L DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATUPAL DBATHS
 

KOSIBOND SACRIFICE
 

SCHETULFD SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMA L MISSING
 

ANIMA L DELETED
 

8 INCLUDES AUTOLYZED ANIMALS
 

TUMOF SUMMAR Y
 

TOTAL ANIMAL S WITH PPIMARY TUMORS*
 

TOTAL PRIMARY TUKOPS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 TOTAL BENIGN TUMORS
 

TO^AL ANIMALS KITH MALIGNANT TUMORS
 TOTAI MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORS*
 

TOTAI SECONDARY TUMORS
 

TOTAI ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAI UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PFIMAPY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

CONTROL (UNTR)
 

02-1055
 

(U8)
 

50
 

3
 

6
 

5
 

34
 

2
 

42
 

70
 

3fl
 

53
 

in
 

17
 

LOW DOSE HIGH DOSE
 

02-0058 02-0059
 

(50)	 <«9)
 

1 <2»)
 

50 50
 

9 it
 it 1
 

5
 

37 39
 

1
 

U1 32
 

75 15
 

2« 26
 

149 32
 

20 7
 23 9
 

2
 

8
 

3 II
 

3 4
 

* PEIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUBORS
 *	 SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-ll
 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH APC
 

TABLE Bl

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH APC 

HIGH DOSE 

05-0059 

50 

1 

48 

48 

(48) 

(47) 

7 (15X) 

5 (11X) 

(48) 

2 (4X) 

(46) 

1 <2X) 

(41) 

1 (2X) 

(41) 

(41) 

1 (2X) 

(45) 

1 <2X ) 

ANIMALS INITIALLY IN STUDY 

ANIKAL S MISSING 

ANIMAL S NECEOPSIED 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY *" 

INTIGOMENTAR Y SYSTEM 

*SKIN 

FIBROSARCOMA 

RESPIRATOR Y SYSTEM 

*LUNG 

ALVEOLAR/BRONCHIOLA H ADENOM A 

ALVEOLAR/BFONCHIOLAR CARCINOM A 

HFHATOPOIETIC SYSTEM 

*MOLTIPLE ORGAN S 

MALIGNAN T LYHPHOMA , NO S 

MALIG.LYMPHOMA , HISTIOCYTIC TYPE 

*SPLEEN 

HEMANGIOM A 

HEMANGIOSARCOM A 

MALIG.LYMPHOMA . HISTIOCYTIC TYPE 

»LYMP H NODE 

PLASMA-CELL TUMOR

 

tMANDIBULAR L. NODE 

MALIG.LYMPHOMA , HISTIOCYTIC TYPE 

•MESENTFHIC L. NODE 

MALIG.LYHPHOBA, HISTIOCYTIC TYPE 

IPEYERS PATCH 

MALIG.LYMPHOHA , HISTIOCYTIC TYPE 

CIRCULATOR Y SYSTEM 

, N015, __ 

CONTROL (UNTR) 

05-0037 

50 

148 

48 

(18) 

(48) 

6 (13%) 

<t8) 

2 (4X) 

2 (IX) 

(47) 

1 (2X) 

(1«) 

(UU) 

(Ui» ) 

(H8) 

LOW DOSE 

05-0058 

50 

1 

48 

47 

(48) 

1 (2X) 

(46) 

6 (13X ) 

3 (7X) 

(48) 

4 <8X) 

(46) 

1 (2X) 

(44) 

(44) 

1 (2X) 

(44) 

(45) 

1 (2X) 

t NUMBER OF ANIMALS WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
TABLE Bl (CONTINUED) 

CONTROL (UNTR) LOU DOSE HIGH DOSE 05-0037 05-0058 05-0059 

DIGESTIVE SYSTEM 

KLIVFR (»8) («6) <«7) HEPATOCELLULAR CARCINOMA 7 (15*) 11 (21*) 6 (13X) HTMANSIOMA 

HEKANGIOSARCOMA 1 <2X) 

1 (2X) 

1 (2*) 

2 (UX) 

tSTOMACH (i»7) («3) (i»6) 

SQUAKO'JS CELL CARCINOMA 1 (2*) 

URINAR Y SYSTEM 

NON? 

ENDOCRINE SYSTEM 

tTHYSOID (i»7) (»2) (U2) FOLLIC'JLAS-CELL ADENOMA 1 (2X) 1 (2*) 

IPANCREATIC ISLETS 

ISL=:T-CELL ADENOMA 

(«8) («5) (U5) 

1 (2X) 

REPRODUCTIVE SYSTEM 

NON5 

NERVOUS SYSTEM 

NONE 

SFECTU SENSE ORGANS 

*EXTEBNAL FAR (148) (US) (K8) 

FIBROSARCOMA 1 (2X) 

nUSCIHOSKElETAL SYSTEM 

*MUSCLE HIP/THIGH (19) (it 8) (48) 

F.HABnOSYOSARCOMA 1 (2X) 

BODY CAVITIES 

...Wi . . _. ._ ., , 

* N'J^BFE OF ANIMAL S WIT H TISSI'E EXAMINE D MICROSCOPICALL Y 

» NUK3E R OF ANIMAL S NECROPSIE D 

B-4 
TABLE Bl (CONCLUDED)
 

CONTROL (UNTR) LOU DOSE HIGH DOSE
 05-0037 05-3058 05-0059
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 NA"-UFAL DEATHS 3 4 6
 

MORIBUND SACRIFICE 1 2
 SCHEDULED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE 42 14 a 36
 ANIMAL MISSING 1 1
 

* INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMAR Y
 

TOTAL ANIMAL S WITH PRIMARY T'JMORS* 17 24 21
 TOTAL PRIMARY TUMORS 21 33 30
 

TOTAL ANIMAL S HITH BENIGN TUMORS 2 7 8
 TOTAL BENIGN TUMORS 3 7 9
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 15 21 17
 

TOTAL MALIGNANT TUMORS 13 23 20
 

TOTU ANIMAL S HITH SECONDARY TUMORS*
 TOTAL SECONDARY TUMORS
 

TOTAL ANIMAL S WITH TUMORS UNCERTAIN­ EEKISN OR MALIGNANT 1
 TOTAL UNCERTAIN TUKORS 1
 

TOTU ANIMAL S WITH TUMORS UNCERTAIN­ FSI.1ARY OF. METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

» PRI1ARY TUMORS: ML TUH08S EXCEPT SECONDABl TUMORS
 * SFCONDARY TUMORS: METASTHTTC TONORS OF TUHOFS INVASIV F INTO AM ADJACENT ORGAN
 

B-5
 
TABLE B2 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH APC 

_
 CONTROL (UNTR) LOW DOSE HIGH DOSE
 06-0037 06-0058 06-0059
 

ANIMALS INITIALLY IN STUDY 50 50 50
 ANIMALS MISSING 1 1
 ANIMALS HECROPSIED 18 45 47
 

ANIMALS EXAMINED HISTOPATHOLOGICALL Y ** 47 45 47
 

INTFGUMENTASY SYSTEM
 

*SUBCUT TISSUE (18) (45) (47)
 

