1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 30 

BIOASSAYOF 

DIARYLANILIDE YELLOW 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 6358-85-6 

NCI-CG-TR-30 

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

Public Health Service 

National Institutes of Health 

BIDASSAY OF
 

DIARYLANILIDE YELLOW
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

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

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-830
 

REPORT ON THE BIOASSAY OF DIARYLANILIDE YELLOW
 

FOR POSSIBLE CARCINOGEN1CITY
 

CARCINOGENSSIS PROGRAM, DIVISION OF CANCER CAUSE AND PREVENTION
 

NATIONAL CANCER INSTITUTE, NATIONAL INSTITUTES OF HEALTH
 

CONTRIBUTORS: This report presents the results of the bioassay of
 

diarylanilide yellow conducted for the Carcinogen Bioassay and Program
 

Resources Branch, Carcinogenesis Program, Division of Cancer Cause
 

and Prevention, National Cancer Institute (NCI), National Institutes
 

of Health, Bethesda, Maryland. This bioassay was conducted by Mason
 

Research Institute, Worcester, Massachusetts, initially under direct
 

contract to the NCI and currently under a subcontract to Tracer
 

Jitco, Inc. (1), prime contractor for the NCI Carcinogenesis Bioassay
 

Program.
 

The experimental design was determined by the NCI Project Officers, Dr. J. H. Weisburger (2,3) and Dr. E. K. Weisburger (2). The
 

principal investigators for the contract were Dr. E. Smith (4) and
 

Dr. A. Handler (4). Animal treatment and observation were supervised
 

by Mr. G. Wade (4) and Ms. E. Zepp (4). Chemical analysis was performed by Midwest Research Institute (5).
 

Histopathology was performed by Dr. R. W. Fleischman (4) at the
 

Mason Research Institute, and the diagnoses included in this report
 

represent the interpretation of this pathologist.
 

Compilation of individual animal survival, pathology, and
 

summary tables was performed by EG&G Mason Research Institute (6);
 

the statistical analysis was performed by Dr. A. Chu (6) and Mr. W.
 

W. Belew (7), using methods selected for the Bioassay Program by Dr.
 

J. J. Gart (8).
 

This report was prepared at METREK, a Division of The MITRE
 

Corporation (7) under the direction of the NCI. Those responsible
 

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

Thomas (7), the task leader, Dr. M. R. Kornreich (7), and the senior
 

biologist, Ms. P. Walker (7). The final report was reviewed by
 

members of the participating organizations.
 

The statistical analysis was reviewed by a member or members of
 

the Mathematical Statistics and A plied Mathematics Section of the
 

NCI: Dr. J. J. Gart (8), Mr. J. i.am (8), Dr. H. M. Pettigrew (8),
 

and Dr. R. E. Tarone (8).
 

iii
 
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 (2), Dr. C.
 

Cueto, Jr. (2), Dr. J. F. Douglas (2), Dr. D. G. Goodman (2), Dr.
 

R. A. Griesemer (2), Di. T. W. Orme (2), Dr. R. A. Squire (9), and
 

Dr. J. M. Ward (2).
 

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

Maryland.
 

2.	 Carcinogenesis Program, Division of Cancer Cause and Prevention,
 

National Cancer Institute, National Institutes of Health,
 

Bethesda, Maryland,
 

3.	 Now with the Nayloi: Dana Institute for Disease Prevention,
 

American Health Foundation, Hammon House Road, Valhalla,
 

New York.
 

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

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

Montana.
 

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

Rockville, Maryland.
 

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

Boulevard, McLean, Virginia.
 

8.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

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

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

iv
 
SUMMARY 

A bioassay of technical-grade diarylanilide yellow for possible
 

carcinogenicity was conducted using Fischer 344 rats and B6C3F1 mice.
 

Diarylanilide yellow was administered in the feed, at either of two
 

concentrations, to groups of 50 male and 50 female animals of each
 

species. The high and low dietary concentrations used in the chronic
 

study for the male and female rats and mice were 5.0 and 2.5 percent,
 

respectively, of the chemical in the feed. After a 78-week treatment
 

period, observation of the rats continued for an additional 28 weeks
 

and observation of the mice continued for an additional 19 weeks for
 

high dose males and low and high dose females and 18 weeks for low
 

dose males. For each species, 50 animals of each sex were placed on
 

test as controls, and fed only the basal diet.
 

The high concentration administered to both species in this study
 

was the maximum recommended in the Guidelines for Carcinogen Bioassay
 

in Small Rodents (Sontag et al., 1976). These guidelines indicate
 

that a chronic dietary level of 5 percent, or 50,000 ppm, should not
 

be exceeded even when no signs of toxicity are observed during sub-


chronic testing, except under special circumstances (e.g., when the
 

compound is a major component of the human diet). No toxic effects
 

were reported during subchronic testing and diarylanilide yellow did
 

not qualify for exception; therefore, the highest permissible concentration (5 percent) was utilized in the chronic bioassay.
 

The dietary concentrations of diarylanilide yellow administered
 

during the chronic bioassay had no significant effect on survival or
 

body weight gain in either species. Except for yellow staining and
 

some isolated neoplasms, the only adverse clinical sign or pathologic
 

lesion observed in treated rats or mice was basophilic cytoplasm
 

changes in hepatocytes of treated rats.
 

In both species the survival in all groups was adequate for
 

statistical analysis of late-appearing tumors.
 

No treatment-related increase in the incidence of neoplasms or
 

nonneoplastic lesions was evident in treated rats or mice. A few unusual findings were observed in both species, including single cases
 

of metastatic chordoma and ost.eogenic sarcoma in rats, and single
 

cases of squamous-cell carcinoma of the ear, infiltrating duct carcinoma of the mammary gland, and subcutaneous mastocytoma in mice.
 

The results of this study did not provide evidence for the carcinogenicity of diarylanilide yellow in Fischer 344 rats or B6C3F1 mice.
 

v 

TABLE OF CONTENTS
 

I. INTRODUCTION
 

II. MATERIALS AND METHODS
 

A. Chemicals
 

B. Dietary Preparation
 

C. Animals
 

D. Animal Maintenance
 

E. Selection of Initial Concentrations
 

F. Experimental Design
 

G. Clinical and Histopathologic Examinations
 

H. Data Recording and Statistical Analyses
 

III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

B. Survival
 

C. Pathology
 

D. Statistical Analyses of Results
 

IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

B. Survival
 

C. Pathology
 

D. Statistical Analyses of Results
 

V. DISCUSSION
 

VI. BIBLIOGRAPHY
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH DIARYLANILIDE YELLOW
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH DIARYLANILIDE YELLOW
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH DIARYLANILIDE
 

YELLOW
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH DIARYLANILIDE
 

YELLOW
 

Page
 

1
 

3
 

3
 

3
 

A
 

4
 

6
 

8
 

8
 

12
 

17
 

17
 

17
 

20
 

20
 

26
 

26
 

26
 

29
 

30
 

33
 

35
 

A-l
 

B-l
 

C-l
 

D-l
 

vii
 
Figure Number
 

1
 

2
 

3
 

4
 

Table Number
 

1
 

2
 

3
 

4
 

5
 

6
 

Al
 

LIST OF ILLUSTRATIONS
 

GROWTH CURVES FOR DIARYLANILIDE YELLOW
 

CHRONIC STUDY RATS
 

SURVIVAL COMPARISONS OF DIARYLANILIDE
 

YELLOts CHRONIC STUDY RATS
 

GROWTb CURVES FOR DIARYLANILIDE YELLOW
 

CHRONIC STUDY MICE
 

SURVIVAL COMPARISONS OF DIARYLANILIDE
 

YELLOW CHRONIC STUDY MICE
 

LIST OF TABLES
 

DESIGN SUMMARY FOR FISCHER 344 RATS—
 

DIARYL\NILIDE YELLOW FEEDING EXPERIMENT
 

DESIGN SUMMARY FOR B6C3F1 MICE—DIARYLANILIDS YELLOW FEEDING EXPERIMENT
 

ANALYSIS OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN MALE RATS TREATED WITH
 

DIARYLANILIDE YELLOW
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN FEMALE RATS TREATED WITH
 

DIARYLANILIDE YELLOW
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN MALE MICE TREATED WITH
 

DIARYLANILIDE YELLOW
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN FEMALE MICE TREATED WITH
 

DIARYLANILIDE YELLOW
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MALE RATS TREATED WITH DIARYLANILIDE YELLOW
 

Page
 

18
 

19
 

27
 

28
 

Page
 

9
 

10
 

21
 

23
 

31
 

32
 

A-3
 

viii
 
LIST OF TABLES (Concluded) 

Table Number 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH DIARYLANILIDE 

YELLOW A ~ 7 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH DIARYLANILIDE 

YELLOW B ~ 3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH DIARYLANILIDE 

YELLOW B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH DIARYLANILIDE YELLOW C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 

DIARYLANILIDE YELLOW c ~ 8 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH DIARYLANILIDE YELLOW D ~ 3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 

DIARYLANILIDE YELLOW D ~ 7 

I. INTRODUCTION
 

Diarylanilide yellow (NCI No. C03269), one member of a family of
 

organic azo pigments known as benzidine yellows, was selected for
 

bioassay by the National Cancer Institute in an attempt to elucidate
 

those dyes and dye intermediates which may be responsible for the
 

increased incidence of bladder cancer observed among workers in the
 

dye manufacturing industry (Wynder et al., 1963; Anthony et al.,
 

1970). The structural relationship of this compound to the documented
 

carcinogen 3,3'-dichlorobenzidine (Occupational Safety and Health
 

Administration, 1973) was also a factor in its selection.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is 2,2'-[(3,3'-dichloroU,1'-biphenyl)­

*
 

4,4'-diyl)-bis (azo)] bis (3-oxo-N-phenyl)-butanamide. It is also
 

called Color Index (C.I.) Pigment Yellow 12 (C.I. 21090), diarylide
 

yellow, and dichlorobenzidine coupled into acetoacetanilide. Diarylanilide yellow is an ingredient in industrial paints, most notably
 

the paint applied to lead pencils (Weisburger, 1976). It is also an
 

ingredient in printing inks and may sometimes be used to color
 

plastics, rubber, linoleum, floor tiles, textiles, and wallpaper
 

(Society of Dyers and Colourists, 1971; Hawley, 1971). According to
 

the U.S. International Trade Commission (1977a), 6.028 x 10 pounds
 

*
 

The CAS registry number is 6358-85-6.
 

1
 
of diarylani1ide yellow were produced in the United States in 1975—
 

the largest quantity of any single pigment produced in that year.
 

U.S. imports of the pigment through principle U.S. customs districts
 

amounted to 62,040 pounds in 1975 (U.S. International Trade Commission, 1977b).
 

The risk of exposure to diarylanilide yellow is greatest among
 

workers in the dye marufacturing industry and at facilities where
 

dyeing of textiles or production of inks, paints, and other commodities containing the pigment takes place. Additional occupational
 

exposure may also occur among users of pigment-containing products
 

(e.c., among printers engravers, lithographers, textile workers,
 

etc. ) .
 

Epiaemioiogical studies suggest a relationship between occupational exposure to pa .nts and increased incidences of cancer of the
 

lung and bladder and between occupational exposure to printing inks
 

and increased incidences of cancer of the liver and bladder (Hoover
 

and Fraumeni, 1975). An increased incidence of bladder cancer has
 

also been observed among textile workers and tailors (Anthony and
 

Thomas, 1970).
 

Exposure of the general population to diarylanilide yellow is
 

likely, due to the large variety of consumer products colored with
 

this pigment. Chroni: ingestion of the dye over long periods of time
 

may result from habitjal holding in the mouth or chewing of wooden
 

pencils.
 

2 
II. MATERIALS AND METHODS
 

A. Chemicals
 

Diarylanilide yellow was purchased from Chemtron Corporation and
 

chemical analysis was performed by Midwest Research Institute. The
 

melting point range (311° to 320°C) suggested the presence of impurities. Thin-layer chromatography was performed utilizing two different
 

solvent systems (methylene chloride and 95:5 chloroform:diethylamine).
 

Each plate was visualized with ultraviolet and visible light. One
 

homogeneous spot was detected on each plate; however, the amounts of
 

compound spotted on each plate (2.4 |ag and 7.2 |j.g) were so low that
 

only major impurities could have been detected by this technique.
 

Infrared and mass spectrometry analyses were not inconsistent with
 

the structure of the compound.
 

Throughout this report the term diarylanilide yellow is used to
 

represent this technical-grade material.
 

B. Dietary Preparation
 

The basal laboratory diet for both treated and control animals
 

(F
 consisted of Wayne Lab-Blox (Allied Mills, Inc.). Diarylanilide
 

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

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

Patterson-Kelly standard model stainless steel twin-shell V-blender.
 

After 20 minutes of blending, the mixtures were placed in double
 

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

weekly and stored for not longer than 2 weeks.
 

3
 
C. Animals
 

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

through contracts of the Division of Cancer Treatment, National Cancer
 

Institute. Animals of both species were supplied by Charles River
 

Breeding Laboratories, Wilmington, Massachusetts. Treated animal
 

groups of both species were received in separate shipments from their
 

corresponding controls.
 

