1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 115 

BIOASSAY OF 

SULFALLATE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 95-06-7 

NCI-CG-TR-115 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

SULFALLATE
 

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) 78-1370
 

REPORT ON THE BIOASSAY OF SULFALLATE
 

FOR POSSIBLE CARCINOGENICITY
 

CARCINOGENESIS TESTING PROGRAM
 

DIVISION OF CANCER CAUSE AND PREVENTION
 

NATIONAL CANCER INSTITUTE, NATIONAL INSTITUTES OF HEALTH
 

FOREWORD; This report presents the results of the bioassay of sulfallate conducted for the Carcinogenesis Testing Program, Division
 

of Cancer Cause and Prevention, National Cancer Institute (NCI),
 

National Institutes of Health, Bethesda, Maryland. This is one of
 

a series of experiments designed to determine whether selected
 

chemicals have the capacity to produce cancer in animals. Negative
 

results, in which the test animals do not have a significantly
 

greater incidence of cancer than control animals, do not necessarily
 

mean the test chemical is not a carcinogen because the experiments
 

are conducted under a limited set of circumstances. Positive results
 

demonstrate that the test chemical is carcinogenic for animals under
 

the conditions of the test and indicate a potential risk to man.
 

The actual determination of the risk to man from animal carcinogens
 

requires a wider analysis.
 

CONTRIBUTORS: This bioassay of sulfallate was conducted by Hazleton
 

Laboratories America, Inc., Vienna, Virginia, initially under direct
 

contract to the NCI and currently under a subcontract to Tracer Jitco,
 

Inc., prime contractor for the NCI Carcinogenesis Testing Program.
 

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

principal investigators for the contract were Dr. M. B. Powers (3),
 

Dr. R. W. Voelker (3), Dr. W. A. Olson (3,4) and Dr. W. M. Weatherholtz (3). Chemical analysis was performed by Dr. C. L. Guyton (3,
 

5) and the analytical results were reviewed by Dr. N. Zimmerman (6);
 

the technical supervisor of animal treatment and observation was Ms.
 

K. J. Petrovics (3).
 

Histopathologic examinations were performed at the Hazleton
 

Laboratories America, Inc. Histopathologic examinations of the rats
 

were done by Dr. R. H. Habermann (3) and reviewed by Dr. R. W. Voelker
 

(3). The histopathologic examinations of the mice were done by Dr.
 

R. W. Voelker (3). The diagnoses included in this report represent
 

the interpretation of these pathologists. Histopathology findings
 

and reports were reviewed by Dr. R. L. Schueler (7).
 

Compilation of individual animal survival, pathology, and summary tables was performed by EG&G Mason Research Institute (8); the
 

statistical analysis was performed by Mr. W. W. Belew (6,9), using
 

iii
 
methods selected for the Carcinogenesis Testing Program by Dr. J. J.
 

Gart (10).
 

This report was prepared at METREK, a Division of The MITRE Corporation (6) under the direction of the NCI. Those responsible for
 

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

(6), task leader Dr. M. R. Kornreich (6,11), senior biologist Ms. P.
 

Walker (6), biochemist Mr. S. C. Drill (6), and technical editor Ms.
 

P. A. Miller (6). The final report was reviewed by members of the
 

participating organizations.
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

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

Cueto, Jr. (1), Dr. J. F. Douglas (1), Dr. D. G. Goodman (1,11),
 

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

Dr. T. W. Orme (1), Dr. R. A. Squire (1,12), Dr. S. F. Stinson (1),
 

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

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

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

3.	 Hazleton Laboratories America, Inc., 9200 Leesburg Turnpike,
 

Vienna, Virginia.
 

4.	 Now with the Center for Regulatory Services, 2347 Paddock Lane,
 

Reston, Virginia.
 

5.	 No wit w h Rhodia, Inc., 23 Belmont Drive, Somerset, New Jersey.
 

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

Boulevard, McLean, Virginia.
 

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

Maryland.
 

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

Rockville, Maryland.
 

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

Golden, Colorado.
 

iv
 
10.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

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

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

Washington, D.C.
 

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay for possible carcinogenicity of sulfallate was conducted using Osborne-Mendel rats and B6C3F1 mice. Sulfallate was
 

administered in the feed, at either of two concentrations, to groups
 

of 50 male and 50 female animals of each species. Twenty mice and 50
 

rats of each sex were placed on test as controls for the bioassay.
 

The time-weighted average high and low dietary concentrations of
 

sulfallate were, respectively, 410 and 250 ppm for male rats, 404 and
 

250 ppm for female rats, 1897 and 949 ppm for male mice, and 1815
 

and 908 ppm for female mice. After the 78-week period of chemical
 

administration, there was an additional observation period of 25 to
 

26 weeks for dosed rats, 33 weeks for control rats, and 12 to 13
 

weeks for dosed and control mice.
 

There were significant positive associations between increased
 

sulfallate concentration and accelerated mortality in both sexes of
 

rats and in female mice. However, adequate numbers of animals in all
 

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

tumors.
 

Statistical analyses of the incidences of mammary adenocarcinomas in female rats, stomach neoplasms (i.e., combination of papillomas NOS, squamous-cell papillomas, and squamous-cell carcinomas)
 

in male rats, combined alveolar/bronchiolar carcinomas and alveolar/
 

bronchiolar adenomas in male mice, and adenocarcinomas of the mammary
 

gland in female mice revealed a significant positive association
 

between dosage and incidence. These associations were all supported
 

by at least one significant Fisher exact comparison.
 

The incidence of toxic tubular nephropathy observed in male rats
 

and in mice of both sexes increased with the concentration of the compound administered. This nonneoplastic lesion was not observed in
 

control animals.
 

Under the conditions of this bioassay dietary administration of
 

sulfallate was carcinogenic to Osborne-Mendel rats and to B6C3F1 mice,
 

inducing mammary gland tumors in females of both species, tumors of
 

the forestomach in male rats, and lung tumors in male mice.
 

VI1
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 4
 

A. Chemicals 4
 

B. Dietary Preparation 4
 

C. Animals 5
 

D. Animal Maintenance 5
 

E. Selection of Initial Concentrations 7
 

F. Experimental Design 8
 

G. Clinical and Histopathologic Examinations 12
 

H. Data Recording and Statistical Analyses 14
 

III. CHRONIC TESTING RESULTS: RATS 19
 

A. Body Weights and Clinical Observations 19
 

B. Survival 21
 

C. Pathology 21
 

D. Statistical Analyses of Results 26
 

IV. CHRONIC TESTING RESULTS: MICE 38
 

A. Body Weights and Clinical Observations 38
 

B. Survival 38
 

C. Pathology 41
 

D. Statistical Analyses of Results 44
 

V. DISCUSSION 56
 

VI. BIBLIOGRAPHY 59
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH SULFALLATE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH SULFALLATE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH SULFALLATE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH SULFALLATE D-l
 

ix
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF SULFALLATE 2 

2 GROWTH CURVES FOR SULFALLATE CHRONIC STUDY 

RATS 20 

SURVIVAL COMPARISONS OF SULFALLATE CHRONIC 

STUDY RATS 22 

GROWTH CURVES FOR SULFALLATE CHRONIC STUDY 

MICE 39 

SURVIVAL COMPARISONS OF SULFALLATE CHRONIC 

STUDY MICE 40 

LIST OF TABLES 

Table Number Page 

DESIGN SUMMARY FOR OSBORNE-MENDEL

SULFALLATE FEEDING EXPERIMENT 

 RATS— 

10 

DESIGN SUMMARY FOR B6C3F1 MICE—SULFALLATE 

FEEDING EXPERIMENT 11 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

SULFALLATE 27 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH SULFALLATE 32 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

SULFALLATE 45 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH SULFALLATE 50 
LIST OF TABLES (Concluded) 

Table Number Page 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH SULFALLATE A~3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH SULFALLATE A-8 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH SULFALLATE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH SULFALLATE B-7 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH SULFALLATE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 

SULFALLATE C-9 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH SULFALLATE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 

SULFALLATE D-9 

xi
 

I. INTRODUCTION
 

Sulfallate (Figure 1) (NCI No. C00453), a chlorinated dithiocarbamate derivative used as a preemergence herbicide on vegetable
 

crops, was selected for bioassay by the National Cancer Institute
 

because of its structural relationship to the known tumorigens
 

selenium diethyldithiocarbamate and potassium bis(2-hydroxyethyl)
 

dithiocarbamate and to a number of suspected tumorigens containing
 

the diethyldithiocarbamate or dimethyldithiocarbamate moieties (innes
 

et al., 1969).
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is diethylcarbamodithioic acid 2-chloro­

*
 2-propenyl ester. It is also called 2-chloro-2-propenyl diethylcarbamodithioate; 2-chloroallyl diethyldithiocarbamate; diethyldithiocarbamic acid 2-chloroallyl ester; CDEC; and CP4742.
 

Sulfallate is a selective preemergence herbicide used to control
 

annual grasses and broadleaf weeds in vegetable and fruit crops.
 

It is also used to control weeds among shrubbery and ornamental
 

plants (Farm Chemicals Handbook, 1976). Sulfallate is especially
 

suited to sandy soils, but it is also effective on peat muck soil-


types (Farm Chemicals Handbook, 1976). Established plants, or those
 

which are vegetatively propagated, are not effectively eradicated by
 

sulfallate administration (Martin and Worthing, 1977).
 

*
 

The CAS registry number is 95-06-7
 
S Cl 

CH3CH2v || | 

/N—C—S—CH2—C=CH2 

CH3CH2' 

FIGURE 1 

CHEMICAL STRUCTURE OF SULFALLATE 
Sulfallate is currently produced in commercial quantities (in
 

excess of 1000 pounds or $1000 in value annually) by only one U.S.
 

company (Stanford Research Institute, 1977); consequently, recent
 

production data are considered proprietary and are withheld from the
 

public.
 

The potential for exposure to sulfallate is greatest for agricultural workers, but may also be considerable for workers in sulfallate
 

production facilities. Residents of agricultural communities may be
 

exposed to airborne residues following spraying operations. The herbicide is readily taken up by plant roots (Martin and Worthing, 1977)
 

and the general population may be exposed via ingestion of residues
 

in food crops. Ingestion is unlikely, however, since sulfallate is
 

hydrolyzed in aqueous solution with a half-life of 47 days at pH 5
 

and 30 days at pH 8 (Martin and Worthing, 1977).
 

Chronic and acute toxicity data in humans are not available;
 

however, sulfallate is considered a moderate skin and eye irritant
 

(Gosselin et al., 1976).
 
II. MATERIALS AND METHODS 

A. Chemicals
 

Technical-grade sulfallate was purchased from Monsanto Chemical
 

Company, St. Louis, Missouri, and analyzed by Hazleton Laboratories
 

America, Inc., Vienna, Virginia. Gas-liquid chromatography (GLC)
 

utilizing the internal standard method demonstrated that the purity
 

was greater than 90 percent. This was in agreement with the manufacturer's specification of purity. GLC total-area analysis revealed
 

the presence of 14 minor components with the major component accounting
 

for over 90 percent of the area. Internal standard and total-area
 

analyses performed 12 months later provided similar results, although
 

total-area analysis indicated the presence of only 7 minor components.
 

The similarity in the results using both methods of GLC analysis over
 

the period of one year indicated a relatively high stability for the
 

compound. Studies performed 24 months after the initial analyses
 

reconfirmed these results.
 

Throughout this report the term sulfallate is used to represent
 

this technical-grade material.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals
 

consisted of 2 percent Duke's corn oil (S. F. Sauer Company, Richmond,
 

Virginia) by weight added to Wayne Lab-Blox meal (Allied Mills, Inc.,
 

Chicago, Illinois). Fresh mixtures of sulfallate in corn oil were
 

prepared each week and stored in the dark. The mixtures of sulfallate
 

4
 
in corn oil were incorporated into the appropriate amount of basal
 

laboratory diet in a twin-shell blender fitted with an accelerator
 

bar.
 

C. Animals
 

Two animal species, rats and mice, were used in the carcinogenicity bioassay. The Osborne-Mende1 rat was selected on the basis
 

of a comparative study of the tumorigenic responsiveness to carbon
 

tetrachloride of five different strains of rats (Reuber and Glover,
 

1970). The B6C3F1 mouse was selected because it has been used by the
 

NCI for carcinogenesis bioassays and has proved satisfactory in this
 

capacity.
 

