National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 142 

1979 

BIOASSAY OF 

p-CRESIDINE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 120-71-8 

NCI-CG-TR-142 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

p-CRESIDINE
 

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) 79-1397
 

REPORT ON THE BIOASSAY OF p-CRESIDINE
 

FOR POSSIBLE CARCINOGENICITY
 

CARCINOGENESIS TESTING PROGRAM
 

DIVISION OF CANCER CAUSE AND PREVENTION
 

NATIONAL CANCER INSTITUTE, NATIONAL INSTITUTES OF HEALTH
 

FOREWORD: This report presents the results of the bioassay of
 

p-cresidine 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 p-cresidine 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), Dr. A. S. Krishna Murthy (3) and Dr. A. Russfield (3) at the
 

Mason Research Institute, and the diagnoses included in this report
 

represent the interpretation of these pathologists. Histopathology
 

findings and reports were reviewed by Dr. R. L. Schueler (6).
 

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

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

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

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

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

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

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

P. Walker (5), biochemist, Mr. S. C. Drill (5), and technical editor
 

Ms. P. A. Miller (5). The final report was reviewed by members of
 

the participating organizations.
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

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

Cueto, Jr. (1), Dr. J. F. Douglas (1), Dr. D. G. Goodman (1,10), Dr.
 

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

T. W. Orme (1), Dr. R. A. Squire (1,11), Dr. S. F. Stinson (1), Dr.
 

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

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

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

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

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

Missouri.
 

5.	 The MITRE Corporation, METREK Division, 1820 Dolley Madison
 

Boulevard, McLean, Virginia.
 

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

Maryland•
 

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

Rockville, Maryland.
 

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

Golden, Colorado.
 

9.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

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

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

Washington, D.C.
 

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland•
 

SUMMARY
 

A bioassay of p-cresidine for possible carcinogenicity was conducted using Fischer 344 rats and B6C3F1 mice. p-Cresidine was administered in the feed, at either of two concentrations, to groups
 

of 50 male and 50 female animals of each species. The dietary concentrations used in the chronic bioassay for low and high dose rats
 

were 0.5 and 1.0 percent, respectively. The time-weighted average
 

concentrations fed to low dose male, low dose female, high dose male
 

and high dose female mice were 0.22, 0.22, 0.46, and 0.44 percent,
 

respectively.
 

All dosed animals, except for high dose male mice, were administered p-cresidine in the diet for 104 weeks and observed for an additional period of up to 2 weeks. All high dose male mice were dead by
 

the end of week 92. For each species, 50 animals of each sex were
 

placed on test as controls and fed only the basal laboratory diet.
 

Mortality rates were dose-related for both sexes of both species. That incidences of certain tumors were higher in low dose than
 

in high dose groups was probably due to accelerated mortality in the
 

high dose groups.
 

In dosed rats of both sexes, statistically significant incidences
 

of bladder carcinomas (combined incidences of papillary carcinomas,
 

squamous-cell carcinomas, transitional-cell papillomas, transitional-


cell carcinomas, and undifferentiated carcinomas) and olfactory neuroblastomas were observed. The combined incidence of neoplastic nodules
 

of the liver, hepatocellular carcinomas, or mixed hepato/cholangio carcinomas was also significant in low dose male rats.
 

In both male and female dosed mice, the incidence of bladder
 

carcinomas (combined incidence of carcinomas NOS, squamous-cell carcinomas, and transitional-cell carcinomas) was significant. The incidence of hepatocellular carcinomas was significant in dosed female
 

mice.
 

Under the conditions of this bioassay, p-cresidine was carcinogenic to Fischer 344 rats, causing increased incidences of carcinomas
 

and of papillomas of the urinary bladder in both sexes, increased
 

incidences of olfactory neuroblastomas in both sexes, and of liver
 

tumors in males. p-Cresidine was also carcinogenic in B6C3F1 mice,
 

causing carcinomas of the urinary bladders in both sexes and hepatocellular carcinomas in females.
 

vii
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 4
 

A. Chemicals 4
 

B. Dietary Preparation 4
 

C. Animals 5
 

D. Animal Maintenance 6
 

E. Selection of Initial Concentrations 9
 

F. Experimental Design 9
 

G. Clinical and Histopathologic Examinations 12
 

H. Data Recording and Statistical Analyses 14
 

III. CHRONIC TESTING RESULTS: RATS 19
 

A. Body Weights and Clinical Observations 19
 

B. Survival 19
 

C. Pathology 22
 

D. Statistical Analyses of Results 26
 

IV. CHRONIC TESTING RESULTS: MICE 42
 

A. Body Weights and Clinical Observations 42
 

B. Survival 42
 

C. Pathology 45
 

D. Statistical Analyses of Results 47
 

V. DISCUSSION 57
 

VI. BIBLIOGRAPHY 60
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH p-CRESIDINE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH p-CRESIDINE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH p-CRESIDINE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH p-CRESIDINE D-l
 

ix
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF p-CRESIDINE 2 

2 GROWTH CURVES FOR p-CRESIDINE CHRONIC STUDY 

RATS 20 

3 SURVIVAL COMPARISONS OF p-CRESIDINE CHRONIC 

STUDY RATS 21 

4 GROWTH CURVES FOR p-CRESIDINE CHRONIC STUDY 

MICE 43 

5 SURVIVAL COMPARISONS OF p-CRESIDINE CHRONIC 

STUDY MICE 44 

LIST OF TABLES 

Table Number Page 

1 DESIGN SUMMARY FOR FISCHER 344 RATS--p-CRESIDINE FEEDING EXPERIMENT 10 

2 DESIGN SUMMARY FOR B6C3F1 MICE—p-CRESIDINE 

FEEDING EXPERIMENT 11 

3 ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

p-CRESIDINE 27 

4 ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH p-CRESIDINE 34 

5 TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN MALE 

MICE TREATED WITH p-CRESIDINE 48 

6 TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF 

PRIMARY TUMORS AT SPECIFIC SITES IN FEMALE 

MICE TREATED WITH p-CRESIDINE 51 
LIST OF TABLES (Concluded) 

Table Number Page 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH p-CRESIDINE A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH p-CRESIDINE A-8 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH p-CRESIDINE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH p-CRESIDINE B-7 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH p-CRESIDINE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH p-CRE­ SIDINE C-8 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH p-CRESI­ DINE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH p-CRE­ SIDINE D-6 

xi
 

I. INTRODUCTION 

p-Cresidine (Figure 1) (NCI No. CO2982), used in the production 

of various azo dyes, was selected for bioassay by the National Cancer 

Institut e in response to the high incidence of bladder cancer observed among dye manufacturing industry workers (Anthony and Thomas, 

•1970; Wynder et al. , 1963), Aromatic amines are one class of chemicals believed to contribute to the increased cancer risk in this 

industry (Wynder et al. , 1963). 

The Chemical Abstracts Service (CAS) Ninth Collective Index 

(1977) name for this compound is 2-methoxy-5-methylbenzenamine.* It 

is also known as 2-methoxy-5-methylaniline; 5-methyl-o-anisidine; 

m-amino-p-cresol methyl ether; MASO; and simply as cresidine . 

p-Cresidine appears to be used solel y as an intermediate for 

dyes and pigments. It is believed to be used in the preparation of 

eleven dyes which are produced commercially in the United States: 

C.I. (Colour Index) Direct Black 17, Direct Blue 67, Direct Blue 126, 

Direct Green 26, Direct Orange 34, Direct Orange 83, Direct Red 79, 

Direct Violet 9, Direct Violet 51, Direct Yellow 41, and Disperse 

Black 2 (Society of Dyers and Colourists, 1971; as cited in Urso, 

1977). 

Precise production figures for p-cresidine are not available; 

however, the Sherwin-Williams Company, which produces p-cresidine , 

*The CAS registry number is 120-71-8, 
FIGURE 1

 

CHEMICAL STRUCTURE OF p-CRESIDINE 

2 
estimates that average use of the compound during the period 1971
 

to 1976 was about 590,000 pounds annually (Compton, 1977).
 

Since p-cresidine is used solely as an intermediate in dye manufacture, potential for exposure is primarily limited to workers in
 

dye production facilities.
 

While adequate chronic toxicity data for p-cresidine are not
 

available, a study of workmen involved in the manufacture and handling of the compound, conducted by the Sherwin-Williams Company,
 

revealed no adverse health effects related to p-cresidine exposure
 

(Compton, 1977).
 
II. MATERIALS AND METHODS
 

A. Chemicals
 

p-Cresidine was purchased from Carroll Products, Wood River Junction, Rhode Island by the NCI for Mason Research Institute, Worcester,
 

Massachusetts and chemical analysis was performed by Midwest Research
 

Institute, Kansas City, Missouri. The experimentally determined melting point of 51.5° to 52.5°C, due to its narrow range and proximity to
 

the literature value of 51° to 52°C (Pollock and Stevens, 1965), suggested a compound of high purity. Thin-layer chromatography using a
 

benzene:ethyl acetate solvent system and visualized with ultraviolet
 

light indicated the presence of three impurities, two less motile
 

and one more motile than the major compound. Infrared analysis was
 

consistent with the structure of the compound. Ultraviolet analysis
 

indicated K m a x at 290 and 237 nm with respective molar extinction
 

coefficients (e) of 3312 and 6820. The literature values are \ ma x =
 

290 and 236 nm with respective e values of 3620 and 7120 (Mehta, 1978).
 

Titration of the amine function with perchloric acid indicated a
 

purity of 98.9 +^ 0.1 percent.
 

Throughout this report the term p-cresidine is used to represent
 

this technical-grade material.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals
 

consisted of Wayne Lab-Blox®(Allied Mills, Inc., Chicago, Illinois).
 
p-Cresidine was administered to the dosed animals as a component of
 

the diet.
 

The brown amorphous chemical was removed from its shipping container and stored in an amber glass stock bottle inside a plastic
 

bucket. The p-cresidine was weighed out in proper amounts from the
 

stock bottle under an exhaust hood. The compound was dissolved with
 

an appropriate amount of acetone (0.016 percent by volume of the
 

final mix) with the help of a mortar and pestle. This liquid mixture
 

was hand-blended in an aluminum bowl with an aliquot of the ground
 

feed until all liquid was absorbed and visual homogeneity was attained.
 

This premix was then placed into a 6 kg capacity Patterson-Kelley
 

standard model twin-shell stainless steel V-blender along with the
 

remainder of the diet. The blender was sealed and operated for 20
 

minutes. The mixture was then emptied into stainless steel trays and
 

allowed to sit under the exhaust hood for 2 and 1/2 hours while 'being
 

stirred with a glass rod every 1/2 hour, allowing evaporation of a
 

large proportion of the acetone.
 

Prepared diets were placed in double clear plastic bags and
 

stored in the dark at 4°C. Diets were prepared weekly and any unused
 

portions were discarded after 2 weeks.
 

C. Animals
 

Two animal species, rats and mice, were used in the carcinogenicity bioassay. Fischer 344 rats and B6C3F1 mice were obtained through
 
contracts of the Division of Cancer Treatment, National Cancer Institute from Charles River Breeding Laboratories, Wilmington, Massachusetts,
 

Dosed and control animals for both species were received in separate
 

shipments.
 

Upon arrival a sample of animals was examined for parasites and
 

other signs of disease. Animals to be used in the chronic study
 

were quarantined by species 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
 

species and sex.
 

D. Animal Maintenance
 

All animals were housed by species in rooms having a temperature
 

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

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

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

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

Rats were housed five per cage by sex. During quarantine and for
 

the first 12 months of study, dosed rats were housed in galvanized
 

steel wire-mesh cages (Fenco Cage Products, Boston, Massachusetts)
 

suspended over newspapers. Control rats were held in galvanized
 

cages for the first 14 months of study. Newspapers under cages were
 

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

of the study, all rats were housed in suspended polycarbonate cages
 

(Lab Products, Inc., Garfield, New Jersey) equipped with disposable
 
nonwoven fiber filter sheets• Clean bedding and cages were provided
 

twice weekly. SAN-I-CEL® corncob bedding (Paxton Processing Company,
 

Paxton, Illinois) was used during the first 6 months of polycarbonate
 

caging. Aspen hardwood chip bedding (American Excelsior Company,
 

Baltimore, Maryland) was used for the remainder of the study. Stainless steel cage racks (Fenco Cage Products) were cleaned once every
 

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

Mice were housed by sex in polycarbonate cages fitted with perforated stainless steel lids (Lab Products, Inc.). Nonwoven fiber
 

filter bonnets were used over cage lids. Dosed mice were housed
 

ten per cage for the first 9 months and five per cage thereafter.
 