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

other signs of disease. The remaining animals were quarantined for
 

2 weeks prior o initiation of test. Animals were assigned to groups
 

and distributed among cages so that average body weight per cage was
 

approximately equal for a given sex and species.
 

D. Animal Maintenance
 

All animals were housed by species in rooms having a temperature
 

range of 23° to 34°C and a range in relative humidity of 10 to 85
 

(R)
 percent. Incoming air was filtered through Tri-Dek 15/40 denier
 

Dacron filters 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 6 weeks cf study, they were kept in galvanized-steel
 

wire—mesh cages suspended above newspapers. Newspapers were replaced
 

daily, and cages and racks washed weekly. From week 6 rats were kept
 

in suspended polycarbonate cages equipped with disposable nonwoven
 

4 
filter sheets. Clean bedding and cages were provided twice weekly.
 

Hardwood chips (Ab-sorb-dri , Wilner Wood Products Co.) were used
 

through the first 3 months of study, then corncob bedding (SAN-I-CEL,
 

Paxton Processing Co.) for the next 12 months, and then another type
 

®


of corncob bedding (Bed-o'Cobs , The Anderson's Cob Division) for
 

the remainder of the bioassay. During the quarantine period Wayne
 

Lab-Blox was supplied in Alpine aluminum feed cups (Curtin Matheson
 

Scientific, Inc.) equipped with stainless steel baffles. The same
 

(H)
 feeding apparatus, containing treated Wayne Lab-Blox , was utilized
 

during the treatment period. The food assembly was replaced weekly.
 

During the observation period following treatment, rats were supplied
 

with pelleted Wayne Lab-Blox on the cage floor.
 

Mice were housed by sex in polycarbonate cages. During quarantine and treatment periods, cages fitted with perforated stainless
 

steel lids (Lab Products, Inc.) were used. During the observation
 

period following treatment, stainless steel wire bar lids (Lab
 

Products, Inc.) were used. Nonwoven fiber filter bonnets were used
 

over cage lids. Treated mice were housed ten per cage for the first
 

12 months of the study and five per cage thereafter. Control mice
 

were housed ten per cage for the first 13 months of study and five
 

per cage thereafter. Cages, lids, filters, and bedding were provided
 

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

week when cage populations were reduced to five. Bedding was of the
 

same type as that used for rats. Reusable filter bonnets and pipe
 

5 
racks were sanitized biweekly throughout the study. During the
 

quarantine and test periods, Wayne Lab-Blox was supplied in Alpine
 

aluminum feed cups equipped with stainless steel baffles. This food
 

assembly was replaced weekly. During the observation period following
 

(S)
 

treatment, mice were supplied with pelleted Wayne Lab-Blox through
 

a food hopper incorporated into the cage lid.
 

Water was available from 250 ml water bottles equipped with
 

rubber stoppers and stainless steel sipper tubes. Bottles were
 

replaced twice weekly. Food and water were available ad libitum to
 

both rats and mice.
 

Treated and control rats used for this bioassay were housed with
 

*
 

other rats treated with fenaminosulf (140-56-7), 2,5-dithiobiurea
 

(142-46-1), m-cresidine (102-50-1), and cupferron (135-20-6). After
 

6 weeks the diarylanilide yellow-treated rats were segregated from
 

all other animals. The treated and control mice utilized in this
 

bioassay were housed with other mice treated with 1-nitronaphthalene
 

(86-57-7).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

diarylanilide yellow for administration to treated animals in the
 

chronic studies, subchronic toxicity tests were conducted with both
 

rats and mice. Rats vere distributed among five groups and mice
 

among six groups, each consisting of five males and five females.
 _
 

CAS registry numbers are given in parentheses.
 

6
 
Diarylanilide yellow was incorporated into the basal laboratory diet
 

and fed ad libitum to four of the five rat groups in concentrations
 

of 0.1, 0.3, 1.0, and 3.0 percent and to five of the six mouse groups
 

in concentrations of 0.03, 0.1, 0.3, 1.0 and 3.0 percent. The
 

remaining group of each species served as a control group, receiving
 

only the basal laboratory diet. The dosed dietary preparations were
 

administered for a period of 8 weeks.
 

A dosage inducing no mortality or body weight gain retardation
 

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

chronic bioassay.
 

No decreases in food consumption or significant weight depression
 

relative to controls were observed in any group. All animals survived
 

until necropsy (week 8). Although the external surfaces of all animals at all concentrations were bright yellow, gross necropsy revealed
 

no abnormalities or organ discoloration other than the mucosal surfaces of the intestinal tract, which appeared bright yellow due to
 

direct contact with the test compound.
 

In the Guidelines for Carcinogen Bioassay in Small Rodents
 

(Sontag et al., 1976) it is indicated that a chronic dietary concentration of 5 percent (50,000 ppm) should not be exceeded. This
 

applies even if the compound causes no toxicity during subchronic
 

testing. An exception can be made under special circumstances, e.g.,
 

if the chemical is a major component of the human diet. Because no
 

toxic symptoms or gross abnormalities were observed clinically or at
 

7 
necropsy in animals receiving the tested concentrations, 5.0 percent
 

was selected as the concentration to be administered to the high dose
 

groups of both species during the chronic bioassay.
 

F.	 Experimental Design
 

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

treated and untreated observation periods) are summarized in Tables 1
 

and	 2.
 

At initiation of the study animals of both species were approximately 7 weeks old. Hi£;h and low dose animals of both species and
 

sexes received concentrations of 5.0 and 2.5 percent, respectively,
 

of the chemical in their food. Animals were treated for 78 weeks,
 

followed by a 28-week observation period for rats and observation
 

periods of 19 weeks for high dose male and low and high dose female
 

mice and 18 weeks for low dose male mice, during which they received
 

the basal laboratory dieit. For both species, control animals were
 

maintained and observed in the same manner as the treated animals.
 

G.	 Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. From the fi.rst day, all animals were inspected twice
 

daily for mortality. Body weights were recorded twice weekly for
 

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

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

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

8 
TABLE 1 

DESIGN SUMMARY FOR FISCHER 344 RATS 

DIARYLANILIDE YELLOW FEEDING EXPERIMENT 

MALE 

INITIAL

GROUP

SIZE

DIARYLANILIDE 

 YELLOW 

 CONCENTRATION 

 (PERCENT) 

OBSERVATION PERIOD 

TREATED UNTREATED 

(WEEKS) (WEEKS) 

CONTROL 50 0 0 109 

LOW DOSE 50 2.5 

0 

78 

28 

HIGH DOSE 50 5.0 

0 

78 

28 

FEMALE 

CONTROL 50 0 0 110 

LOW DOSE 50 2.5 

0 

78 

28 

HIGH DOSE 50 5.0 

0 

78 

28 

9
 
TABLE 2 

DESIGN SUMMARY FOR B6C3F1 MICE 

DIARYLANILIDE YELLOW FEEDING EXPERIMENT 

MALE 

INITIAL

3ROUP

SIZE

DIARYLANILIDE 

 YELLOW 

 CONCENTRATION 

 (PERCENT) 

OBSERVATION PERIOD 

TREATED UNTREATED 

(WEEKS) (WEEKS) 

CONTROL 50 0 0 97 

LOW DOSE 50 2.5 

0 

78 

18 

HIGH DOSE 50 5.0 

0 

78 

19 

FEMALE 

CONTROL 50 0 0 98 

LOW DOSE 50 2.5 

0 

78 

19 

HIGH DOSE 50 5.0 

0 

78 

19 

10 
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, or gross lesions taken from sacrificed animals and, whenever
 

possible, from animals found dead.
 

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

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

bile duct, pancreas, esophagus, stomach, small intestine, large intestine, kidney, urinary bladder, pituitary, adrenal, thyroid, parathyroid, pancreatic islets, testis, prostate, brain, uterus, mammary
 

gland, and ovary.
 

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

with hematoxylin and eosin prior to microscopic examination. An
 

occasional section was subjected to special staining techniques for
 

more definitive diagnosis.
 

A few tissues were not examined for some animals, particularly
 

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

11
 
preclude histopathologLc 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
 

missing; animals dying from natural causes were not statistically
 

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

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

12
 
equality and used Tarone's (1975) extensions of Cox's methods for
 

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

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

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

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

histologically. However, when macroscopic examination was required
 

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

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

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

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

13
 
be less than or equal t:> 0.05/k. In cases where this correction was
 

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

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

The Cochran-Armita^e test for linear trend in proportions, with
 

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

Under the assumption of a linear trend, this test determined if the
 

slope of the dose-response curve is different from zero at the one-


tailed 0.05 level of significance. Unless otherwise noted, the direction of the significant trend was a positive dose relationship.
 

This method also provides a two-tailed test of departure from linear
 

trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

14
 
observed tumor were computed as in Saffiotti et al. (1972). The week
 

during which animals died naturally or were sacrificed was entered as
 

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

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

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

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

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

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


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

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

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

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

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

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

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

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

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

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

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

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

15
 
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 turners by the test chemical which could not be
 

detected under the conditions of this test.
 

16
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

The body weight patterns for control and treated rat groups of
 

both sexes were generally equivalent throughout the treatment period
 

(Figure 1).
 

All the treated rats, both male and female, appeared bright
 

yellow in color. In addition, the conjunctivas were faintly yellow
 

as were most organs and internal mucosal surfaces. The only other
 

clinical sign recorded for male or female rats was a hard crusted
 

lesion on the back of one male control animal.
 

B.	 Survival
 

The estimated probabilities of survival for male arid female rats
 

in the control and diarylanilide yellow-treated groups are shown in
 

Figure 2.
 

For both male and female rats the Tarone test detected no statistically significant positive association between dosage and mortality. In the males survival was quite high, as 74 percent of the
 

high dose, 84 percent of the low dose, and 64 percent of the control
 

rats survived until the end of the study, despite the sacrifice of
 

five high dose and five control rats in week 78. In the females, 66
 

percent of the high dose, 80 percent of the low dose, and 72 percent
 

of the control rats survived until the end of the study, despite the
 

sacrifice of five high dose and five control rats in week 78.
 

17
 
TIME ON TEST (WEEKS) 

FIGURE 1 

GROWTH CURVES FOR DIARYLANILIDE YELLOW CHRONIC STUDY RATS 

18 
TIME ON TEST (WEEKS) 

FIGURES 

SURVIVAL COMPARISONS OF DIARYLANILIDE YELLOW CHRONIC STUDY RATS 

19 
In both sexes, survival was adequate for meaningful statistical
 

analyses of tumor incidence.
 

C. Pathology
 

Histopathologic findings on neoplasms in rats are tabulated in
 

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

tabulated in Appendix C (Tables Cl and C2).
 

With a few exceptions, the same variety of neoplasms occurred
 

sporadically and randomly in the chemically treated and control
 

groups. No particular organ or system seemed to be the target of
 

this chemical. Sporadic and unusual neoplasms that occurred in the
 

treated but not in control animals were as follows: a metastatic
 

chordoma of unknown origin occurred in the lung of 1/49 of the low
 

dose males, and 1/49 of the low dose females had an osteogenic
 

sarcoma.
 

The incidence and variety of nonneoplastic degenerative, proliferative, and inflammatory lesions were similar in the control and
 

the chemically treated rats, except for treatment-related basophilic
 

cytoplasm changes in hepatocytes of treated males and females.
 

The results of this histopathologic examination did not provide
 

evidence for the carcinogenicity of diarylanilide yellow in Fischer
 

344 rats.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

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

20
 
TABLE 3 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT 

SPECIFIC SITES IN MALE RATS TREATED WITH DIARYLANILIDE YELLOW3

 

I 

21 

TOPOGRAPHY :MORPHOLOGY
 

Skin: Fibroma or Basal-cell Carcinoma
 

P Values0 >d
 

Relative Risk (Control)6
 

Lover Limit
 

Upper Limit
 

Weeks to First Observed Turoor
 

Hematopoletic System: Leukemia or Malignant Lymphotna
 

P Values0 >d
 

Relative Risk (Control)6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Adenoma
 

P Values0 'd
 

Departure from Linear Trend
 

Relative Risk (Control)6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma
 

P Values0 >d
 

Relative Risk (Control)6
 

Lover Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW HIGH 

CONTROL DOSE DOSE 

2/50(0.04) 4/50(0.08) 1/50(0.02) 

N.S. N.S. N.S. 

2.000 0.500 

0.301 0.009 

21.320 9.290 

85 91 106 

10/50(0.20) 2/50(0.04) 1/50(0.02) 

P ­ O.OOl(N) P ­ 0.014(N) P ­ 0.004(N) 

0.200 0.100 

0.022 0.002 

0.877 0.662 

78 99 103 

7/45(0.16) 12/43(0.28) 5/45(0.11) 

N.S. N.S. N.S. 

P ­ 0.041 

1.794 0.714 

0.723 0.193 

4.856 2.414 

78 106 106 

3/50(0.06) 3/47(0.06) 5/49(0.10) 

N.S. N.S. N.S. 