LEIOMYOSARCOMA 1 <2X)
 

RESPIRATORY SYSTEM
 

ILUNG («6) (45) (47)
 

HEPJTOCELLULAR CARCINOMA, METAST 1 <2X) 1 (2X)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 3 (7X) 7 (16%) 2 (4X)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 1 (2X) 3 (6X)
 

HEMATOPOIETIC SYSTEM 

*MULTIPLE ORGANS (48) (45) (47) 

MALIGNANT LYMPHOMA, NOS 1 (2X) 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 2 (U%) 4 (9X) 1 (2X) 

»SPLEEN (46) (44) (47) 

HEMANGIOMA 1 <2X) 

H^MANGIOSARCOMA 1 (2X) 

KALIG.LYMPHOHA, HISTIOCYTIC TYPE 1 (2X) 

tSUBMANDIBULAR L.KODE (39) (34) (38) 

KALIG. LYMPHOMA, HISTIOCYTIC TYPE 1 (3X) 

IMESSNTERIC L. NODE (39) (34) (38) 

MALIGNANT LYMPHOMA, NOS 1 (3X) 

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

tLIVER (47) (45) (47) 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 <2X) 

»PEYERS PATCH 

,.,__«.UIG..LYMPHOM4i_HISTIOCYTIC_TYPE.,i 

(44) __, (45) (46) 

._l_J2Si___ ... 

» NUMBE R OF ANIMAL S WITH TISSUF EXAMINE D MICROSCOPICALLY 

* NUHBS B OF ANIMAL S NECROPSIED 

**i-.XCLUDES PARTIALLY AUTOLYZEIi ANIMALS 

B-6 
TABLE B2 (CONTINUED) 

CONTROL (ONTR ) LOW DOSE HIGH DOSE 

06-0037 Ob-0058 16-0059 

*THYMU S (31) (39) (12) 

MALIGNAN T LYMPHOMA , NOS 1 (3*) 

CIECULATOR Y SYSTE M 

NON E 

DIGESTIV E SYSTE M 

*LIV5R (if?) (15) (17) 

HEPATOCELLULAR CARCINOMA 1 (2X) 2 (1«) 3 (6X) FIERGSAPCOM A 1 (2X) 

#STOBAC H (Ull) (13) (15) 

SQUAKOU S CEIL PAPILLOM A 1 (2*) 

tCOLON («") (11) (13) LETOMYOSARCOM A 1 (3*) 

OEINAB Y SYSTE M 

NON E 

ENCCCRTN E SYSTE M 

tFITUITAB Y (12) (33) (11) CARCINOM A ,NOS 1 (2X) ADENOMA , NO S 2 <5X) 1 (11*1 2 (5«) 

tADRSNA L (U5) (12) (16) PHEOCHROMOCYTOHA 1 (2%) 

ITHYROI D (13) (31) (15) FOLLICU1AR-CELL ADENOM A 1 (2*) FOLIICIJLAR-CELL CARCINOM A 1 <2X ) 

REPBODUCTIV E SYSTK M 

IUTERU S (15) (13) (16) 

-WIOMYOSARCOM.A, . . . ,-..„ l.IJSJ.. •'..»._-;.*.-.*.._.__ _— _-.-—.»,. .„—_ — »—„-__. , 

# NUMBFR OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NHM3ER OF ANIMALS NECROPSIED
 

B-7
 
TABLE B2 (CONTINUED) 

CONTROL (UNTR) LOU DOSE HIGH DOSE 

06-0037 06-0058 06-0059 

FNDOMETRIA L STROMA L POLYP 3 (7X)

 

HEHANGIOM A 1 (2«)

 

• OVAR Y (U5 ) (115) <U2 )

 

TUBULA R ADENOM A 1 (2*)

 

NERVCU S SYSTE M 

NON E 

SPECIA L SENS E ORGAN S 

*HAPDERIA N GLAN D (148) («5) (<»7) 

PAPILLAR Y ADENOM A 1 (2*) 

MUSC<JLOSK1*LETA L SYSTEM 

NON E 

BOT Y CAVITIE S 

MOD* 

A L L Olil^P SYSTEMS 

MOMS 

ANIMA L DISPOSITIO N SUMMAR Y 

ANTMAL K INITIALL Y I N STUDY 50 50 50

 NATURA L DEATH S 6 5 3

 MORIBUN D SACRIFICE 2

 

SCHECULE D SACRIFICE 5 5

 

ACCIDENTALL Y KILLE D 1

 

TERMINA L SACRIFICE 37 «3 U1

 ANIMA L MISSIN G 1 1

 

*.-isf.vmg.vsmx35JB.,aiU8Jus „ * NUf!3"R CF ANIMAL S HIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE E OF ANIMAL S NECROPSIE D 

B-8 
TABLE B2 (CONCLUDED)
 

CONTROL (UNTR) LOW DOSE HIGH DOSE
 06-0037 06-0058 06-0059
 

TUMOR SUMMAR Y
 

TOTAL ANIMAL S WITH PRIMARY TUMOPS* 20 17 11
 

TOTAI PRIMARY TUMORS 20, 20 19
 

TOTAL ANIMALS WITH BENIGN TUMORS 11 11 6
 

TOTAL BENIGN TUMORS 11 12 7
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 1 1 8 11
 

TOTAL MALIGNANT TUMORS 13 8 12
 

TOTAL ANIMAL S WITH SECONDARY TUMORS* 1 1
 

TOTAL SECONDARY TUMORS 1 1
 

TOTAL ANIMAL S WITH TUMORS UNC5RTAIN­ BESIGN OP MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMAL S WITH TUMORS UNCEFTAIN-
 PETMAfcY OR METASTATIC
 TOTAI UNCERTAIN TUMORS
 

* PRIMAR Y TOHOPS: ALL TUNORS EXCEPT SECONDARY TUM03S
 * SECONDARY TUMORS: HETASTATIC TUMORS OR TUMOF.S INVASIVE INTO AN ADJACENT ORGAN
 

B-9
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH APC
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS TREATED WITH APC 

ANIMALS INITIALLY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECEOPSIED
 

ANIMALS EXAMINED HI STOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

EPIDFRMAL INCLUSION CYST
 

INFLAMMATION, NECROTIZING
 

INFLAMMATION, ACOTE FOCAL
 

ABSCESS, SOS
 

RESPIRATORY SYSTEM
 

tLUNG/BRONCHUS
 

ABSCF.SS, NOS
 

(tLUNG
 

BRONCHOFN5UMONIA, NOS
 

HYPERPLASIA, ALVEOLAR EPITHELIUM
 

HEMATOPOIETIC SYSTEM
 

IBONE MARROH
 

KYELOFIBROSIS
 

#SPLEEN
 

INFARCT HEMORRHAGIC
 

HYPERPLASIA, NOS
 

HEMATOPOIESIS
 

tLYMPH NODE
 

FIBROSIS
 

tPANCREATIC L. NODE
 

INFLAMMATION, ACUTE/CHRONIC
 

#ILHOCOLIC LYMPH NODE
 

- M-B£H*DENQPA,THY__ ,_ ,_
 

t NUMBE R OF ANIMAL S WITH TISSUE

* NUMBE R OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED AMMALS 

CONTROL (UNTR)
 

01-0055
 

50
 

2
 

47
 ** U7
 

(D7)
 

1 <2X)
 

1 (2%)
 

1 (2%}
 

(K7)
 

1 (2X)
 

(47)
 

(U6)
 

(«7)
 

1 (2X)
 

(12)
 

<K2)
 

1 (2X)
 

(«2)
 

J.i25I.__.-.
 