Control mice were changed from ten per cage to five per cage after
 

11 months. Clean cages, lids, and bedding were provided three times
 

per week when cage populations were ten and twice per week when cage
 

populations were five. SAN-I-CEL® was used for the first 12 months
 

for control mice, and for the first 10 months for dosed mice. Bedo-Cobs® corncob bedding (The Andersons Cob Division, Maumee, Ohio)
 

was used for 8 months subsequent to utilization of SAN-I-CEL®. Aspen
 

bedding was then used until completion of the study for all groups of
 

mice. Reusable filter bonnets and pipe racks were sanitized every 2
 

weeks throughout the study.
 

Water was available from 250 ml water bottles equipped with rubber stoppers and stainless steel sipper tubes. Bottles were replaced
 
twice weekly and, for rats only, water was supplied as needed between
 

changes. Food and water were available ad libitum.
 

During the period of chemical administration, animals were supplied Wayne Lab-Blox®containing the appropriate concentrations of
 

p-cresidine and acetone. The basal laboratory diet was supplied to
 

control animals during the same period. Meal was distributed from
 

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

Massachusetts) to rats while in galvanized-steel cages. While in
 

polycarbonate cages, rats were fed from stainless steel gangstyle
 

hoppers (Scientific Cages, Inc., Bryan, Texas). Mice were fed from
 

Alpine® feed cups for the first 12 months (dosed groups) or 14 months
 

(controls) of study. Stainless steel gangstyle feed hoppers were
 

used for the remainder of the test. Food hoppers were changed on the
 

same schedule as were cages. Food was replenished daily in Alpine®
 

feed cups.
 

Dosed and control rats were housed in a room with other rats
 

receiving diets containing* acetylaminofluorene (53-96-3); 2,3,5,6­

tetrachloro-4-nitroanisole (2438-88-2); 4-chloro-o-phenylenediamine
 

(95-83-0); lH-benzotriazole (95-14-7); and 4-chloro-m-phenylenediamine (5131-60-2).
 

Dosed and control mice were housed in a room with other mice
 

receiving diets containing 2-methyl-l-nitroanthraquinone (129-15-7);
 

aniline hydrochloride (142-04-1); acetylaminofluorene (53-96-3); fenaminosulf (140-56-7); and 4-chloro-m-phenylenediamine (5131-60-2).
 

CAS registry numbers are given in parentheses.
 
E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations
 

of p-cresidine for administration to dosed animals in the chronic
 

studies, subchronic toxicity tests were conducted with both rats
 

and mice.
 

Animals of each species were distributed among three groups,
 

each consisting of five males and five females. p-Cresidine was
 

administered to two of the three groups of each species at dietary
 

concentrations of 1.0 and 3.0 percent for 8 weeks. The third group
 

of each species served as a control group, receiving only the basal
 

laboratory diet.
 

As deaths occurred among all groups except male rats receiving
 

3.0 percent p-cresidine in the feed, the high concentration chosen
 

for the chronic study was 1.0 percent p-cresidine for both rats and
 

mice.
 

F. Experimental Design
 

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

treated and untreated observation periods, and time-weighted average
 

concentrations) are summarized in Tables 1 and 2.
 

All animals were approximately 6 weeks old at the time they
 

were placed on test. Dosed rats and mice shared the same median date
 

of birth, while controls were born approximately 7 weeks earlier and
 

placed on test 7 weeks before dosed animals.
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

p-CRESIDINE FEEDING EXPERIMENT
 

INITIAL p-CRESIDINE OBSERVATION PERIOD 

GROUP CONCENTRATION TREATED UNTREATED 

SIZE (PERCENT) (WEEKS) (WEEKS) 

MALE 

CONTROL 50 0 0 106 

LOW DOSE 50 0.5 104 

0 1 

HIGH DOSE 50 1.0 104 

0 1 

FEMALE 

CONTROL 50 0 0 106 

LOW DOSE 50 0.5 104 

0 2 

HIGH DOSE 50 1.0 104 

0 2 

10
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

p-CRESIDINE FEEDING EXPERIMENT
 

INITIAL p-CRESIDINE OBSERVATION PERIOD TIME-WEIGHTED 

GROUP CONCENTRATION TREATED UNTREATED AVERAGE 

SIZE (PERCENT) (WEEKS) (WEEKS) CONCENTRATION8 

MALE 

CONTROL 50 0 0 97 0 

LOW DOSE 50 0.5 21 0.22 

0.15 83 

0 2 

HIGH DOSEb 50 1.0 21 0.46 

0.3 71 

FEMALE 

CONTROL 50 0 0 97 0 

LOW DOSE 50 0.5 21 0.22 

0.15 83 

0 2 

HIGH DOSE 50 1.0 21 0.44 

0.3 83 

0 2 

3. Σ(concentration X weeks received)
 

Time-weighted average concentration = Σ(weeks receiving chemical)
 

b All animals in this group were dead by the end of week 92·
 

11
 
When it was noted that a trace acetone odor remained in the dosed
 

feed, acetone controls were placed on test in addition to the untreated
 

controls. However, these acetone controls were not entirely comparable to dosed animals because one set of acetone controls had an inadequate number of animals (only 10 animals of each sex and species) and
 

were treated for parasites with piperazine adipate. A second set of
 

acetone controls was not utilized in comparisons with dosed animals
 

because they were obtained from a different supplier than the other
 

groups.
 

The initial concentrations of p-cresidine used for both rats and
 

mice were 1.0 and 0.5 percent. Throughout this report those animals
 

initially receiving the former concentration are referred to as the
 

high dose groups, while those initially receiving the latter concentration are referred to as the low dose groups. These concentrations
 

were utilized throughout the study for rats; however, after 21 weeks,
 

the concentrations administered to mice were decreased to 0.3 and
 

0.15 percent for the high and low dose groups, respectively. p-Cresidine was administered in the diet to dosed animals for a period of
 

104 weeks (except for high dose male mice which were all dead by week
 

92), followed by an observation period of up to 2 weeks.
 

For both species, control animals had the basal laboratory diet
 

available ad libitum during the bioassay.
 

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
 

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

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

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

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

bioassay and for three consecutive days each month thereafter. The
 

presence of tissue masses and lesions was determined by monthly
 

observation and palpation of each animal.
 

A necropsy was performed on each animal regardless of whether it
 

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

bioassay. The animals were euthanized by carbon dioxide inhalation,
 

and were immediately necropsied. The histopathologic examination consisted of gross and microscopic examination of major tissues, organs,
 

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

from animals found dead.
 

Tissues were preserved in 10 percent buffered formalin, embedded
 

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

to microscopic examination. An occasional section was subjected to
 

special staining techniques for more definitive diagnosis.
 

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

spleen, lymph nodes, thymus, heart, salivary gland, liver, gallbladder
 

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

kidney, urinary bladder, seminal vesicle, pituitary, adrenal, thyroid,
 

parathyroid, testis, prostate, brain, uterus, mammary gland, and ovary.
 

13
 
A few tissues were not examined for some animals, particularly
 

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

preclude histopathologic interpretation. Thus, the number of animals
 

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

animals that were placed on experiment in each group.
 

H.	 Data Recording and Statistical Analyses
 

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

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

as recommended by the International Union Against Cancer (Berenblum,
 

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

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

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

14
 
missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect
 

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

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

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

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

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

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

histologically. However, when macroscopic examination was required
 

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

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

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

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

15
 
with those for a control group, a correction to ensure an overall
 

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

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

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

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

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

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

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

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

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

16
 
procedures for analyses of the incidence of tumors (Fisher exact tests,
 

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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

during which animals died naturally or were sacrificed was entered
 

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

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

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

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

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

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


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

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

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

is defined as p^/Pc where pt is the true binomial probability of the
 

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

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

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

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

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

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

larger proportion in the treated group than in the control.
 

17
 
The lower and upper limits of the confidence interval of the
 

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

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

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

would be within the interval calculated from the experiment. When
 

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

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

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

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

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

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

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

detected under the conditions of this test.
 

18
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Male and female dosed rats had mean group body weights lower
 

than those of their respective controls (Figure 2), especially the
 

weights of the high dose male group which were markedly decreased
 

throughout the study. Fluctuations in the growth curve may be due to
 

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

may be subject to wide variations.
 

Subcutaneous masses on various parts of the body were observed
 

in 2 control, 3 low dose, and 1 high dose males and in 3 control, 7
 

low dose, and 1 high dose females. Cutaneous masses or nodules were
 

detected in 2 control and 3 low dose males and in 2 low dose and 1
 

high dose females. Skin lesions were observed in 1 control male.
 

Other isolated clinical observations included penile swelling in 1
 

high dose male, swelling of a single nipple in 1 low dose female, and
 

severe emaciation in 1 low dose male.
 

B. Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and p-cresidine-dosed groups are shown in Figure 3.
 

For both male and female rats there were significant positive associations between dosage and mortality.
 

For males five rats from the control group were sacrificed in
 

week 78. Sixty-two percent (31/50) of the high dose, 96 percent
 

19
 
TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR p-CRESIDINE CHRONIC STUDY RATS 

20 
TIME ON TEST (WEEKS) 

FIGURE 3 

SURVIVAL COMPARISONS OF p-CRESIDINE CHRONIC STUDY RATS 

21 
(48/50) of the low dose, and 94 percent (47/50) of the control rats
 

survived on test at least 75 weeks.
 

For female rats five of the controls were sacrificed in week 78.
 

Seventy-six percent (38/50) of the high dose, 98 percent (49/50) of
 

the low dose, and 96 percent (48/50) of the control rats survived
 

on test at least 75 weeks.
 

C. Pathology
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

summarized in Appendix C (Tables Cl and C2).
 

Both male and female rats fed p-cresidine showed compound-


related effects in the urinary bladder and in the epithelium of the
 

nasal cavity.
 

No neoplasms of the bladder were found in the control rats. In
 

contrast, both male and female rats fed p-cresidine developed numerous
 

dose-related neoplasms or hyperplasias in the bladder. The incidences
 

of these lesions are tabulated in the table below:
 

22
 
MALE Control 

Low 

Dose 

High 

Dose 

Number of animals with tissues 

examined histopathologically (48) (48) (47) 

Urinary Bladder 

Hyperplasia 0 11(23%) 2(4%) 

Undifferentiated Carcinoma 0 0 1(2%) 

Papillary Carcinoma 0 0 4(9%) 

Squamous-Cell Carcinoma 0 1(2%) 6(13%) 

Transitional-Cell Papilloma 0 14(29%) 3(6%) 

Transitional-Cell Carcinoma 0 15(31%) 31(66%) 

Leiomyosarcoma 0 0 2(4%) 

FEMALE 

Number of animals with tissues 

examined histopathologically (47) (49) (46) 

Urinary Bladder 

Hyperplasia 0 11(22%) 1(2%) 

Neoplasm NOS 0 0 2(4%) 

Papillary Carcinoma 0 0 1(2%) 

Squamous-Cell Carcinoma 0 9(18%) 7(15%) 

Transitional-Cell Papilloma 0 6(12%) 2(4%) 

Transitional-Cell Carcinoma 0 22(45%) 33(72%) 

Leiomyosarcoma 0 2(4%) 3(7%) 

Hyperplasia was defined as the presence of more than three layers
 

of epithelial cells lining the fully inflated bladder. Nuclei of
 

these cells were histologically normal.
 

The neoplasms were classified as papillomas when they consisted
 

of histologically benign, regularly arranged, transitional epithelium
 

covering a papillary core composed of fibrovascular tissue. Tumors
 

were diagnosed as transitional-cell carcinomas when nuclei of the
 

transitional cells became pleomorphic and hyperchromatic and mitotic
 

figures became frequent. They were classified as squamous-cell
 

23
 
carcinomas when, in addition to nuclear criteria of malignancy, the
 

tumor cells showed cytoplasmic keratinization or developed intercellular bridges. There were no sharp boundary lines between these categories which formed a continuous spectrum of progressively malignant
 

change. The carcinomas were invasive and a few of these tumors
 

metastasized to distant organs. A few rats fed p-cresidine developed
 

leiomyosarcomas of the bladder wall which tended to occur in association with malignant tumors of the bladder epithelium. There were
 

neoplasms of the transitional-cell epithelium of the renal pelvis in
 

1/49 low dose female rats and in 2/49 low dose males. One high dose
 

female developed an undifferentiated carcinoma of the renal parenchyma.
 