1.064 1.701 

0.149 0.351 

7.570 10.420 

78 96 106 
TOPOGRAPHY: MORPHOLOGY
 

Thyroid: C-Cell Adenoma or Carcinoma
 

P Values0 >d
 

Relative Risk (Control)6
 

Lower Limit
 

Upper Limit
 

Pancreatic Islets: Adenoma
 

P Values0 >d
 

Relative Risk (Control)6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Test Is: Interstitial-Cell Tumor
 

P Values0 >d
 

Relative Risk (Control)6
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 

(CONCLUDED) 

LOU HIGH 

CONTROL DOSE DOSE 

3/37(0.08) 5/47(0.11) 1/48(0.02) 

N.S. N.S. N.S. 

1.312 0.257 

0.275 0.005 

7.994 3.055 

105 96 ICC 

1/47(0.02) 2/47(0.04) 5/46(0.11) 

N.S. N.S. JI.S. 

2.000 5.109 

0.108 0.603 

115.500 235.900 

109 106 93 

42/50(0.84) 44/48(0.92) 39/49(0.80) 

N.S. N.S. N.S. 

1.091 0.948 

0.922 0.782 

1.240 1.161 

78 96 78 

22 

Dosed groups received concentrations of 2.5 and 5.0 percent in feed.
 

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

C

Beneath the incidence of the control is the probability level for the Cochran-Armitage test for dose-related trend in proportions when it Is below
 

0.05; otherwise N.S.- not significant. Departure from linear trend is noted when it is below 0.05 for any comparison. Beneath each dose group
 

incidence is the probability level for the Fisher exact (conditional) test for the comparison of that dose group to the control group when it la
 

below 0.05, otherwise N.S.- not significant.
 

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

e

Relative risk of the treated group versus the control group is shown along with the lower and upper limit of the 95% confidence interval for that
 

relative risk.
 
8

 

TABLE 4 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT 

SPECIFIC SITES IN FEMALE RATS TREATED WITH DIARYLANILIDE YELLOW8 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Hematopoietlc System: Leukemia
 

P Values0'*1
 

Relative Riak (Control)'
 

Lower Limit
 

Upper Limit
 

Weeka to First Observed Timor
 

Pituitary: Adenomab
 

P Values0 >d
 

Departure from Linear Trend
 

Relative Risk (Control)8
 

Lover Limit
 

Upper Limit
 

Weeka to Flrat Observed Tumor
 

Adrenal: Pheochromocytoma
 

P Values0 >d
 

Relative Rlak (Control)*
 

Lover Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Carcinoma*1
 

P Values0 >d
 

Relative Risk (Control)8
 

Lover Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

7/49(0.14)
 

N.S.
 

96
 

17/39(0.44)
 

N.S.
 

P - 0.027
 

78
 

3/49(0.06)
 

N.S.
 

110
 

2/45(0.04)
 

N.S.
 

110
 

4/49(0.08)
 

N.S.
 

0.571
 

0.130
 

2.096
 

104
 

26/44(0.59)
 

N.S.
 

1.356
 

0.852
 

2.188
 

94
 

2/49(0.04)
 

N.S.
 

0.667
 

0.058
 

5.564
 

106
 

2/42(0.05)
 

N.S.
 

1.071
 

0.081
 

14.190
 

98
 

4/48(0.08)
 

N.S.
 

0.583
 

0.133
 

2.137
 

87
 

14/42(0.33)
 

N.S.
 

0.765
 

0.411
 

1.416
 

85
 

1/47(0.02)
 

N.S.
 

0.348
 

0.007
 

4.144
 

74
 

1/46(0.02)
 

N.S.
 

0.489
 

0.009
 

9.060
 

106
 

23 
24 

TABLE 4
 

(CONCLUDED)
 

LOW HIGH
 

TOPOGRAPHY :HORPHOLOGY CONTROL DOSE DOSE
 

Mammary Gland: Pibroadenoma 12/49(0.24) 9/49(0.18) 10/48(0.21)
 

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

Relative Risk (Control)' 0.750 0.851
 

Lower Limit 0.308 0.364
 

Upper Limit 1.757 1.757
 l.SJB
 

Week* to Flrat Observed Tumor 103 86 73
 

Uterus: Endometrlal Stromal Polyp 6/46(0.13) 13/49(0.27) 7/47(0.15)
 

P Value«c>d N.S. N.S. N.S.
 

Relative Rlak (Control)8 2.034 1.142
 

Lower Limit 0.794 0.356
 

Upper Limit 5.984 3.807
 

Weeks to First Observed Tumor 78 94 78
 

T>osed groups received concentrations of 2.5 and 5.0 percent in feed.
 

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

Beneath the incidence of the control is the probability level for the Cochran-Armltage test for dose-related trend in proportions when it is below
 

0.05; otherwise N.S. - not significant. Departure from linear trend is noted when it is below 0.05 for any comparison. Beneath each dose group
 

Incidence is the probability level for the Fisher exact (conditional) test for the comparison of that dose group to the control group when it is
 

below 0.05, otherwise N.S. - not significant.
 

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

ilelatlve risk of the treated group versus the control group is shown along with the lower and upper limit of the 95Z confidence interval for that
 

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

diarylanilide yellow-dosed groups of either sex is included.
 

None of the statistical tests for rats of either sex indicated a
 

significant positive association between dosage and tumor incidence.
 

To provide additional insight, 95 percent confidence intervals
 

on the relative risk have been estimated and entered in the tables
 

based upon the observed tumor incidence rates. In many of the intervals shown in Tables 3 and 4, the value one is included; this indicates the absence of statistically significant results. It should
 

also be noted that many of the confidence intervals have an upper
 

limit greater than one, indicating the theoretical possibility of a
 

significantly increased rate of tumor incidence induced in rats by
 

diarylanilide yellow that could not be established under the conditions of this test.
 

25
 
IV. CHRONIC TESTING RESULTS: MICE 

A. Body Weights and Clinical Observations 

No difference s between body weight gain patterns of high dose 

groups and low dose groups were evident in male or female mice during 

the 78-week treatment period (Figure 3). The control animals for 

both sexes began to experience marked weight gain beginning in week 

36 when compared to ths treated mice. 

All the treated mice, both male and female, acquired a yellow 

discoloration of the hair coat during treatment. Because of the 

normal darker color of the B6C3F1 mice, the external appearance of 

the mice was not as strikingly affected as that of the rats, which 

are normally white. However, internal discoloration was as apparent 

in the mice as it was in the rats. 

B. Survival 

The estimated probabilities of survival for male and female mice
 

in the control and diarylanilide yellow-treated groups are shown in
 

Figure 4.
 

For both male and female mice the Tarone test did not detect a
 

statistically significant positive association between dosage and
 

mortality. In the male groups, 74 percent of the high dose, 88
 

percent of the low dose, and 84 percent of the control mice survived
 

until the end of the study, despite the sacrifice of five high dose
 

mice in week 78 and five control mi^e in week 79. In the female
 

groups 68 percent of the high dose, 86 percent of the low dose, and
 

26
 
TIME ON TEST (WEEKS) 

FIGURES 

GROWTH CURVES FOR DIARYLANILIDE YELLOW CHRONIC STUDY MICE 

27 
TIME ON TEST (WEEKS) 

FIGURE 4 

SURVIVAL COMPARISONS OF DIARYLANILIDE YELLOW CHRONIC STUDY MICE 

28 
80 percent of the control group survived until the end of the study,
 

despite the sacrifice of five high dose mice in week 78 and five
 

control mice in week 79.
 

In both sexes survival was adequate for meaningful statistical
 

analyses of tumor incidence.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are tabulated in
 

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

tabulated in Appendix D (Tables Dl and D2).
 

There appeared to be no dose- or sex-related increase in the
 

incidence of neoplasms or toxic changes in the treated versus the
 

control groups.
 

With a few exceptions, the same variety of neoplasms occurred
 

sporadically and at random in the chemically treated and control
 

groups. No particular organ or system seemed to be the target of
 

this chemical. Sporadic and unusual problems that occurred in the
 

treated but not in control animals were as follows: in the integumentary system, one mastocytoma affected the subcutaneous tissue of
 

a high dose female; one squamous-cell carcinoma of the ear affected
 

a low dose male; and one infiltrating duct carcinoma of the mammary
 

gland affected one low dose female.
 

The incidence and variety of nonneoplastic degenerative, proliferative, and inflammatory lesions were similar in control and
 

chemically treated mice.
 

29
 
The results of this histopathologic examination did not provide
 

evidence for the carcinogenicity of diarylanilide yellow in B6C3F1
 

mice.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 5 and 6. The analysis for every type
 

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

diarylanilide yellow-dosed groups of either sex is included.
 

None of the statistical tests for mice of either sex indicated a
 

significant positive association between the administration of diarylanilide yellow and an increased tumor incidence in B6C3F1 mice.
 

To provide additional insight, 95 percent confidence intervals
 

on the relative risk have been estimated and entered in the tables
 

based upon the observed tumor incidence rates. In many of the intervals shown in Tables 5 and 6, the value one is included; this indicates the absence of statistically significant results. It should
 

also be noted that many of the confidence intervals have an upper
 

limit greater than one, indicating the theoretical possibility of a
 

significantly increased rate of tumor incidence induced in mice by
 

diarylanilide yellow that could not be established under the conditions of this test.
 

30
 
31 

TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH DIARYLANILIDE YELLOW3
 

LjVSW
 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE DOSE
 

Lung: Alveolar/ Bronchlolar Adenoma or Carcinoma 7/47(0.15) 5/49(0.10) 4/49(0.08)
 

P Values0 >d N.S. N.S. N.S.
 

Relative Risk (Control)6 0.685 0.548
 

Lower Limit 0.184 0.125
 

Upper Limit 2.329 2.008
 

Weeks to First Observed Tumor 97 96 78
 

Hematopoletlc System: Leukemia or Malignant Lyraphoraa 1/50(0.02) 3/49(0.06) 3/49(0.06)
 

P Valuesc>d N.S. N.S. N.S.
 

Relative Risk (Control)6 3.061 3.061
 

Lower Limit 0.256 0.256
 

Upper Limit 157.400 157.400
 

Weeks to First Observed Tumor 97 96 97
 

Liver: Hepatocellular Carcinoma 15/49(0.31) 11/49(0.22) 4/46(0.09)
 

P ValuesC>d P • 0.006(N) N.S. P - 0.007(N)
 

Relative Risk (Control)6 0.733 0.284
 

Lower Limit 0.341 0.074
 

Upper Limit 1.528 0.814
 

Weeks to First Observed Tumor 94 96 86
 

^osed groups received concentrations of 2.5 and 5.0 percent in feed.
 

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

c

Bene«th the Incidence of the control Is the probability level for the Cochran-Armltage test for dose-related trend In proportions when It Is below
 

0.05; otherwise N.S. - not significant. Departure from linear trend is noted when It is below 0.05 for any comparison. Beneath each dose group
 

incidence is the probability level for the Fisher exact (conditional) test for the comparison of that dose group to the control group when It Is
 

below 0.05, otherwise N.S. - not significant.
 

''A negative trend (N) Indicates a lower Incidence In a treated group than in a control group.
 

"Relative risk of the treated group versus the control group Is shown along with the lower and upper limit of the 95* confidence interval for that
 

relative risk.
 
TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH DIARYLANILIDE YELLOW*
 

LOW HIGH 

TOPOGRAPHY :MOaPHO LOGY CONTROL DOSE DOSE

 

Lung: Alveolar/Bronchiolar Adenoma or Carcinoma 4/50(0.08) 3/49(0.06) 1/48(0.02)

 

P Values0'

d N.S. N.S. N.S.

 

Relative Risk (Control)6 0.765 0.260

 

Lower Limit 0.118 0.005 

Upper Limit 4.288 /. DUO

 

Weeks to First Observed Tumor 79 96 97 

Hematopoletic System: Leukemia or Malignant Lymphoma*" 6/50(0.12) 3/50(0.06) 6/50(0.12)

 

P Values0 >(J N.S. N.S. N.S.

 

% 

Relative Risk (Control)6 0.500 1.000

 

Lower Limit 0.085 0.287 

Upper Limit 2.200 3.489

 

Weeks Co First Observed Tumor 97 84 68 

Doaed group* received concentrations of 2.5 and 5.0 percent in feed.
 

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

°Beneath the incidence of the control is the probability level for the Cochran-Armitage test for dose-related trend in proportions when It is below
 

0.05; otherwise N.S. - not significant. Departure from linear trend is noted when it is below 0.05 for any comparison. Beneath each dose group
 

incidence is the probability level for the Fisher exact (conditional) test for the comparison of that dose group to the control group when it is
 

below 0.05, otherwise N.S. - not significant.
 

A negative trend (N) Indicates a lower incidence In a treated group than in a control group.
 

8

Relatlve risk of the treated group versus the control group is shown along with the lower and upper limit of the 95% confidence interval for that
 

relative risk.
 
V. DISCUSSION
 

Under the conditions of this bioassay, adequate numbers of
 

chemically treated rats and mice survived for meaningful statistical
 

analysis of the incidence of late-developing tumors. However, exposure to diarylanilide yellow did not result in a positive association
 

between dietary concentration and increased incidence of any tumor in
 

either species.
 