LOW DOSE HIGH DOSE
 

01-0058 01-0059
 

50 50
 

50 49
 

50 49
 

(50) (49)
 

1 (2X)
 

(50) (49)
 

(50) («9)
 

1 (2X)
 2 (IX)
 

(49)	 (47)
 

1 (2X)
 

(50) (49)
 

1 (2X)
 

1 (2X)
 

(HI)	 (12)
 

1 (2X)
 

(41) (U2)
 

(41) («2)
 

 EXAMINE D MICROSCOPICALLY 

C-3
 
TABLECl (CONTINUED) 

CONTRO L (ONTR ) LOW DOSE 

01-0055 01-0058 

CIRCULATOR Y SYSTE M 

KHEAR T (U7 ) (50) 

?"EIfRTEaiTI S 1 <2« ) 

#BYOCARDI0 M («7) (50) 

DFGENERATION , NO S 10 (21*) 17 (34«) 

ttCARDIA C VALV E (U7 ) (50) 

INFLAMMATION , CHRONI C 

*PULMONAB Y ARTER Y (U7 ) (50) MINE F ALIZATIO N 1 (2%) 

DIGESTIV E SYSTE M 

tLIVER (47) (50) INFLAMMATION , NECEOTIZIN G 1 (2X) FEGENERATION . HYALIN E 1 (2X) 

NECROSIS , FOCAL 1 <2X) METAMORPHOSI S FATT Y 3 (6«) FOCAL CELLULA R CHANG E 12 (26X) 1 (2X) HYPERPLASIA , FOCAL 1 (2*) 

ULIVER/CFNTSILOBULA R (K7 ) (50) NECROSIS , NOS 3 (6%) 

*BILE DUCT 

HYPEPPLASIA , NO S 

(47) 

4 (9X) 

(50) 

*FANCRFA S (45) (47) 

INFLANHATION ,

FIBROSIS 

 ACUTE/CHRONIC 1 (2X) 

1 (2X) 

iPANCREATI C

ATROPHY ,

 ACINU S 

 NO S 

(45) 

3 (7*) 

(47) 

ATROPHY , FOCAL 

HYPERPLASIA , NOS 

1 <2X ) 

1 (2X) 

•STOMACH (47) (49) 

ULCEF , NO S 1 (2X) INFLAMMATION , ACDTE 1 (2*) HYPEfPLASIA , EPITHELIAL 1.12SL 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-4
 

HIGH DOSE 

01-0059 

(49) 

(49) 

14 (29*) 

(49) 

1 (2X) 

(19) 

(49) 

1 (2%) 

3 (6X) 

(49) 

1 (2X) 

(49) 

(47) 

(47) 

2 (4X) 

(47) 

,_2_l«Sl 
TABLECl (CONTINUED)
 

CONTROL (UNTR ) LOW DOSE 

01-0055 01-005B 

UPINAR Y SYSTE M 

IKIMTY (Ul ) (50) 

CYST, NOS 1 (2X) 

GLOMERULONEPHRITIS , NOS 1 (2X) 

INFLAMMATION , CHRONIC 39 (83«) 12 (8UX ) 

*UfINAR Y BLADDER (H7 ) (U9 ) 

HYPFRPLASIA , EPITHELIA L 1 <2X ) 

ENDCCRIN F SYSTE M 

tPITUITAF Y (17) (17) 

COLLOID CYST 

HYPERPLASIA , NO S 1 <2X ) 

HYPFRPLASIA , FOCAL 3 (6*) 3 (6X) 

• ADRENA L (K7 ) (U9 ) 

HYPEFPL'SIA , FOCAL 1 (2X) 

*ADRENA L CORTK X (U7 ) (19) 

HYPERPLASIA , FOCAL 6 (12J) 

tADHENA L MEDULL A (H7 ) (19) 

PYPFP.PLASIA , NODULA R 3 (6X) 1 (2X) 

HYPFPPLASIA , NOS 

HYPERPLASIA , FOCAL 2 (1«> 

tTHYSOID (16) (18) 

FOLLICULA R CYST, NOS 

HYPERPLASIA , C-CELL 1 (9X) 1 (2X) 

tPARATHYHOI D (21) (27) 

HYPEEPLASIA , NOS 1 (1*1 

REPRODUCTIV E SYSTEM 

*MAHMAE Y GLAN D 

GALACTOCEL E 

HYPERPLASIA ,

HYPERPLASIA ,

 NO S 

 FOCAL 

(17) 

1 (2X) 

1 <2X ) 

(50) 

*PENIS (17) (50) 

._...I.NFLAN»MIONt .ACUTE_NlCRQTIZING_ _ 

* NUMBER OF ANIMALS WITH TISSUE FXARINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECHOPSIED
 

C-5
 

HIGH DOSE 

01-0059 

(19) 

10 (82%) 

(18) 

1 (2%) 

(11) 

1 (2X > 

5 (11*) 

(18) 

(18) 

5 (10X) 

(18) 

2 (IX) 

1 (2X) 

(16) 

1 (2X) 

1 (2X) 

(25) 

(19) 

1 (2X) 

<U9 ) 

LJ2X). 
TABLECl (CONTINUED) 

CONTRO L (UNTR ) LOW DOSE HIG H DOSE 

01-0055 01-0058 01-0059 

*PBFPUTIA L GLAN D («7) (50) (K9 ) INFLAMMATION , ACUT E 1 (2%)

 

tPROSTAT F (U6 ) («!)) («9) 

INFLAMMATION , ACUTE 1 (15X) 5 (11X) 1 (2«) INFLAMMATION , ACUT E FOCA L 3 (7*) 1 <2X ) INFL A MMATION , ACUTE/CHRONI C 2 <t« > 

INFLAMMATION , CHRONI C 1 (2%) 1 (2X) 

#TESTTS (it?) (K9 ) (i»9) DEGENERATION , NO S 1 (2%) NFCROSIS , DIFFUS E 1 (2%) 

HTFSTIS/TUBUL E («7) (i»9) (49) DEGENEF1TION , NO S 1 <2X ) 11 (22X ) 10 (20X) 

NEPVCT S SYSTE M 

SON? 