Rats and mice of both sexes were obtained through contracts with
 

the Division of Cancer Treatment, National Cancer Institute. The
 

Osborne-Mende1 rats were procured from the Battelle Memorial Institute, Columbus, Ohio, and the B6C3F1 mice were obtained from the
 

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

Upon receipt, animals were quarantined for at least 10 days, observed
 

for visible signs of disease or parasites, and assigned to the various dosed and control groups.
 

D. Animal Maintenance
 

All animals were housed by species in temperature- and humidity-


controlled rooms. The temperature range was 20° to 24°C and the
 

relative humidity was maintained between 45 and 55 percent. The
 

air conditioning system in the laboratory provided filtered air at
 
a rate of 12 to 15 complete changes of room air per hour. Fluorescent lighting was provided on a 12-hour-daily cycle.
 

The rats were individually housed in suspended galvanized-steel
 

wire-mesh cages with perforated floors. Mice were housed by sex in
 

groups of ten in solid-bottom, polypropylene cages equipped with
 

filter tops. Sanitized cages with fresh bedding (Sanichips®, Pine-


wood Sawdust Company, Moonachie, New Jersey) were provided once each
 

week for mice. Rats received sanitized cages with no bedding with
 

the same frequency. Food hoppers were changed and heat-sterilized
 

once a week for the first 10 weeks and once a month thereafter.
 

Fresh heat-sterilized glass water bottles and sipper tubes were
 

provided three times a week. Food and water were available ad
 

libitum.
 

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

containing chlorobenzilate (510-15-6); DDT (50-29-3); and TDE (72­

54-8). Control rats were housed in a room with other rats receiving
 

diets containing trifluralin (1582-09-8); dioxathion (78-34-2);
 

dicofol (115-32-2); nitrofen (1836-75-5); endosulfan (115-29-7); and
 

mexacarbate (315-18-4).
 

All mice, including controls, used in the sulfallate chronic
 

bioassay were housed in the same room as other mice receiving diets
 

containing trifluralin (1582-09-8); dioxathion (78-34-2); methoxychlor (72-43-5); DDE (72-55-9); TDE (72-54-8); dicofol (115-32-2);
 

*
 CAS registry numbers are given in parentheses.
 
chlorobenzilate (510-15-6); acetylaminofluorene (53-96-3); clonitralid
 

(1420-04-8); DDT (50-29-3); pentachloronitrobenzene (82-68-8);
 

nitrofen (1836-75-5); endosulfan (115-29-7); mexacarbate (315-18-4);
 

amitrole (61-82-5); and safrole (94-59-7).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

sulfallate for administration to dosed animals in the chronic
 

studies, subchronic toxicity tests were conducted with both rats
 

and mice. Animals of each species were distributed among six groups,
 

each consisting of five males and five females. Sulfallate was
 

premixed with a small amount of the laboratory diet. The mixture was
 

then incorporated into the laboratory diet with 2 percent corn oil
 

and fed ad libitum to five of the six rat groups and five of the six
 

mouse groups in concentrations of 215, 464, 1000, 2150, and 4640 ppm.
 

The sixth group of each species served as a control group, receiving
 

only the basal laboratory diet. The dosed dietary preparations were
 

administered for a period of 6 weeks, followed by a 2-week observation
 

period during which all animals were fed the basal laboratory diet.
 

A concentration inducing no mortality and resulting in a depression in mean group body weight of approximately 20 percent relative
 

to controls was selected as the high concentration for administration
 

in the chronic study. When weight gain criteria were not applicable,
 

mortality data alone were utilized.
 
Mean body weight depression was observed at the lowest administered concentration in male rats and at 464 ppm and above in female
 

rats. At a level of 464 ppm the depression in mean body weight as
 

compared to controls was 5 percent in male rats and 19 percent in
 

female rats. At 1000 ppm the depression in mean body weight was 42
 

percent in males, and 40 percent in females. No deaths were observed
 

at these levels. The high concentration selected for administration
 

to both male and female rats in the chronic bioassay was 500 ppm.
 

In the 8-week mouse subchronic study, weight gain patterns were
 

erratic. At a concentration of 464 ppm mean body weight gain in the
 

male mice was only 19 percent of that gained by the controls. In the
 

female mice receiving 464 ppm, mean body weight gain was 177 percent
 

of that gained by controls. At 1000 ppm an increase in the mean body
 

weight gain of 244 percent in male mice, and 168 percent in the
 

female mice was observed. At 2150 ppm mean body weight gain in male
 

mice was only 56 percent of that in controls, while in females
 

receiving the same concentration no increase or depression in mean
 

body weight was noted when compared to controls. No deaths were
 

observed at any of these concentrations. The high concentrations
 

selected for administration to male and female mice in the chronic
 

bioassay were 2000 and 1200 ppm, respectively.
 

F. Experimental Design
 

The experimental design parameters for the chronic bioassay (species, sex, group size, actual concentrations administered, duration
 
of treated and untreated observation periods, and the time-weighted
 

average concentrations) are summarized in Tables 1 and 2.
 

The dosed and control rats were all approximately 6 weeks old
 

at the time they were placed on test. The dosed rats were approximately 3 weeks younger than the controls and were started on test
 

approximately 3 weeks after the controls. The concentrations of
 

sulfallate initially utilized for male and female rats were 500 and
 

250 ppm. Throughout this report those rats initially receiving the
 

former concentration are referred to as the high dose groups, while
 

those initially receiving the latter concentration are referred to as
 

the low dose groups. In week 8 of the study, administration of
 

sulfallate to the high dose female rats ceased for 1 week, followed
 

by 4 weeks of feeding at the previous concentration of 500 ppm. This
 

cyclic pattern of administration continued for the remainder of the
 

dosing period. This same method of total sulfallate intake reduction
 

was employed for the high dose male rats beginning in week 12. The
 

dosing period was 78 weeks followed by an observation period during
 

which the rats received only the basal laboratory diet and water.
 

Although a 32-week final observation period was originally planned,
 

surviving rats were sacrificed after 25 to 26 weeks of observation.
 

The dosed and control mice were all approximately 6 weeks old at
 

the time the experiment began. The concentrations initially administered to the male mice were 2000 and 1000 ppm. Throughout this
 

report those males initially receiving the former concentration are
 

9
 
TABLE 1
 

DESIGN SUMMARY FOR OSBORNE MENDEL-RATS
 

SULFALLATE FEEDING EXPERIMENT
 

INITIAL 

GROUP

SIZE

MALE 

CONTROL 50

LOW DOSE 50

HIGH DOSE 50

FEMALE 

CONTROL 50

LOW DOSE 50

HIGH DOSE 50

 SULFALLATE
 

 CONCENTRATION 3


 0
 

 250
 

0
 

 500
 

500°
 

0
 

0
 

 25C
 

0
 

 500
 

500C
 

0
 

OBSERVATION PERIOD
 

TREATED UNTREATED
 

 (WEEKS) (WEEKS)
 

0 111
 

78
 

26
 

11
 

53 14
 

25
 

0 111
 

78
 

26
 

7
 

56 15
 

26
 

a
 .. .
 Concentrations given in parts per million.
 

2 (concentration X
 

Time-weigl ited average concentration = r^r
 

Q
 

TIME-WEIGHTED
 

AVERAGE CONCENTRATION OVER A^
 

78-WEEK PERIOD
 

0
 

250
 

410
 

0
 

250
 

404
 

weeks received)
 

These dosages were cyclically administered with a pattern of 1 dose-


free week followed by 4 weeks (5 days per week) of administration at
 

the dosage indicated.
 

10
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

SULFALLATE FEEDING EXPERIMENT
 

TIME-WEIGHTED 

INITIAL OBSERVATION PERIOD AVERAGE CONCENGROUP SULFALLATE TREATED UNTREATED TRATION OVER A 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) 78-WEEK PERIODb 

MALE
 

CONTROL 20 0 0 91	 0
 

LOW DOSE 50 1000 62 949
 

ioooc 12 4
 

0 13
 

HIGH DOSE 50 2000 62 1897
 

2000° 12 4
 

0 13
 

•
 

FEMALE
 

CONTROL 20 0 0 91	 0
 

LOW DOSE 50 600 8 908
 

1000 54
 

1000° 12 4
 

0	 12
 

HIGH DOSE 50	 1200 8 1815
 

2000 54
 

2000° 12 4
 

0	 12
 

a
Concentrations in parts per million.
 

b	 2 (concentration X weeks received)
 J. A, lilt, Wt^J-gLiU-WU Cl V <L- J. d £2 *_ \_.WLL^^lll_i,lil--t-Wl.l	 ~7 O 1
 

'These dosages were cyclically administered with a pattern of 1 dose-


free week followed by 4 weeks (5 days per week) of administration at
 

the dosage indicated.
 

11
 
referred to as the high dose group, and those initially receiving the
 

!

.-Ttter concentration are referred to as the low dose group. Female
 

mice received initial concentrations of 1200 and 600 ppm. Throughout
 

this report those female mice initially receiving the former concentration are referred to as the high dose group, while those initially
 

receiving the latter concentration are referred to as the low dose
 

group. During week 9, the high and low concentrations administered
 

to the female mice were increased to 2000 and 1000 ppm, respectively.
 

In week 63 of the study, administration of sulfallate to both males
 

and females ceased for 1 week followed by 4 weeks of dietary administration. This cyclic pattern of sulfallate administration continued
 

for the remainder of the dosing period. The 78-week dosing period
 

was followed by a 12- to 13-week observation period during which the
 

mice received only the basal laboratory diet and water.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. Body weights, food consumption, and data concerning
 

appearance, behavior, signs of toxic effects, and incidence, size,
 

and location of tissue masses were recorded at weekly intervals for
 

the first 10 weeks and at monthly intervals thereafter. From the
 

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

presence of tissue masses was determined by observation and palpation
 

of each animal.
 

12
 
During the course of this bioassay several pathology protocols
 

were in effect, each for different periods of time. The minimum protocol required that, if possible, certain tissues were to be taken
 

and examined histopathologically from all control animals, from any
 

animal in which a tumor was observed during gross examination, and
 

from at least 10 grossly normal males and 10 grossly normal females
 

from each dosed group. In addition, any tissues showing gross
 

abnormalities were to be taken and examined histopathologically.
 

Under later protocols, some tissues were taken from additional dosed
 

animals. The number of animals in each group from which a tissue
 

was examined is indicated in Appendices A through D.
 

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 exsanguination under sodium
 

pentobarbital anesthesia, and were immediately necropsied. The histopathologic examination consisted of gross and microscopic examination
 

of major tissues, organs, and gross lesions taken from sacrificed animals and, whenever possible, from animals found dead.
 

Tissues were preserved in 10 percent buffered formalin, embedded
 

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

to microscopic examination. An occasional section was subjected to
 

special staining techniques for more definitive diagnosis.
 

Slides were prepared from the following tissues from selected
 

animals: brain, pituitary, spinal cord, eye, adrenal, thyroid,
 

13
 
mesenteric lymph node, heart, lung, spleen, liver, kidney, stomach,
 

small intestine, large intestine, pancreas, urinary bladder, prostate
 

or uterus, testis with epididymis or ovary, skin with mammary gland,
 

muscle, bone marrow, and tissue masses. .
 

A few tissues were not examined for some animals, particularly
 

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

preclude histopathologic interpretation. Thus, the number of animals
 

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

animals that were placed on experiment in each group.
 

H.	 Data Recording and Statistical Analyses
 

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

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

as recommended by the Internationa] 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.
 

14
 
Probabilities of survival were estimated by the product-limit
 

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

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

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

missing; animals dying from natural causes were not statistically
 

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

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

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

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

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

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

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

histologically. However, when macroscopic examination was required
 

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.
 

15
 
As a part of these analyses, the one-tailed Fisher exact test (Cox,
 

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

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

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

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

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

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

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

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

noted, the direction of the significant trend was a positive dose 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
 

16
 
tumor was found, comparisons were based exclusively on animals that
 

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

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

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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

during which animals died naturally or were sacrificed was entered as
 

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

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

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

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

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

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


tailed test) were also noted.
 