No nasopharyngeal neoplasms were found in the control groups.
 

The incidence of neoplasms observed in the nasal cavity is summarized
 

in the following table:
 

Low High 

MALES Control Dose Dose 

Number of animals 

necropsled (48) (50) (47) 

Nasal Cavity 

Neoplasm NOS, Malignant 0 0 1(2%) 

Carcinoma NOS 0 0 1(2%) 

Adenoma NOS 0 1(2%) 0 

Neuroblas toma 0 0 1(2%) 

Olfactory Neuroblastoma 0 1(2%) 20(43%) 

24
 
Low High
 

FEMALES Control Dose Dose
 

Number of animals
 

necropsied (50) (50) (49)
 

Nasal Cavity
 

Squamous-Cell Carcinoma 0 0 1(2%)
 

Squamous-Cell Carcinoma, Invasive 0 0 0
 

Adenocarcinoma NOS 0 0 1(2%)
 

Olfactory Neuroblastoma 0 0 11(22%)
 

The rats fed p-cresidine developed dose-related olfactory neuroblastomas. These tumors consisted of masses of cells having scant
 

cytoplasm and large, ovoid nuclei with prominent chromatin. They
 

were divided into lobules by very fine fibrovascular septae. In many
 

tumors, rosettes, pseudorosettes, or palisade arrangement of cells
 

suggested a neural origin. They frequently exhibited osteoclastic
 

activity and local invasion, destroying the cribriform plate of the
 

skull and invading the forebrain. Some tumors showed focal hemorrhagic necrosis. In addition, there was an adenoma, an adenocarcinoma, a squamous-cell carcinoma, and a polyp involving the nasal
 

cavity of dosed rats. Males appeared to be more susceptible to
 

nasal tumors than females.
 

p-Cresidine feeding had an equivocal effect on the liver of the
 

rats. Males receiving the low dose of the compound developed several
 

(10/49 [20 percent]) neoplastic nodules plus a few hepatic carcinomas.
 

However, neoplastic nodules were seldom seen in high dose males (1/46
 

[2 percent]) and low dose females (4/48 [8 percent]) and were not seen
 

at all in high dose females. Only a few carcinomas were observed in
 

25
 
dosed male rats. The criteria used for the diagnosis of these
 

lesions were those of Squire and Levitt (1975).
 

No tumors of the preputial or clitoral glands were seen in controls; 3/50 (6 percent) occurred in low dose males, 3/50 (6 percent)
 

in low dose females, and 2/47 (4 percent) in high dose males. There
 

were gliomas in 2/48 (4 percent) low dose females, and in 3/43 (7
 

percent) high dose males. As the high dose of p-cresidine shortened
 

the survival of both sexes by inducing massive bladder tumors, an
 

effect on other organ systems may have been obscured.
 

Based on the results of this pathologic examination, p-cresidine
 

was considered to be carcinogenic in Fischer 344 rats because it induced neoplasms of the urinary bladder and olfactory neuroblastomas
 

of the nasal cavity. Neoplastic nodules of the liver may have been
 

induced in the male rat.
 

D.	 Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

rats are summarized in Tables 3 and 4. The analysis is included for
 

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

tumors were observed in at least one of the control or p-cresidinedosed groups and where such tumors were observed in at least 5 percent
 

of the group.
 

In both male and female rats, the combined incidence of undifferentiated carcinomas, papillary carcinomas, squamous-cell carcinomas,
 

transitional-cell papillomas, or transitional-cell carcinomas of the
 

26
 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH p-CRESIDINEa
 

TOPOGRAPHY: MORPHOLOGY
 

Subcutaneous Tissue: Fibroma^
 

P Values0
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Nasal Cavity and Nasopharynx: Malignant
 

Neoplasm NOS; Carcinoma NOS; or
 

Adenocarcinoma NOS^
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Nasal Cavity: Olfactory Neuroblastoma
 

or Neuroblastoma^
 

P Values0
 

Departure from Linear Trende
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

3/48(0.06)
 

N.S.
 

105
 

0/48(0.00)
 

P = 0.035
 

0/48(0.00)
 

P < 0.001
 

P - 0.001
 

LOW
 

DOSE
 

2/50(0.04)
 

N.S.
 

0.640
 

0.055
 

5.345
 

105
 

0/50(0.00)
 

N.S.
 

1/50(0.02)
 

N.S.
 

Infinite
 

0.051
 

Infinite
 

84
 

HIGH
 

DOSE
 

1/47(0.02)
 

N.S.
 

0.340
 

0.007
 

4.058
 

105
 

3/47(0.06)
 

N.S.
 

Infinite
 

0.615
 

Infinite
 

67
 

21/47(0.45)
 

P < 0.001
 

Infinite
 

6.974
 

Infinite
 

48
 
TABLE 3 (Continued ) 
TABLE 3 (Continued) 

TOPOGRAPHY: MORPHOLOGY CONTROL
 

Liver: Hepatocellular Carcinoma, or
 

Mixed Hepato/Cholangio Carcinomab 0/48(0.00)
 

Ρ Valuesc N.S.
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Neoplastic Nodule, Hepatocellular
 

Carcinoma, or Mixed Hepato/Cholangio
 

Carcinoma13 0/48(0.00)
 

Ρ Values0 N.S.
 

Departure from Linear Trende Ρ < 0.001
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW
 

DOSE
 

3/49(0.06)
 

N.S.
 

Infinite
 

0.590
 

Infinite
 

105
 

13/49(0.27)
 

Ρ < 0.001
 

Infinite
 

3.938
 

Infinite
 

105
 

HIGH
 

DOSE
 

1/46(0.02)
 

N.S.
 

Infinite
 

0.056
 

Infinite
 

78
 

2/46(0.04)
 

N.S.
 

Infinite
 

0.309
 

Infinite
 

78
 
TABLE 3 (Continued ) 

o
 
___ 

TOPOGRAPHY:MORPHOLOGY
 

Adrenal: Pheochromocytoma"
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Carcinoma, or C-Cell
 

Adenoma"
 

P Values0
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pancreatic Islets: Islet-Cell Adenoma,
 

or Islet-Cell Carcinoma"
 

P Values0
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (Continued)
 

CONTROL
 

4/46(0.09)
 

N.S.
 

78
 

5/43(0.12)
 

N.S.
 

95
 

0/44(0.00)
 

N.S.
 

LOW
 

DOSE
 

7/48(0.15)
 

N.S.
 

1.677
 

0.459
 

7.336
 

102
 

2/41(0.05)
 

N.S.
 

0.420
 

0.042
 

2.398
 

105
 

2/48(0.04)
 

N.S.
 

Infinite
 

0.272
 

Infinite
 

105
 

HIGH
 

DOSE
 

3/44(0.07)
 

N.S.
 

0.784
 

0.121
 

4.367
 

80
 

2/40(0.05)
 

N.S.
 

0.430
 

0.043
 

2.455
 

93
 

2/40(0.05)
 

N.S.
 

Infinite
 

0.327
 

Infinite
 

97
 
_ __ 

TOPOGRAPHY: MORPHOLOGY
 

Preputial Gland: Adenoma NOS, or
 

Carcinomas NOSb
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Testis: Interstitial-Cell Tumorb
 

P Values0
 

Departure from Linear Trende
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Brain and Brain Stem: Glioma NOS, or
 

Oligodendroglioma"
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (Continued ) 

CONTROL
 

0/48(0.00)
 

N.S.
 

37/48(0.77)
 

P - 0.003(N)
 

P < 0.001
 

78
 

0/46(0.00)
 

P = 0.032
 

LOW
 

DOSE
 

3/50(0.06)
 

N.S.
 

Infinite
 

0.578
 

Infinite
 

79
 

45/47(0.96)
 

P = 0.008
 

1.242
 

1.038
 

1.354
 

79
 

0/48(0.00)
 

N.S.
 

HIGH
 

DOSE
 

1/47(0.02)
 

N.S.
 

Infinite
 

0.055
 

Infinite
 

97
 

23/46(0.50)
 

P = 0.006(N)
 

0.649
 

0.471
 

0.916
 

52
 

3/43(0.07)
 

N.S.
 

Infinite
 

0.645
 

Infinite
 

56
 
TABLE 3 (Concluded)
 

aTreated groups received doses of 0.5 or 1.0 percent in feed.
 

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

cThe 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 percent confidence interval on the relative risk of the treated group to the control group.
 

eThe 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 p-CRESIDINEa
 
TABLE 4 (Continued ) 
TABLE 4 (Continued)
 
TOPOGRAPHY: MORPHOLOGY
 

Thyroid: C-Cell Adenomab
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma^
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polypi
 

P Values0
 

Departure from Linear Trende
 

Relative Risk (Control)^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (Continued)
 

CONTROL
 

0/43(0.00)
 

N.S.
 

6/50(0.12)
 

P = 0.045(N)
 

106
 

2/48(0.04)
 

N.S.
 

P = 0.050
 

106
 

LOW
 

DOSE
 

4/42(0.10)
 

N.S.
 

Infinite
 

0.954
 

Infinite
 

103
 

4/50(0.08)
 

N.S.
 

0.667
 

0.147
 

2.635
 

106
 

8/47(0.17)
 

P = 0.042
 

4.085
 

0.872
 

37.831
 

80
 

HIGH
 

DOSE
 

0/40(0.00)
 

N.S.
 

1/49(0.02)
 

N.S.
 

0.170
 

0.004
 

1.328
 

79
 

4/45(0.09)
 

N.S.
 

2.133
 

0.322
 

22.646
 

82
 
TABLE 4 (Concluded)
 

aTreated groups received doses of 0.5 or 1.0 in feed.
 

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

cThe 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 percent confidence interval on the relative risk of the treated group to the control group.
 

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

group when P < 0.05.
 
urinary bladder was increased in both dosed groups compared to the
 

control group—appearing as early as week 51 in the males and week 59
 

in the females. For both sexes, the Cochran-Armitage test indicated
 

a significant (P < 0.001) positive association between dose and incidence. For both sexes this was supported by significant (P < 0.001)
 

Fisher exact test results for both dose levels compared to the control. Based on these results the administration of p-cresidine was
 

associated with the increased incidence of carcinomas of the urinary
 

bladder in both male and female rats.
 

Both the incidence of olfactory neuroblastomas in females and
 

the combined incidence of neuroblastomas or olfactory neuroblastomas
 

in males were increased in the high dose groups compared to the
 

controls—appearing as early as week 48 in the males and week 62 in
 

the females. For both sexes the Cochran-Armitage test indicated a
 

significant (P < 0.001) positive association between dose and incidence. For both sexes these results were supported by significant
 

(P < 0.001) Fisher exact tests comparing the high dose group to the
 

control group. For males the combined incidence of nonspecific
 

neoplasms, malignant carcinomas, or adenocarcinomas of the nasal
 

cavity or nasopharynx was significant for the Cochran-Armitage test,
 

but not for the Fisher exact tests. Based on these results, the
 

administration of p-cresidine was associated with the increased incidence of neuroblastomas and olfactory neuroblastomas of the nasal
 

cavity in both male and female rats.
 

39
 
In male rats the combined incidence of neoplastic nodules of the
 

liver, hepatocellular carcinomas, or mixed hepato/cholangio carcinomas was increased in the low dose group compared to the control. The
 

tumors were found after termination of the study in week 105. The
 

Fisher exact test indicated a significantly (P < 0.001) higher incidence in the low dose group than in the control. The high dose
 

Fisher exact comparison and the Cochran-Armitage test, however, were
 

not significant, but these tests must be discounted due to the early
 

deaths from cancer in the high dose group. Based on these results,
 

the administration of p-cresidine was probably associated with the
 

increased incidence of liver neoplasms in the male rats.
 