SPECIA L SENS E ORGAN S 

*EY E (U7 ) (50) (119) TNFLA1MATIO N , CHRONI C 1 (2X) CATAFAC T 3 (6%) 

*EYE/PETIN A (it 7) (50) (19) DEGENERATION , NO S 1 <2«) 2 (IX) 

MUSCIJLOSKF.LFTA L SYSTE M 

NON E 

BOD Y CAVITIE S 

*MES3NTFB Y («7) (5^ ) (U9 ) 

fSUSTEaiTIS 1 (2X) 

A L L CTHE F SYSTEM S 

NON 3 

» NUMBEP OF ANIMALS WITH TISSUE EXAMINED BICBOSCOPICALLY
 

* N'JKSFR OF ANIMAL S NECROPSI5D
 

C-6
 
TABLE Cl (CONCLUDED) 

t NUMBE R O F ANIMAL S WIT H TISSUE FXAHINE n BICROSCOPICALL Y 

» NUKBE P OF ANIMAL S NECBOPSIE T 

C-7 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS TREATED WITH APC 

CONTROL (UNTR) LOW DOSE HIGH DOSE 

02-0055 02-0058 02-0059 

ANIMAL S INITIAL!,* IN STUDY 50 50 950 ANIMAL S MISSIN G 2 ANIMAL S NECPOPSIED «8 50 19 

ANIMAL S EXAMINE D HISTOPATHOLOGICALL Y ** 47 50 49 

INTEGUMENTAB Y SYSTEM 

NON E 

RESPIRATOR Y SYSTE M 

#LONG/BEOHCHOS (17) (<18) (15) BRONCHIECTASI S 1 <2X) 

*LUNG (It?) (18) (15) BRONCHOPNEOMONI A SUPPURATIV E 1 (2X> 

HEMATOPOIETIC SYSTEM 

tBONE NARRO H (45) («6) (16) MYELOSCLEROSI S 1 (2X) HYPERPLASIA , HEMATOPOIETIC 2 {««) 1 (2X) 

#SPLSEN (47) (18) (45) HYPERPLASIA , NOS 1 (2*) 

HYPERPLASIA , HEMATOPOIETIC 1 (2%) 

HEMATOPOIESIS 1 (2X) 2 («X) 

CIRCULATOR Y SYSTEM 

tHEART (17) (18) (16) THROMBUS , MURA L 1 (2X) 

tMYOCARDIU M (K7 ) (18) (46) DEGENERATION , NOS 7 (15X) 5 (1IOX) 5 (11X) 

tENDOCAPDIU M (U7) (18) (46) iNFiAM'iATjoN,. ^CHRONIC . „„. »— -v^.-.«_—_ >__.. -_...,l.i2il__. «._..*_..-««.......««._.«^ 

t NUMBFR OP ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

**tXCLUDES PARTIALLY AUTOLYZED ANIMALS 

a 50 ANIMA L WER E INITIALLY IN THE STUDY, BUT 1 ANIMA L HAS FOUND TO BE A MALE ANIMA L 

T-f U i 0 
DIGESTIVE SYSTEM
 

*LIVER
 

DEGENERATION, NOS
 

METAMORPHOSIS FATTY
 

FOCAL CELLULAR CHANGE
 

HFPATOCYTONEGALY
 

HYPERPLASIA, FOCAL
 

ANGIECTASTS
 

*LIVER/CFNT3ILOBULAR
 

NFCROSIS, NOS
 

tLIVER/PFRIPOETAL
 

INFLAMMATION, CHRONIC
 

*BILE DUCT
 

INFLAMMATION, CHRONIC
 

HYPERPLASIA, NOS
 

*PANCREAS
 

INFLAMMATION, CHRONIC
 

tPANCREATIC ACINUS
 

ATROPHY, NOS
 

tSTOMACH
 

INFLAMMATION, FOCAL
 

URINARY SYSTEM
 

tKIDNEY
 

HYDRONFPHROSIS
 

INFLAMMATION, CHRONIC
 

POSTMORTEM CHANGE
 

NECROSIS, MEDULLARY
 

((KIDNEY/CORTEX
 

CYST, NOS
 

*KIDNEY/PFLVIS
 

SECRCSIS, MEDULLARY
 

..... HXPERP,LASIAt... EPITHELIAL
 

TABLE C2 (CONTINUED) 

CONTROL (DNTR) LOU DOSE
 02-0055 02-0058
 

<K7) (17)
 

1 (2%)
 

2 (14%)
 

25 (53*) 26 (55X)
 

2 (IX)
 

1 (2*)
 

(17)	 (47)
 

1 (2X)
 

(17) (17)
 

("8) (50)
 

1 (2X)
 

2 <HX) 1 (2X)
 

(16)	 («6)
 

1 (2«)
 

(Ifi) (16)
 

8 (17K)
 

(16) (16)
 

(147) (17)
 

29 (62%) 11 (30*)
 

1 (2X)
 

(17)	 (17)
 

1 (2X)
 

(17) (17)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIF.D
 

C-9
 

HIGH DOSE
 02-0059
 

(15)
 

22	 (149X)
 

1 (2*)
 

(45)
 

(15)
 

1 (2X)
 

(49)
 

(44)
 

(44)
 

(45)
 

1 (2X)
 

(16)
 

1 <2X)
 

20 (43X)
 

1 (2X)
 

(46)
 

(46)
 

3 (7X)
 

... .....«?_iasi _-.
 
TABLE C2 (CONTINUED) 

CONTRO L (UNTR) LOK DOSE HIGH DOSE 

02-0055 02-0058 02-0059 

tUEINAR Y BLADDE R (»7) (47) (46) 

CALCULDS , NOS 1 (2*) 

ENDCCHIN F SYSTEd 

*PITUITAPY (U6 ) (««) (13) 

COLLOI D CYST 1 (2*) 

HYPEEPLA.SIA , NOS 1 (2*) 

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

• ADRENA L CORTE X (17) (48) (4U ) 

CYST, NOS 1 (2%) 

HEMOPRHAGI C CYST 1 (2*) 

DEGENERATION , NO S 3 (6%) 

METAHORPHOSIS FATTY 1 <2X) 2 (5X) 

HYFEPPLASIA , NODULA R 2 (UX ) 

HYPEFPLASIA , FOCAL 3 (6*) 3 |6X) 7 (16X) 

IADR3NA L MEDULL A <«7 ) (43) (44) 

THROMBOSIS , NOS 1 (2*)

 

HYPEFPLASIA , FOCAL 1 (2*) 1 (215)

 

#THYSOI D (lib) (46) (43) 

HYPEEPLASIA , C-CELL « (9%) 2 (4*) 2 (5X) 

REPRODUCTIV E SYSTE M 

*«AMMAR Y GLAN D (48 ) (50) (49) 

DILATATIOS/DUCTS 3 (fiX) 

GALACTOCELE 7 (15X)

 

HYPEFPLASIA , NO S » (flX)

 

*MAMMAR Y DUCT (»8) (50) (49) 

FIBBOSIS 2 <«X )

 

*CLITOEAL GLAN D (US ) (50) (49) 

CYSTIC DUCTS 1 (2X) 

*VAGIN A («8) (50) (49) 

INFLAMMATION , ACBTE 1 (21)

 