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
 

17
 
^
 and the proportion in a control group corresponds to a relative risk
 

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

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

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

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

would be within the interval calculated from the experiment. When
 

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

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

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

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

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

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

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

detected under the conditions of this test.
 

18
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Distinct dose-related mean body weight depression was evident in
 

male and female rats from the beginning of the bioassay and generally
 

continued throughout the study (Figure 2). Fluctuations in the growth
 

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

the mean body weight may be subject to wide variations.
 

During the first 6 months of the study no clinical signs were
 

observed except for occasional hunched or thin appearance, squinted
 

or reddened eyes, and labored respiration in a few rats in each group.
 

From week 30 to cessation of chemical administration in week 78, an
 

unusually high incidence of signs characteristic of eye irritation
 

(i.e., red-tinged lacrimation; swollen, protruding, or squinted eyes;
 

brown crust around the eyes) were observed in the dosed groups,
 

particularly in the females.
 

Respiratory signs characterized by labored respiration, wheezing
 

and/or nasal discharge were noted at a low incidence in all groups
 

including the controls. The incidence of this observation increased
 

as the animals aged. Other signs usually associated with aging, which
 

were noted at comparable rates in control and dosed rats, included
 

body sores, alopecia, reddish discharge or brown crust around body
 

orifices, and abdominal urine stains. Observations in one or two rats
 

in each group included transient or sporadic incoordination, salivation, head tilt, and tremors.
 

19
 
750 750 

-600 

-450 

-300 

-150 

750 

TIME ON TEST (WEEKS) 

120 

750 

-600 

-450 

— 300 

— 150 

TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR SULFALLATE CHRONIC STUDY RATS 

20 

120 
Tissue masses, palpable nodules and/or swollen areas were observed at a slightly greater frequency among the dosed rats than
 

among the controls.
 

B.	 Survival
 

The estimated probabilities of survival for male and female rats
 

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

For both males and females the Tarone test indicated a significant
 

(P < 0.001) positive association between dosage and mortality.
 

There were adequate numbers of male and female rats at risk from
 

late-developing tumors. In the males 54 percent (27/50) of the high
 

dose, 74 percent (37/50) of the low dose, and 84 percent (42/50) of
 

the control rats survived on test at least 90 weeks. In the females
 

60 percent (30/50) of the high dose, 86 percent (43/50) of the low
 

dose, and 90 percent (45/50) of the control group survived on test
 

at least 90 weeks. For both sexes the surviving dosed rats were
 

sacrificed in weeks 103 and 104 while the surviving control rats were
 

sacrificed in week 111.
 

C.	 Pathology
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

summarized in Appendix C (Tables Cl and C2).
 

Long-term dietary intake of sulfallate in rats was associated
 

with increased incidences of neoplasia, especially of squamous-cell
 

21
 
PROBABILITY OF SURVIVAL PROBABILITY OF S

o o o o o 

o CD o i* a> 

! I I I 

I 

s

3) 

O 

O 

•o 

1 

v> 

O 

O 

Z 

m m m m 

7S 

C/l O

I 

30 

O 

J

' ,rsH > 

3)

3 > 

C/) 

I T o o o o o o o O) o 
carcinomas, in several organ systems. These tumors were seen in the
 

skin and adnexal tissues, the esophagus and forestotnach, and the
 

thyroid gland in bo^h sexes and in the mammary gland in females.
 

The incidence of esophageal and gastric lesions are tabulated
 

below for dosed rats. None of the neoplasms were observed in control
 

animals.
 

Males Females
 

Low High Low High
 

Dose Dose Dose Dose
 

Esophagus
 

Papilloma 0/26 1/21 0/44 0/28
 

Squamous-Cell Carcinoma 0/26 1/21 0/44 1/28
 

Forestomach
 

Papilloma 0/30 3/25 0/43 2/35
 

Squamous-Cell Carcinoma 0/30 2/25 0/43 0/35
 

Acanthosis 7/30 7/25 1/43 9/35
 

Hyperkeratosis 7/30 6/25 1/43 6/35
 

The incidences of acanthosis and hyperkeratosis shown in this table
 

were those in animals without stomach tumors. Microscopically, the
 

papillomas appeared as pedunculated growths showing marked localized
 

acanthosis and hyperkeratosis and projecting into the lumen. The
 

squamous-cell carcinomas, on the other hand, consisted of areas of
 

acanthosis and hyperkeratosis with a downward proliferation of anaplastic basal cells invading lamina propria and submucosa. Basal
 

cells appeared hyperbasophilic with numerous mitotic figures and grew
 

in disorganized sheets of varying-sized cells.
 

23
 
In the integument, squamous-cell carcinomas were observed in
 

2/48 (4 percent) low dose males, 1/45 (2 percent) high dose males,
 

1/50 (2 percent) low dose females and 1/48 (2 percent) high dose
 

females. Squamous-cell carcinomas also occurred in the external ear
 

of 1/50 (2 percent) low dose females and 2/48 (4 percent) high dose
 

females. None of these neoplasms occurred in control animals.
 

Primary squamous-cell carcinomas of the thyroid occurred in 2/21
 

(10 percent) high dose males. A squamous-cell carcinoma metastasized
 

to the thyroid from the stomach in one high dose male. Microscopically, both tumors were composed of proliferating anaplastic epithelial
 

cells. One was a small discrete lesion consisting of neoplastic cells
 

sequestered in collagenous tissue and the other, a large proliferating
 

structure producing masses of keratin and destroying the normal architecture of the thyroid.
 

Mammary adenocarcinomas were observed in 0/50 control females,
 

7/50 (14 percent) low dose females, and 11/48 (23 percent) high dose
 

females. The first mammary carcinoma was observed in the high dose
 

group in week 39. These tumors were characterized microscopically
 

by the presence of irregularly formed acini lined by anaplastic
 

epithelial cells, often several cells in thickness, with papillary
 

projections into the lumen. Some were highly cellular, composed
 

of tightly packed epithelial cells growing in sheets or forming tiny
 

acini and filled with eosinophilic amorphous material. Broad bands
 

24
 
of collagenous connective tissue intersected these tumors and often
 

massive areas of ischemic necrosis were seen containing islands of
 

viable neoplastic tissue around blood vessels. Metastatic lesions
 

were similar in morphology to those observed at the primary site.
 

A variety of other neoplastic lesions was also observed in
 

the dosed and control animals. These lesions probably represented
 

spontaneously occurring tumors.
 

The incidence of inflammatory, degenerative, and hyperplastic
 

lesions in the control rats was similar to that of the dosed rats
 

with the exception of the following: in the kidney, minimal changes
 

of toxic tubular nephropathy occurred in 1/31 (3 percent) low dose
 

males, 9/22 (41 percent) high dose males, and 1/28 (4 percent) high
 

dose females. This change was characterized microscopically by
 

the presence of megalocytic tubular epithelial cells having abundant
 

eosinophilic cytoplasm and large hyperchromatic nuclei. These
 

occurred in varying numbers affecting tubules of the inner cortex.
 

Often this change was present in conjunction with spontaneous chronic
 

interstitial nephritis; however, it was felt to be a toxic change
 

rather than part of the spectrum of changes seen in lesions of
 

the kidneys of aging rats.
 

In the eye, inflammatory and degenerative changes were observed
 

in somewhat greater frequency in dosed rats, especially females,
 

than in control rats.
 

25
 
Based on the results of this histopathologic examination, evidence was provided for the carcinogenicity of sulfallate in Osborne-


Mendel rats under the conditions of this study. Administration of
 

the compound was associated with an increased incidence of mammary
 

adenocarcinomas in females and squamous-cell carcinomas of the skin,
 

adnexal tissues, esophagus, and stomach in males and females.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

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

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

tumors were observed in at least one of the control or sulfallatedosed groups and where such tumors were observed in at least 5 percent of the group.
 

In females mammary adenocarcinomas were observed in 7/50 (14 percent) of the low dose rats, and in 11/48 (23 percent) of the high
 

dose rats, but not among the control rats. The Cochran-Artnitage test
 

indicated a significant (P = 0.001) positive association between
 

dosage and the incidence of mammary adenocarcinomas. The results of
 

the Fisher exact tests supported this finding with significant comparisons of both low dose (P = 0.006) and high dose (P < 0.001) to
 

the control. In historical control data collected by Hazleton
 

Laboratories for the NCI Carcinogenesis Testing Program, only 9/350
 

(3 percent) of the untreated female Osborne-Mendel rats had mammary
 

adenocarcinomas.
 

26
 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH SULFALLATE*
 

LOW 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE 

Stomach: Squamous-Cell Carcinoma 0/46(0.00) 0/30(0.00) 

P Values0 N.S. N.S. 

Relative Risk (Control) _— 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Stomach: Papilloma NOS, Squamous-Cell 

Papilloma, or Squamous-Cell Carcinoma^3 0/46(0.00) 0/30(0.00) 

P Values0 P ­ 0.003 N.S. 

Departure from Linear Trend P = 0.034 

Relative Risk (Control) 

Lower Limit —— 

Upper Limit 

Weeks to First Observed Tumor 

Kidney: Mixed Tumor Malignant 0/47(0.00) 2/31(0.06) 

P Values0 N.S. N.S. 

Departure from Linear Trend P = 0.037 

Relative Risk (Control) Infinite 

Lower Limit 0.450 

Upper Limit Infinite 

Weeks to First Observed Tumor 77 

HIGH
 

DOSE
 

2/25(0.08)
 

N.S.
 

Infinite
 

0.548
 

Infinite
 

66
 

5/25(0.20)
 

P - 0.004
 

Infinite
 

2.346
 

Infinite
 

66
 

0/22(0.00)
 

N.S.
 
__ _ 

___ 

TOPOGRAPHY: MORPHOLOGY
 

Pituitary: Chromophobe Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Carcinoma 

NJ P Values0 

oo 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Adenoma or
 

Follicular-Cell Carcinomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

CONTROL
 

4/41(0.10)
 

N.S.
 

108
 

4/48(0.08)
 

N.S.
 

106
 

5/48(0.10)
 

N.S.
 

106
 

LOW
 

DOSE
 

2/30(0.07)
 

N.S.
 

0.683
 

0.065
 

4.406
 

85
 

1/27(0.04)
 

N.S.
 

0.444
 

0.009
 

4.158
 

104
 

6/27(0.22)
 

N.S.
 

2.133
 

0.593
 

7.888
 

95
 

HIGH
 

DOSE
 

3/20(0.15)
 

N.S.
 

1.538
 

0.244
 

8.040
 

103
 

2/21(0.10)
 

N.S.
 

1.143
 

0.109
 

7.172
 

103
 

2/21(0.10)
 

N.S.
 

0.914
 

0.092
 

4.987
 

103
 
_ __ 

___ 

_ __ 

___ 

TOPOGRAPHY: MORPHOLOGY
 

Thyroid: Squamous-Cell Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Carcinoma
 

P Values0
 

fsj J
 

10

 Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma*3
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

CONTROL
 

0/48(0.00)
 

N.S.
 

0/48(0.00)
 

N.S.
 

0/48(0.00)
 

P = 0.034
 

LOW
 

DOSE
 

0/27(0.00)
 

N.S.
 

2/27(0.07)
 

N.S.
 

Infinite
 

0.528
 

Infinite
 

104
 

4/27(0.15)
 

P = 0.014
 

Infinite
 

1.660
 

Infinite
 

104
 

HIGH
 

DOSE
 

2/21(0.10)
 

N.S.
 

Infinite
 

0.683
 

Infinite
 

1/21(0.05)
 

N.S.
 

Infinite
 

0.123
 

Infinite
 

101
 

2/21(0.10)
 

N.S.
 

Infinite
 

0.683
 

Infinite
 

59 

101 
TABLE 3 (CONTINUED) 

TOPOGRAPHY : MORPHOLOGY CONTROL 

Brain: Astrocytoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

0/47(0.00) 

N.S. ___ 

Weeks to First Observed Tumor 

OJ 

o 

Subcutaneous Tissue: Hemangiosarcoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

4/49(0.08) 

P = 0.014(N) 

Weeks to First Observed Tumor 72 

Spleen: Hemangiosarcoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

4/47(0.09) 

N.S. ___ 

90 

LOW
 

DOSE
 

2/27(0.07)
 

N.S.
 