The high concentration administered to both species in the
 

chronic bioassay was the highest permissible as indicated by the
 

Guidelines for Carcinogen Bioassay in Small Rodents (Sontag et al.,
 

1976). These guidelines indicate that a dietary concentration
 

greater than I percent should not be administered except under
 

special circumstances (e.g., when the compound is a major component
 

of the human diet). As human exposure to diarylanilide yellow does
 

not warrant special exemption, the 5 percent limit applied. Dietary
 

administration of diarylanilide yellow had no significant effect on
 

survival or body weight gain in rats or mice of either sex. The only
 

clinical observation associated with chemical treatment was bright
 

yellow staining of the fur and mucosal surfaces in both species and
 

the only sign of toxicity observed during the histopathologic examination was basophilic cytoplasm changes in treated rats.
 

In rats, no treatment-related increase in the incidence of neoplasms, nonneaplastic lesions, or toxic effects was evident with the
 

exception of basophilic changes in hepatocyte cytoplasm in treated
 

33
 
males and females. There were, however, two unusual findings: metastatic chordoma in 1/49 low dose males, and an osteogenic sarcoma
 

in 1/49 low dose females.
 

In mice, no treatment-related increase in the incidence of neoplasms, nonneoplastic lesions, or toxic effects was evident. There
 

were, however, three unusual findings: squamous-cell carcinoma of
 

the ear in 1/49 low dose males, an infiltrating duct carcinoma of the
 

mammary gland in 1/50 low dose females, and a mastocytoma of the subcutaneous tissue in 1/50 high dose females.
 

The results of this bioassay did not provide evidence for the
 

carcinogenicity of diatylanilide yellow in Fischer 344 rats or B6C3F1
 

mice.
 

34
 
VI. BIBLIOGRAPHY
 

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

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

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

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

J. Wiley & Sons, New York, 1971.
 

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

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

Union Against Cancer, Geneva, 1969.
 

Chemical Abstracts Service, The Chemical Abstracts Service (CAS)
 

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

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

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

Co., Ltd., London, 1970.
 

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

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

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

Tests, Confidence Limits, and Adjustments for Stratification."
 

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

Hawley, G.G., The Condensed Chemical Dictionary, 8th edition.
 

Van Nostirand Reinhold Company, New York, 1971.
 

Hoover, R. and J. Fraumeni, "Cancer Mortality in U.S. Counties with
 

Chemical. Industries." Environmental Research 9:196-207, 1975.
 

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

Observations." Journal of the American Statistical Association
 

_53_: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
 

Researcti 7:230-248, 1974.
 

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

Co., New York, 1966.
 

Occupational Safety and Health Administration, "Occupational Safety
 

and Health Standards 1910.93c Carcinogens." Federal Register
 

J38_:974, 1973.
 

35
 
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, Third edition 4^3272­

3305, 1971. ~
 

Sontag, J.M., N.P. Page, and U. Saffiotti, Guidelines for Carcinogen
 

Bioassay in Small Rodents. Carcinogen Program, Division of
 

Cancer Cause and Prevention, National Cancer Institute, Bethesda,
 

Maryland, NCI-CG-TR-1. DREW Publication No. (NIH) 76-801,
 

February 1976.
 

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

62^679-682, 1975.
 

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

United States Production and Sales, 1975. USITC Publication
 

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

U.S. International Trade Commission, Imports of Benzenoid Chemicals
 

and Products, 1975. USITC Publication 806, U.S. Government
 

Printing Office, Washington, D.C., 1977b.
 

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

Cancer Institute, Bethesda, Maryland. Personal communication,
 

December 17, 1976.
 

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

36
 
Review of the Bioassay of Diarylanilide Yellow*
 

for Carcinogenicity
 

by the Date. Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

September 26, 1977
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976 under the authority of the
 

National Cancer Act of 1971 (P.L. 92-218). The purpose
 

of the Clearinghouse is to advise on the National Cancer
 

Institute's bioassay program to identify and 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
 

organic 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 NCI bioassay reports on chemicals studied for
 

carcinogenicity. In this context, below is the edited
 

excerpt from tne minutes of the Subgroup's meeting at
 

which Diarylanilide Yellow was reviewed.
 

The primary reviewer said that the compound is used
 

as a dye coatiig for yellow lead pencils. It could be a
 

public health concern from the standpoint of people ingesting
 

the dye by chewing on their pencils. Diarylanilide Yellow
 

belongs to the chemical class of diazobenzidines. Some
 

members of this class are reduced by hepatic enzymes to
 

free amines which may be carcinogenic. It was noted that
 

certain bladder carcinogens were not identified until they
 

were tested in appropriate animal models.
 

The primary reviewer said that the conclusion drawn
 

in the bioassay report was that the study did not provide
 

evidence for the carcinogenicity of Diarylanilide Yellow
 

in either rats or mice. He pointed out, however, that the
 

incidence of pituitary chromophobe adenomas in the treated
 

*	 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.
 

37
 
rats was statistically significant when compared to the
 

controls. A staff pathologist commented that in this
 

particular laboratory, the pituitary tumors were sub­ classified. If they were considered simply as pituitary
 

adenomas NOS, or had the control pituitary adenomas been
 

sub-classified, they would not have been statistically
 

significant.
 

The primary reviewer also noted a finding of a single
 

squamous-cell ear carcinoma in a mouse and that this lesion
 

was unreported among the historical control animals. In
 

addition, he commented on a number of other "odd tumors"
 

found in the treated animals. The primary reviewer was
 

critical of the report for not pointing out these tumors
 

in the treated animals since it could mislead readers to
 

believe that there should be no concern about the dye.
 Another Subgroup member noted a significant increase in
 

the incidence of leukemias and lymphomas in the treated
 

male rats, as well as a decrease in the incidence of
 

hepatocellular carcinomas in the treated male mice. He
 

said that consideration should be given to this phenomenon
 

in evaluating the biological potential of the test compound.
 Another Subgroup member commented that a survival analysis
 

would be necessary to determine whether there was a true
 

reduction in tumor incidence among the treated animals.
 

A motion was, made that Diarylanilide Yellow was not
 

carcinogenic under the conditions of test. It was further
 

moved that metabolism studies be done to determine if the
 

compound is reduced to a free amine and if so, consideration
 

be given to a retest in an animal model appropriate for
 

studying bladder carcinogenesis. The motion was seconded
 

and accepted unanimously.
 

Members present were:
 

Gerald N. Wogan (Chairman), Massachusetts Institute of
 

Technology
 

Arnold L. Brown, Mayo Clinic
 

Lawrence Garfinkel, American Cancer Society
 

Joseph H. Highland, Environmental Defense Fund
 

George Roush, Jr., Monsanto Company
 

Sheldon Samuels, Industrial Union Department, AFL-CIO
 

Michael Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

John Weisburger, American Health Foundation
 

Sidney Wolfe, Health Research Group
 

38
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH DIARYLANILIDE YELLOW
 

TABLE Al

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH DIARYLANILIDE YELLOW 

.
 CONTROL(ONTR) LOS DOSE HIGH DOSE
 

01-0160 01-0195 01-0200
 

ANIMALS INITIALLY IN STUDY 50 50 50
 ANIMALS NECBCPSIED 50 50 50
 ANIMALS EXAMINED HISTOFATHOLOGICALLY ** 50 K9 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (50) (SO) (50)
 BASAL-CELL CARCINOMA 1 (2%)
 FIBROMA 2 («%) U (8»)
 

*SUBCUT TISSUE (50) (50) (50)
 

UNCIFFEBENTIA1ED CARCINOMA 1 (2%)
 SAECOBA, NOS 1 (2%)
 

FIBROMA 1 (2*)
 FIBECSARCOMA 1 (2S)
 

RESflBATOBY SYSTEM
 

tLUNG <!|9) (19) (50)
 

ALVEOLAE/BBONCHIOLAK ADFNOMA 1 (2*) 2 I"**)
 

ALVEOLAR/BSONCHIOLAE CARCINOMA 1 (2%) 1 (2*)
 

CHOELORA «£TA STATIC 1 <2X)
 

HEMATOPOIETIC SYSTEM
 

•MULTIPLE	 OEGANS (50) (50) (50)
 

LEUKEMIA, NOS 1 (2%)
 

HYELCMONOCYTi: LEUKEMIA 9 (18*) 1 (2J)
 

tSPLEEN (50) (U9) (U8)
 

ANGIOSABCOMA 1 (2*)
 

MYELOMONOCYTIC LEUKEMIA 1 <2<)
 

tKESENTERIC L. NODE (U9) (Ub) (16)
 

MALIG.LYNPHOFA. LYMPHOCYTIC TYPE 1 (2%)
 

CIBCULATOEY SYST!M
 

•HEART	 (U8) C«9) (50)
 , ,. . „,. ,.^_ _____„__
 SARCOjJAi_NOSJ._BET_A^TiU£ _ 1 12il_ _
 

* NUMEZ R OF ANIMI.LS KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

"EXCLUDES PARTIALLV AUTOLYZED ANIMALS 

A-3 
TABLE Al (CONTINUED) 

CONTROL(ONTB) LOW DOSE HIGH DOSE 01-0160 01-0195 01-0200 

DIGESTIVE SYSTEM 

tLIVER («9) («9) (50) 

HEPATOCEILULAR CARCINOMA 1 (2S) 

UHINABY SYSTEM 

•KIDNEY (50) («8) (50) 

SARCOMA, NOS 1 (2*) 

HEPHROBLASTOMA 1 (2%) 

EHDOCBINE SYSTEM 

f PITUITARY (45) C«3) («5) 

ADENOMA, NOS 

CHROMOPHOBE ADENOMA 5 (11X) 

2 («%) 12 (28%) 5 (11*) 

tADRENAL (50) (HI) (l»9) 

PHEOCHROBOCYTOHA 3 (6%) 3 (6X) 5 (10S) 

tTHYROIC (37) C*7) (U8) 

FOLLICULAfi-CELL CARCINOMA 

C-CELL ADENOMA 

C-CE1L CARCINOMA 

1

1

2

 (3%) 

 (3S) 

 (5S) 

3 <6X) 

2 (US) 

1 (2S) 

•PANCREATIC ISLETS (1*7) (U7) («6) 

ISLET-CELL ADENOMA 1 (2%) 2 (UX) 5 (11%) 

BEPBODUCTIVE SYSTEM 

•MAMMARY GLAND (50) (50) (50) 

ADENOCARCINOtlA, NOS 1 (2X) 

FIBRCADENOMA 1 (2%) 

*PREPUTIAL GLAND (50) (50) (50) 

CARCINOMA. NOS 2 (MS) 

SQUAMOOS CELL CARCINOMA 1 (2%) 

ADENOMA, NOS 1 (2%) 

*TESTIS (50) C*8) («9) 

INTEESTITIAL-CELL_TDMOR l»2 JBttXl «U I92SJ 32 J80X) 

t SOMBER OF ANIMAL S SITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

A-4 
TABLE Al (CONTINUED) 

CONTROL (ONTB) LOU DOSE HIGH DOSE 

01-0160 01-0195 01-0200 

BERVCOS SYSTEM 

*BRAIN (SO) (l»7) (U9) 

GLIOMA. NOS 1 (2») 

ICEEEB8AL CCHTEX (50) («7) (<t9) 

GLIOMA. NOS 1 (2») 

SPECIAL SENSE ORGANS 

NONE 

BUSCDLOSKELETAL S (STEM 

NCNE 

BODY CAVITIES 

*PERITONEUK (50) (50) (50) 

MESOTHELIOMA, MALIGNANT 1 (2*) 

ALL OTHEE SYSTEMS 

•MULTIPLE ORGAN S (50) (50) (50) 

MESOTHELIOMA, NOS 1 (2X) 

ANIMAL BISECSITICN SUMMARY 

ANIMALS INITIAILY IN STUDY 50 50 50 

NATURAL EEATUi 5 6 6 

MORIBUND SACHIFICE 8 2 2 

SCHEDULED SACRIFICE 5 5 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 32 U2 37 

ANIMAL BISSII.G 

2 INCLUDES AUTOLCZEfi ANIfiiiS _ _ 

* NUMEE B O F ANIMU S HITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMHL S NECROPSIED 

A-5 
TABLE Al (CONCLUDED) 

CONTROL (ONTR) LOH DOSE HIGH DOSE
 01-0160 01-0195 01-0200
 

TUECE SUMMAP Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS* U9 U6 U3
 

TOTAL PRIMARY TUMOflS 76 77 68
 

TOTAL ANIMALS KITH BENIGN TUMORS U6 1*6 39
 

TOTAL BENIGN TUMORS 57 69 59
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 16 7 8
 

TOTAL HALIGNANT TUMORS 19 8 8
 

TOTAL ANIHALS WITH SECONDARY TUMOEb* 1 1
 

TOTAL SECONDARY TUMORS 1 1
 _
 

TOTAL ANIMAL? WITH TUKORS UNCERTAIN
 

BENIGN OR MALIGNANT 1
 

TOTAL UNCERTAIN TUMORS 1
 

_
 

TOTAL ANIMAL S WITH TUMORS UNCERTAIN
 FFIMAR1 CE METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT 5"CONDARY TDHORS
 * SECONDARY TUMORS: METASTATIC TUMOFS OR TUNORS INVASIVE INTO AN ADJACENT ORGAN
 

A-6 
TABLE A2 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH DIARYLANILIDE YELLOW 

HIGH DOSE
 

02-0200
 

50
 

i»8
 

U8
 

(«8)
 

(«8)
 

(48)
 

1 (2X)
 

(48)
 

1 (2»)
 

1 (2X)
 

(48)
 

2 C*X)
 

(48)
 

2 (4X)
 

.