#UTEHU S (U7 ) (48) (46 ) 

amgOHSTRA. ,_ ,»- ,-,J,.J12Ll- - B.HISl- ..".-laSJL 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NDMBFF OF ANIMALS NECROPSIED
 

C-10
 
PYOMFTP A 

HUTZHUS/ENIJOMETRIUM 

INFLAMMATION ,

INFLAMMATION ,

HYPFPPLASIA ,

HYPERPLASIA ,

tOVASY/OVIDUC T 

INFLAMMATION ,

INFLAMMATIO N

INFLAMMATION ,

tOVAB Y 

INFLAMMATION ,

• OVARY/FOLLICL E 

HYPFRPLASIA ,

NERVOU S SYSTE M 

NON E 

 ACUTE 

 CHRONIC 

 CYSTIC 

 STROMA L 

 AC'ITE 

 ACTIVF CHRONI C 

 CHRONI C 

 CHP.ONIC 

 NOS 

SPECIA L SENS E ORGAN S 

*EYF 

CATARAC T 

*EY r/COFN" A 

INFLAMMATION , ACUTE 

*FYE/P.ETNA 

DEr.ENFRATION , NO S 

MUSCULOSKEIETA L SYSTFrl 

*BON E 

OSTEOSCL3ROSI S 

*AECOMIHA L MUSCLF , 

TEGFliEIATIO N , SOS 

BODY CAVITIE S 

*PLFUR A 

INFLAMMATION , CHRONIC 

TABLE C2 (CONTINUED) 

CONTROL (UNTR) 

02-0055 

(17) 

9 (19*) 

1 (2X) 

3 (6%) 

1 (2%) 

(U7 ) 

3 (6X) 

1 (2%) 

1 (2*) 

(U6 ) 

1 (2%) 

(16) 

1 (2% ) 

(18) 

(US ) 

(18) 

(18) 

1 (2%) 

(18) 

(18) 

1 .t?S) , 

LOW DOSE 

02-3058 

(13) 

3 (6*) 

8 (17X ) 

(18) 

(18) 

(13) 

(50) 

1 (2X ) 

(50) 

(50) 

1 (2*) 

(50) 

(50) 

(50) 

HIGH DOSE 

02-0059 

1 (2X) 

(16) 

2 (1«) 

1 (2X) 

(16)

1 (2*) 

(15) 

(15) 

(19) 

7 (1U») 

(19) 

1 (2X) 

(19) 

7 (1UX) 

(19) 

(19) 

1 (2X) 

(19) 

* NUMB"? OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE R CF ANIMALS NECROPSIF.D
 

C-ll
 
TABLE C2 (CONCLUDED) 

*EFICAKDIUM
 

INFLAMMATION, CHRONIC
 

ALL CTHEf. SYSTEMS
 

NONE
 

SPECIAL KORFHOLOGY SUMMARY
 

NO LESION REPORTED
 

ANIMAL BISSINS/NO NECROPSY
 

AUTO/NECROPSY/HI STO PFRF
 

A'JTO/NECSOPSY/NO HISTO
 

CONTROL (UNTP)
 

02-0055
 

(«8)
 

1 (2*)
 

2
 

1
 

LOU DOSE HIGH DOSE
 02-0058 02-0059
 

(50) (49)
 

1 3
 

1
 

t NUMBER OF ANIMALS KITH TISSUE EXAHINED MICROSCOPICALLY
 

* NBMBEE OF ANIMALS NECROPSIED
 

C-12
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH APC
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE TREATED WITH APC 

CONTROL (UNT8) 

05-0037 

LOW DOSE 

05-0058 

ANIMALS INITIALLY IN STUDY 

ANIMALS MISSING 

ANIMALS NECROPSIED 

50 

48 

50 

1 

48 ANIMALS FXAMINED HISTOPATHOLOGICALLY *» «fl 47
 

INTF.GOMENTARY SYSTEM
 

*SKIN (l»8) (48)
 

INFLAMMATION, NOS
 

ABSCESS, NOS 1 <2X)
 FIBROSIS 1 (2X)
 ALOPECIA 1 (2%)
 

*SUBCUT TISSUE (US) (48)
 

INFLAMMATION, CHRONIC
 

NECROSIS, NOS 1 (2X)
 

RESEIFATCPY SYSTEM
 

tLUNG/BPONCHUS (U8) (46)
 

INFLAMMATION, NOS 1 (2X)
 

INFLAMMATION, FOCAL 1 (2%)
 

#LUNG (US) (46)
 

INFLAMMATION, NOS 1 (2%)
 

INFLAMMATION, INTERSTITIAL 14 (29X) 1 (2X)
 

HYPEPPLASIA, EPITHELIAL 2 (i»%)
 

ILUNG/ALVEOLI (49) (46)
 

INFLAMMATION, FOCAL 2 («*)
 

FIBROSIS, FOCAL 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

#BONE BARROW (47) (45)
 

HYPERPLASIA, HEMATOPOIETIC
 

*SPLEFN (47) (46)
 

HEMOR EHA.G;C _c T§!„„_. „,
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

**hXCLUDEE PARTIALLY AUTOLYZED ANIMALS 

D-3 

HIGH DOSE
 

05-0059
 

50
 

1
 

48
 

48
 

(48)
 

1 (2*)
 

(48)
 

1 (2»)
 

(47)
 

(47)
 

1 (2X)
 

(47)
 

(45)
 

1 (2X)
 

(46)
 

._ 1_1J8I._
 
TABLE Dl (CONTINUED)
 

CONTROL (ONTR) L OW DOSE HIGH DOSE 

05-0037 05-0058 05-0059 

INFLAMMATION , NO S 1 (2%) 

HYPBFPLASIA , NO S 2 (IX) 1 (2X) HYPERPLASIA , HEMATOPOI5TIC 1 (««) 3 <7X) 3 (7X) HYPEFPLASIA , BRYTHROI D 2 («*) HYPEF.PLASIA , RETICULU M CELL 1 <2X )

 HYPF.FPLASIA, LYNPHOID 2 (««)

 F3MATOPOIESI S 1 (2X)

 

tLYMP H NODE (UK ) («») (HI)

 PSflOESHAGIC CYST 1 (2X)

 INFLAMMATION , NO S 13 (30X)

 

DEGENERATION , CYSTIC 1 12%)

 HYPEFPLASIA , NO S 2 <5X)

 HYPEFPLASIA , HEMATOPOIE1IC 1 (2X)

 HYPEFPLASIA , LYMPHOID 2 (5X)

 MYFLCI D METAPLASI A 2 (5X)

 

IMEDIASTINAL L. NODE (tt) (»«) (11)

 NSCBCSIS, NOS 1 <2X )

 

*ABDOMINA L LYMPH NODE (""*) (<"») (111)

 HYPFFPLASIA , LYMPHOI D 1 (2X)

 

tPANCBEATIC L. NODE (1«) (a «) (HI)

 INFLAMMATION , NO S 1 <2X)

 

tMESENTFFIC L. NODE (It) (tt) (11) H3HOFRHAG 3 1 (2X) INFLAMMATION , NO S 9 (2CX) 1 (2X) lYMPHUDENOPAlH Y 26 (59X) 18 (tax) HYPERPLASIA , LYKPHOI D 1 <2< ) 

*THYMUS (3K) (35) (31)

 NECROSIS , NOS 1 (3X)

 

CIRCULATOR Y SYSTE M 

tHEART/VENTRICLE («8) (<I7) (t7)

 M2LANI N 2 (tXJ

 

•MYOCARDIUM (U8) (« 7) (17)

 INFLAMMATION , INTEESTITIAL 2 (IX)

 FIBROSIS 5 (10X)

 

*BLOOD VESSE L (1*8) (48) («8,

 

. I.NFLAMf!A.TIONA,NOS._,,. .,.._ _ . i.JfiSL.