In male rats an increase in the combined incidence of gliomas and
 

oligodendrogliomas of the brain or brain stem were observed in the
 

high dose group. The Cochran-Armitage test indicated a significant
 

(P - 0.032) positive association between dosage and incidence. This
 

was not supported, however, by significant Fisher exact tests.
 

In males for lung neoplasms and for the incidence of leukemia or
 

malignant lymphomas the Cochran-Armitage test was significant in the
 

negative direction, but the Fisher exact tests were not significant
 

under the Bonferroni criterion. Similarly, in the female rats the
 

results did not impute any real importance to adrenal cortical adenomas, to mammary fibroadenomas, to endometrial stromal polyps, or
 

to leukemias or malignant lymphomas.
 

The possibility of a negative association was observed between
 

chemical administration and the incidence of pituitary adenomas in
 

40
 
female rats and the incidence of interstitial-cell tumors of the
 

testis in males. This may have been due to the fact that survival
 

was markedly better in the control and low dose groups than in the
 

high dose group.
 

In summary, the administration of p-cresidine was associated
 

with the incidences of carcinomas of the urinary bladder and of
 

olfactory neuroblastomas in both males and females, and possibly
 

with the incidence of liver neoplasms in males.
 

41
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

The mean body weights for dosed mice of both sexes were consistently lower than those for the control groups (Figure 4). Fluctuations in the growth curve may be due to mortality; as the size of
 

the group diminishes, the mean body weight may be subject to wide
 

variations.
 

Only two dosed mice exhibited clinical signs. These were 1 low
 

dose male with a hard mass in the abdominal cavity and 1 low dose
 

female with a swollen, infected eye.
 

B. Survival
 

The estimated probabilities of survival for male and female
 

mice in the control and p-cresidine-dosed groups are shown in
 

Figure 5. For male mice and for female mice the Tarone tests showed
 

significant positive associations between dosage and mortality.
 

For males five of the control mice were sacrificed in week 78.
 

High early mortality was observed in the high dose group, with 50
 

percent (25/50) of the males dead by week 52 and 90 percent (45/50)
 

dead by week 73. With 27 of these 45 mice observed to have a tumor
 

of the urinary bladder, it seemed that mortality was associated with
 

tumor incidence. Fifty percent (25/50) of the low dose and 98 percent (49/50) of the control mice survived on test for at least 75
 

weeks.
 

42
 
TIME ON TEST (WEEKS) 

TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR p-CRESIDINE CHRONIC STUDY MICE 

43 
TIME ON TEST (WEEKS) 

FIGURE 5 

SURVIVAL COMPARISONS OF p-CRESIDINE CHRONIC STUDY MICE 

44 
For female mice five of the controls were sacrificed in week 78.
 

Twenty-eight percent (14/50) of the high dose, 78 percent (39/50) of
 

the low dose, and 90 percent (45/50) of the control mice survived on
 

test for at least 75 weeks.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

summarized in Appendix D (Tables Dl and D2).
 

p-Cresidine feeding produced a compound-related effect in the
 

urinary bladder. No neoplasms of the urinary bladder occurred in the
 

control mice. In the mice fed p-cresidine, there was a dose-related
 

spectrum of bladder neoplasms which resembled rat bladder neoplasms
 

previously described in this report. The hyperplasias and neoplasms
 

of the urinary bladder in the mice are summarized in the table
 

below:
 

MALE
 

Number of animals with tissues
 

examined histopathologically
 

Urinary Bladder
 

Hyperplasia
 

Dysplasia
 

Squamous-Cell Carcinoma
 

Transitional-Cell Carcinoma
 

Leiomyosarcoma
 

Control
 

(50)
 

0
 

0
 

0
 

0
 

0
 

Low
 

Dose
 

(42)
 

0
 

0
 

27(64%)
 

13(31%)
 

2(5%)
 

High
 

Dose
 

(35)
 

3(9%)
 

1(3%)
 

28(80%)
 

3(9%)
 

4(11%)
 

45
 
FEMALE 

Number of animals with tissues 

examined histopathologically (45) (47) (48) 

Urinary Bladder 

Hyperplasia 0 2(4%) 0 

Dysplasia 0 1(2%) 0 

Neoplasm NOS 0 0 1(2%) 

Neoplasm NOS, Malignant 0 1(2%) 0 

Carcinoma NOS 0 0 1(2%) 

Squamous-Cell Carcinoma 0 21(45%) 43(90%) 

Transitional-Cell Carcinoma 0 20(43%) 0 

Sarcoma NOS 0 0 1(2%) 

Leiomyosarcoma 0 2(4%) 2(4%) 

The bladders of those few mice that did not have bladder neoplasms
 

showed epithelial hyperplasia such as that seen in rats or dysplasia
 

which was characterized by nuclear hyperchromatism and irregularity.
 

In contrast to those in the rats, the bladder tumors in the mice more
 

frequently invaded adjacent structures or metastasized to distant organs
 

and were more frequently squamous in type rather than transitional-


cell. The mouse bladder tumors tended to be large, often filling the
 

lumen of the organs and obstructing the ureters. In consequence, kidneys of the tumor-bearing animals had some degree of hydronephrosis.
 

As in the rats, transitional-cell tumors occurred in the renal pelvis
 

of two p-cresidine-dosed females.
 

There were no tumors of the nasal cavity in control mice. Of
 

the mice fed p-cresidine two developed sarcomas, one an adenocarcinoma, and one a squamous-cell carcinoma of the nasal cavity. However,
 

no olfactory neuroblastomas were observed.
 

46
 
The incidence of hepatocellular carcinomas in low dose female
 

mice (13/45 [29 percent]), but not in the high dose female group
 

(6/43 [14 percent]), was increased relative to controls (0/46). The
 

incidence of liver tumors in dosed male mice was not increased. The
 

early mortality resulting from fulminating tumors of the bladder in
 

both sexes may have obscured a dose-related response of the liver.
 

Based upon this pathologic examination, p-cresidine was carcinogenic in B6C3F1 mice because it induced carcinomas of the urinary
 

bladder in both sexes and hepatocellular carcinomas in female mice.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 5 and 6. The analysis is included for
 

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

tumors were observed in at least one of the control or p-cresidinedosed groups and where such tumors were observed in at least 5 percent of the group. Because of the high mortality noted in mice of
 

both sexes, these analyses were based on those mice surviving at
 

least 52 weeks or at least until the development of the first tumor
 

at the site of interest, whichever occurred first.
 

In both male and female mice the combined incidence of carcinomas NOS, squamous-cell carcinomas, or transitional-cell carcinomas
 

of the urinary bladder was increased in the dosed groups compared to
 

the control groups. These tumors were observed as early as week 44
 

47
 
TIME-ADJUSTED ANALYSES OF

SPECIFIC SITES IN MALE

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar/Bronchiolar Carcinoma,
 

or Alveolar/Bronchiolar Adenoma**
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma^
 

P Values0
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma-»*
 

P Valuesc
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 5 

 THE INCIDENCE OF

 MICE TREATED WITH

CONTROL
 

7/50(0.14)
 

N.S.
 

97
 

4/50(0.08)
 

P = 0.047(N)
 

97
 

10/50(0.20)
 

N.S.
 

_
 

60
 

 PRIMARY TUMORS AT 

p-CRESIDINEa>f 

LOW
 

DOSE
 

2/39(0.05)
 

N.S.
 

0.366
 

0.039
 

1.788
 

106
 

0/42(0.00)
 

N.S.
 

0.000
 

0.000
 

1.278
 

11/41(0.27)
 

N.S.
 

1.341
 

0.574
 

3.146
 

65
 

HIGH
 

DOSE
 

1/21(0.05)
 

N.S.
 

0.340
 

0.008
 

2.374
 

92
 

0/22(0.00)
 

N.S.
 

0.000
 

0.000
 

2.370
 

3/32(0.09)
 

N.S.
 

0.469
 

0.088
 

1.648
 

44
 
TABLE 5 (Continued ) 

TOPOGRAPHY: MORPHOLOGY CONTROL 

Liver: Hepatocellular Carcinoma, or 

Hepatocellular Adenoma*5

 >f 15/50(0.30) 

P Values0 P = 0.028CN) 

Relative Risk (Control)*1 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 60 

Urinary Bladder: Squamous-Cell Carcinoma 

of Transitional-Cell Carcinoma*5

*f 0/50(0.00) 

P VaLuesc P < 0.001 

Departure from Linear Trende P < 0.001 

Relative Risk (Control)d 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

LOW 

DOSE 

11/41(0.27) 

N.S. 

0.894 

0.418 

1.836 

65 

40/42(0.95) 

P < 0.001 

Infinite 

17.734 

Infinite 

51 

HIGH 

DOSE 

3/32(0.09) 

P = 0.024(N) 

0.313 

0.062 

0.991 

44 

31/31(1.00) 

P < 0.001 

Infinite 

20.738 

Infinite 

44 
TABLE 5 (Concluded)
 

aTreated groups received time-weighted average doses of 0.22 or 0.46 percent in feed.
 

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

cThe 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 percent confidence interval on the relative risk of the treated group to the control group.
 

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

group when P < 0.05.
 

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

the first tumor of interest was observed earlier than 52 weeks in any group of this sex and species,
 

where the analyses were based upon all animals that survived until or past the date that the first
 

tumor was observed.
 
TABLE 6
 

TIME-ADJUSTED ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH p-CRESIDINEa>f
 

LOW HIGH
 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE DOSE
 
TOPOGRAPHY: MORPHOLOGY
 

Liver: Hepatocellular Carcinoma">*
 

P Values0
 

Departure from Linear Trende
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma, or
 

Hepatocellular Adenoma*5>^
 

P Values0
 

Departure from Linear Trende
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Urinary Bladder: Carcinoma NOS,
 

Squamous-Cell Carcinoma, or
 

Transitional-Cell Carcinomab

>f
 

P Values0
 

Departure from Linear Trende
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 6 (Continued)
 

CONTROL
 

0/45(0.00)
 

P = 0.028
 

P < 0.001
 

0/45(0.00)
 

P = 0.030
 

P < 0.001
 

0/45(0.00)
 

P < 0.001
 

P < 0.001
 

LOW
 

DOSE
 

13/44(0.30)
 

P < 0.001
 

Infinite
 

4.126
 

Infinite
 

53
 

14/44(0.32)
 

P < 0.001
 

Infinite
 

4.484
 

Infinite
 

53
 

41/46(0.89)
 

P < 0.001
 

Infinite
 

14.258
 

Infinite
 

53
 

HIGH
 

DOSE
 

6/40(0.15)
 

P = 0.009
 

Infinite
 

1.809
 

Infinite
 

44
 

6/40(0.15)
 

P = 0.009
 

Infinite
 

1.809
 

Infinite
 

44
 

44/46(0.96)
 

P < 0.001
 

Infinite
 

16.126
 

Infinite
 

40
 
TOPOGRAPHY:MORPHOLOGY
 

Urinary Bladder: Transitional-Cell
 

Carcinoma, Neoplasm NOS, Neoplasm MOS
 

Malignant, Carcinoma NOS, or Squamous-


Cell Carcinomab>f
 

P Values0
 

Departure from Linear Trende
 

ReLative Risk (Control)**
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Adenomab
 

P Values0
 

Relative Risk (Control)4
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 6 (Continued)
 

CONTROL
 

0/45(0.00)
 

P < 0.001
 

P < 0.001
 

1/28(0.04)
 

N.S.
 

97
 

LOW
 

DOSE
 

42/46(0.91)
 

P < 0.001
 

Infinite
 

14.784
 

Infinite
 

53
 

2/31(0.06)
 

N.S.
 

1.806
 

0.100
 

103.026
 

94
 

HIGH
 

DOSE
 

45/46(0.98)
 

P < 0.001
 

Infinite
 

17.153
 

Infinite
 

40
 

0/30(0.00)
 

N.S.
 

0.000
 

0.000
 

17.204
 
TABLE 6 (Concluded)
 

a

Treated groups received time-weighted average doses of 0.22 or 0.44 percent in feed.
 

•^Number of tumor-bearing an ima Is/number of animals examined at site (proportion).
 

cThe 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 percent confidence interval on the relative risk of the treated group to the control group.
 

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

group when P < 0.05.
 