Infinite
 

0.520
 

Infinite
 

104
 

0/48(0.00)
 

N.S.
 

0.000
 

0.000
 

1.100
 

4/29(0.14)
 

N.S.
 

1.621
 

0.324
 

7.967
 

101
 

HIGH
 

DOSE
 

1/22(0.05)
 

N.S.
 

Infinite
 

0.115
 

Infinite
 

82 "*
 

0/45(0.00)
 

N.S.
 

0.000
 

0.000
 

1.171
 

2/20(0.10)
 

N.S.
 

1.175
 

0.111
 

7.334
 

101
 
TABLE 3 (CONCLUDED)
 

Treated groups received time-weighted average doses of 250 or 410 ppm in feed.
 

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

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

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

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

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

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

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

in the control group.
 

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

The probability level of the test for departure from linear trend is given beneath the con­ trol group when P < 0.05.
 
_ __ 

___ 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH SULFALLATE*
 

TOPOGRAPHY : MORPHOLOGY
 

Liver: Neoplastic Nodule
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

U) Stomach: Papilloma NOS
 

ho
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

1/50(0.02)
 

N.S.
 

—_
 

Ill
 

0/50(0.00)
 

N.S.
 

_—
 

15/50(0.30)
 

N.S.
 

100
 

LOW
 

DOSE
 

3/47(0.06)
 

N.S.
 

3.191
 

0.267
 

163.836
 

104
 

0/43(0.00)
 

N.S.
 

9/44(0.20)
 

N.S.
 

0.682
 

0.293
 

1.484
 

82
 

HIGH
 

DOSE
 

2/31(0.06)
 

N.S.
 

3.226
 

0.176
 

183.955
 

104
 

2/35(0.06)
 

N.S.
 

Infinite
 

0.426
 

Infinite
 

4/26(0.15)
 

N.S.
 

0.513
 

0.135
 

1.408
 

91 

103 
___ 

_ __ 

_ __ 

TOPOGRAPHY : MORPHOLOGY
 

Thyroid: Follicular-Cell Adenoma or
 

Follicular-Cell Carcinoma13
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma or C-Cell
 UJ Carcinoma15
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

CONTROL
 

1/50(0.02)
 

N.S.
 

Ill
 

5/50(0.10)
 

N.S.
 

Ill
 

0/50(0.00)
 

P = 0.001
 

LOW
 

DOSE
 

3/44(0.07)
 

N.S.
 

3.409
 

0.285
 

174.727
 

103
 

3/44(0.07)
 

N.S.
 

0.682
 

0.111
 

3.288
 

104
 

7/50(0.14)
 

P = 0.006
 

Infinite
 

1.944
 

Infinite
 

88
 

HIGH
 

DOSE
 

*
 

0/28(0.00)
 

N.S.
 

0.000
 

0.000
 

32.810
 

1/28(0.04)
 

N.S.
 

0.357
 

0.008
 

2.942
 

104
 

11/48(0.23)
 

P < 0.001
 

Infinite
 

3.459
 

Infinite
 

39
 
___ 

___ 

___ 

___ ___ 

TOPOGRAPHY : MORPHOLOGY
 

Mammary Gland: Fibroadenoina
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polyp
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Brain: Astrocytoma
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

CONTROL
 

15/50(0.30)
 

N.S.
 

P = 0.039
 

87
 

4/49(0.08)
 

N.S.
 

Ill
 

0/50(0.00)
 

N.S.
 

LOW
 

DOSE
 

21/50(0.42)
 

N.S.
 

1.400
 

0.784
 

2.544
 

89
 

3/46(0.07)
 

N.S.
 

0.799
 

0.122
 

4.463
 

104
 

1/45(0.02)
 

N.S.
 

Infinite
 

0.060
 

Infinite
 

103
 

HIGH
 

DOSE
 

10/48(0.21)
 

N.S.
 

0.694
 

0.310
 

1.483
 

77
 

2/28(0.07)
 

N.S.
 

0.875
 

0.083
 

5.631
 

77
 

2/26(0.08)
 

N.S.
 

Infinite
 

0.570
 

Infinite
 

76 
TABLE 4 (CONCLUDED)
 

TOPOGRAPHY: MORPHOLOGY CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Subcutaneous Tissue: Hemangiosarcoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

3/50(0.06) 

P = 0.034(N) ___ 

0/50(0.00) 

N.S. 

0.000 

0.000 

1.664 

0/48(0.00) 

N.S. 

0.000 

0.000 

1.730 

Weeks to First Observed Tumor Ill 

Spleen: Hemangiosarcoma 

P Values0 

0/50(0.00) 

N.S. 

3/45(0.07) 

N.S. 

1/28(0.04) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Infinite 

0.669 

Infinite 

Infinite 

0.096 

Infinite 

Weeks to First Observed Tumor 86 104 

Treated groups received time-weighted average doses of 250 or 404 ppm in feed.
 

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

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

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

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

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

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

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

in the control group.
 Q
 

The probability level of the test for departure from linear trend is given beneath the control group when P < 0.05.
 
In male rats when the incidences of C-cell adenomas or C-cell
 

carcinomas of the thyroid were combined, the Cochran-Annitage test
 

indicated a significant (P = 0.034) positive dose-response association.
 

The Fisher exact test comparing low dose to control supported this
 

finding with a significant (P = 0.014) result, but the comparison of
 

high dose to control was not significant. Only 27 thyroids in the
 

low dose group and 21 thyroids in the high dose group were examined
 

microscopically. Squamous-cell carcinomas, follicular-cell adenomas,
 

and follicular-cell carcinomas were also observed, but not in statistically significant proportions. In the historical controls 32/352
 

(9 percent) of the untreated male Osborne-Mendel rats had either a
 

follicular-cell adenoma or a follicular-cell carcinoma of the thyroid
 

and 12/352 (3 percent) had either a C-cell adenoma or a C-cell
 

carcinoma. No conclusion has been drawn relative to the thyroid
 

C-cell neoplasms in the males because of the difficulty in interpreting
 

the lack of a statistically significant high dose result and tae
 

absence of information as to whether the dosed animals selected for
 

histopathologic examination represented a random sampling of the
 

dosed groups.
 

In males a significant number of stomach neoplasms were observed
 

in the high dose rats. When incidences were combined so that the
 

numerator represented males with either a papilloma NOS,a squamouscell papilloma, or a squamous-cell carcinoma, the Cochran-Armitage
 

test indicated a significant (P = 0.003) dose-response association.
 

36
 
The departure from linear trend was significant (P = 0.034), principally because the neoplasms were only observed in the high dose rats.
 

The Fisher exact test comparing high dose to control was also significant (P = 0.004). In the historical control data none of the 352 rats
 

had a tumor of these types, compared to the 5/25 (20 percent) observed
 

in the high dose males for this bioassay.
 

Based upon these statistical results the administration of sulfallate was associated with increased incidences of mammary adenocarcinomas in female rats and of stomach neoplasms in male rats.
 

The possibility of a negative association between administration
 

and incidence was observed for hemangiosarcomas of the subcutaneous
 

tissue in both males and females, but in both cases the Fisher exact
 

tests were not significant.
 

37
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Distinct dose-related mean body weight depression was evident
 

throughout the study for male mice and after week 10 for female mice
 

(Figure 4). Fluctuations in the growth curve may be due to mortality;
 

as the size of the group diminishes, the mean body weight may be subject to wide variations.
 

Physical appearance and behavior of dosed mice during the first
 

year of the study gave no evidence of compound-related effects. Signs
 

often observed in group-housed laboratory mice, particularly in the
 

males, were observed at a comparable rate in dosed and control mice.
 

These common signs included sores on the body and/or extremities; a
 

hunched appearance; localized alopecia; penile, vulvar or anal irritation; and rough or stained fur.
 

A hunched or thin appearance was observed with greater frequency
 

in the dosed groups than in the controls from week 62 to termination
 

of the study in week 90. The incidence of palpable nodules, tissue
 

masses and/or swollen areas of the body was slightly greater in the
 

dosed mice than in the controls.
 

B.	 Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and sulfallate-dosed groups are shown in Figure 5. For
 

males no dose-mortality association was demonstrated, but for females
 

38
 
i 1 r T

 

45 60 75 90 105 120

 

TIME ON TEST (WEEKS) 

120 

TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR SULFALLATE CHRONIC STUDY MICE 

39 
PROBABILITY OF SURVIVAL PROBABILITY OF SUR

c3 0 O 0 0 c3 O P O 

i: 3 k) -f 0> 00 C3 cD kj ^ 13) 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

o — 0— 

-

i - 1 

1 

1 r j

c/> a'~ 1 1 Ul

 

c i r n i i

n

- I 

X —j 

3D 

s i 1 I < 

1 1 ? ' I i ^ 

s- 

O °~ 

O 

S 11 -O ­ |r 

3D 1 rr 

§ H f t _ d S- 

^ 7 

GO =• i r 

0 °-n ° Li 

r

1

 •

m 

c 25 H C3 r~ G m _H !' m -n 30 c/5 o> _ CO 05 _ 

> m H ° f* • H ° 

_-" ^.;" s 

m —4 m m i s - 

^n ^ ^ • 7;

o 52 

mf 

c/) 

I -j_ ^f ~

jf**

* |* •vj 

30 °> ••*1^^^ iJ"i* Ul 

O 

J .../ -"' | 0 

CO 

1 g- i11"

1

 ,"-;"

;

 r g- J

O 

m -n 

m 

$ r~ o — o— CJl i~ 01 ^m

m <: «J o 

O m 

m 

0 1 | t j 1 | i | i o i | i | i | 

D O O O o —' c3 o o p 

3 k) *. in bo o c3 fo 4i (31 
the Tarone test indicated a significant (P < 0.001) positive association between dosage and mortality.
 

For males sufficient numbers survived sufficiently long to be at
 

risk from late-developing tumors, as 92 percent (46/50) of the high
 

dose, 86 percent (43/50) of the low dose, and 80 percent (16/20) of
 

the control mice survived on test at least 75 weeks. For females the
 

survival was also adequate as 44 percent (22/50) of the high dose,
 

74 percent (37/50) of the low dose, and 20/20 of the control mice
 

survived on test at least 75 weeks.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

summarized in Appendix D (Tables Dl and D2).
 

Long-term dietary administration of sulfallate to mice was
 

associated with increased incidences of neoplastic lesions affecting
 

several organs and tissues (i.e., skin and subcutaneous tissue, lung,
 

liver, stomach, and mammary gland).
 

Mammary adenocarcinomas occurred in 33/48 (69 percent) low dose
 

females and 27/46 (59 percent) high dose females. In 10 of the 33
 

low dose females and 16 of the 27 high dose females the adenocarcinomas were accompanied by squamous metaplasia. No mammary adenocarcinomas were observed in the corresponding controls. Microscopically,
 

the morphology of these mammary carcinomas consisted of numerous
 

41
 
irregular acini and papillary structures which were lined by anaplastic epithelial cells sometimes in multiple layers. In a number of
 

mammary tumors, large areas of squamous differentiation were observed.
 

Neoplasms were usually intersected by dense fibrous connective tissue,
 

and large cystic areas of necrosis were noted. When mammary adenocarcinomas metastasized, the metastatic lesions observed in other organs
 

were similar in morphology to the primary tumors.
 

Squamous-cell carcinomas of the skin or subcutaneous tissue,
 

frequently adjacent to the preputial gland, occurred in 3/46 (7 percent) low dose males, 6/49 (12 percent) high dose males, and 2/48 (4
 

percent) low dose females. Basal-cell carcinomas of the skin occurred
 

in 1/49 (2 percent) high dose males and 1/46 (2 percent) high dose females. None of these types of tumors were recognized in control mice.
 