.._-__ -.-- ____^-_ ._
 

ANIMALS INITIALLY IN STUDY
 

ANIHALS KISSING
 

ANIMALS NECROPStED
 

ANIMALS EXAMINED HISTOPATBOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

•SKIN
 

EPITHELIAL TUMOR, NOS, BENIGN
 

*SUECUT TISSUE
 

FIBBOMA
 

RESFIEATORY SYSTEM
 

tLUNG
 

ALVECLAK/BIIONCHIOLAR CARCINOMA
 

PHEOCHEOHOCYTOHA, HETASTATIC
 

HEMATOEOIETIC SYSTEM
 

*MDITIPLE ORGANS
 

LEUKEMIA, NOS
 

CYELCMONGCfTIC LEUKEMIA
 

ISPLEEN
 

HYELCMONOCfTIC LEOKFMIA
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

iLIVER
 

NEQFLASTIC NODULE
 

HEPATgCELIULAB CARCINOMA
 

CONTROL (UNTH)


C2-0160


50


1
 

49


** 49


(49)


1 12X)
 

(149)


2 («*)
 

(l»9)


(<*9)


1 (2%)
 

6 (12X)


(U7)


C»8)


1 12S1_


 LOB DOSE
 

 02-0195
 

 50
 

 49
 

 49
 

 (49)
 

 (U9)
 

 (1»8)
 

1 (2X)
 

 (U9)
 

4 (8%)
 

 (49)
 

 (U9)
 

t KUKEE E OF AMMALS HITR TISSUE EXAMINE E MICROSCOPICALLY 

* NUMBE R OF AMBALS NECFOPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMAI.S 

A-7 
HIGH DOSE
 

02-0200
 

(48)
 

1 (2%)
 

(42)
 

14	 (33%)
 

(47)
 

1 (2%)
 

(46)
 

1 (2«)
 

1 (2%)
 

(47)
 

2 (4%)
 

(48)
 

2 (4%)
 

1 (2%)
 

10 (21%)
 

(48)
 

1 (2%)
 

(47)
 

7 (15%)
 

1 (2%)
 

(47)
 

1 (2%)
 

(48)
 

1 (2%)
 

_	 _ i_12Sj_
 

URINARY SYSTEM
 

IKIDNEY
 

TUEULAR-CHLL ADENOCARCINOM A
 

ENDOCRINE SYSTFM
 

tPITUITARY
 

ADENOMA. NOS
 

CHFOBOPHOBE ADENOMA
 

IADRENAL
 

FHEOCHPOMOCY10KA
 

PHEOCHBOMOCYTOHA, MAIIGNANT
 

tTHYROID
 

FOLUCULAR-CELL CARCINOM A
 

C-CELL CARCINOMA
 

•PANCREATIC	 ISLETS
 

ISIET-CELL ADENOMA
 

HEERCDUCTIVE SYSTEH
 

•MAMMARY	 GLAND
 

ADENOMA, NOS
 

ACZNCCAtCINOKA, NOS
 

PAPILLARY ADENOCAKCI <OMA
 

FIBRCADENOMA
 

•CLITORAL GLAND
 

ADENOMA, NOS
 

tDTERUS
 

ENDOMETKIAL STRCMAL POLYP
 

ENEOHETPIAL STROMAL SARCOMA
 

tCERVIX tITERI
 FIBROSAfiCOMA
 

tOVARY
 

1HECOMA
 

__SBAlLJll°SArCE.L.L_Tafl£R
 

t NUKBE R OF ANIMAL S WITH TISSUE

* NUMBER OF ANIMALS NECPOPSIED 

TABLE A2 (CONTINUED) 

CONT80L(UNTR)
 

02-0160
 

(48)
 

(39)
 

15 (38%)
 

2 (5S)
 

(U9)
 

3 ((,%)
 

(U5)
 

2 (4%)
 

(49)
 

C*9)
 

12 (2«X)
 

(U9)
 

1 (2%)
 

C»6)
 

6 (13%)
 

(U6)
 

(l»7)
 

LOB DOSE
 

02-0195
 

(«9)
 

(44)
 

26 (59%)
 

(49)
 

1 (2X)
 

1 (2%)
 

(«2)
 

2 (5*)
 

(«6)
 

1 (2J)
 

(49)
 

1 (2S)
 

9 (18%)
 

(»9)
 

(49)
 

13 (27*)
 

(«9)
 

(47)
 

 EXAMINE D MICROSCOPICALLY 

A-8 
GRANULOSA-CILL CA8CINOHA
 

HEBVCOS SYSTEM
 

tEBAIN
 

ASTBCCYTOHA
 

SPECIAL SENSE OIGANS
 

•ZYHBAL'S GLANt
 

CERUHINOUS CARCINOMA
 

HDSCULOSKELETAL SYSTEM
 

•BONE
 

CSTECSARCOMA
 

BODY CAVITIES
 

*EODY CAVITIES
 

HESOTHELIOMJ , HALIGNANT
 

•ABDOMINAL	 CAVJTY
 

SABCOHA, N05
 

ALL CTHEP SYSTEI.S
 

NCNE
 

ANIHAL DISPOSITION SDHMABY
 

ANIMALS INITIALLY IN STUDY
 NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TEEBINAL SACRIFICE
 

ANIMAL MISSING
 

8_IN£IUDES_AUTSI1Y2ID_MI!1ALS
 

TABLE A2 (CONTINUED) 

CONTROL (ONTB) LOH DOSE HIGH DOSE 

02-0160 02-0195 ( ->0200 

1 (2«) 

(»9) (<»8) («8) 

1 (2%) 

(U9) (49) (48) 

1 (2*) 

(19) C»9) (48) 

1 (2*) 

(U9) (49) (48) 

1 (2*) 

(49) (49) (48) 

1 (2%) 

50 50 50 

2 6 6 

6 4 6 

5 5 

36 40 33 

1 

* NDMBEB OF ANIMALS SITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANItlALS NECROPSIED
 

A-9
 
TABLE A2 (CONCLUDED)
 

TUHCB SUMMAR Y
 

TOTAL ANIMALS BITH PRIMARY TUMORS*


TOTAL PRIMAR Y TUMORS


TOTAL ANIMALS WITH BENICN TUMORS


TCTAL BENIGN TDHOBS


TOTAL ANIMALS WITH MALIGNANT TUHORS


TOTAL MALIGNANT TUMOSIJ


TOTAL ANIMALS WITH SECONDARY TUMORS*
 

TOTAL SECONDARY TUKOR >
 

TOTAL ANIMALS WITH TOMO *S UNCERTAIN­ BENIGN OF MALIGNANT
 

TOTAL UNCERTAIN TUBOR3
 

TOTAL ANIMALS WITH TUMORS UNCZBTAINFEIMAHY CB METASTATIC
 

TOTAL UNCERTAIN TUMOR 3
 

CONTROL (UNTH)
 

02-0160
 

 32
 

 54
 

 28
 

 42
 

 10
 

 12
 

LOW DOSE HIGH DOSE
 

02-0195 02-0200
 

UO 34
 

59 55
 

36 26
 

50 38
 

8 13
 

9 14
 

1
 

1
 

3
 

3
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUHORS
 * SECONDARY TUMORS: METASTATIC TUMORS OR TUHORS INVASIVE INTO AN ADJACENT ORGAH
 

A-IO
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH DIARYLANILIDE YELLOW
 

TABLEB1

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH D1ARYLANILIDE YELLOW 

.. _
 

AHIHAIS INITIJILY IK STUDY
 

ANIMALS MISSI1G
 ANIMALS NECBCrSIIE
 

ANIMALS EXAMI1ED HISTOFATHCLOGICALLY


INTEGUMENTARY SYSTEM
 

*SKIN
 

SQUAMOUS CELL CARCINOMA
 

*SUECUT TISSIE
 

HEMANGIOSJ1SCCMA
 

HESPIKATORY SYSTEM
 

tLUNG
 

HEPATOCELIULAR CARCINOMA, METAST
 

ALVEOLAR/ERONCHIOLAR ADENOMA
 

AIVEOLAE/EHONCHIOLAB CARCINOMA
 

HEHATOFOIETIC SYSTEM
 

*KULTIPLE ORGANS
 

MALIGNANT LYMPHOMA, NOS
 

MALIG.LYMEHOMA, LYMPHOCYTIC TYPE
 

HALIG.LYKEHOMA. HISTIOCYTIC TYPE
 

*SPLEEN
 

HEMANGIOSARCOMA
 

ANGIOSARCCMA
 

*BESENTERIC 1. NODE
 

MALIG.LYMEHOMA, HISTIOCYTIC TYPE
 

tPEYERS PATCH
 

HALIG. LYKEHOHA, HISTIOCYTIC TYPE
 

CIBCQLATORY SYSTEM
 

NCNE
 

CONTROL(DNTR)
 

05-0160
 

50
 

50
 

** U9
 

(50)
 

(50)
 

(47)
 

2 (US)
 

t (9*)
 

3 (6X)
 

(50)
 

1 (2*)
 

(«9)
 

1 (2»)
 

(«0)
 

(«9)
 

LOB DOSE HIGH DOSE
 05-0195 05-0200
 

50 50
 

1
 

«9 49
 

49 49
 

(49)	 (49)
 

1 (2X)
 

(49)	 (49)
 

1 (2«)
 

(«9) (49)
 

1 (2%) 1 (2%)
 

4 (8X) 3 <6X)
 

(49) (49)
 

1 (2%) 1 (2%)
 

1 (2%) 1 (2%)
 

(4b) (47)
 

1 (2»)
 

1 (2*) 1 (2%)
 

(35)	 (43)
 

1 (3X)
 

(48)	 (49)
 

1 (2%)
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D fllCBOSCOPICALLY 

* KUKBE R OF AMPULS NECROPSIE D 

**EXCLUDES PARTI UiY AUTOLYZED ANIMALS 

B-3 
TABLE Bl (CONTINUED) 

CONTROL (DNTR)

05-0160

 LOW DOSE

 05-0195

 HIGH DOSE 

 05-0200 

DIGPSIIVF SYSTEM 

• LIVER

HEFATOCELLULAB CARCIN01A

HEMANGIOSABCCMA

ANGIOSARCOMA

•STCMACH

ADENOMATOUS FOLYP, NOS

 (U9)

 15 (31*)

 (49)

 1 (2%) 

 (49)

 11 (22%)

 1 (2*) 

 (47)

 C*6) 

4 (9X) 

1 (2X) 

 (46) 

URINARY SYSTEM 

NCNE 

ENDOCRINE SYSTEM 

• PITUITARY

CHEOMOFHOBE ADENOMA

• ADRENAL

FHECCHROMOCYTOMA

• THYROID

FOLLICULAfc-CELL CARCI1IOMA

 (42)

 (47)

 (42)

 (36)

 2 (6X) 

 (45)

 (34)

 2 (6%) 

 (40) 

 (45) 

1 (2%) 

 (43) 

REFFODUCTIVE SYSTEM 

• TESTIS

INTERSTIIIAL-CELL TUBOR

SEBINOMA/DYSGERMINOMA

 (49)

 1 (2%) 

 (49) (48) 

1 (2%) 

NEFVCUS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

NONE 

BUSCULCSKELETAL SYSTEH 

NCNE _ _ 

* NUMEEK OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECRCPSIED 

B-4
 
TABLE Bl (CONCLUDED)
 

CONTROL (ONTB) LOB DOSE HIGH COSE
 05-0160 05-0195 05-0200
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISECSITICN SUMMARY
 

ANIMALS INITIALLY I!i STUDY 5C 50 50
 

NATURA L DEATHS 3 5 5
 

MORIBUND SACRIFICE 1 2
 

SCHEDDLED SACRIFICE 5 5
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE 42 UU 37
 

ANIBAL MISSING ' 1
 

3 INCLUDES AUTOLYZED ANIMALS
 

TDeCR SUMMAF V
 

TOTAL ANIMALS WITH PRIMARY TDMOSS* 20 21 13
 

TOTAL PRIMAR Y TUMORS 26 26 17
 

TOTAL ANIMSLS WITH BENIGN TOMOES b 3 1
 

TOTAL BEMGN TUMORS 6 j 2
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 18 20 12
 

TOTAL MAIIGNANT TUMORS 20 21 15
 

TOTAL ANIMJLS WITH SECONDARY TUMORS* 2
 

TOTAL SECONDARY TUMORS 2
 

TOTAL ANIM;LS WITH TUMORS UNCERTAIN­ BENIGN OB PALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMAL S WITH TUMORS UNCERTAIN­ ERIMARY OR MITASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUflORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

f SECONDARY TUMORS: METASTATIC TUMORS OF TUMORS INVASIVE INTO A I. ADJACENT ORGAN
 

B-5
 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH DIARYLANILIDE YELLOW 

*__
 

CONTROL (UNTH) LOW DOSE HIGH DOSE
 

06-0160 06-0195 06-0200
 

ANIMALS INITIALLY IN STDCf 50 50 50
 

ANIMALS NECROESIED 50 50 50
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY ** 50 «9 US
 