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

the first tumor of interest was observed earlier than 52 weeks in any group of this sex and species,
 

where the analyses were based upon all animals that survived until or past the date that the first
 

tumor was observed.
 
in the males and week 40 in the females. For both sexes, the Cochran-


Armitage test indicated a significant (P < 0.001) positive association between dose and incidence. These were supported by significant
 

(P < 0.001) Fisher exact test results for both dose levels compared
 

to the control.
 

In female mice an increased incidence of hepatocellular carcinomas was observed in the dosed groups compared to the control. The
 

Cochran-Armitage test indicated a significant (P = 0.028) positive
 

association between dose and incidence. This was supported by significant Fisher exact test results of both the high (P - 0.009) and
 

low (P < 0.001) dose groups to the control. The departure from linear trend was also significant (P * 0.001) since tumor incidence was
 

greater in the low dose group than in the high dose group. In historical control data compiled by this laboratory for the NCI Carcinogenesis Testing Program 19/275 (7 percent) of the female untreated
 

control B6C3F1 mice had a hepatocellular carcinoma or a hepatocellular adenoma. For the male mice, however, the Cochran-Armitage test
 

indicated a significant (P • 0.028) negative association between
 

dose and the combined incidence of hepatocellular carcinomas and
 

hepatocellular adenomas. The Fisher exact test comparing high dose
 

to control also indicated a significant (P • 0.024) negative association. The historical control rate for untreated male B6C3F1
 

mice was 88/275 (32 percent) with these lesions. It was considered,
 

55
 
however, that the shorter survival in the high dose than in the control and low dose groups accounted for the apparently negative trend.
 

Based on these results the administration of p-cresidine was
 

associated with the increased incidence of hepatocellular carcinoma
 

in female mice and of carcinomas of the urinary bladder in both male
 

and female mice*
 

56
 
V. DISCUSSION
 

Mortality rates were dose-related for both sexes of both species.
 

Accelerated mortality of high dose animals during the latter half of
 

the study was associated with bladder tumors. That incidences of
 

certain tumors were higher in low dose than in high dose groups was
 

probably due to this accelerated mortality.
 

Addition of p-cresidine to the diet was associated with increased
 

incidences of neoplasms at several sites in rats. Male rats, having
 

a higher incidence of neoplasms than females in the urinary bladder,
 

nasal cavity, liver, and brain, appeared to be more sensitive to the
 

tumorigenic effects of p-cresidine.
 

The urinary bladder was a target organ of p-cresidine carcinogenicity in rats. Bladder neoplasms were observed in dosed rats, but not
 

in their controls. For both male and female rats the incidence of
 

carcinomas (combined incidence of papillary carcinomas, squamous-cell
 

carcinomas, transitional-cell papillomas, transitional-cell carcinomas
 

and undifferentiated carcinomas) of the urinary bladder was dose-


related and was significantly higher in each dosed group than in the
 

corresponding control group. An elevated incidence of hyperplasia
 

was also observed in urinary bladders of low dose rats of both sexes.
 

Dose-related increases in the incidence of neoplasms were also
 

observed in nasal cavities of rats. The incidence of olfactory
 

neuroblastomas in females and the combined incidence of neuroblastoma
 

57
 
and olfactory neuroblastoma of the nasal cavity in males were both
 

significantly associated with p-cresidine dosage. The incidence of
 

these neoplasms in high dose groups of both sexes was significantly
 

higher than in controls. The combined incidences of carcinomas and
 

adenocarcinomas of the nasal cavity or nasopharynx were significantly
 

dose-related in male rats.
 

Among low dose male rats, the incidence of liver tumors (the
 

combined incidence of neoplastic nodules, hepatocellular carcinomas,
 

or mixed hepato/cholangio carcinomas) was significantly higher than
 

in controls.
 

The urinary bladder was a target organ for carcinogenicity
 

in mice as well as in rats. Although the spectrum of dose-related
 

bladder neoplasms in mice was similar to that in rats, the bladder
 

tumors in mice were more invasive and more frequently metastasized.
 

In both male and female mice, the incidence of carcinomas (combined
 

incidence of carcinomas NOS, squamous-cell carcinomas, and transitional-cell carcinomas) of the urinary bladder was significantly
 

associated with p-cresidine dosage and was significantly greater in
 

each dosed group than in controls. The bladders of those few mice
 

without bladder neoplasms showed evidence of hyperplasia or dysplasia.
 

No bladder neoplasms were found in control mice.
 

The incidence of hepatocellular carcinomas in each dosed female
 

mouse group was significantly greater than in controls, and this
 

58
 
increased incidence was significantly associated with concentration
 

of p-cresidine in the diet.
 

Under the conditions of this bioassay, p-cresidine was carcinogenic to Fischer 344 rats, causing increased incidences of carcinomas
 

and of papillomas of the urinary bladder in both sexes, increased
 

incidences of olfactory neuroblastomas in both sexes, and of liver
 

tumors (hepatocellular carcinomas, hepato/cholangio carcinomas and
 

neoplastic nodules) in males. p-Cresidine was also found to be carcinogenic in B6C3F1 mice, causing carcinomas of the urinary bladders
 

in both sexes and hepatocellular carcinomas in females.
 

59
 
VI. BIBLIOGRAPHY
 

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

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

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

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

J. Wiley & Sons, New York, 1971.
 

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

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

Union Against Cancer, Geneva, 1969.
 

Chemical Abstracts Service. The Chemical Abstracts Service (CAS)
 

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

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

Compton, E.D., Group Director, Environmental Control, Chemicals,
 

The Sherwin-Williams Company, Cleveland, Ohio. Letter to Dr. J.
 

Donoso, The MITRE Corporation, METREK Division, McLean, Virginia,
 

May 31, 1977.
 

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

Co., Ltd., London, 1970.
 

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

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

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

Tests, Confidence Limits, and Adjustments for Stratification."
 

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

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

Observations." Journal of the American Statistical Association
 

51:457-481, 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.
 

Mehta, N., Sadtler Research Institute, Philadelphia, Pennsylvania.
 

Personal communication, 24 August 1978.
 

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

Co., New York, 1966.
 

60
 
Pollock, J.R.A. and R. Stevens, editors, Dictionary of Organic
 

Compounds, 4th edition. Volume 1. Oxford University Press,
 

New York, 1965.
 

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

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

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

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

Yorkshire, England, p. 4706, 1971.
 

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

of Specific Hepatocellular Lesions in Rats." Cancer Research
 

35.:3214-3223, 1975.
 

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

.62:679-682, 1975.
 

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

Industries Center, Stanford Research Institute, Menlo Park, California. Personal communication, June 8, 1977.
 

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

61
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH p-CRESIDINE
 

TABLE Al

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE 

RATS TREATED WITH p-CRESIDINE 

HIGH DOSE
 

01-0265
 

50
 

47
 

47
 

(47)
 

(47)
 

1 (2%)
 

(47)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

20 (43%)
 

(47)
 

1 (2%)
 

(45)
 

1 (2%)
 

(47)
 

1 (2%)
 

(45)
 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY**
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

TRICHOEPITHELIOMA
 

*SUBCUT TISSUE
 

FIBROMA
 

FIBROSARCOMA
 

LIPOSARCOMA
 

RESPIRATORY SYSTEM
 

*NASAL CAVITY
 

NEOPLASM, NOS, MALIGNANT
 

CARCINOMA,NOS
 

ADENOMA, NOS
 NEUROBLASTOMA
 

OLFACTORY NEUROBLASTOMA
 

^NASOPHARYNX
 

ADENOCARCINOMA, NOS
 

#LUNG
 

HEPATOCELLULAR CARCINOMA, METAST
 

ALVEOLAR/BRONCHIOLAR ADENOMA
 

FIBROSARCOMA, METASTATIC
 

HEMATOPOIETIC SYSTEM
 

^MULTIPLE ORGANS
 

MALIGNANT LYMPHOMA, NOS
 

UNDIFFERENTIATED LEUKEMIA
 

•SPLEEN
 

MYELOMONOCYTIC LEUKEMIA
 

CONTROL(UNTR)
 

'01-0220
 

50
 

48
 

48
 

(48)
 

(48)
 

3 (6%)
 

2 (4%)
 

(48)
 

(48)
 

(48)
 

3 (6%)
 

1 (2%)
 

(48)
 

1 (2%)
 

2 (4%;
 

(48)
 

4 (8%)
 

LOW DOSE
 

01-0260
 

50
 

50
 

49
 

(50)
 

2 (4%)
 

(50)
 

2 (4%)
 

2 (4%)
 

1 (25O
 

(50)
 

1 (2%)
 

1 (2%)
 

(50)
 

(49)
 

(50)
 

1 (2%)
 

(49)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

A-3
 
TABLE Al 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE
 01-0220 01-0260
 

•LYMPH NODE	 (43) (48)
 

MALIGNANT LYMPHOMA, NOS
 

•SUBMANDIBULAR	 L.NODE (43) (48)
 

OLFACTORY NEUROBLASTOMA, METASTA
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

•SALIVARY GLAND	 (45) (48)
 

ACINAR-CELL ADENOMA 1 (2%)
 

•PAROTID GLAND	 (45) (48)
 

OLFACTORY NEUROBLASTOMA, METASTA
 

•LIVER	 (48) (49)
 

NEOPLASTIC NODULE 10 (20%)
 HEPATOCELLULAR CARCINOMA 2 (45O
 

MIXED HEPATO/CHOLANGIO CARCINOMA 1 (2%)
 

•STOMACH	 (48) (49)
 

SQUAMOUS CELL PAPILLOMA 2 (4%)
 

BASAL-CELL CARCINOMA 1 (2JO
 

URINARY SYSTEM
 

•KIDNEY/PELVIS	 (48) (49)
 

TRANSITIONAL-CELL PAPILLOMA 1 (2%)
 

TRANSITIONAL-CELL CARCINOMA, INV 1 (2%)
 

•URINARY BLADDER	 (48) (48)
 UNDIFFERENTIATED CARCINOMA
 

PAPILLARY CARCINOMA
 

SQUAMOUS CELL CARCINOMA 1 (2%)
 

TRANSITIONAL-CELL PAPILLOMA 14 (295O
 

TRANSITIONAL-CELL CARCINOMA 15 (31JO
 

LEIOMYOSARCOMA
 

*PROSTATIC URETHRA (48) (50)
 

UNDIFFERENTIATED CARCINOMA, INVA
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 01-0265
 

(38)
 

1 (3X)
 

(38)
 

1 (3%)
 

(43)
 

(43)
 

1 (2%)
 

(46)
 

1 (2%)
 1 (2%)
 

(46)
 

(46)
 

(47)
 

1 (2%)
 

4 (9%)
 

6 (13X
 

3 (6%)
 

31 (663
 

2 (4%)
 

(47)
 

1 (2%)
 

A-4
 
TABLE Al 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 01-0220 01-0260 01-0265
 

ENDOCRINE SYSTEM
 

•PITUITARY	 (45) (48) (38)
 

ADENOMA, NOS 10 (22%) 1 (2%) 3 (8%)
 

CHROMOPHOBE ADENOMA 4 (8%)
 

BASOPHIL ADENOMA 4 (8%)
 

•ADRENAL	 (46) (48) (44)
 

CORTICAL ADENOMA 3 (6%)
 

PHEOCHROMOCYTOMA 4 (9%) 7 (15%) 3 (7%)
 

•THYROID	 (43) (41 ) (40)
 

FOLLICULAR-CELL ADENOMA 1 (2%)
 

C-CELL ADENOMA 3 (7%) 1 (2%) 1 (3%)
 

C-CELL CARCINOMA 2 (5%) 1 (2%) 1 (3%)
 

•PARATHYROID	 (25) (21 ) (15)
 

ADENOMA, NOS 1 (4%)
 

•PANCREATIC	 ISLETS (44) (48) (40)
 

ISLET-CELL ADENOMA 1 (2%) 2 (5%)
 

ISLET-CELL CARCINOMA 1 (2%)
 

REPRODUCTIVE SYSTEM
 

*PREPUTIAL GLAND (48) (50) (47)
 

CARCINOMA,NOS 2 (4%) 1 (2%)
 PAPILLOMA, NOS 1 (2%)
 

ADENOMA, NOS 1 (2%)
 