Papillomas of the forestomach occurred in 4/40 (10 percent) low
 

dose males, 3/43 (7 percent) high dose males, and 1/43 (2 percent)
 

low dose females. Squamous-cell carcinomas of the forestomach were
 

observed in 2/40 (5 percent) low dose males, 1/43 (2 percent) high
 

dose males, and 1/38 (3 percent) high dose females. No stomach tumors were observed in control mice. Microscopically, the papillomas
 

occurred as pedunculated growths composed of hyperkeratosis and
 

acanthosis of the superficial epithelium which was supported on a
 

stalk of well-vascularized fibrous connective tissue. The squamouscell carcinomas demonstrated invasiveness, documented by the presence
 

of small nests of anaplastic epithelial cells in the submucosa and in
 

42
 
some instances, local vascular emboli. A few male and female mice of
 •
 /
 

the high and low dose groups had hyperkeratosis and acanthosis in the
 

forestomach.
 

x
 

Hepatocellular carcinomas occurred in 7/20 (35 percent) control
 

males, 22/45 (49 percent) low dose males, 24/49 (49 percent) high
 

dose males, 0/19 control females, 5/47 (11 percent) low dose females, and 8/46 (17 percent) high dose females. The hepatocellular
 

carcinomas varied greatly in morphology; some were relatively well-


differentiated and composed of proliferating basophilic hepatocytes
 

forming liver plates one or more cells in thickness, and these
 

growths compressed adjacent hepatic parenchyma. The anaplastic
 

carcinomas consisted of proliferating large intensely basophilic
 

staining hepatocytes forming pseudoacini and thick blunted liver
 

plates which often projected into ectatic sinusoids. Metastases
 

appeared essentially similar to the primary liver tumor.
 

Alveolar/bronchiolar adenomas occurred in 11/43 (26 percent) low
 

dose males, 9/45 (20 percent) high dose males, 1/19 (5 percent) control females, 11/45 (24 percent) low dose females, and 2/37 (5 percent)
 

high dose females. Alveolar/bronchiolar carcinomas were observed in
 

3/43 (7 percent) low dose males, 9/45 (20 percent) high dose males,
 

4/45 (9 percent) low dose females, and 2/37 (5 percent) high dose
 

females.
 

Subcutaneous fibrosarcomas occurred in 5/46 (11 percent) low dose
 

males and 2/46 (4 percent) high dose females. Metastatic fibrosarcoma
 

43
 
occurred in the cervical lymph node of 1/41 (2 percent) low dose
 

males. A fibrosarcoma involving muscles of the leg occurred in 1/46
 

(2 percent) low dose males. None of these tumor types were observed
 

in control mice.
 

A variety of other neoplastic lesions, apparently unrelated to
 

chemical administration, was seen throughout all groups.
 

A variety of nonneoplastic lesions was also observed in both
 

dosed and control groups, regardless of experimental regimen. However, 26/42 (62 percent) low dose males, 35/43 (81 percent) high dose
 

males, 10/44 (23 percent) low dose females and 26/37 (70 percent)
 

high dose females had evidence of minimal toxic tubular nephropathy.
 

This change was characterized by the presence of megalocytic tubular
 

epithelial cells, with abundant eosinophilic cytoplasm and large
 

hyperchromatic nuclei. These occurred in varying numbers in the
 

inner renal cortex and in the absence of other changes.
 

Based on the results of this histopathologic examination, evidence was provided for the carcinogenicity of sulfallate in B6C3F1
 

mice under the conditions of this experiment. Administration of the
 

compound was associated with an increased incidence of mammary adenocarcinomas in females, squamous-cell carcinomas of the skin, adnexal
 

tissues, and stomach in males and females, lung tumors in males and
 

females, and hepatocellular carcinomas in females.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

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

44
 
___ 

TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH SULFALLATE*
 

TOPOGRAPHY: MORPHOLOGY
 

Skin, Subcutaneous Tissue, or Preputial
 

Gland: Squamous-Cell Carcinoma''
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Subcutaneous Tissue: Fibroma
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Subcutaneous Tissue: Fibrosarcoma
 

P Values
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/20(0.00)
 

N.S.
 

2/20(0.10)
 

P = 0.043(N)
 

91
 

0/20(0.00)
 

N.S.
 

P = 0.008
 

LOW
 

DOSE
 

3/46(0.07)
 

N.S.
 

Infinite
 

0.272
 

Infinite
 

71
 

3/46(0.07)
 

N.S.
 

0.652
 

0.083
 

7.437
 

91
 

5/46(0.11)
 

N.S.
 

Infinite
 

0.571
 

Infinite
 

53
 

HIGH
 

DOSE
 

6/49(0.12)
 

N.S.
 

Infinite
 

0.680
 

Infinite
 

91
 

0/49(0.00)
 

N.S.
 

0.000
 

0.000
 

1.372
 

0/49(0.00)
 

N.S.
 
TABLE

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar /Bronchiolar Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar Adenoma
 

or Alveolar/Bronchiolar Carcinoma'3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant Lymphoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

5 (CONTINUED) 

CONTROL
 

0/20(0.00)
 

P = 0.009
 

0/20(0.00)
 

P = 0.004
 

0/20(0.00)
 

N.S.
 

LOW
 

DOSE
 

3/43(0.07)
 

N.S.
 

Infinite
 

0.291
 

Infinite
 

90
 

14/43(0.33)
 

P = 0.002
 

Infinite
 

2.125
 

Infinite
 

79
 

3/46(0.07)
 

N.S.
 

Infinite
 

0.273
 

Infinite
 

91
 

HIGH
 

DOSE
 

9/45(0.20)
 

P = 0.028
 

Infinite
 

1.223
 

Infinite
 

87
 

17/45(0.38)
 

P = 0.001
 

Infinite
 

2.508
 

Infinite
 

65
 

3/49(0.06)
 

N.S.
 

Infinite
 

0.256
 

Infinite
 

47
 
___ 

__ _ 

TOPOGRAPHY : MORPHOLOGY
 

Circulatory System: Hemangiosarcoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Adenoma
 

or Hepatocellular Carcinoma**
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 5 (CONTINUED) 

CONTROL
 

0/20(0.00)
 

P = 0.024
 

7/20(0.35)
 

N.S.
 

91
 

7/20(0.35)
 

N.S.
 

91
 

LOW
 

DOSE
 

1/46(0.02)
 

N.S.
 

Infinite
 

0.024
 

Infinite
 

69
 

22/45(0.49)
 

N.S.
 

1.397
 

0.719
 

3.275
 

69
 

22/45(0.49)
 

N.S.
 

1.397
 

0.719
 

3.275
 

69
 

HIGH
 

DOSE
 

6/49(0.12)
 

N.S.
 

Infinite
 

0.679
 

Infinite
 

87
 

24/49(0.49)
 

N.S.
 

1.399
 

0.693
 

3.159
 

69
 

24/49(0.49)
 

N.S.
 

1.399
 

0.728
 

3.273
 

69
 
___ _ __ 

TABLE 5 (CONTINUED)
 

TOPOGRAPHY : MORPHOLOGY CONTROL
 

Stomach: Squamous-Cell Carcinoma 0/19(0.00)
 

P Values0 N.S.
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Stomach: Squamous-Cell Carcinoma,
 

Squamous-Cell Papilloma, or Papilloma
 

NOSb 0/19(0.00)
 CX3
 

P Values N.S.
 _.__
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Kidney: Tubular-Cell Adenoma 0/20(0.00)
 

P Values N.S.
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW
 

DOSE
 

2/40(0.05)
 

N.S.
 

Infinite
 

0.147
 

Infinite
 

91
 

6/40(0.15)
 

N.S.
 

Infinite
 

0.796
 

Infinite
 

91
 

0/42(0.00)
 

N.S.
 

HIGH
 

DOSE
 

1/43(0.02)
 

N.S.
 

Infinite
 

0.024
 

Infinite
 

91
 

4/43(0.09)
 

N.S.
 

Infinite
 

0.428
 

Infinite
 

91
 

3/43(0.07)
 

N.S.
 

Infinite
 

0.291
 

Infinite
 

86
 
TABLE 5 (CONCLUDED)
 

Q
 

Treated groups received time-x^eighted average doses of 949 or 1897 ppm in feed.
 

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

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

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

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

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

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

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

in the control group.
 

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

6
 

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

group when P < 0.05.
 
_ __ 

___ 

TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH SULFALLATE*
 

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar /Bronchiolar Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ul Lung: Alveolar /Bronchiolar Adenoma or
 

o
 Alveolar/Bronchiolar Carcinoma"3
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant Lymphoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/19(0.00)
 

N.S.
 

1/19(0.05)
 

N.S.
 

P = 0.008
 

—__
 

91
 

5/20(0.25)
 

P = 0.038(N)
 

91
 

LOW
 

DOSE
 

4/45(0.09)
 

N.S.
 

Infinite
 

0.408
 

Infinite
 

73
 

13/45(0.29)
 

P = 0.032
 

5.489
 

0.942
 

226.304
 

73
 

4/48(0.08)
 

N.S.
 

0.333
 

0.078
 

1.439
 

81
 

HIGH
 

DOSE
 

2/37(0.05)
 

N.S.
 

Infinite
 

0.157
 

Infinite
 

85
 

4/37(0.11)
 

N.S.
 

2.054
 

0.227
 

98.234
 

54
 

3/46(0.07)
 

P = 0.049(N)
 

0.261
 

0.046
 

1.228
 

54
 
TOPOGRAPHY: MORPHOLOGY
 

Circulatory System: Hemangiosarcoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower 'Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma NOS 

P Values0 

e 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 6 (CONTINUED) 

CONTROL
 

1/20(0.05)
 

N.S.
 

79
 

0/19(0.00)
 

P = 0.038
 

0/20(0.00)
 

N.S.
 

P < 0.001
 

LOW
 

DOSE
 

0/48(0.00)
 

N.S.
 

0.000
 

0.000
 

7.942
 

5/47(0.11)
 

N.S.
 

Infinite
 

0.534
 

Infinite
 

83
 

23/48(0.48)
 

P < 0.001
 

_—_
 

Infinite
 

3.268
 

Infinite
 

39
 

HIGH
 

DOSE
 

3/46(0.07)
 

N.S.
 

1.304
 

0.114
 

66.966
 

60
 

8/46(0.17)
 

N.S.
 

Infinite
 

0.990
 

Infinite
 

67
 

11/46(0.24)
 

P = 0.012
 

Infinite
 

1.506
 

Infinite
 

46
 
___ 

TABLE 6 (CONCLUDED) 

LOW HIGH
 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE DOSE
 

Mammary Gland: Adenocarcinoma with
 

Squamous Metaplasia" 0/20(0.00) 10/48(0.21) 16/46(0.35)
 

P Values0 P = 0.002 P = 0.022 P - 0.001
 

Relative Risk (Control) Infinite Infinite
 

Lower Limit 1.292 2.296
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 41 29
 

Mammary Gland : Adenocarcinoma with
 

Squamous Metaplasia or Adenocarcinoma NOSb 0/20(0.00) 33/48(0.69) 27/46(0.59)
 

NJ
 

Ul 

P Values0 P = 0.001 P < 0.001 P < 0.001
 

Departure from Linear Trend P < 0.001 _—
 

Relative Risk (Control) Infinite Infinite
 ———
 Lower Limit 4.830 4.061
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 39 29
 

Treated groups received time-weighted average doses of 908 or 1815 ppm in feed.
 

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

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

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

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

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

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

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

in the control group.
 

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

6

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

group when P < 0.05.
 
every type of malignant tumor in either sex where at least two such
 

tumors were observed in at least one of the control or sulfallatedosed groups and where such tumors were observed in at least 5
 

percent of the group.
 

For males when the incidences were combined so that the numerator
 

of the incidence represented mice with either an alveolar/bronchiolar
 

adenoma or an alveolar/bronchiolar carcinoma, the Cochran-Armitage
 

test indicated a significant (P = 0.004) positive association between
 

dosage and incidence. The Fisher exact test supported these findings
 

with significant comparisons of both high dose (P = 0.001) and low
 

dose (P = 0.002) to control. In historical control data collected by
 

this laboratory for the NCI Carcinogenesis Testing Program, 13/386 (3
 

percent) of the untreated male B6C3F1 mice had an alveolar/bronchiolar
 

adenoma; 2/386 (0.01 percent) had alveolar/bronchiolar carcinomas.
 

Alveolar/bronchiolar neoplasms were also observed in female
 

mice, but not in statistically significant incidences. The Fisher
 

exact test comparing the combined incidence of 13/45 (29 percent) in
 

the low dose to the 1/19 (5 percent) in the control resulted in a
 

probability level of P = 0.032, a marginal result which was not
 

significant under the Bonferroni criterion. It must be noted,
 

however, that the power of this test was low due to the small sample
 

size in the control and early deaths from tumors in the high dose
 

females. In the historical data, 6/380 (1.5 percent) of the untreated
 

female B6C3F1 mice had alveolar/bronchiolar adenomas, while 4/380 (1
 

percent) had alveolar/bronchiolar carcinomas.
 