INlfGUHENTARY SYSTEH
 

*St)ECDT TISSUE (50) (50) (50)
 

PIBRCSARCOMA 1 (2%)
 

HEKANGIOSARCOMA 1 (2%)
 

RESPIRATORY SYSTEH
 

»IUNG (50) C»9) (48)
 

HEPATOCELLULAR CARCINOHA. SETAST 1 (2S)
 

ALVEOLAR/BKONCHIOLAR ADENOMA 1 (2%) 1 (2*) 1 (2%)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 3 (6») 2 (a*)
 

HEHATOEOIETIC SYSTEM
 

*HULTIPLE CFGANS (50) (50) (50)
 

MALIGNANT LYMPBOHA, NOS 3 (6S) 1 (2%)
 

HALIG.LYBPHOMA, ONEIFPER -TYPE 1 (2%)
 

HALIG.LYMPHOHA, LYHPHOCYTIC TYPE 2 ("«%)
 

BALIG.LYHEHOMA. HISTIOCYTIC TYPE 1 (2%) 1 (2S)
 

UNDIFFERENTIATED LEUKEMIA 1 (2*)
 

•SUBCUTANEOUS TISSUE (50) (50) (50)
 

HAST-CELL TUMOB 1 (2X)
 

ISPLEEN («9) (l»9) (47)
 

HEMANGIOSARCOMA 1 (2X)
 

MALIG.LYHPHOMA, HISTJOCYTIC TYPE 1 (2X)
 

tHANDIBULAR L. NODE (40) (36) (45)
 

MALIGNANT LYHPHOHA, IIOS 1 (3%)
 

*MESENTERIC L. NODE (aoj (36) (45)
 

BiiISSiNI_IXi3PfloaAJ._!HXEp_TiE£ 1_L2SI_


* NUMBE R OF ANIMALS BITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZID ANIMALS 

­

B-6 
TABLE B2 (CONTINUED) 

CONTROL (ONTR)

06-0160

 L0¥ DOSE

 06-0195

 HIGH DOSE 

 06-0200 

IEFYERS PATCU

MALIGNANT LY HPHOHA, NOS

tJIJDHOK

MAHG. LYMPHO3A, HISTIOCYTIC TYPE

 <«9)

 1 (2*) 

 («9)

 <«6)

 («6)

 1 (2%) 

 (l»7) 

 (U7) 

CIHCDLATCKY SYSTEM 

NCNE 

DIGESTIVE SYSTEM 

tLIVER

HEPATOCELLOL\B CARCINOMA

 (149)

 2 («%) 

 («7) (U6) 

ORINABY SYSTEM 

NONE 

ENDOCRINE SYSTEM 

*PITUITARY

ADENOMA, NOS

CHROKOPHOBE ADENOMA

• ADRENAL

EHEOCHROMOCYIOMA

 (U2)

 (a7)

 (39)

 1 (3X) 

 (1»6)

 1 (2X) 

 (l*U) 

1 (2«) 

(US) 

REPBCDOCTIVE SYSTEM 

*BAMMARY GLAND

INFILTRATING DUCT CARCINOMA

IOTEROS

ENDOBETRIAL STROMAL SARCOMA

 (50)

 («9)

 (50)

 1 (2%) 

 (17)

 1 (2«) 

 (50) 

 (U6) 

NEEVOOS SYSTEM 

NCNE 

SPECIAL SENSE ORGANS 

NONE

t NUMBE R

* NHMBE E

 OF

 OF

 ANIMUL S

 ANIMAL S

 HITH TISSUE

 NECROPSIED 

 EXAMINE D

 .

 MICROSCOPICALLY 

_ _ . . 

B-7
 
TABLE B2 (CONCLUDED)
 

*CONTROL (UNT 

06-0160 

B) LOU DOS3

06-0195

 HIGH DOSE 

 06-0200 

BUSCDLOSKEiriAL SYSTEH 

NONE 

BODY CAVITIES 

NONE 

ALL CTHEP SYSTEMS 

NONE 

ANIMAL EISEOSITICN SUMHAEY 

ANIKALS INITIALLY IN STUDY 

NATUFAL EEATHd 

MORIBUND SACRIFICE 

SCHECULEE SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACBIFICE 

ANIMAL MISSING 

50 

3 

2 

5 

HO 

50

6

1

U3

 50 

8 

3 

5 

 31* 

3 INCLUDES AUTCLYZED ANIMALS 

TUMOR SUMMAR Y 

TOTAL ANIMALS KITH PRII1ABY TUMCBS* 

TOTAL PEIMABY TUMOBS 

12 

15 

10

10

 9 

9 

TOTAL ANIMALS WITH BENIGN TUMORS 

TOTAL BENIGN TDMORS 

1 

1 

3

3

 2 

2 

TOTAL ANIMALS WITH MALIGNANT TUMORS 

TOTAL MALIGNANT TUMORS 

11 

1H 

7

7

 6 

6 

TOTAL ANIMALS HITH SEC3NDABY TUKORS* 

TOTAL SECONDARY TDHOBS 

1 

1 

TOTAL ANIMALS SITH TUM3HS UNCERTAINBENIGN OR MALIGNANT 

TOTAL UNCERTAIN TUMORS 

1 

1 

TOTAL ANIMALS KITH TUMORS UNCERTAIN­ FFIMABY CE MITASTATIC 

TOTAL UNCERTAIN TUMORS 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUHORS
 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUHORS ISVASIVE INTO AN ADJACENT ORGAN
 

B-8
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH DIARYLANILIDE YELLOW
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH DIARYLANILIDE YELLOW

 

* NOMEER OF ANIMAL S WITH TISSOE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED AKIMALS 

C-3 
TABLECl (CONTINUED)
 

... ...
 CONTHOL(ONTE) LOW DOSE
 

01-0160 01-0195
 

tHESENTERIC L. NODE (149) (t6)
 

HYPESPLASIA, PLASHA CELL 1 (2%)
 

CIRCULATORY SYSTEM
 

#HEART It8) (149)
 

PEPIARTERITIS
 

•MYOCARDIUM	 (<*U) (149)
 

INFLAMMATION, FOCAL 1 (2%)
 

INFLAMMATION, INTERSTITIAL 1 (2%)
 

FIBRCSIS 2 (UX)
 

DEGENERATION, NOS 1 (2%)
 

DIGESTIVE SYSTEM 

tSALIVARY GLAND (50) (147) 

HYPEFPLASIA, INTRADUOTAL 1 (2») 

*LIVER (149) (149) 

INFLAMMATION, CHRONi: FOCAL 1 (2S) 

NECROSIS, FOCAL 3 (6%) 

NECROSIS, DIFFUSE 

NECROSIS, HEMORRHAGi: METAMORPHOSIS FATTY 1 (2%) 

BASOPHILIC CYTO CHAN3E 5 (10%) 

ECSINOPHIL1C CYTO CHANGE 1 <2X) 

HYFEFPLASIA, FOCAL 1 <2X) 

tLIVIP/CENTRHOBULAR (49) (149) 

CONGESTION, PASSIVE 1 (2%) 

»BILE DUCT 

HYPEFPLASIA, NOS 

(50) 

2 C4%) 

(50) 

3 (6*) 

*PANCREAS (U7) (147) 

INFLAMMATION, NOS 

INFLAMMATION, CKRONIC 

2 («%) 

i I1**) 

INFLAMMATION, CHRONIC FOCAL 1 (2%) 

tSTCMACh 

HYFEEKEBATOSIS 

ACANTHOSIS ._

(49) 

1 (2%) 

_ 1.J2J1 ... 

(148) 

* NUMBIR OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECFOPSIED
 

HIGH DOSE
 

01-0200
 

(46)
 

(50)
 

2 (<4%)
 

(50)
 

(47)
 

(50)
 

1 (2%)
 

2 («%)
 

11 (22%)
 

(50)
 

(50)
 

(146)
 

2 (14%)
 

(48)
 

C-4
 
TABLE Cl (CONTINUED)
 

"" *
 

CONTROL (DNTR) LOB DOSE
 01-0160 01-0195
 

tGASTRIC MUCOSA <U9) (U8)
 

NECEOSIS, F3CAL 1 (2*)
 

iEEYERS PATCH (1*9) C»8)
 

HYPERPLAEIA, NOS 1 (2%)
 

ICOtON C*8) (U8)
 

NEMATODIASIS 2 (««)
 

OKINARY SYSTEM 

tKIDNEY (50) (U8) 

CYST, NOS 1 (2X) 

CONGESTION, NOS 1 (2%) 

GLCMEBULCNEPHRITIS. NOS 14 (8%) 

INFLAMMATION, CHRONIC 

GLOHERULCNEPHRITIS, CHRONIC 

1 (2S) 

5 (10%) 

NEEHROPAIHY 3 (6%) 

NEPHROSIS, NOS 35 (70*) 

ENECCRINE SYSTEM
 

#EITDITARY (us) <U3)
 

CYST, NOS 1 (2*)
 

CONGESTION, NOS 1 (2%)
 

HYPEPPLASIA, CHROMOPH03E-CELL 1 (2*)
 

ANGIECTASIS 1 (21.)
 

IADRENAL CORTEX (50) (U7)
 

HYPERPLAEIA, FOCAL 1 (2J)
 

•ADRENAL	 MEDULLA (50) (U7)
 

HYPERPLASIA, NOS
 

HYPERPLASIA, FOCAL 1 (2%)
 

ITHYROIE (37) (U7)
 

CYSTIC FOLLICLES 1 (3S)
 

HYPEKPLASIA, FOCAL
 

HYPERPLAEIA, C-CELL 2 (5X) 2 (U%)
 

tPANCREATIC ISLETS (U7) (U7)
 

HYPERPLASIA, NOS
 

IIPEfiEiASIA^.FOCAi 1_1211_
 — — —
 t NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 <M-0200
 

(U8)
 

(U9)
 

(47)
 

3 (6%)
 

(50)
 

9 (18%)
 

(45)
 

2 (U%)
 

(U9)
 

(U9)
 

1 (2X)
 

(48)
 

1 (2%)
 

1 (2%)
 

(U6)
 

1 (2%)
 „____„ —
 

C-5
 
TABLE Cl (CONTINUED) 

CONTROL (CNTB)

01-0160

 LOB DOSE 01-0195 HIGH DOSE 01-0200 

REPRODUCTIVE SYSTEM 

•PREPUTIAL GLAND ABSCFSS, NOS (50) (50) (50) 

1 (2%) 

*EECSTATS

INFLAMMATION, NOS

INFLAMMATION, ACUTE INFLAMMATION, ACUTE FOCAL

 (1*8)

 3 (6X) 

 (1*7)

 3 (6%)

 8 (17%)

 (U8) 

6 (13X) 

 11 (23X) 

tTESTIS

HYDBOCELE FEEIVASCULITIS

CIGENERATION, N0£

CALCIFICATION, NC'S

CALCIFICATION, FCiCAL

ATEOPHK, NOS

HYPEEPLASIA, INTKBSTITIA L CELL

 (50)

 1 (2X) 

3 (6X) 

1 (2X) 

 11 (22X) 

4 (8X)

 (U8)

 15 (31%) 

1 (2X)

 (<*9) 

1 (2X) 

2 (UX) 

•EPIDIDYMIS

GHANULOMA, NOS

 (50) (50) (50) 

1 (2X) 

NEKVCUS SYSTEM 

NCNE 

SPECIAL SENSE ORGANS 

NCNE 

MOSCOLOSK2LETAL SYSTEM 

NCNE 

BOCY CAVITIES 

•PLEURA

FIBROSIS, DIFFUSE

 (50)

 1 (2%) 

 (50) (50) 

ALL C1HER SYSTEMS 

CMENTDM S1EATITIS _ __ _ _ 1 _ __ 

* NUMEEH OF UNINAL S WITH TISSUE EXAMINED

* NUMBEE OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

C-6 
TABLE Cl (CONCLUDED) 

TM"TM TM»*"~TM~— «­*— » W IJ •TM _IL LL I—

KECBCSIS, FM 

 Ul n _. IT -I- ^•••B 

CONTROL (DNTB)

01-0160

 LOB DOSE

 01-0195

1 

 HIGH DOSE 

 01-0200 

SPECIAL (tOBPHOL'JGI SUMHARY 

SC LESION ECFOETED ADTO/NECROPSlf/HISTO PERF AUTO/BECROP3Y/NO HISTO 

1

1 

1 

1 

C-7 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH DIARYLANILIDE YELLOW

 