•PROSTATE	 (45) (48) (46)
 

TRANSITIONAL-CELL CARCINOMA, INV 1 (2%)
 

•TESTIS	 (48) (47) (46)
 

INTERSTITIAL-CELL TUMOR 37 (77%) 45 (96%) 23 (50%
 

NERVOUS SYSTEM
 

•BRAIN/MENINGES	 (46) (48) (43)
 

HEPATOCELLULAR CARCINOMA, METAST 1 (2%)
 

•BRAIN	 (46) (48) (43)
 NEOPLASM, NOS, METASTATIC 1
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-5
 
TABLE Al 

(CONTINUED) 

CONTROL(UNTR) 

01-0220 

LOW DOSE 

01-0260 

CARCINOMA, NOS, NETASTATIC 

OSTEOSARCOMA, METASTATIC 

GLIOMA, NOS 

OLFACTORY NEUROBLASTOMA, INVASIV 

OLFACTORY NEUROBLASTOMA, METASTA 

1 (2%) 

#BRAIN STEM 

GLIOMA, NOS 

(46) (48) 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

*SKULL (48) (50)
 

OSTEOSARCOMA 1 (2%)
 

BODY CAVITIES
 

*BODY CAVITIES (48) (50)
 

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

ALL OTHER SYSTEMS
 

^MULTIPLE ORGANS (48) (50)
 C-CELL CARCINOMA, METASTATIC 1 (2%)
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50
 NATURAL DEATHS 8 2
 

MORIBUND SACRIFICE 5 9
 

SCHEDULED SACRIFICE 5
 ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 32 39
 ANIMAL MISSING
 

9 INCLUDES AUTOLYZED ANIMALS
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 

01-0265
 

1 (2%)
 

2 (5%)
 

2 (5%)
 

5 (12%)
 

(43)
 

1 (2%)
 

(47)
 

(47)
 

2 (4%)
 

(47)
 

50
 

25
 

21
 

4
 

A-6
 
TABLE Al
 

(CONCLUDED)
 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 

01-0220 01-0260 01-0265
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 42 49 46
 

TOTAL PRIMARY TUMORS 74 131 116
 

TOTAL ANIMALS WITH BENIGN TUMORS 40 48 28
 

TOTAL BENIGN TUMORS 61 91 37
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 11 21 46
 

TOTAL MALIGNANT TUMORS 12 29 76
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2 3 11
 

TOTAL SECONDARY TUMORS 2 3 14
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT 1 11 2
 

TOTAL UNCERTAIN TUMORS 1 11 3
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

A-7
 
TABLE A2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE 

RATS TREATED WITH p-CRESIDINE 

CONTROL(UNTR) 

02-0220 

LOW DOSE 

02-0260 

ANIMALS INITIALLY IN STUDY 

ANIMALS NECROPSIED 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 

50 

50 

50 

50 

50 

49 

INTEGUMENTARY SYSTEM
 

*SKIN (50) (50)
 

SEBACEOUS ADENOCARCINOMA 1 (2%)"
 

FIBROMA 1 (2%)
 

*SUBCUT TISSUE (50) (50)
 

SARCOMA, NOS 1 (2%) 1 (2JO
 

FIBROSARCOMA 1 (2%)
 

RESPIRATORY SYSTEM
 

*NASAL CAVITY (50) (50)
 

SQUAMOUS CELL CARCINOMA
 

ADENOCARCINOMA* NOS
 

OLFACTORY NEUROBLASTOMA
 

*LUNG (50) (48)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (2JJ)
 

TRANSITIONAL-CELL CARCINOMA, MET 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

^MULTIPLE ORGANS (50) (50)
 MALIGNANT LYMPHOMA, NOS 2 (4%)
 

•SPLEEN	 (50) (49)
 

MYELOMONOCYTIC LEUKEMIA 2 (4«)
 

CIRCULATORY SYSTEM
 

NONE
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

HIGH DOSE
 

02-0265
 

50
 

49
 

48
 

(49)
 

(49)
 

(49)
 

1 (2Ji)
 

1 (2%)
 

11 (22%)
 

(48)
 

1 (25O
 

(49)
 

(47)
 

A-8 
TABLE A2 

(CONTINUED) 

DIGESTIVE SYSTEM
 

*MOUTH
 

SQUAMOUS CELL PAPILLOMA
 

•SALIVARY GLAND
 

ACINAR-CELL ADENOMA
 

tLIVER
 

NEOPLASTIC NODULE
 

•PANCREAS
 

TRANSITIONAL-CELL CARCINOMA, MET
 

•STOMACH
 

SQUAMOUS CELL CARCINOMA
 

URINARY SYSTEM
 

•KIDNEY
 

UNDIFFERENTIATED CARCINOMA
 

•KIDNEY/PELVIS
 

TRANSITIONAL-CELL CARCINOMA
 

•URINARY BLADDER
 

NEOPLASM, NOS
 PAPILLARY CARCINOMA
 

SQUAMOUS CELL CARCINOMA
 

TRANSITIONAL-CELL PAPILLOMA
 

TRANSITIONAL-CELL CARCINOMA
 

LEIOMYOSARCOMA
 

*URETHRA
 

SQUAMOUS CELL PAPILLOMA
 

ENDOCRINE SYSTEM
 

•PITUITARY
 

ADENOMA, NOS
 

CHROMOPHOBE ADENOMA
 

•ADRENAL
 

CORTICAL ADENOMA
 

CONTROL(UNTR) 02-0220 LOW DOSE 02-0260 HIGH DOSE 02-0265 

(50) (50) (49) 

1 (2%) 

(46) (48) (43) 

1 (2%) 

(50) (48) (48) 

4 (8%) 

(46) (49) (43) 

1 (2%) 

(49) (48) (46) 

1 (2%) 

(49) (49) (46) 

1 (2%) 

(49) (49) (46) 

1 (2%) 

(47) (49) (46) 

2 (4%) 

1 (2%) 

9 (18%) 7 (15%) 

6 (12%) 2 (4%) 

22 (45%) 33 (72%) 

2 (4%) 3 (7%) 

(50) (50) (49) 

1 (2%) 

(40) (45) (40) 

16 (40%) 2 (4%) 6 (15%) 

1 (3%) 9 (20%) 1 (3%) 

(48) (48) (44) 

5 (10%) 1 (2%) 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-9
 
TABLE A2 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 

02-0220 02-0260 02-0265
 

PHEOCHROMOCYTOMA	 6 (13%) 1 (2%) 1 (2%)
 

•THYROID	 (43) (42) (40)
 

FOLLICULAR-CELL CARCINOMA 2 (5%)
 

C-CELL ADENOMA 4 (10%)
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND (50) (50) (49)
 

ADENOCARCINOMA, NOS 1 (2%)
 

FIBROADENOMA 6 (12%) 4 (8%) 1 (2%)
 

*CLITORAL GLAND (50) (50) (49)
 

CARCINOMA,NOS 2 (4%)
 

ADENOMA, NOS 1 (2%)
 

•UTERUS	 (48) (47) (45)
 

ADENOCARCINOMA, NOS 1 (2%)
 

FIBROMA 2 (4%)
 

ENDOMETRIAL STROMAL POLYP 2 (4%) 8 (17%) 4 (9%)
 

ENDOMETRIAL STROMAL SARCOMA 2 (4%)
 

CARCINOSARCOMA 1 (2%)
 

•UTERUS/ENDOMETRIUM	 (48) (47) (45)
 

ADENOCARCINOMA, NOS 1 (2%)
 

ADENOCA/SQUAMOUS METAPLASIA 1 (2%)
 

NERVOUS SYSTEM
 

•BRAIN	 (50) (48) (46)
 

SQUAMOUS CELL CARCINOMA, METASTA 2 (4%)
 ADENOCARCINOMA, NOS, INVASIVE 1 (2%)
 GLIOMA, NOS 1 (2%)
 OLIGODENDROGLIOMA 1 (2%)
 OLFACTORY NEUROBLASTOMA, INVASIV 1 (2%)
 OLFACTORY NEUROBLASTOMA, METASTA 3 (7%)
 

•CEREBRAL CORTEX	 (50) (48) (46)
 

OLFACTORY NEUROBLASTOMA, METASTA 1 (2%)
 

SPECIAL SENSE ORGANS
 

*EAR CANAL (50) (50) (49)
 

SQUAMOUS CELL CARCINOMA 1 (2%)
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-IO
 
TABLE A2 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 02-0220 02-0260 02-0265
 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

"ABDOMINAL CAVITY 

TRANSITIONAL-CELL CARCINOMA, INV 

(50) (50) (49) 

1 (2%) 

ALL OTHER SYSTEMS 

NONE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 

NATURAL DEATH3 

MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIMAL MISSING 

50 

7 

2 

5 

36 

50 

3 

6 

50 

30 

20 

a INCLUDES AUTOLYZED ANIMALS 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED

* NUMBER OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

A-ll
 
TABLE A2
 

(CONCLUDED)
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 

TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORSt
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

CONTROL(UNTR)
 02-0220
 

27
 

42
 

23
 

33
 

9
 

9
 

LOW DOSE HIGH DOSE
 02-0260 02-0265
 

48 44
 

93 79
 

35 17
 

43 17
 

36 42
 

46 60
 

2 9
 2 11
 

4 2
 4 2
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

A-I2
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH p-CRESIDINE
 

TABLE Bl

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE 

MICE TREATED WITH p-CRESIDINE 

CONTROL(UNTR) LOU DOSE HIGH DOSE
 05-0220 05-0260 05-0265
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

ANIMALS MISSING 1 3
 

ANIMALS NECROPSIED 50 47 41
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY ** 50 43 37
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

*NASAL CAVITY (50) (47) (41 )
 

ADENOCARCINOMA, NOS 1 (2%)
 

SARCOMA, NOS 1 (2%)
 

FIBROSARCOMA 1 (2%)
 

• LUNG	 (50) (40) (37)
 

SQUAMOUS CELL CARCINOMA, METASTA 4 (10%) 2 (5%)
 

TRANSITIONAL-CELL CARCINOMA, MET 1 (3%)
 

HEPATOCELLULAR CARCINOMA, METAST 2 (5%)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 5 (10%) 1 (3%)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 2 (4%) 1 (3%) 1 (3%)
 

HEMATOPOIETIC SYSTEM
 

"MULTIPLE ORGANS (50) (47) (41 )
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2%)
 

•SPLEEN	 (50) (37) (33)
 HEMANGIOMA 1 (2%)
 

HEMANGIOSARCOMA 2 (4%) 1 (3%)
 

#MESENTERIC L. NODE (44) (26) (16)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2%)
 

#LIVER (50) (41 ) (37)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2%)
 

CIRCULATORY SYSTEM
 

NONE
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

* * EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
TABLE Bl 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 05-0220 05-0260 05-0265
 

DIGESTIVE SYSTEM
 

*LIVER (50) (41 ) (37)
 

HEPATOCELLULAR ADENOMA 5 (10%)
 

HEPATOCELLULAR CARCINOMA 10 (20%) 11 (27%) 3 (8%)
 

*PANCREAS (50) (30) (31 )
 

SQUAMOUS CELL CARCINOMA, INVASIV 1 (3%)
 

*DUODENAL GLAND (50) (30) (29)
 

ADENOMA, NOS 1 (3%)
 

URINARY SYSTEM
 

#URINARY BLADDER (50) (42) (35)
 

SQUAMOUS CELL CARCINOMA 27 (64%) 28 (80%)
 

TRANSITIONAL-CELL CARCINOMA 13 (31%) 3 (9%)
 

LEIOMYOSARCOMA 2 (5%) 4 (11%)
 

*PROSTATIC URETHRA (50) (47) (41 )
 

SQUAMOUS CELL CARCINOMA, INVASIV .2 (4%)
 

TRANSITIONAL-CELL CARCINOMA, INV 1 (2%)
 

ENDOCRINE SYSTEM
 

#THYROID (39) (31 ) (29)
 

FOLLICULAR-CELL ADENOMA 1 (3%)
 FOLLICULAR-CELL CARCINOMA 1 (3%)
 

REPRODUCTIVE SYSTEM
 

•PROSTATE	 (41 ) (37) (26)
 

SQUAMOUS CELL CARCINOMA, INVASIV 6 (16%) 2 (8%)
 