53
 
A high incidence of dosed female mice with an adenocarcinoma
 

accompanied by squamous metaplasia of the mammary gland was noted.
 

These tumors were noted as early as week 29 of the study. The
 

Cochran-Armitage test indicated a significant (P = 0.002) dose-


response association and the Fisher exact tests indicated significant
 

comparisons of both low dose (P = 0.021) and high dose (P = 0.001) to
 

control. Additionally, a high incidence of dosed female mice with
 

mammary adenocarcinomas NOS in the absence of glandular squamous
 

metaplasia was observed. The Fisher exact tests were significant in
 

comparing both the low dose (P < 0.001) and the high dose (P = 0.012)
 

to the control group.
 

For males the Cochran-Armitage test indicated a significant (P =
 

0.024) positive association between dosage and the incidence of
 

hemangiosarcomas, but the Fisher exact tests were not significant.
 

Similarly, for females the Cochran-Armitage test was significant (P =
 

0.038) for the incidence of hepatocellular carcinomas, but the
 

Fisher exact tests did not support this finding. In the historical
 

data, 8/389 (2 percent) of the untreated males had a hemangiosarcoma
 

or a hemangioma, while 8/411 (2 percent) of the untreated female
 

B6C3F1 mice had a hepatocellular carcinoma.
 

The Cochran-Armitage test also indicated significant negative
 

associations between dosage and the incidence of malignant lymphomas
 

in females and the incidence of fibromas of the subcutaneous tissue
 

in males. None of these results, however, were supported by Fisher
 

exact tests under the Bonferroni criterion.
 

54
 
Based upon these statistical results the administration of
 

sulfallate was associated with the increased incidence of alveolar/
 

bronchiolar neoplasms in male mice and of mammary adenocarcinomas NOS
 

and of mammary adenocarcinomas with squamous metaplasia in female
 

mice.
 

55
 
V. DISCUSSION '
 

Under the conditions of this bioassay there were significant
 

positive associations between increased sulfallate concentration '• .
 

and accelerated mortality in both sexes of rats and in female mice.
 

In spite of early deaths with tumors, adequate numbers of animals
 

in all groups survived sufficiently long to be at risk from late-


developing tumors.
 

Among female rats, there was a statistically significant positive
 

association between dosage and the incidence of mammary adenocarcinomas. The incidence in each dosed group was significantly higher than
 

the incidence in the control group.
 

Among male rats, there was a statistically significant positive
 

association between dosage and the incidence of stomach tumors (i.e.,
 

combined incidences of squamous-cell carcinomas, squamous-cell papillomas, and papillomas NOS). The incidence of stomach tumors in the
 

high dose group was significantly higher than in the male rat control
 

group.
 

An increased incidence of thyroid tumors was observed among
 

male rats (i.e., 5/48 [10 percent] controls, 10/27 [37 percent] low
 

dose, and 7/21 [33 percent] high dose). These thyroid tumors included
 

squamous-cell carcinomas (both primary and metastatic), follicularcell adenomas, follicular-cell carcinomas, C-cell adenomas, and
 

C-cell carcinomas. In historical controls, 8/160 (5 percent) of the
 

untreated male Osborne-Mendel rats had thyroid tumors, all of which
 

56
 
were either follicular-cell adenomas or follicular-cell carcinomas.
 

Statistical calculations based on observed incidences of thyroid
 

tumors are of limited usefulness, however, since there is no indication that the 27 thyroids from low dose rats and 21 thyroids from
 

high dose rats were randomly selected for histopathologic examination. It is evident, nevertheless, that more thyroid tumors did
 

occur among dosed rats than among controls.
 

The occurrence of rare tumors of the esophagus (i.e., one papilloma and one squamous-cell carcinoma in high dose males and one
 

squamous-cell carcinoma in a high dose female) was observed, supporting
 

the finding of carcinomas of the forestomach. No esophageal tumors
 

were observed in control rats. In addition, a few squamous-cell carcinomas of the skin were observed in dosed but not in control rats.
 

Among female mice, the incidences of mammary adenocarcinomas were
 

significantly higher in both high and low dose groups than in controls. Many of these adenocarcinomas were accompanied by squamous
 

metaplasia.
 

When the incidence of alveolar/bronchiolar adenomas was combined
 

with the incidence of alveolar/bronchiolar carcinomas, there was a
 

statistically significant positive association between dosage and
 

tumor incidence for male mice. The combined incidence of these lung
 

tumors was significantly higher in each dosed group than in the male
 

mouse control group. When the incidences of alveolar/bronchiolar
 

57
 
adenomas or alveolar/bronchiolar carcinomas were analyzed separately,
 

the incidence of alveolar/bronchiolar adenomas in the low dose group
 

was significantly higher than that in the male mouse control group,
 

but other comparisons did not indicate statistically significant results. The incidences of these neoplasms in low dose females were
 

also increased but limited survival among high dose females and the
 

small size of the female control group contributed to the lack of
 

statistical significance in the occurrence of these tumors in female
 

mice.
 

Dietary administration of sulfallate to male and female mice
 

appeared to be associated with increased occurrences of uncommon neoplasms in several organs. These included squamous-cell and basal-


cell carcinomas of the skin, squamous-cell papillomas of the stomach
 

and squamous-cell carcinomas of the forestomach. None of these neoplasms occurred in statistically significant incidences.
 

The incidence of toxic tubular nephropathy observed in male rats
 

and in mice of both sexes increased with the concentration of the compound administered. This nonneoplastic lesion was not observed in
 

control animals.
 

Under the conditions of this bioassay, dietary administration of
 

sulfallate was carcinogenic to Osborne-Mendel rats and to B6C3F1 mice,
 

inducing mammary gland tumors in females of both species, tumors of
 

the forestomach in male rats, and lung tumors in male mice.
 

58
 
VI. BIBLIOGRAPHY
 

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, B.C., 1977.
 

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

Co., Ltd., London, 1970.
 

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

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

Farm Chemicals Handbook, Meister Publishing Company. Willoughby,
 

Ohio, 1976.
 

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

Tests, Confidence Limits, and Adjustments for Stratification."
 

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

Gosselin, R.E., B.C. Hodge, R.P. Smith, and M.N. Gleason, Clinical
 

Toxicology of Commercial Products, 4th edition. The Williams
 

and Wilkins Company, Baltimore, Maryland, 1976.
 

Innes, J.R.M., B.M. Ulland, M.G. Valerio, L. Petrucelli, L. Fishbein,
 

E.R. Hart, A.J. Pallotta, R.R. Bates, H.L. Falk, J.J. Gart, M.
 

Klein, I. Mitchell, and J. Peters, "Bioassay of Pesticides and
 

Industrial Chemicals for Tumorigenicity in Mice: A Preliminary
 

Note." Journal of the National Cancer Institute 42 (6): 1101-1114,
 

1969.
 

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

Observations." Journal of the American Statistical Association
 

5_3:457-481, 1958.
 

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

"Carcinogenesis Bioassay Data System." Computers and Biomedical
 

Research 7:230-248, 1974.
 

Martin, H., and C.R. Worthing, editors, Pesticide Manual, Fifth edition. British Crop Protection Council, 1977.
 

59
 
Miller, R.G., Simultaneous Statistical Inference. McGraw-Hill Book
 

Co., New York, 1966.
 

Reuber, M.D., and E.L. Glover, "Cirrhosis and Carcinoma of the Liver
 

in Male Rats Given Subcutaneous Carbon Tetrachloride." Journal
 

of the National Cancer Institute 44:419-423, 1970.
 

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.
 

Stanford Research Institute, 1977 Directory of Chemical Producers,
 

U.S.A., Menlo Park, California, 1977.
 

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

62:679-682, 1975.
 

60
 
Review of the Bioassay of Sulfallate* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DREW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

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

academia, industry, organized labor, public interest groups,
 State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

Subgroup of the Clearinghouse is charged with the responsi­ bility of providing a peer review of reports prepared on
 

NCI-sponsored bioassays of chemlc:- 3 studied for carcinogenic­ ity. It is in this context that tiie below critique is given
 

on the bioassay of Sulfallate for carcinogenicity.
 

The reviewer agreed with the conclusion in the report
 

that Sulfallate was carcinogenic in both rats and mice,
 under the conditions of test. After a brief description of
 

the experimental design, the reviewer noted that the dosages
 

had to be adjusted downward during the course of the chronic
 

study and that the compound had to be administered on an
 

intermittent schedule. She pointed out a number of neoplastic
 

lesions, in both the treated rats and mice, that were found
 

in increased but nonstatistically significant rates. She
 

opined that some of the tumor types may not have been
 

statistically significant because of the small number of
 

control animals. Despite the experimental shortcomings,
 the reviewer said the conclusion was still valid that
 

Sulfallate was carcinogenic in both sexes of treated rats
 

and mice. She added that Sulfallate would appear to pose a
 

potential carcinogenic risk to humans. The reviewer moved
 

61
 
that the report on the bioassay of Sulfallate be accepted as
 

written. The motion was approved without objection.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical U.S.A.
 Michael B. Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

*U.S. GOVERNMENT PRINTING OFFICE:1978 260-899/3m 1-3 62 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH SULFALLATE
 

TABLE A1

 

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

ANIMAL S INITIALL Y IN STUD Y 

ANIMAL S NECEOPSIE D 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY '»*

INTEGUMENTAR Y SYSTEM 

PAPILLOMA , NOS 

St^'AMOUS CELL CAPCINGM A 

FIBECSAECOM A 

HEMANGIOFEEICYTOMA, MALIGNANT
 

*SUBCUT TISSUE
 

SUUAMOUS CELL CARCINOMA
 FIBROMA
 

FIBRCSARCCMA
 

FIBFCUS HISTIOCYTOMA, MALIGNANT
 

HEMANGIOSARCOMA
 

RESPIRATCRY SYSIEK
 

*LUN G 

CARCINOMA , NOS, METASTATIC 

ALVECLAR/BROtiCHIOLA R ADENOM A 

MIXI D TUMOR, MALIGNAN T 

HEKANGIOSAhCOttA , HE1ASIATIC 

HEMATOEOIETIC SYSTEM
 

'MULTIPLE ORGANS
 

liALIGdAN T LYMPHOMA , NO S 

MALIG.LYKPHOflA , LYMPHOCY1IC

JSPLEE N 

SABCOMA , NOS 

HEMANGIOM A 

HEMANGIOSAECOM A 

*CERVICAL LYMPH NODE
 

t NUMBE R OF ANIMAL S WIT H TISSUE

* NUMBE R OF ANIMALS NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

 TYP E 

CONTROL (VEH ) 

01-M001 

50 

1*9 

 49 

(149) 

1 <2J)
 

(49)
 

1 (2*)
 

1 (2*)
 

4 (3*)
 

(149) 

1 (2%) 

2 (14%) 

(49) 

1 (2*) 

(47) 

4 (9%) 

(45) 

LOW DOSE 

01-M016 

50 

148 

38 

(1*8) 

1 (2X) 

2 («») 

1 <2X) 

(48)
 

1 (2JS)
 2 <i»X)
 

1 (2%)
 

(28) 

1 (4X) 

1 («X) 

(48) 

(29) 

1 (3%) 

4 (14%) 

(25) 

 EXAMINE D MICROSCOPICALLY 

A-3 

HIGH DOSE 

01-M017 

50 

45 

30 

(45) 

1 (2%) 

(45)
 

1 <2S)
 

1 (2*)
 

(21) 

(45) 

<2X) 

(20) 

1 15%) 

2 (10*) 

(22) 

1 (5X1 
CIKCUIA1CBY SYS1EH 

»HEAR T 

HEKANGIOSARCOM A 

HEMASGIOSAKCOKA , ME1ASTATIC 

DIGESTIV E SYSTZ H 

KSUBMAJCILLAR Y GLAU D 

SARCOMA , NOS 

FldSCSAHCOilA 

0LIVER 

HEMANGIOSARCOM A 

HEMANGIOSARCOMA , MEIASTATIC 

tESOPHAGU S 

PAPILLOBA, NOS 

SQUAKOU S CELL CARCINOM A 

ISTOMACH 

PAPILLOKA , DOS 

SQUAilOlJS CELL tAPILLOBA 

SQUAMOU S CELL CASCINOMA 

BEINAR Y SYSlEfl 

#KIDNI Y 

LIPOMA 

MIXE D TUMOR , MALIGNAN T 

HTKANGIObARCOM A 

tURINAR Y tiLADDER 

PAPILLOHA, NOS 

ENBOCRIN ! SYSTE M 

tPITUITAE Y 

CABCINOMA.NO S 

CHEOMOPH06E ADENOM A 

MEN1NGICM A 

#ADRE^A L 

CORTICAL ADENOM A __ 

t iNUBBE E OF ANIMAL S HUH TISSUE

* NUMEL R OF ANIMAL S NECROPSIED 

TABLE A1 (CONTINUED) 

CONTROL (VKH ) LOW DOSE 

01-M001 C1-MC1 6 

(t7) (28) 

1 (2*) 

1 (2%) 

(4) 

1 (25%) 

1 (25*) 

(U 9) (31) 

1 (2X) 

(D (26) 

(46) (30) 

(47) (31) 

2 (UX ) 

2 (6X) 

1 (2X) 

(46) (27) 

j (7X) 

(41) (30) 

1 <3X) <» (10X) 2 <7X) 

(»6) (28) 

^~« _.«.— 1 .1*81. 