ANIKAtS INITIALLY IN STODY
 

ANIHALS MISSING
 

ASIBALS NECROPSIED
 

ANIBALS EXAMINED HISTOFATHOLOGICALLY
 

INTEGUBENTAR't SYSTEM
 

NONE
 

RESPIRATORY 5YSTEH
 

ILUNG/BBONCHDS
 

EHCNCHIECTASIS
 

ILUNG
 

CONGESTION, ACUTE PASSIVE
 

INFLAMMATION, FOCAL GR ANHLOMATOU
 

HEBATOPOIET1C SYSTEM
 

*SPLEEN
 

HEflOSIDIJROSIS
 

HYPEEPLKSIA, HEBATOPOIETIC
 

HYFEBPLASIf, EBYTHEOID
 

CIRCULATORY SYSTEK
 

* HEART
 

PERIARTERITIS
 

*MYCCARtIU«
 

FIBROSIS
 

» AORTA
 

PERIAR1ERITIS
 

DIGESTIVE !iYSTEM
 

* LIVER
 

Ii!FLAM.3ATIONi_ACUTE/C.HfiONI£
 

» NUBBE R 0 ? ANIMAL S WITH TISSUE

* NUBBE R OP ANIBAL S NECROPSIE D 

**EXCLUDES IARTIALLY AUTOLYZED ANIMALS 

CONTROL (UNTR) LOi DOSE HIGH DOSE 

02-016C 02-0195 C2-0200 

50 50 50 

1 

U9 49 US 

** U9 49 «8 

(U9) (U8) («8) 

1 (2%) 

(l»9) (H8) (U8) 

1 <2S) 

1 (2S) 

(17) («9) («8) 

3 (6%) 

2 («*) 

2 ("»%) 

(08) («8) (U8) 

1 (24) 

(US) (U8) C*8) 

1 (2X) 

(U9) (U9) (U8) 

1 (2%) 

(K8) (19) (<*8) 

l_12il 

 EXAMINE D MICROSCOPICALLY 

C-8
 
"
 

INFLAMMATION, CHBONIC FOCAL
 

NECROSIS, FOCAL
 

METAMORPHOSIS FATTY
 

BASOFHILIC CYVO CHANGE
 

HIPEFPLASIA, 1'OCAL
 

*EILE DUCT
 

HYFERPLASIA, 1IOS
 

HYPEKPLASIA, 1'OCAL
 

*EANCREAS
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

tSTCMACh
 

INFLAMMATION, NOS
 

IGASTRIC SUBKUCOSA
 

EDEMA, NOS
 

tFEYERS PATCH
 

HYPERPLASIA, DOS
 

* COLON
 

NEKATODIASIS
 

PARASITISM
 

URINARY SYSTEM
 

f KIDNEY
 

CYST, NOS
 

FOLYCYSTIC KI 3NEY
 

GLCMERULCNEPHUTIS. NOS
 

NEPHROPATHY
 

NEPHBOSIS, NOS
 

tKIDNEY/CORTEX
 

METAMORPHOSIS FATTY
 

ENDOCRINE SYSTEM
 

tPITDITARY
 

CYST, NOS
 

_HIPE££iA£IA^._~H£2flO£a2BE^££iI


TABLE C2 (CONTINUED) 

CONTROL (ONTH)
 

02-0160
 

1 (2%)
 

1 (2%)
 

1 (2*)
 

2 (<»%)
 

1 (2%)
 

<«9)
 

2 («%)
 

1 (2S)
 

(U9)
 

(U9)
 

1 (2»)
 

C*9)
 

1 (2*)
 

(i»9)
 

2 (U%)
 

(19)
 

1 (2%)
 

(«8)
 

a (8*)
 

29 (60%)
 

<t8)
 

1 (2X)
 

(39)
 

LOU DOSE
 

02-0195
 

3 (6*)
 

12 (86»)
 

C«9)
 

{«6)
 

2 (<•*)
 

1 (2X)
 

(U7)
 

(«7)
 

(U7)
 

(U5)
 

3 (7»)
 

(U9)
 

a (8%)
 

(U9)
 

(an)
 

1 <2X)
 _1 i2JL
 

HIGH DOSE
 

02-0200
 

1 (2%)
 

3 (6%)
 

UO (83S)
 

(U8)
 

(«7)
 

(U8)
 

C*8)
 

(U7)
 

(148)
 

3 (6%)
 

(48)
 

1 (2%)
 

1 (2%)
 

3 (6»)
 

(1*8)
 

(«2)
 

J_^iJ
 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-9
 
tACRENAL
 

HETAMORPHOSIS FATTY
 

tADRENAL KECULLA
 

CYST. NOS
 

HYPEBPLASIA. NOS
 

tTHYBOID
 

HYPEEPLASIA, EPITHELIAL
 

HYFEEPLAS1A. C-CELL
 

BEFFODUCTIVE SYSTEM
 

•BABMARY GLAND
 

DILATATION/DUCTS
 

GALACTCCELE
 

*HAHHARY DUCT
 

HYPEEPLASIA, CYSTIC
 

*VAGINA
 

EPIDERMAL INCLUSIO^ CYST
 

IDTERUS
 

HYDROMETRA
 

HEMATOHA, NOS
 

IUTERUS/ENDOBETRIDH
 

INFLAMMATION. ACUTi:
 

AESCESS, NOS
 

HYPERPLASIA. EPITHELIAL
 

HYFEEPLASIA. CYSTIC
 

tOVARY/OVIDUCT
 

INFLABMATION, ACUTE
 

fOVARY
 

ABSCESS, NOS
 

INFLAMMATION, CHBONIC
 

NEBVCUS SYSTEM
 

NCNI_ __ _
 

t NUHEE H OF ANIMAL S WIT H TISSUE

* KUMBER OF ANIMALS NICROPSIED 

TABLE C2 (CONTINUED) 

CONTROL (UNTB) LOB DOSE
 02-0160 02-0195
 

(149) (U9)
 

(49) (U9)
 

C»5) («2)
 

1 <2X)
 

2 (US) 1 (2«)
 

(l»9) («9)
 

1 (2X)
 

5 (10%)
 

(i»9) (U9)
 

1 (2%)
 

(19)	 (U9)
 

1 (2S)
 

(U6) («9)
 

1 (2%) 1 (2X)
 

1 (2%)
 

(<*6) (<«9)
 

1 (2X)
 

(U6) (<*9)
 

1 (2X)
 

(«7) (U7)
 

1 (2*)
 

1 (2»)
 

 EXAMINE D MICROSCOPICALLY 

HIGH DOSE
 02-0200
 

(47)
 

2 (U«)
 

(l»7)
 

1 (2%)
 

1 (2*)
 

(U6)
 

U (9*)
 

C*8)
 

6 (13%)
 

(i»8)
 

(18)
 

(1*7)
 

2 (<*%)
 

(47)
 

1 (2%)
 

1 (2%)
 

7 (15%)
 

(47)
 

1 (2%)
 

(48)
 

C-10 
TABLE C2 (CONCLUDED) 

CONTROL (UHTR)

02-0160

 LOB DOSE

 02-0195

 HIGH DOSE 

 02-0200 

SPECIAL SENSE OHC.AHS 

*EYE

CATARACT

*LENS CAPSULE

CALCIFICATION, NOS

 (149)

 (i|9)

 1 (2%) 

 (49)

 (19)

 (U8) 

1 (2%) 

 (U8) 

HUSCULOSKELE1AL iiYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL CTHEP SYSTEHfi 

CMENTUK 

NECROSIS, FAT 3 

SPECIAL HOEPHOLOC.Y SOBHABY 

NO LESION BEl:OBTED

ANIMAL HISSIIIG/NC NECROPSY

AUTOLYSIS/NO NECROPSY

t M1BBER OF ANIMALS HUH TISSDE EXAMINED

* NUMBEK OF ANIMALS NECROPSIED 

2 

1 

 MICROSCOPICALLY 

1 2 

C-ll
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH DIARYLANILIDE YELLOW
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH DIARYLANILIDE YELLOW 

-1 —"•" TM"~'"-1 -" T" "­ ' " "' ~TM '" ~~ TM-~l ' 

CONTHOL(UNTB) LOB DOSE 05-0160 05-0195 

ANIMALS INITIALLY IN STOEY 50 50 

ANIMALS HISSING 

ANIMALS NECRCPSIEI 50 U9 

ANIMALS EXAMINED hISTOPATHOLOGICALLY ** 49 49
 

INTEGDMENTARY SYSTEM
 

*SKIN (50) (49)
 

ABSCESS, NOS 1 (21)
 

INFLAMMATION, ACUTE/CHEONIC 1 (2X)
 

*SUBCCT TISSUE (50) (49)
 

HEMATOMA, NOS 1 (2%)
 

INFLAMMATION, NECBOTIZING
 

INFLAMMATION, HCDTt FOCAL 1 (2*)
 

ABSCESS, NOS 1 (2*)
 

BESPIEATORY SYSTEM
 

HUNG <«7) (49)
 

INFLAMMATION, \iOS 1 (2X)
 

*IUNG/ALVEOLI (U7) (49)
 

HYPERTROPHY, NOS 1 <2S)
 

HEMATOPCIETIC SYST1!M
 

tSPLEEN (1*9) (46)
 

INFARCT, NOS
 

HYPEPPLASIA, RIITICULUM CELL 3 (7S)
 

ERYTHBOPOIESI5 2 ("»%)
 

tRANDIBULAfi L. NODE (HO) (35)
 

ATROPHY, NOS
 

tflESENTZRIC L. NO!E («0) (35)
 

HYPERPLASIA, RITICULUM CELL 1 (3»)
 

tRENAL LYMPH NODE (10) (35)
 

i!I£I£PiASJAx_acs _ 2_i5ii
 — TMr — — — - *


t NUMBER OF ANIMALS HITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECFOPSIED 

**EXCLUDES PARTIALLY AJTOLYZED ANIMALS 

HIGH DOSE
 05-0200
 

50
 

1
 

49
 

49
 

(49)
 

1 (2«)
 

(49)
 

1 (2%)
 

(49)
 

(49)
 

(47)
 

1 (2%)
 

(43)
 

1 (2X)
 

(43)
 

(43)
 

-— —^—
—


D-3 
CIRCDLATCRY SYSTEH
 

tHEARl
 PERIARTERITIS
 

• AORTA
 PEBIARTERITIS
 

•VF.SICAL	 ARTERY
 

FERIVASCULITIS
 

DIGESTIVE SYSTEH
 

ISUEBAXILLARY GLAND
 

ATROFHY, NOS
 

ILIVER
 

NECROSIS, FOCAL
 

BETArOREHOSIS FATTY
 

ANGIECTASIS
 

tUVER/KUPFFER CELL
 

HYPEFPLASIA, NOS
 

tPANCREAS
 

CYSTIC DOCTS
 

INFLAHHATICN, FOCAI.
 

INFLABBATICN, ACUTL/CHRONIC
 

PERIVASCULITIS
 

EEGENERATICN, CYST"C
 

NECROSIS, FOCAL
 

NECROSIS, FAT
 

tEANCREATIC ACINUS
 

DEGENERATION, NOS
 

tEEYERS PATCH
 

INFLABBATION, ACUTE
 

HYPERPLASIA, LYHPHOID
 

tCOLON
 

KEBATODIASIS
 

DRINABY SYSTEM
 

tKIDNEY
 

HYERONEPHROSIS
 

TABLE Dl (CONTINUED) 

CONTROL (DNTR)
 

05-0160
 

(<*9)
 

(50)
 

(50)
 

(47)
 

(U9)
 

1 (2%)
 

1 (2S)
 

(U9)
 

1 (2%)
 

(U6)
 

1 (2X)
 

1 (2%)
 

1 (2%)
 

<U6)
 

(<49)
 

1 (2%)
 

1 (2»)
 

C*8)
 

(U9)
 

2 (M)
 

LOH DOSE HIGH DOSE
 

05-0195 05-0200
 

(49)	 (49)
 

1 (2%)
 

(49)	 (49)
 

2 (4%) 1 (2%)
 

(49)	 (49)
 

1 (2*)
 

(46)	 (49)
 

1 (2%)
 

(49)	 (46)
 

2 (4*)
 

(49) (46)
 

C»6) (48)
 

1 (2%)
 

1 (2X)
 

1 (2X)
 

1 (2S)
 

(46)	 (48)
 

1 (2%)
 

(48) (49)
 

1	 (2X)
 

(43)	 (45)
 

1 (2*)
 

(49) (49)
 

I HUHEER OF ANIflALS HI TH TISSUE EXAMINE D BICROSCOPICALLY 

* SUBBE F OF ANIBAL S NI.CROPSIED 

D-4 
PYELONEPHRITIS, NOS


PYELONEPHRITIS, FOCAL


INFLAMMATION, CNTEESTITIAL


INFLAMMATION, CHRONIC


tURINARY BLADDER


INFLAMMATION, HCUTF./CHRONIC


INFLAMMATION, SHBONIC


ENEOCBINE SYSTEM
 

tEITOITARY


CYST, NOS
 

tTHYROIC
 PERIARTERITIS
 

HYPEPPLASIA, FOCAL


REEEODUCTIVE SYSTEM
 

*PREPUIIAL GLAND


DILATATION, NOS


*PROSTATE


INFLAMMATION, ACUTF
 

*SEMINAL VESICLI


INFLAMMATION MITH FIBROSIS
 

*IESTIS


DEGENERATION, NOS
 

tTESTIS/TUBDLE


NECROSIS, FOCAL


NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NCNE
 

HUSCUIOSKELETAL S1STEM
 

NCNE
 

TABLED1 (CONTINUED) 

CONTROL (ONTB)


05-0160


 1 (2X)
 