TRANSITIONAL-CELL CARCINOMA, INV 3 (8%) 1 (4%)
 LEIOMYOSARCOMA, INVASIVE 1 (3%) 1 (4%)
 

*SEMINAL VESICLE (50) (47) (41 )
 

TRANSITIONAL-CELL CARCINOMA, INV 1 (2%)
 

^COAGULATING GLAND (50) (47) (41 )
 

SQUAMOUS CELL CARCINOMA, INVASIV 1 (2%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

B-4
 
TABLE Bl 

(CONTINUED) 

TRANSITIONAL-CELL CARCINOMA, INV
 

•TESTIS
 

EMBRYONAL CARCINOMA
 

NERVOUS SYSTEM
 

fBRAIN
 

SARCOMA, NOS, METASTATIC
 

SPECIAL SENSE ORGANS
 

*HARDERIAN GLAND
 

CYSTADENOMA, NOS
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

^ABDOMINAL CAVITY
 

SQUAMOUS CELL CARCINOMA,


SQUAMOUS CELL CARCINOMA,


 INVASIV
 

 METASTA
 

TRANSITIONAL-CELL CARCINOMA, INV
 

LEIOMYOSARCOMA, INVASIVE
 

^•PERITONEAL CAVITY
 

SQUAMOUS CELL CARCINOMA, INVASIV
 

ALL OTHER SYSTEMS
 

"MULTIPLE ORGANS
 

SQUAMOUS CELL CARCINOMA, INVASIV
 

SQUAMOUS CELL CARCINOMA, METASTA
 

TRANSITIONAL-CELL CARCINOMA, INV
 

ADENOCARCINOMA, NOS, INVASIVE
 

LEIOMYOSARCOMA, INVASIVE
 

CONTROL(UNTR)
 05-0220
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

(50)
 

LOU DOSE
 05-0260
 

1 (2%)
 

(41 )
 

(36)
 

(47)
 

1 (2%)
 

(47)
 

4 (9%)
 

1 (2£)
 

1 (2%)
 

(47)
 

(47)
 

4 (9K)
 2 (4%)
 

1 (2%)
 

1 (2%)
 

HIGH DOSE
 05-0265
 

(37)
 

(32)
 

1 (3%)
 

(41 )
 

(41 )
 

3 (7%)
 

1 (2%)
 

(41 )
 

2 (5%)
 

(41 )
 

1 (2%)
 2 (5%)
 

2 (5%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

B-5
 
TABLE Bl
 

(CONCLUDED)
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

3 INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORS#
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 TOTAL UNCERTAIN TUMORS
 

CONTROL(UNTR)
 05-0220
 

50
 

3
 

5
 

42
 

22
 

33
 

12
 

13
 

1 6
 

20
 

LOW DOSE
 05-0260
 

50
 

37
 

5
 

7
 

1
 

42
 

60
 

3
 

3
 

42
 

57
 

27
 

38
 

HIGH DOSE
 05-0265
 

50
 

40
 

7
 

3
 

32
 

40
 

32
 

40
 

14
 

18
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

B-6
 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE 

MICE TREATED WITH p-CRESIDINE 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-7
 
TABLE B2 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE
 06-0220 06-0260
 

•ABDOMINAL	 LYMPH NODE (38) (38)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (3%)
 

#MESENTERIC L. NODE (38) (38)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (3%)
 

#LIVER (46) (45)
 MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

•LIVER	 (46) (45)
 

SQUAMOUS CELL CARCINOMA, METASTA
 

HEPATOCELLULAR ADENOMA 1 (2%)
 

HEPATOCELLULAR CARCINOMA 13 C 29%)
 

•STOMACH	 (46) (40)
 

ADENOCARCINOMA, NOS 1 (3%)
 

URINARY SYSTEM
 

•KIDNEY/PELVIS	 (46) (45)
 

TRANSITIONAL-CELL CARCINOMA 1 (2%)
 

•URINARY BLADDER	 (45) (47)
 

NEOPLASM, NOS
 

NEOPLASM, NOS, MALIGNANT 1 (2%)
 

CARCINOMA,NOS
 

SQUAMOUS CELL CARCINOMA 21 (45%)
 

TRANSITIONAL-CELL CARCINOMA 20 (43%)
 

SARCOMA, NOS
 LEIOMYOSARCOMA 2 (4%)
 

ENDOCRINE SYSTEM
 

•PITUITARY	 (33) (32)
 ADENOMA, NOS 1 (3%)
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 06-0265
 

(28)
 

(28)
 

(43)
 

1 (2%)
 

(43)
 

1 (2%)
 

6 (14%)
 

(36)
 

(45)
 

1 (2%)
 

(48)
 

1 (2%)
 

1 (2%)
 

43 (90%)
 

1 (2%)
 

2 (4%)
 

(26)
 

B-8
 
TABLE B2 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE
 06-0220 06-0260
 

•ADRENAL/CAPSULE	 (40) (42)
 ADENOMA, NOS 1 (3S)
 

#THYROID (29) (32)
 

FOLLICULAR-CELL ADENOMA 1 (3%) 2 (6%)
 

REPRODUCTIVE SYSTEM
 

#UTERUS (45) (46)
 

HEMANGIOSARCOMA 1 (25O
 

tOVARY (40) (41 )
 

PAPILLARY ADENOMA 1 (3%)
 HEMANGIOMA 1 (2%)
 

NERVOUS SYSTEM
 

tBRAIN (44) (45)
 

SQUAMOUS CELL CARCINOMA, INVASIV
 

SPECIAL SENSE ORGANS
 

*HARDERIAN GLAND (47) (47)
 

PAPILLARY ADENOMA 1 (2%)
 

PAPILLARY ADENOCARCINOMA 1 (2%)
 

PAPILLARY CYSTADENOMA, NOS
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

^ABDOMINAL CAVITY (47) (47)
 

SQUAMOUS CELL CARCINOMA, INVASIV 2 (4%)
 

^PERITONEAL CAVITY (47) (47)
 

SQUAMOUS CELL CARCINOMA, INVASIV
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 0 6-0265
 

(39)
 

(37)
 

(36)
 

(35)
 

(39)
 

1 (3%)
 

(48)
 

1 (25O
 

(48)
 

2 (4%)
 

(48)
 

2 (4%)
 

B-9
 
TABLE B2
 

(CONCLUDED)
 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 06-0220 06-0260 06-0265
 

ALL OTHER SYSTEMS
 

^MULTIPLE ORGANS (47) (47) (48)
 

SQUAMOUS CELL CARCINOMA, INVASIV 2 (4X)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (2%)
 

LEIOMYOSARCOMA, INVASIVE 1 (2J4)
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

NATURAL DEATH3 4 21 41
 

MORIBUND SACRIFICE 3 4 8
 

SCHEDULED SACRIFICE 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 36 25 1
 

ANIMAL MISSING 2
 

3 INCLUDES AUT0LY2ED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 14 45 45
 

TOTAL PRIMARY TUMORS 15 83 59
 

TOTAL ANIMALS WITH BENIGN TUMORS 5 7 2
 

TOTAL BENIGN TUMORS 6 7 2
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 9 45 44
 

TOTAL MALIGNANT TUMORS 9 lf>
 56
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 6 9
 

TOTAL SECONDARY TUMORS 7 11
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT 1
 

TOTAL UNCERTAIN TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

B-IO
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH p-CRESIDINE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN MALE RATS TREATED WITH p-CRESIDINE 

CONTROL(UNTR)
 

01-0220
 

ANIMALS INITIALLY IN STUDY 50
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY *'

k

 48
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

EPIDERMAL INCLUSION CYST
 

*SUBCUT TISSUE
 

ABSCESS, NOS
 

RESPIRATORY SYSTEM
 

*NASAL CAVITY
 

HYPERPLASIA, EPITHELIAL
 POLYP
 

*NASAL TURBINATE
 

INFLAMMATION, SUPPURATIVE
 

•LUNG/BRONCHUS
 CALCIFICATION, NOS
 

*LUNG
 

BRONCHOPNEUMONIA, ACUTE
 

INFLAMMATION, ACUTE FOCAL
 

PNEUMONIA, CHRONIC MURINE
 METAPLASIA, NOS
 

•LUNG/ALVEOLI
 

CALCIFICATION, FOCAL
 

HEMATOPOIETIC SYSTEM
 

*SPLEEN
 

INFARCT, NOS
 

(48)
 

1 (2%)
 

(48)
 

3 (6%)
 

(48)
 

(48)
 

1 (25S)
 

(48)
 

(48)
 

(48)
 

(48)
 

1 (2%)
 

LOW DOSE
 

01-0260
 

50
 

50
 

49
 

(50)
 

(50)
 

(50)
 

1 (25S)
 

(50)
 

(49)
 

(49)
 

3 (6%)
 

(49)
 

(49)
 

HIGH DOSE
 

01-0265
 

50
 

47
 

47
 

(47)
 

(47)
 

(47)
 

5 (11%)
 

(47)
 

(45)
 

1 (2%)
 

(45)
 

1 (2%)
 

2 (4%)
 

2 (4%)
 

(45)
 

1 12%)
 

(45)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 ** EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

•C-3
 
TABLE Cl 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 01-0220 01-0260 01-0265
 

ATROPHY, NOS 1 (2JO
 

HEMATOPOIESIS 1 (2%)
 

#THYMUS (32) (33) (23)
 

HYPERPLASIA, NOS 1 (3JS)
 

CIRCULATORY SYSTEM
 

•HEART	 (48) (49) (45)
 

PERIARTERITIS 1 (2%)
 

CALCIFICATION, FOCAL 1 (2%)
 

•MYOCARDIUM	 (48) (49) (45)
 

DEGENERATION, NOS 6 C12%) 1 (2%)
 

CALCIFICATION, FOCAL 2 (4%)
 

*BLOOD VESSEL (48) (50) (47)
 

MEDIAL CALCIFICATION 2 (4%)
 

*CORONARY ARTERY (48) (50) (47)
 

CALCIFICATION, FOCAL 1 (2X)
 

DIGESTIVE SYSTEM
 

•SALIVARY	 GLAND (45) (48) (43)
 

HYPERPLASIA, INTRADUCTAL 1 (2%)
 

DYSPLASIA, NOS 1 (2%)
 

#LIVER (48) (49) (46)
 METAMORPHOSIS FATTY 2 (4%) 1 (2%)
 BASOPHILIC CYTO CHANGE 5 (10%) 1 (2%)
 CLEAR-CELL CHANGE 1 (2%) 1 (2X)
 

HYPERPLASIA, FOCAL 3 (6%) 1 (2%)
 

•PANCREAS	 (44) (48) (40)
 

INFLAMMATION, NOS 2 C 5%)
 

•PANCREATIC	 ACINUS (44) (48) (40)
 ATROPHY, NOS 1 (3X)
 

•STOMACH	 (48) (49) (46)
 

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

CALCIFICATION, FOCAL 2 (4%)
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-4
 
HYPERPLASIA, BASAL CELL
 

iGASTRIC MUCOSA
 

CALCIFICATION, NOS
 

•GASTRIC	 MUSCULARIS
 

INFLAMMATION, NECROTIZING
 CALCIFICATION, NOS
 

URINARY SYSTEM
 

•KIDNEY
 

HYDRONEPHROSIS
 

HEMORRHAGE
 

GLOMERULONEPHRITIS, NOS
 

INFLAMMATION, INTERSTITIAL
 

NEPHROPATHY
 

NEPHROSIS, NOS
 

NECROSIS, MEDULLARY
 CALCIFICATION, NOS
 

CALCIFICATION, FOCAL
 

HYPERPLASIA, TUBULAR CELL
 

HYPERPLASIA, EPITHELIAL
 

•KIDNEY/MEDULLA
 

AMYLOIDOSIS
 

•RENAL PAPILLA
 

INFLAMMATION, ACUTE NECROTIZING
 

•KIDNEY/TUBULE
 

CAST, NOS
 

FIBROSIS
 

DEGENERATION, NOS
 

CALCIFICATION, FOCAL
 

•KIDNEY/PELVIS
 

HYPERPLASIA, EPITHELIAL
 

•URINARY BLADDER
 

INFLAMMATION, ACUTE FOCAL
 

HYPERPLASIA, EPITHELIAL
 

HYPERPLASIA, PAPILLARY
 

POLYP, INFLAMMATORY
 

TAELE Cl 

(CONTINUED) 