 EXAMINE D MICROSCOPICALLY 

A-4 

rfIGH DOSE 

01-M017 

(21) 

(22) 

1 (5X) 

(21) 

1 (5X) 

1 (5X) 

(25) 

2 (8X) 

1 (*«) 2 (8X) 

(22) 

(21) 

(20) 

3 (15X) 

1 (5X) 

(20) 

.1.,J5St.- _ 
PHEOCHROKOCYTOH A 

UTriYKOID 

SQUAMOU S CEL L CARCINOM A 

SQUAMOU S CtLL CARCINOMA,

FOLIICULAR-CEL L ADENOM A 

FOLLICULAE-CELL CARCINOM A 

C-CELL ADENOM A 

C-CELL CARCINOM A 

• PARATHYROI D 

ADENOMA , NO S 

*PANCREATI C ISLETS 

ISLET-CELL ADENOM A 

REPRODUCTIVE SYSTEM 

*KAMMAR Y GLAN D 

ADENCCAECINOMA ,

FIbPOADENCM A 

tPROSTATE 

HEUANblOSARCOMA ,

*SEHINA L VESICLE 

HEMANOIOSAtCOMA ,

NERVOUS SYSTEM 

*bRAIH 

FIbl-CSARCOfi A 

GLIOCA . NOS 

ASIRCCYTCMA 

SPECIAL SENS E ORGAN S 

NONE 

 NO S 

 METASTA1IC 

 MEIASIATIC 

MUbCULCSKELIIA L SYSTE M 

'SKELETAL MUSCLE 

£lbROSARCOMA_ _ . 

* NUMBE R OF ANIMALS HTH TISSUE

* MiiBER OF ANIMAL S NECBOESItP 

TABLE A1 (CONTINUED) 

CONTRO L (VEH ) 

01-M001 

(48) 

 METASTA 

3 (6X) 

4 (S%) 

(46) 

1 (2X ) 

(46) 

1 (2*) 

(49) 

2 (U %) 

1 (2%) 

(34) 

1 (3X) 

(49) 

1 (2%) 

(47) 

1 (2*) 

(49) 

J_J2XJ 

 EXAMINED MICROSCOPICALL i 

A-5 

LO« DOSE HIG H DOSE 

01-M016 01-M017 

1 (4X) 1 (5X) 

(27) (21)

2 (10X) 

1 (5X) 

-5 (19*) 

1 (4X) 2 (IDS) 

3 (11*) 1 (5X) 

2 (7%) 1 (51) 

(3) (D 

(26) (21) 

(48) (45) 

(9) (1*) 

(48) (45) 

(27) (22) 

1 (4S) 

1 (5X) 

2 (1%) 1 (5X) 

(48) (45) 

. 
•MUSCLE

 OF THORAX

HEMANGIOSABCOMA

BODY CAVITIES 

• ABDOMINA L CAVIT Y

LIPOHA

"TUNICA VAGINALI S

MESOTHELIOMA , NOS

A L L OTHE B SYSTEMS 

•MULTIPLE CKGANS

HEMASGICSASCOM A

ANIrtA L EISPCSlTIOil SUKHAE Y 

ANIMALS INITIALLY IN STUDY

NATURA L DEATHS

MORIBUN D SACH1FICE

SCHEDULE D SACRIFICE 

ACCIDENTALLY KILLED 

TERMINA L SACRIFICE

ANIMA L HISSING 

* NUMBER OF ANIMALS WITH TISSUE

* NUMBER UF ANIMALS NECBOFSIED 

TABLE A1 (CONTINUED) 

CONTROL (VEH) LOW DOSE HIG H DOSE 

01-M001 01-8016 C1-M017 

 (19) (US) (15) 

1 (2X) 

 (19) (18) (15) 

1 (24) 

 (19) (18) (15) 

1 (2%) 2 (IX) 

 (19) (18) (15) 

1 (2X ) 

 50 50 50 

 21 30 37 

2 1 1 

 21 19 12 

 EXAMINE D MICROSCOPICALLY 

A-6 
TABLE A1 (CONCLUDED)
 

cZOSTRO L (VEH ) LOW DOSE HIGH DOSE 

01-M001 01-MOI b 01-M017 

TUMO R SUMMAR Y 

TOTA L ANIMAL S WITH PRIMAR Y TUMORS * 28 26 19 

TOTA L PRIBAR Y TUMOR S 41 3B 32

 

TOTA L ANIMAL b KITH BEMG N TUMOR S 13 1U 9 

TOTA L BENIG N TUMORS 17 16 10

 

TOTA L ANIftAL S KIT H MALIGNAN T TUMOR S 18 19 14

 

TOTAL MALIGNAN T TUMOR S 2<J 21 20 

TOTAL ANIMAL S N ITU SECONDAR Y TUMORS * I 1 1 

TOTAL SECONDAR Y TUMORS 6 1 1 

TOTA L ANIMAL S WIT H TUMORS UNCERTAIN ­

BENIGN OR MALIGNAN T 1 2

 

TOTA L UNCERTAI N TUMOR S 1 2 

TOTAL ANIMALS WITH TUMORS UNCERTAIN ­

PRIMARY OR METASTATIC
 

TOTAL UNCEPTUN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUBOES
 

* SECONDAFY TUMO&S: METASTATIC TUMORS OB TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-7
 
TABLE A2 

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

CONTBOL(VEH ) LOW DOSE HIGH DOSE 

01-F001 C1-FC16 01-F017 

ANIBAL S INITIALLY IN STUDY 50 50 50 

ANIBAL S NECROPSIED 50 50 18 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 50 47 39 

INTIGUNENTAEY SYSTFM 

*SKIN (50) (50) (48) 

SgUABOUS CELL CARCINOMA 1 (2*) 

•SUECUT TISSUE (50) (50) (18)
 

SQOAKOUS CELL CARCINOMA 1 (2X)
 EASAL-CEIL CARCISOKA 1 (2%)
 FIBROMA 2 (4X) 2 (KX)
 

FIBSCSARCOMA 1 (2*)<­

IIPOBA 1 (2X) 1 (2X)
 

HEHANGIOSABCOHA 3 (6*)
 

BESUBATCRY SYSTEM 

*LUNG (50) (44) (29)
 

SQUAHOUS CELL CARCINOHA, WETASTA 1 (3%)
 

ADEHOCARCINOMA, NOS. KETASTATIC 2 (7X)
 

HEMATOPOIETIC SYSTEH
 

*MULTIPLE ORGANS (50) (50) (48)
 

MALIG.LYdPHOMA , HISTIOCYTIC TYPF 2 (4X) 1 (2X) 

tSPLEEN (50) (45) (28)
 

SAFCOHA, NOS, METASTATIC 1 (4X)
 

HEBANGIOSARCOHA 3 (7X) 1 (4X)
 

tCEHTICAL LYHP H MODE (48) (43) (26) 

SQUAMOUS CELL CARCINOMA , BETASTA 1 (2X) 

BSLIG.LYBPHOBA, IYBPHOCYTIC TYPI? 1 (2X) 

CISCDLATORY SYSTEH 

* NOBBER OF ANIBALS WITH TISSUE EXABINE D BICROSCOPICALLY 

* NOBBER OF AMIHALS KECPOPSIED 

<- MDLTIPLE OCCURRENCE OF MORPHOLOGY IN THE SAK E ORGA N TISSBES IS COUNTED ONCE ONLY 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-8 
DIGESTIVE SYS1IM
 

(LIVER
 

NEOPLASTIC NODULE
 

(ESOPHAGUS
 

SQUAMOUS CELL CARCINOMA
 

(STOMACH
 

PAPILLOMA, NOS
 

(DUODENUM
 

HEMANGIOSARCOHA
 

*ANAL CP.NAL
 

HEHANGIOSARCOMA
 

UBINABY SYSTEH
 

(KIDNEY
 

IIPCBA
 

MIXED TUMOR, MALIGNANT
 

HAHARTCNA+
 

(URINARY ELADDEE
 

FIBROSARCOMA
 

ENECCEINE SYSTEM
 

(PITUITARY
 

CHROMOPHCBE ADENOMA
 

(ADFENAL
 

CORTICAL ADENOMA
 

(THYROID
 

SQDAMOU S CELL CAPCINOMA 

TABLE A2 (CONTINUED) 

CONTROL (VEH) 

01-F001 

(50) 

1 (2%) 

(50) 

(50)
 

1 (2%)
 

(50)
 

(50)
 

1 (2X)
 

1 (2*)
 

1 (2X)
 

(49)
 

(50)

15 (SOX) 

(50) 

(50) 

LOW DOSE 

01-F016 

3 (6%) 

(43)
 

(43)
 

(50)
 

1 (2X)
 

(t2)
 

C»t)

9 (20X) 

(U 5) 

(tU) 

HIGH DOSE 

01-F017 

(3D 

2 (6*) 

(28) 

(35)
 

2 (6X)
 

(28)
 

(48)
 

(28)
 

(28)
 

1 («*>
 

(26) 

1 (15*) 

(30) 

1 (3*) 

(28) 

1 (*%) 

ADENOMA , NOS 

FOLLICULAR-CELL ADENOMA 

FOLLICULAR-CELL CAPCINOMA (2X)

C-CELL ADENOMA (10%)

 

C-CELL CARCINOMA 

* NUMBEP OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

1

2

1

2 

(2*) (5*) 

(2X) (5X) 1 (HX) 

+ THIS IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TUMOR OF THE KIDNEY AND CONSISTS OF
 

PROLIFERATIVE LIPOCfTES, TUBULAR STRUCTURES, FIBROBLASTS, AND VASCULAR SPACES IN VARYING PROPORTIONS.
 