 

» NUMBF R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUHBFR OF ANIMALS NECROPSIED 

D-4 
'PULMONARY ABTERY
 

MINERALIZATION
 

DIGESTIVE SYSTEM
 

•SALIVARY	 GLAND
 

INFLAMMATION, NOS
 

PERIVASC'JLAR CUFFING
 

*LIVEF
 

NECROSIS, FOCAL
 

METAMORPHOSIS FATTY
 

HYPEBPLASIA, NODULAR
 

HYPERPLASTIC NODULE
 

HYPERPLASIA, FOCAL
 

HYPEFPLASIA, DIFFUSE
 

ANGIFCTASIS
 

MYFLOID METAPLASIA
 

iLIVER/HEPAIOCYTES
 

DEGENERATION, NOS
 

*GAILBLADDE8
 

INFLAMMATION, FOCAL
 

IPANCREAS
 

CYSTIC DUCTS
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

INFLAMMATION, ACUTE/CHRONIC
 

DEGENERATION, CYSTIC
 

METAMORPHOSIS FATTY
 

*PANCSEATIC DUCT
 

HYPERPLASIA, NOS
 

tPANCREATIC ACINUS
 

ATROPHY, NOS
 

HYPERTROPHY, FOCAL
 

HYPEFPLASIA, FOCAL
 

(tSTONACH
 

INFLAMMATION,. NOS
 

»	 NUMBER OF ANIMALS WITH TISSU!
 

* NUMBER OF ANIMALS NECROPSIEC
 

TABLE Dl (CONTINUED) 

CONTROL (UNTR) LOW DOSE
 05-0037 05-0058
 

(48)	 (1»8)
 

2 (IX)
 

(K7) (46)
 

2 (UX)
 

1 (2X)
 

C*8) (U6)
 

13 (27%)
 

3 (6X) 1 (2«)
 

2 (UX)
 

1 (2X)
 

1 (2X) 1 (2X)
 1 (2*)
 

1 (2X)
 

1 (2X)
 

(K8) (U6)
 

1 (2X)
 

(U8) (US)
 

1 (2X)
 

(«8) (145)
 

1 (2X)
 7 (15X)
 

1 <2X)
 

1 (2%)
 1 (2%)
 

1 (2X)
 

(148)	 (U5)
 

1 (2X)
 

(U8) (145)
 

1 (2X)
 1 (2X)
 

1 (2X)
 

(U7) (143)
 

13 J2SSL,.,,
 

EXAMINE D MICROSCOPICALL Y 

D-5 

HIGH DOSE
 05-0059
 

(148)
 

(45)
 

(47)
 

1	 (2X)
 

(47)
 

(48)
 

(45)
 

(45)
 

(45)
 

(46)
 
TABLE Dl (CONTINUED) 

CONTRO L (UNTR ) L OW DOSE HIGH DOSE 

05-0037 05-0058 05-0059 

ULCER , NO S 1 (2X) 

INFLAMMATION , FOCAL 1 (2X) INFLAMMATION , INTERSTITIA L 1 (2X) 

HYPFRPLASIA , NO S 1 (2X ) HYPEFPLASIA , FOCAL 1 (2X) 

HYPEFKFRATOSI S 3 (6X ) ACANTHOSI S 3 (6%) 

*GASrP.IC BUCOS A (U7 ) (143) (146) 

HYPFFPLASIA , FOCAL 1 (2X) 

»PEYEE S PATC H («8 ) (it 5) (15) 

HYPERPLASIA , NOS 2 (tX) 1 (2X) HYPEFPLASIA , LY»PHOI D 2 (H*) 

*ILE!)M (US ) (U5) (15) 

HSMOBF.HAG E 1 (2*) 

INFLAMMATION , NOS 2 (1%) 

•COLON (»$} (145) (38) 

PARASITIS M 1(2%) 

HRINAR Y SYSTE M 

HKIDMF Y (U7) (147) (16) 

CYST , NOS 1 (2X) 1 (2*) 

GLOKFFULON2PHRITIS , NO S 6 (13X ) 9 (19X ) 7 (15X) 

INFLAMMATION , NO S 1 (2X) INFLAMMATION , INTERSTITIA L 2 3 (U9X ) 1 (2X) FIBRCSIS , DIFFUS E 1 (2X ) 

*KIDNFY/TOBUL E («7) (t7) (16) NECROSIS, FOCAL 1 (2X) 

*KirN"Y/P!LVI S (K7 ) (147) (146) 

INFLAMMATIO N , HCUTF/CHSONI C 1 (2X) 

tURINAR Y BLADDE R (U8 ) (147) (11) INFLAMMATION , NO S 14 (8X) 

HYPEFFLASIA , EPITHELIA L 9 (19X) 

ENDOC3TN ENEOCRIN F 3YSTE SEHB 

iitP.nPL.ASIA,

IPITUITAR Y

H.IP.F.FPLA.SIAJ. ,NOS ^ . 

(12 

-...-.J.J7.SJ.

(42))

...,.,]Mil ,.„„

(30) ) (35) 

...l.jtJS).. .„, 

(35) 

__... I-jtJS)... 

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

* NUMBE E OF ANIMAL S NECROPSIST 

D-6 
TABLE Dl (CONTINUED)
 

CONTROL (ONTR) LOW DOSE HIGH DOSE 

05-0037 05-0058 05-0059 

HYPEFPLASIA , FOCAL 3 (7X) 

tADF'NA L CORTE X (15) (i*6) (t6) 

MODBL3 1 (2X)

 HYPEPTROFHY , FOCA L 1 (2*)

 HYPEPPLASIA , NO S 1 (2X)

 

*ADPENA L MEDULL A (15) (46) (1*6) 

DEGENERATION , NO S 1 (2X) HYPEFPLASIA , NODULA R 1 (2X) 

*THYROI D (U7 ) (12) (1*2)

 

LYKPHOCYTIC INFLAMMATOR Y INFILTR 1 (2X)

 HYPEFPLASIA , PAPILLAR Y 1 (2%)

 HYPIFPLASIA , FOLLICULAR-CEL L 1 <2*>

 

UPANCPEATI C ISLETS (1*9) («5) (U5)

 

HYPE.RPLASIA , NOS 2 («%) 

REPRODUCTIV E SYSTE M 

*PFFFUTIA L GLAN D (U8) (U8) (1*8)