CONTROL(UNTR)
 

01-0220
 

(48)
 

(48)
 

(48)
 

35 (73%)
 

(48)
 

(48)
 

(48)
 

(48)
 

(48)
 

LOW DOSE
 

01-0260
 

8 (16%)
 

(49)
 

(49)
 

(49)
 

1 (2%)
 

26 (53%)
 

(49)
 

(49)
 

(49)
 

(49)
 

1 (2%)
 

(48)
 

11 (23%)
 

HIGH DOSE
 

01-0265
 

(46)
 

1 (2%)
 

(46)
 

1 (2%)
 

1 (2%)
 

(46)
 

3 (7%)
 

1 (2%)
 

3 (7%)
 

1 (2%)
 

14 (30%)
 

3 (7%)
 

3 (7%)
 3 (7%)
 

1 (2%)
 

2 (4%)
 

(46)
 

1 (2%)
 

(46)
 

1 (2%)
 

(46)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(46)
 

(47)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 3 (6%)
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-5
 
TABLE Cl 

(CONTINUED) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-6
 
TABLE Cl
 

(CONCLUDED)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-7
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE RATS TREATED WITH p-CRESIDINE 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-8 
TABLE C2 

(CONTINUED) 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-9
 
TABLE C2 

(CONTINUED) 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-IO
 
TABLE C2
 

(CONCLUDED)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

Oil
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH p-CRESIDINE
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN MALE MICE TREATED WITH p-CRESIDINE 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 05-0220 05-0260 05-0265
 

ANIMAL S INITIALL Y I N STUD Y 50 50 50
 

ANIMAL S MISSIN G 1 3
 

ANIMAL S NECROPSIE D 50 47 41
 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* * 50 43 37
 

INTEGUMENTAR Y SYSTE M
 

*SUBCU T TISSU E (50) (47) (41 )
 

ABSCESS , NO S 1 (2%)
 

RESPIRATOR Y SYSTE M
 

4LUNG/BR0NCHU S (50) (40) (37)
 

INFLAMMATION , NO S 1 (2X)
 

HEMATOPOIETI C SYSTE M
 

tSPLEE N (50) (37) (33)
 

ATROPHY , NO S 17' (46K)
 

#MESENTERI C L . NOD E (44) (26) (16)
 

HEMATOPOIESI S 1 (2S)
 

CIRCULATOR Y SYSTE M
 

#MYOCARDIU M (50) (40) (36)
 MINERALIZATION 1 (35«)
 

DEGENERATION, NOS 1 (3JJ)
 

HCORONARY ARTERY (50) (47) (41 )
 

MEDIAL CALCIFICATION 1 (2%)
 

^PULMONARY ARTERY (50) (47) (4 1 )
 

MEDIAL CALCIFICATION 1 (25 O
 

DIGESTIVE SYSTEM
 

#LIVER (50) (41 ) (37 )
 

NECROSIS. COAGULATIVE 1 (2%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-3 
•PANCREATIC	 ACINUS
 

ATROPHY, NOS
 

HYPERTROPHY, FOCAL
 

•GASTRIC MUCOSA
 

CALCIFICATION, NOS
 

•GASTRIC	 MUSCULARIS
 

CALCIFICATION, NOS
 

URINARY SYSTEM
 

tKIDNEY
 

HYDRONEPHROSIS
 

•URINARY	 BLADDER
 

HYPERPLASIA, EPITHELIAL
 

DYSPLASIA, NOS
 

ENDOCRINE SYSTEM
 

•PANCREATIC	 ISLETS
 

INFLAMMATION, NOS
 

HYPERPLASIA, ADENOMATOUS
 

REPRODUCTIVE SYSTEM
 

•TESTIS
 

ATROPHY, NOS
 

#TESTIS/TUBULE
 

MINERALIZATION
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

TABLE Dl 

(CONTINUED) 

CONTROL(UNTR)
 05-0220
 

(50)
 

1 (2%)
 

1 (2%)
 

(50)
 

(50)
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

3 (6X)
 

1 (2%)
 

(50)
 

1 (25J)
 

(50)
 

2 (4%)
 

LOW DOSE
 05-0260
 

(30)
 

(32)
 

(32)
 

(43)
 

29 (67%)
 

(42)
 

(30)
 

(41 )
 

(41 )
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-4
 

HIGH DOSE
 05-0265
 

(31)
 

(33)
 

1 (3%)
 

(33)
 

1 (3%)
 

(36)
 

31 (86%)
 

(35)
 

3 (9X)
 

1 (3%)
 

(31 )
 

(37)
 

(37)
 
HIGH DOSE
 05-0265
 

(41 )
 

1 (25*)
 

(41 )
 

1
 

3
 

2
 

2
 

6
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

MPERITONEUM
 

INFLAMMATION, ACUTE/CHRONIC
 

ALL OTHER SYSTEMS
 

^MULTIPLE ORGANS
 

ATROPHY, NOS
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 

ANIMAL MISSING/NO NECROPSY
 

NECROPSY PERF/NO HISTO PERFORMED
 

AUTO/NECROPSY/NO HISTO
 AUTOLYSIS/NO NECROPSY
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED


* NUMBER OF ANIMALS NECROPSIED
 

TABLE Dl
 

(CONCLUDED)
 

CONTROL(UNTR)
 05-0220
 

(50)
 

(50)
 

22
 

LOW DOSE
 05-0260
 

(47)
 

(47)
 

1 (2%)
 

1
 

1
 

4
 

2
 

 MICROSCOPICALLY
 

D-5
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE MICE TREATED WITH p-CRESIDINE 

HIGH DOSE
 

06-0265
 

50
 

48
 

48
 

(48)
 

(44)
 

(39)
 

3 (8%)
 

(38)
 

2 (5%)
 

2 (5%)
 

(28)
 

(42)
 

1 (2%)
 

(42)
 

1 (2%)
 

ANIMALS INITIALLY IN STUDY
 

ANIMALS MISSING
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE
 

INFLAMMATION, NOS
 

RESPIRATORY SYSTEM
 

• LUNG
 

THROMBOSIS, NOS
 

INFLAMMATION, ACUTE NECROTIZING
 

HEMATOPOIETIC SYSTEM
 

•BONE MARROW
 

MYELOFIBROSIS
 

MYELOPOIESIS
 

•SPLEEN
 

ATROPHY, NOS
 

HYPERPLASIA, RETICULUM CELL
 

HYPERPLASIA, LYMPHOID
 

HEMATOPOIESIS
 

•MEDIASTINAL	 L.NODE
 

HYPERPLASIA, LYMPHOID
 

CIRCULATORY SYSTEM
 

•HEART
 

PERIARTERITIS
 

•MYOCARDIUM
 

CALCIFICATION, NOS
 

CONTROL(UNTR)
 

06-0220
 

50
 

2
 

47
 ** 47
 

(47)
 

1 (2%)
 

(46)
 

(39)
 

(45)
 

1 (25O
 

(38)
 

(45)
 

(45)
 

LOW DOSE
 

06-0260
 

50
 

47
 

47
 

(47)
 

(47)
 

1 (2%)
 

1 (2%)
 

(44)
 

14 (32%)
 

(44)
 

9 (20%)
 

2 (5%)
 

2 (5%)
 

(38)
 

1 (3%)
 

(47)
 

(47)
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-6 
TABLE D2 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 06-0220 06-0260 06-0265
 

*ARTERY (47) (47) (48)
 

INFLAMMATION, NOS 1 (2%)
 

*CORONARY ARTERY (47) (47) (48)
 

MEDIAL CALCIFICATION 1 (25i)
 

*MEDIASTINAL ARTERY (47) (47) (48)
 

MEDIAL CALCIFICATION 1 (2K)
 

DIGESTIVE SYSTEM
 

#LIVER (46) (45) (43)
 

ABSCESS, CHRONIC 1 (2%)
 

METAMORPHOSIS FATTY 1 (2X) 2 (4%)
 

ANGIECTASIS 1 (2%)
 

•PANCREAS	 (45) (42) (33)
 

CYSTIC DUCTS 1 (2%)
 

•PANCREATIC	 ACINUS (45) (42) (33)
 

ATROPHY, NOS 1 (2%)
 

•STOMACH	 (46) (40) (36)
 

ULCER, NOS 1 (3%)
 

HYPERKERATOSIS 1 (3%)
 

•GASTRIC	 MUCOSA (46) (40) (36)
 

INFLAMMATION, ACUTE FOCAL 1 (3%)
 

CALCIFICATION, FOCAL 1 (3%)
 

•GASTRIC	 MUSCULARIS (46) (40) (36)
 

CALCIFICATION, FOCAL 1 (3%)
 

URINARY SYSTEM
 

•KIDNEY	 (46) (45) (45)
 HYDRONEPHROSIS 18 (40?O 37 (82X)
 GLOMERULONEPHRITIS, NOS 1 (2%) 1 (2%)
 PYELONEPHRITIS, ACUTE 1 (2K) 1 (2JO
 PYELONEPHRITIS, CHRONIC 1 (2%)
 

NEPHROPATHY 1 (2%)
 NEPHROSIS, NOS 1 ( 7 *£ )
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-7
 
GLOMERULOSCLEROSIS, NOS
 

INFARCT, NOS
 

CALCIFICATION, FOCAL
 

•URINARY BLADDER
 

HYPERPLASIA, EPITHELIAL
 

DYSPLASIA, NOS
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLAND
 

HYPERPLASIA, NOS
 

tUTERUS
 

HYDROMETRA
 

tUTERUS/ENDOMETRIUM
 

INFLAMMATION, ACUTE
 

HYPERPLASIA, CYSTIC
 

•OVARY/OVIDUCT
 

DEGENERATION, NOS
 

fOVARY
 THROMBOSIS, NOS
 PLASMA-CELL INFILTRATE
 

NERVOUS SYSTEM
 

*BRAIN
 

HYDROCEPHALUS, NOS
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

TABLE D2 

(CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE 

06-0220 06-0260 06-0265 

1 (2%) 

1 

1 (2%) (2%) 

(45) (47) 

2 (4%) 

(48) 

1 (2%) 

(47 ) 

1 (2%) 

(47 ) (48) 

(45 ) (46 ) (36) 

1 (2%) 

(45 ) (46 ) (36) 

1 (2%) 

14 (.30%) 1 (3X) 

(45 ) (46) (36) 

1 (2%) 

(40 ) (41 ) (35) 

1

1

 (23) 

 (2%) 

(44) (45) (39) 

1 (2%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-8
 
TABLE D2
 

(CONCLUDED)
 

CONTROL(UNTR)
 06-0220
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

^MULTIPLE ORGANS (47)
 

PERIARTERITIS
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 

ANIMAL MISSING/NO NECROPSY
 

AUTO/NECROPSY/HISTO PERF
 

AUTOLYSIS/NO NECROPSY
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED


* NUMBER OF ANIMALS NECROPSIED
 

30
 

2
 

1
 

1
 

 MICROSCOPICALLY
 

LOW DOSE HIGH DOSE
 06-0260 06-0265
 

(47)	 (48)
 

1 (25O
 

1
 

1
 

3 2
 

D-9
 

Review of the Bioassay of p-Cresidine* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance vith 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 p-Cresidine for Garcinogenicity.
 

The reviewer agreed with the conclusion in the report
 

that p-Cresidine was carcinogenic in both rats and mice,
 

under the conditions of test. He said that the nature of
 

the tumors observed clearly indicate that they were treatment-


related. He noted that the p-Cresidine was dissolved in
 

acetone but that there was no acetone control group. This
 

shortcoming, however, did not interefer with the conclusion
 

on the carcinogenicity of p-Cresidine. Based on the results
 

of the-study, he said that p-Cresidine would appear to pose
 

a carcinogenic risk to humans. The reviewer moved that the
 

report on the bioassay of p-Cresidine be accepted as written.
 

The motion was approved without objaction.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical U.S.A.
 

Michael B. Shimkin, University* of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

63
 

"frU.S. GOVERNMEN T PRINTIN G OFFICE * A 979-28 1 "217/308 3
 


DHEW Publication No. (NIH) 79-1397