(49)


 (1*2)


 (1*2)


 1 (2%)
 

(50)


 1 (2%)
 

 (U9)


 (50)


 («9)


 (U9)


 1 (2%)
 

 LOH DOSE
 

 05-0195
 

1 (2X)
 

1 (2S)
 

1 (2S)
 

 (1*9)
 

1 (2%)
 

1 (2*)
 

 (36)
 

 (3U)
 

 («9)
 

 («8)
 

 (U9)
 

 («9)
 

 («9)
 

HIGH DOSE
 

05-0200
 

(U9)
 

(tO)
 

2 (5%)
 

(i*3)
 

1 12%)
 

(1*9)
 

(1*6)
 

1 (2*)
 

(1*9)
 

1 (2%)
 

C*8)
 

1 (2»)
 

(1*8)
 

t NDMEER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NDMBEH OF ANIMALS NECROPSIED
 

D-5 
TABLE Dl (CONCLUDED)
 

CONTROL (ONTR) LOI DOSE HIGH DOSE
 

05-0160 05-0195 05-0200
 

BOEY CAVITIES
 

*ABDOHINAL CAVITY (50) (U9) <«9)
 

ADHESION, NOS 1 (2S)
 

*EERITONEUH (50) («9) (W9)
 

INFLAHHATION, ACUTE 1 (21)
 

»HESENTERY (50) <U9) («9)
 

STEATIT1S 1 (2%)
 ABSCESS, NOS 1 (2*)
 

ALl CTHEP SI STEMS
 

ADIEOSE TJ SSUE
 

S1EATIT3 S 1
 NECROSIS, FAT 2
 

CHENTUB
 

STEATI1 :s
 1
 

NECROSI5, NOS 1
 

NECROSIS, FAT 1
 

SPECIAL MORPHOLOGY SOHHARY
 

17 18 31
 

ANIHAL KISSISG/NO NECROPSY
 

NO LESION EEFORTED
 1
 

ADTO/NECROPSY/HISTO PERF 1 2
 

AUTO/NECROFSY/NO HISTO 1
 

ADTOLYSIS/NO NECROPSY 1
 

* KUC.EER OF ANIHALS HITH TISSUE EXAMINED BICHOSCOPICALLY
 

* NUHBER Of ANIMALS NECROPSIED
 

D-6 
TABLE D2

 

SUMrlARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE -MICE 

TREATED WITH DIARYLANILIDE YELLOW 

COHTROL(UHTB) LOW DOSE
 06-0160 06-0195
 

ANIMALS INITIALIY I» STUDY 50 50
 ANIMALS NECHOPSIED 50 50
 ANIBAIS EXAMINE! HISTOFATHOLOGICALtY ** 50 49
 

INTEGUMENTARY SYSTEM
 

NONE
 

HESPIEATOBY SYSTEM
 

NONE
 

HEMATOFOIETIC SYSTEM
 

tEONE BAHROH («9) («9)
 

HYELOFIBBOSIS 2 («%)
 

tSPLEEN («9) C*9)
 

HYPERPLASIA, HEflATOPOIETIC
 

HYPEEPLASIA. BETICULUH CELL 1 (2*)
 

HYPEEPLASIA, LYMPHOID 1 (2%) 1 (2S)
 

HEMATOPOIESIS 1 <2S)
 

EBYTHHOPCIESIS 1 (2%)
 

tflANDIBULAR L. NODE («0) (36)
 

INFLAMMATION, NOS
 

HYPEEPLASIA, PLASMA CELL 1 (3X)
 

fERCNCHIAL LYMfH NODE <«0) (36)
 

INFLABMA1ION, ACUTE
 

•MEDIASTINAL L. NODE (UO) (36)
 

HYPERPLASIA, NOS 1 (3%)
 

tLUMBAR LYRPH NODE (<«0) (36)
 

HYPERPLASIA, NOS 1 (3%)
 

•HESENTERIC	 L. NODE («0) (36)
 

__HI£EEEIiSIA^_JJ2£
 

t SOMBER OF ANIFAL S SITH TISSUE EXAMINE D MICROSCOPICALLY 

* NOHEE K OF ANItAL S NECROPSIED 

**EXCHJDES PARTIALLY AUTOLYZED ANIMALS 

HIGH DOSE
 

06-0200
 

50
 

50
 

«8
 

(«7)
 

1 (2%)
 

(«7)
 

1 <2%)
 

1 (2%)
 

(«5)
 

1 (2%)
 

C*5)
 

1 (2»)
 

(U5)
 

(«5)
 

(«5)
 

!_L2SL_.
 

D-7 
TABLE D2 (CONTINUED) 

CONTROL (USTB) LOi DOSE
 

06-0160 06-0195
 

« RENAL 1 YHPH NODE (<*0) (36)
 

HYPEliPLASIA. NOS 1 (3S)
 

HYPEl'PLASIA, PLASHA CELL 1 <3S)
 

CIRCULATORY SYSTEM
 

tMYCCARDIUM (50) (U9)
 

INFLAMMATION, FOCAL
 

INFLAMMATION, ACOTE DIFFUSE 1 (2X)
 

* AORTA (50) (50)
 

PERIARTERITIS
 

DIGESTIVE SYSTEM
 

ISALI7AFY GLAND C»8) C»8)
 

ABSCESS, NOS 1 (2*)
 

NECEOSIS, NOS 1 (2%)
 

*LIVER (19) (17)
 

ECTOEIA
 

INFLAMMATION, NOS
 

NECItCSIS, NOS 1 (2X)
 

NECltOSIS, FOCAL 1 (2%) 1 (2*)
 

INFhRCT, NOS 1 (2%)
 

BASOPHILIC CYTO CHANGE 1 (2*)
 

HTP'IRPLASIA, BETICULUM CELL
 

HEMATOFOIESIS 1 (2%)
 

»EILE DUCT (50) (50)
 

INFLAMMATION, CHRONIC FOCAL 2 (1%)
 

tPANCREAS (U7) (US)
 

CYSIIC DQCTS 1 (2%)
 

INFLAMMATION, NOS
 

INFLAMMATION, ACUTE/CHRONIC 1 (2%)
 

INFLAMMATION, CHRONIC FOCAL
 

ABSCESS, CHRONIC 1 (2%)
 

HYPEBPLASIA, FOCAL 1 (2%)
 

tFANCFEATIC ACINUS (U7) («8)
 

DEGENERATION, NOS 1 (2*)
 

f STOMJCH (US) (145)
 

INI'LAHMATICN, NO S _ ____ _
 

* NUMBO OF ANIMAL S HITh TISSUE EXAMINE D MICHOSCOPICALLY * NUMEI;F OF ANIMALS NECROPSIES 

HIGH DOSE
 06-0200
 

(US)
 

C*7)
 

2 ("*%)
 

(50)
 

1 (2X)
 

(U6)
 

(U6)
 

1 (2%)
 

2 («%)
 

1 (211)
 

1 (2%)
 

(50)
 

C»3)
 

1 (2%)
 

1 (2%)
 

C»3)
 

(UK)
 

i_i2SI_.
 

D-8 
TABLE D2 (CONTINUED) 

. .
 

INFLAMMATION. FOCAL
 

INFLAMMATION, ACUTE FOCAL
 

INFLAMMATION, CHRONIC
 

IFEYERS PATCH
 

HYPERPLASIA, LYMPHOID
 

tCCLON
 

NEflATODIASIS
 

URINARY SYSTEM
 

tKIDNEY
 

GLOMEPULCNEPIiRITIS, NOS
 

LYC.PHOCYTIC :.NFILTRATE
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, CHRONIC
 

INFLAMMATION, CHRONIC FOCAL
 

GLCBERDLCSCL11ROSIS, NOS
 

• URINARY ELADDEii
 

INFLAMMATION,. ACUTE
 

INFLAMMATION, CHEONIC FOCAL
 

ID. ELADDER/SUBM JCOSA
 

INFLAMMATION, CHRONIC
 

INFLAMMATION, CHRONIC FOCAL
 

PEPIVASCULIT tS
 

*U. ELADDER/BUSC JLARIS
 

CALCIUM DEPOSIT
 

ENECCRINE SYSTEM
 

IPITOITARY
 CYST. NOS
 

HYPEKPLASIA. CHKONOPHO BE -CELL
 

ITHYROID
 

HYPEFPLASIA, EPITHELIAL
 

HYPEKPLAEIA. FOCAL
 

HYPEFPLASIA, C-CELL
 

HYPEJPLAlIA^.POLLICULAa-CELL
 

CONTROL (UNTB) LOW DOSE HIGH DOSE 06-0160 06-0195 06-0200 

1 (2X) 

1 (2*) 

1 <2«) 

(49) (<»6) (47) 

1 (2%) 

(50) C»D (45) 

1 (2%) 

(49) («9) (47) 

4 (8%) 

1 (2X) 

1 (2%) 

2 («%) 

1 (2%) 

1 (2S) 

(50) (44) (45) 

1 (2%) 

1 (2%) 

(50) (4U) (45) 

1 <2X) 

16 (32*) 

1 <2S) 

(50) («4) (45) 

1 (2S) 

(t2) 139) (44) 

2 (5*) 

1 <3J) 

("»D (41) (44) 

1 (2S) 

1 (2X> 

2 (5%) 

1_JL2SI_ . l-l&L .— —-.— ——«———­

* NUMBER 0" ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

» NUMBE R OF ANIMALS NECROPSIED
 

D-9 
TABLE D2 (CONTINUED)
 

111

 " •"«"

—TM—g»TM-»-

""»~i"> i-mr-- Jr ­_ JT -J.IT ­_ •••••M~HWMrjH«-« 

CONTBOL(DNTH) LOU DOSE 

06-0160 06-0195 

REPRODUCTIVE SYSTEM 

tOTERDS (<»9) (47)

HYDROBETR A 5 (10») ^ (15%) 

ABSCESS, NOS 1 (2%) 

NECHOJIS, FAT 1 (2*) 

CALCIMCATION, NOS 1 <2S) 

* UTERUS/ KNDOMETRIOH (U9) (47) 

INFLAMMATION. SUPPURATIVE 2 («%) 

INFLAMMATION, ACUTE 1 (2%) 

HYPERl'LASIA , NOS 1 (2S) 

HIPEfcl'LASIA, CYSTIC 32 (65%) 34 (72%) 

«OVARY/07IDUCT <«9) (47) 

INFLA1HATION, NOS 

ABSCE3S. NOS 1 (2%) 

INFLASMATION, CHRONIC 

IOVABY/PABOVARIA N (49) (47) 

STEATITIS 

ABSCESS, NOS 2 (4% ) 

INFLAHHATION, ACUTE/CHRONIC 1 (2%) 

*OVARY («8) (42) 

CYST, NOS 6 (13X) 7 (17%) 

INFLAMMATION . NOS 

INFLAMMATION , SUPPUBATIV E 1 (2%) 

ABSCISS, NOS 

INFLAMMATION . ACUTE/CHRONIC 2 (5%) 

INFLAMMATION , CHRONIC 1 (2%) 

HYPE1IPLASIA, EPITHELIAL 1 (2*) 

HEHVCUS liYSTEW 

«ERAIN/KENINGE S (49) (46 ) 

LYMP!10CYT1C INFILTRATE 

IERAI N (49) (46) 

HYBRDCEPHALUS, NOS 1 (2X) 

SPECIAL SENSE ORGAN S 

*EYE/LACRIHAL GLAN D (50) (50) 

iJlPFRPLA£Ii4._JjaS_ 1 J2J1_

t NOHEEB OF ANIMALS WITH TISSUE EXAMINED MICBOSCOPICALLY
 

* NUMBEF OF ANIMALS NECROPSIED
 

. 

HIGH DOSE 

06-0200 

(46) 

7 (15%) 

(46) 

3 (7%) 

1 (2%) 

24 (S2%) 

(46) 

1 (2%) 

1 (2%) 

1 (2%) 

(»6) 

1 (2%) 

(46) 

7 (15%) 

2 (4%) 

1 (2%) 

(47) 

2 (4%) 

(47) 

(50) 

D-IO
 
TABLE D2 (CONCLUDED) 

CONTROL (UNTB)

Ofa-0160

 LOH DOSE

 06-019S

 HIGH DOST 

 jb-0200 

HUSCULOSKELETAL SYSTEM 

DOME 

BOEY CAVITIES 

•MEDIASTINUM INFLAMMATION, ACDTE

NECROSIS, NO:;

•PERITONEUM

INFLAMMATION, NOS

ABSCESS, NOS

 (50)

 (50)

 (50)

 1 (2%)

i (2*) 

 (50)

 (50) 

 (50) 

1 (2X) 

1 (2%) 

ALL OTHEK SYSTEM 

•MULTIPLE ORGAN S

AMYLOIDOSIS

CMENTUK 

PIRIVASCULITJS

 (50)

 1 <2X) 

1 

 (50) (50) 

SPECIAL MORPHOLOGY SUMMARY 

NO LFSION REIOBTED 2 

AUTO/NECROPSY/HISTO PERF 2 

AOTO/NECBOPSY/NO HISTO

t NOHEER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

5 

1

3 

2 

D-ll 


DHEW Publication No. (NIH) 78-830