 

ABSCESS , NO S 2 (US) 

UTESTIS (H7) (U7) (1*6) 

FIEROSI S 1 (2S) TEGENEPATION , NO S 1 (2«) 

•TFSTIS/TUBOLF (147) (U7 ) (K6 > 

DEGENERATION , NO S U (9») 11 (23X ) 1 (2*) 

NEPVCU S SYSTE M 

tCE»EBRA L COSTr.X (1*8) <U6 ) (U7)

 

KINEFALIZATIO N 3 (6%)

 

SPECU L SEHS E ORGAN S 

NON E 

BUSCOLOSKF.LPTAL SYSTEM 

NOS E 

» NUCEEF OF ANIMAL S HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUKBE ? OF ANIMAL S NECROPSIEC
 

D-7
 
BODY CAVITIES
 

NONE
 

AL1 CTHEh SYSTEMS
 

NONE
 

SPECIAL KOBPHOLOGY SUMMARY
 

NO LFSION PEPOBTED
 

ANIMA L MISSING/NO NECROPSY
 

AUTO/HECROPSY/NO HISTO
 

A'JTOLYSIS/NO NECROPSY
 

# NUK3FK 01 ANIMALS WITH TISSUF

* NUMB? B OF ANIMAL S NECROPSIE D 

TABLE DI (CONCLUDED) 

CONTROL (ONTR) LOW DOSE
 

05-0037 05-0058
 

6
 

1
 

1
 

2 1
 

 EXAMINE D MICBOSCOPICALLY 

D-8 

HIGH DOSE
 05-0059
 

13
 

1
 

1
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE TREATED WITH APC 

ANIMALS INITIALLY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE
 

MINERALIZATION
 

FIBROSIS
 

RESPIRATORY SYSTEM
 

•LUNG/BRONCHUS
 

INFLAMMATION, FOCAL
 

»LUN<3
 

INFLAMMATION, INTERSTITIAL
 

HYPEEPLASIA, EPITHELIAL
 

HYPEPPLASIA, ADENOMATOUS
 

HYPERPLASIA, ALVEOLAR EPITHELIUM
 

HEMATOPOIETIC SYSTEM
 

*BONE MARROW
 

MYELOFIBROSIS
 

MYELOSCLEROSIS
 

tSPLEEN
 

HEHORRHAGIC CYST
 

HYPEPPLASIA, HEMATOPOIETIC
 

HYPEFPLASIA, ERYTHROID
 

HYPERPLASIA, RETICULOM CELL
 

HYPERPLASIA, LYMPHOID
 

HEMATOPOIESIS
 

MYELOPOIESIS
 

tLYMPH NODE
 

... C.l3.1»J!25-_. -«.
 

CONTROL (UNTR)
 

06-0037
 

50
 

U8
 ** 1(7
 

0*8)
 

1 (2X)
 

1 (2»)
 

(U6)
 

1 (2X)
 

l«6)
 

10 (22X)
 

3 (7X)
 

<«5)
 

1 (2*)
 

(«6)
 

16 (35X)
 

6 (13X)
 

10 (22X)
 

1 <2X>
 

1 <2X)
 

(39)
 j.im
 

LOW DOSE HIGH DOSE
 

06-0058 06-0059
 

50	 50
 

1 1
 

i»5 «7
 U5 17
 

(U5)	 (17)
 

(U5)	 («7)
 

(«5)	 (17)
 

1 (2X)
 

1 (2X)
 

(UU) (17)
 

2 (5X)
 

.1 (7X)
 

<"»<»> («7)
 

1 (2X)
 

7 (16X) 2 (»X)
 

1 (2X)	 3 <6*>
 2 («X)
 

(3U)	 (38)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIBALS NECROPSIED
 

**l'.:;CLUi,EC PAKTIALLY AUTOLYZEi) AlilliALS 

D-9
 
TABLE D2 (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE 06-0037 06-0053 06-0059 

INFLAMMATION , NO S 15 (36X) 

HYPEPPLASIA , NO S 1 <3X )

 

PETICULOCYTOSIS 1 (3%)

 

HYPERPLASIA , HEM ATOPOIETIC 2 <5X) 

MYELCID METAPLASIA 1 (3X)

 

tSUESANDIBULA R L.NOP E (3^ ) (3t) (38) 

HYPFSPLA SIA, RETICULUM CFLL 1 (3*) 

tMEDIASTINAL I. NODE (39) (3t) (38)

 

HYFEEPLASIA , LYMPHOI D 1 (3*)

 

*PANCREATIC L.NODE (39) (3t) (38) 

HYPEPPLASIA , LYMPHOI D 1 (3%) 

tMESIlNTFRIC L. NODE (39) (314) (3S) 

LYMPHADENOPATH Y 3 (9X) « (11X) 

*THY!1(JS (31) (39) (12) 

HYPERPLASIA , LYMPHOID 1 (3») 1 (2X) 

CIECUIATORY SYSTEM 

IHEART/VENTRICLE («6) (U 5) (H7)

 

MELANI N « (9»)

 

DIGESTIVE SYSTE M 

ISALIVAF Y GLAN D («5) (it 3) (U7 )

 

INFLAMMATION, NOS 2 (t*)

PSRIVASCULA R CUFFIN G K (9X)

 

*LIVER (U7) (»5) (U7) 

INFLAMMATION , NO S 1 (2*) 

DEGENERATION , NOS 1 (2*) 

NECROSIS , FOCAL 22 C*7X) 

HEPATOCYTOMEGAL Y 1 (2*) 2 («*) 

HYPEFTROPHY , FOCAL 1 (2*) 

HYPERPLASTI C NODUL E 1 (2*) 1 (2*) 

HYPEf.PLASIA, FOCAL 1 (2X) 1 (2X) 

ANGIFCTASI S 1 (2X) 2 (UX ) 3 (6X) 

HEBATOPOIESIS 3 (6*) 

*GALLELADDE R (US ) (<»5) («7) 

......I.M.A.MM,AT.ION^.N2S ,..,.,......„ J.^gX.._._.. 

* NUKBEK OF ANIMALS WITH TISSUF EXAMINED MICROSCOPICALLY
 

* HOB3FS OF ANIMALS NECROPSIFD
 

D-10
 
TABLE D2 (CONTINUED) 

CONTROL (BHTR) LOW DOSE HIGH DOSE 

06-0057 06-0058 06-0059 

*BIL H DUCT <U8 ) (45) (U7)

 

INFLAMMATION , NO S 1 (2X)

 

• PANCREA S (UK ) C*t) (H7) 

DILATATION/DUCT S 1 (2X) 

CYSTIC DUCTS 1 (2%) 

INFLAMMATION , NOS 5 (11%) 

FIBROSrS 1 (2«) 

P3FIARTERITI S 1 (2%) 

• PANCREATI C DUCT (tilt) («t) (117)

 

LYMPFOCYTIC INFLAMMATOR Y INFILTP 1 (2X)

 

• PANCREATI C ACINU S (1«) (UK ) (17) 

ATROPHY , NO S 1 <2*> 

• STOI1ACH (tttt) (H 3) (K5) 

INFLAMMATION , NO S 7 (16») 

IILCFR, NOS 1 (2X) 

INFLAMMATION , FOCAL 1 <2X ) 1 <2X) 

ULCEF , FOCAL 1 (2%) 

INFLAMMATION , ACUTE FOCAL 1 (2%) 

r.RANOLOMA , NO S 1 (2*) 

HYPFFPLASIA , NO S 1 (2X) 

HYPEFPLASIA , EPITHELIA L 1 (2%) 

HYPF.fPLASIA , ADENOMATOO S 1 (2«) 

HYPEFKERA T OSI S 1 <2X) 

ACANTHOSI S 1 (2%) 

• GASTRIC MUCOS A (»«) <t3) (45)

 

HYFFFPLASIA , FOCAL 1 (2«)

 

• PAYERS PHTC H (Hit) <«5) (46) 

HYPFRPLASIA , NOS 1 (2X)

 

tlLFU K {««) (15) (46) 

S.1YLOIPOSIS 1 (2X) 

URINAR Y EYSIE M 

HKIDNT Y (U6 ) (U5) (47)

 

CYST, NOS 1 (2*)

 

GLO«ERULONEPHRITIS t NO S ....i^jjasiTM.. l_iHJ.._TM

 

» NtJHBr R OP ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALLY 

* NtJKBF P OF ANIMAL S NECROPSIED 

D-ll 
TABLE D2 (CONTINUED) 

CONTROL (UNTR) LOW DOSE 

06-0037 06-3058 

INFLAMMATION , INTERSTITIAL 16 (35%) 2 (IX) 

NSPHROPATH Y 1 (2X) 

iKIDNEY/PELVI S C*6) (15)

 

LYMPhOCYTIC INFLAMMATOR Y INFILTR 

iUPINAR Y BLADDER (16) (11)

 

INFLAMMATION , NO S 14 <9X) 

HYPERPLASIA, EPITHELIAL 10 (22%)

 

F.NDCCBIN E SYSTE M 

»PITUITAF Y ("2) (38)

 

HYPEEPLASIA, FOCAL 6 <1*X)

 

MDF'NAl CORTSX (15) (12) 

NOD01E 3 (7X) 

MFTAKORPHOSIS FATTY 1 (2X) 

HYPEFPLASIA . FOCAL 1 (2X) 

• THYROI D (13) (31)

 

FOLLICHLAR CYST, NOS 1 (2X)

 

INFLAMMATION , NO S 1 (2X)

 

HYFFRPLASIA , FOLLICtlLAR-CELL 

REPRODUCTIV E SYSTE M 

*MAMMAK Y 3LAN D (18) (15)

 

GALACTOCELE 1 (2%)

 

HYFEFPLASIA , NOS 1 (8X) 

»UTE3U S (15) (13) 

LYKPHANGI'SCTAST. S 

HYDFOMiTTR A 1 (2%) 7 (16X) 

INFLAKJIATION , NO S 1 (211) 

PYOMFTR A 1 (2«)

 

INFLAMMATION , ACUT E 

ABSCFSS, HOS 3 <7X)

 

FI3P.CSIS 1 (2%)

 

• UTESUS/FNDOJ1ETPIU M (15) (13) 

INFLAMMATION , NOS 10 (22*) •4 <9K) 

I.SFLAM.H.ATI.qSU.SUP.PURATIV? . . ,. ...-HJ881 . 

•	 NUKBEf OF ANIMALS KITH TISSUS EXAMINED MICROSCOPICALLY
 

* NUKSEF OF ANIMALS NECROPSIFD
 

HIGH DOSE 

06-0059 

5 (11*) 

(17) 

1 <2X) 

(«6) 

(11) 

(16) 

(15) 

1 <2X) 

(17) 

(16)

1 (2*) 

9 (20X) 

1 (2X) 

(16) 

D-12 
TABLE D2 (CONTINUED)
 

CONTROL (UNTR) LOW DOSE
 06-0037 06-00'S8
 

INFLAMMATION, ACUTE
 

ABSCESS, NOS
 

HYPEF.PLASIA, NOS U (9*)
 

HYFFFPLASIA, CYSTIC 18 (40X) 33 (77X)
 

HYPFF?LASIA, ADENOMATOUS 1 (2X)
 

HYPEFFLASIA, STROMAL
 

METAPLASIA, SQUAMOUS 1 (2%)
 

ffUTFSUS/MYOMETRIUM (U5) (13)
 

LYflPHOCYTIC INFLAMMATORY INFILTF ' (2X)
 

*OVARY/OVIDUCT («5) (13)
 

INFLAMMATION, NOS 5 (11X)
 

INFLAMMATION, SUPPURATIVE 2 (5*)
 INFLAM1ATION, CHRONIC 1 (2X)
 

HOVARY (U5) ft 5)
 

CYST, NOS 3 (7*)
 

INFLAMMATION, NOS t (9»)
 

LYf.PFOCYTIC INFLAMMATORY INFILTR 1 (2%)
 

INFLAMMATION, SUPPURATIVS 10 (22%)
 

ABSCESS, NOS t (9%)
 

DEGENERATION, CYSTIC 1 (2%)
 

HYPEFPLASIA, NOS 1 <2X)
 

NFBVCBS SYSTEM
 

*BEAIN <<ti») (15)
 

LYMPHOCYTIC INFLAMMATORY INFILTR
 

INFLAMMATION, ACUTE/CHRONIC
 

SPECIAL SENSE ORGANS
 

NONE
 

KUSCOLOSKELETAL SYSTEM 

*BONS (48) (15) 

PESORPTION 3 (6X) 

BOtY CAVITIES
 

,.«ONE ,„. _. ., ^_ __,
 

» NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBFR OF ANIMALS NECROPSIED
 

D-13
 

HIGH DOSE
 06-0059
 

1 (2X)
 

1 (2*)
 

23 (50%)
 

1 <2X)
 

1 (2«)
 

(16)
 

(16)
 

(42)
 

(45)
 

1 <2X)
 

1 <2X)
 

(47)
 
ALL OTHER SYSTEMS
 

OMENTUM
 

NECROSIS, FAT
 

SPECIAL KOFPHOLOGY SUMKAK J
 

NO LESION REPORTED
 

ANIMA L MISSING/NO NECROfSY
 

AOTO/N^CROPSY/HISTO PERF
 

A'JTO/NECROPSY/NO HISTO
 

AUT01YSI5/NO NECROPSY
 

» NUMBE R 0 ? ANIMAL S WIT H TISSUE

* NUMBF R O F ANIMAL S NECROPSIE D 

TABLE D2 (CONCLUDED)
 

CONTROL (ONTR) LOW DOSE
 06-0037 06-3053
 

1
 

1 2
 

1
 

2
 

1
 

2 it
 

 EXAMINF D MICROSCOPICALL Y 

D-14 

HIGH DOSE
 06-0059
 

6
 

1
 

2
 

DHEW Publication No. (NIH) 78-1317