A-9
 
TABLE A2 (CONTINUED)
 

CONTROL (VEH) LOU DOSE 

01-F001 01-F016 

IPANCREATIC ISLETS (50) (44) 

ISLET-CELL ADENOM A 2 (5X) 

REPRODUCTIV E SYSTEM 

*MAHHARY GLAND (50) (50) 

ADENOMA, NOS 1 (2X) 

ADENOCARCINOMA , NOS 7 (14X) 

FIBROADENOMA 15 (30*) 21 (42X) 

•UTERUS (49) (46) 

FIBROSARCOBA 1 (2*) 

ENDOBETRIAL STROMAL POLYP 4 (8*) 3 (7*) 

*OVAR Y (49) (44)

 

CARCINOMA, NOS 1 <2X)

 

IUTEOM A 1 (2%) 

GRANOLOSA-CELL TUMO E 1 (2*) 1 (2*) 

HEMANGIOSARCOHA 1 (2*) 

NERVCU S SYSTE M 

* BRAI N (50) (45) 

ASTROCYTOM A 1 (2K) 

SPECIAL SESSE ORGAN S 

*EYF (50) (50) 

MALIGNAN T MELANOM A 1 (2%) 

FIBROSARCOMA 

*EXT5PNA L EAS (50) (50) 

SQUAKOOS CELL CARCINOM A 1 (2%) 

MOSCULOSKELETA L SYSTEM 

*MUSCLE OF HEAD (50) (50) 

SARCOMA , NOS 

FIBROSARCOMA 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMEER OF ANIMALS NECBOPSIED
 

A-IO
 

HIGH DOSE 

01-F017 

(28) 

(48) 

11 (23*) 

1" (2U) 

(28) 

2 (7*) 

(28) 

(26) 

2 (8X) 

(48) 

1 (2X) 

(48)

2 (4*) 

(48) 

1 (2X) 

1 12%) 
TABLE A2 (CONCLUDED)
 

CONTROL (VEH) LOW DOSE HIGH DOSE
 

01-F001 01-F016 01-F017
 

BODY CAVITIES
 

*AEDOMINJL VISCERA (50) (50) (48)
 

HEMANGIOSARCOMA 1 (2*)
 

ALL OIHES SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50
 

NATUFAL DEATHS It 15 33
 

MOBIBOND SACRIFICE 2 2
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 36 33 15
 

ANIMAL MISSING
 

S INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS HITH PEIHARY TUHOBS* 36 37 30
 

TOTAL PRIMARY TUMOFS 56 72 46
 

TOTAL ANIMALS KITH BENIGN TUMOES 31 32 16
 

TOTAL BENIGN TUMOFS 1!) 45 20
 

TOTAL ANIMALS HITH MALIGNANT TUMORS 9 18 23
 TOTAL MALIGNANT TUMORS 10 23 24
 

TOTAL ANIMALS HITH SECONDARY TUMORS* 1 4
 TOTAL SECONDARY TUMOES 1 U
 

TOTAL ANIMALS HITH TUMORS UNCERTAINBENIGN OR MALIGNANT 2 •4 2
 TOTAL UNCERTAIN TUMOES 2 4 2
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

A-ll
 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH SULFALLATE
 

TABLE B1

 

SUMMARY OF 1 HE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH SULFALLATE 

CONTROL (VEH) LOB DOSE HIGH DOSE 

02-M019 02-M020 02-M02 1 

ANIMALS INITIALLY IN STUDY 20 50 50
 

ANIMALS MISSING 1 1
 

ANIMALS NECROPSIED 20 16 19
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 20 46 49
 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (116) (49)
 

SQUAMOOS CELL CARCINOMA 3 (7*) 1 (2%)
 

BASAL-CELL CARCINOMA 1 (2X)
 

FIBROSARCOMA 1 (2%)
 

*SUBCUT TISSUE (20) (46) (49)
 

SQIIAMOUS CELL CARCINOMA 3 (6%)
 

EASAL-CELL CARCINOMA 1 (2%)
 

SEBACEOUS ADENOMA 1 (2%)
 FIBROMA 2 (1T%) 3 (7»)
 

FIBEOSARCOMA 5 (1U)
 

HEMANGIOSARCOMA 1 (2%) 2 (14%)
 

RFSFIEATOEY SYSTEM
 

tLUNG (20) (43) (45)
 

SQUAMOUS CFLL CARCINOMA 1 (2X)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (2%)
 HEPATOCELLULAR CARCINOMA, METAST 1 (2X) 1 (2%)
 

ALVEOLAR/BEONCHIOLAR ADENOMA 11 (26X) 9 (20*)
 

ALVEOLAR/BRONCHIOLAR CABCINOHA 3 (7%) 9 (20X)
 HEMANGIOSARCOMA, METASTATIC 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

*MUITIPLE ORGANS (20) (46) (49) 

MALIGNANT LYMPHOMA, NOS 1 (2*) 2 (4«) 

MALIG.LYMPHOMA, UNDIFFER-TYPE 1 (255) 

tCERVICAL IYMPH NODE (16) (41) (42) 

FIBROSARCOMA. MEIASIATIC
 

* NUMBE R OF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NHMEF R OF ANIMAL S NECBOPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
TABLE BI (CONTINUED)
 

CONTROL (VEH) LOW DOSE
 

02-M019 02-M020
 

•MESENTERIC L. NODE (16) (11)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2X)
 

• THYMUS (14) (40)
 

MALIGNANT LYMPHOMA, NOS
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

• LIVER (20) (45)
 

HEPATOCFLLULAR CARCINOMA 7 (35X) 22 (49X)
 

• STOMACH (19) (10)
 

PAPILLCMA, NOS
 

SQUAMOOS CELL PAPILLOMA 4 (10X)
 

SgOAKOUS CELL CARCINOMA 2 (5X)
 

*ANUS (20) (16)
 

FIBROSARCOMA. HETASTATIC 1 (2X)
 

UBINABY SYSTEM
 

•KIDNEY	 (20) (42)
 

HEPATOCEILULAR CARCINOMA. METAST 2 (5X)
 

TUBULAR-CELL ADENOMA
 

SARCOMA, NOS
 

HEMANGIOSABCONA
 

ENDOCBINF SYSTEM
 

•ADRENAL	 (16) (40)
 

CORTICAL ADENOMA 1 (3X)
 

•THYROID	 (17) (38)
 

FOLLICULAR-CELL CARCINOMA
 

REPRODUCTIVE SYSTEM
 

*PENIS (20) (46)
 

SQUAMOUS CELL CARCINO
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

B-4
 

HIGH DOSE
 

02-M021
 

(12)
 

(13)
 

1 <2X)
 

(19)
 

21 (49X)
 

(13)
 

3 (7X)
 

1 (2X)
 

(19)
 

(13)
 

3 (7X)
 

1 (2X)
 

3 (7X)
 

(41)
 

(41)
 

1 (2X)
 

(49)
 

1 (2*)
 
TABLE Bl (CONTINUED) 

CONTHOL(VEH ) LCH DCSE HIG H DOSE 

02-H019 G2-HC20 02-M021 

*PHEPU1IAL GLAND	 (20 (46 ) ) (49)
 

SQUAHOU S CELL CARCINOM A 2 (t») 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE CHGANS
 

•HARDEEIAN	 GLAND (20) («6) («9)
 

ADENCMA, NOS 1 (2*)
 

BUSCUICSKELETAL SYSTEM
 

*MUSCLE OF LEG (20) (U6) (U9)
 

FIBEObARCOKA, METASTATIC 1 (2X)
 

BODY CAVITIES
 

•AdDCMlNAL	 VISCERA (20) (<tb) (49)
 

HEMANGIOSABCOHA 1 (2X)
 

*MESENTER Y (20) («6) (19) 

SAHCCflA. NOS 1 (2J) 

A LL OTH£S SYSTEMS 

NON E 

ANIMA L DISFCSI1ION SUBMAE I 

ANIKAI S INITIALL Y IN STUDY 20 50 50
 

NATUBA L DEATHS 16 19
 

M08IBUSC SACRIFICE 1
 

SCHEDULE D SACRIFICE 

ACCIDENTALL Y KILLE D 

lERMINAi. SACRIFICE 32 30
 

A&IhA L MISSING 1 1
 

I NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMEiH OF AKIMALS NECBOFSIED
 

B-5
 
TABLE Bl (CONCLUDED)
 

CONTROL (VEH) LOW DOSS HIGH DOSE
 

02-M019 02-M020 02-M021
 

TUMOR SUMMARY
 

TOT9L iNIHALS HUH PRIMARY TUMORS* 8 37 39
 TOTAL PBIMAEY TUMORS 9 59 73
 

TOTAL ANIMALS WITH BENIGN TUMORS 2 16 14
 

TOTAL BENIGN TUMORS 2 19 17
 

TOTAL ANIMALS WITH MALIGNANT TUMO»S 7 32 34
 

TOTAL MALIGNANT TUMOBS 7 40 56
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 5 3
 

TOTAL SECONDARY TUMORS 6 3
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMOBS
 

TOTAL ANIMALS WITH TUMORS UNCEBTAIN­ PRISAPY OB HETASTATIC
 TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

I SECONDARY TUMORS: ME1ASTATIC TUMORS OR TUMORS INVASIVE INTO A N ADJACENT OSGAN
 

B-6
 
TABLE B2

 

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

CONTROL (VEH) LOW DOSE HIGH DOSE 

02-F019 02-7022 02-F023 

ANIMALS INITIALLY IN STUDY 20 50 50
 

ANIMALS NECBOPSIED 20 48 46
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 19 48 46
 

INTEGUMENTARY SYSTEM
 

*SUECUT TISSUE (20) (48) (46)
 SOOAMOUS CBLL CARCINOMA 2 (4*)
 

EASAL-CELL CARCINOMA 1 (2X)
 

FIBFOSABCOMA 2 (4X)
 

HEHANGIOSAHCOMA, MiTASTATIC 1 (5X)
 

RESEIBATORY SYSTEM
 

iLUNS (19) (15) (37)
 ADENOCAECINOMA, NOS, HETASTATIC 3 (7X) 1 (3*)
 

ALVEOLAB/BRONCHIOLJIR ADENOMA 1 <5X) 11 (214*) 2 <5X)
 

ALVEOLAR/ERONCHIOLAR CARCINOMA it (9X) 2 (5*)
 

ADENOCA/SQUAHOUS METAPLASIA, MET 3 (7X) 3 (8X)
 

HEMATOPOIETIC SYSTEM
 

'flULTIPLE ORGANS (20) (48) (46)
 

MALIGNAN T LYMPHOMA, NOS 2 (4X)
 

MALIG.LYHPHOMA. LYMPHOCYTIC TYPE It (2li*' 1 (2X) 1 (2*)
 

ISPLSEN (19) (45) (42)
 

HEMANGIOSARCOHA 1 (5%J
 

MALIGNAN T LYMPHOMA, NOS 1 (2X)
 

HALIG.LYHPHOHA, LYMPHOCYTIC TYPE 1 (2<)
 

tLYMPH NODE (18) (42) (35) MALIGNAN T LYMPBOMA, NOS 1 (3X)

 

ICERVICAL LYMPH NODE (18) (42) (35) 

ADENOCA/SQUAHOUS METAPLASIA, MET 1 (2X) 

OSTEOSARCOMA. HETASIATIC 1 _£$! 

* NUMBE R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY * NDMEE R OF ANIHALS NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-7
 
TABLE B2 (CONTINUED) 

CONTROL (VEH) LOB DOSE 

02-F019 02-F022 

*KICNEY (19) 

NALIG.LYHPHOBA , LYBPHOCYTIC TYPE 1 (5X) 

CIRCOLATOEY SYSTEB
 

NONE
 

DIGESTIVE SYSTEM
 

•LIVER	 (19) (47)
 

HEPATOCEILULAR CAFCINOBA 5 (11X)
 

HEMAKGIOSARCOMA
 

ISTOMACH (19) (43)
 

SQDABODS CELL PAPILLOMA 1 <2X)
 

SSUAMOUS CELL CARCINOMA
 

tSMALL INTESTINE (19) (45)
 

HEPATOCELLULAR CARCINOMA, BETAS!
 

URINARY SYSIHM
 

•KIDNEY	 (19) 

HEMANGIOSA8COMA, METASTATIC 1 (5X) 

ENDOCEINI SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEB
 

*BABHARY GLAND (20) (48)
 

CARCINOMA, NOS 1 <2X)
 

ADENOMA, NOS
 

ADENOCAPCINOBA, NOS 23 ("48X)
 

ABENOCA/SQOABOBS METAPLASIA 10 (21X)
 

FIBEOSARCOMA 1 (2X)
 

FIBROADENOMA 1 (2X)
 

*VAGINA (20) (48)
 

SOD1HOUS CELL CARCINOHA ,._.. 1 (2X)
 

* NDHBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMEEE OF ANIMALS NECROPSIED
 

B-8
 

HIGH DOSE
 

02-F023
 

(37)
 

(46)
 

8 (17X)
 

1 (2X)
 

(38)
 

1 (3X)
 

(37)
 

1 (3X)
 

(37)
 

(H6)
 

1 (2X)
 

11 (21%)
 

16 (35X)
 

1 (2X)
 

(46)
 
TABLE B2 (CONTINUED)
 

CONTROL (VEH) LOB DOSE
 

02-F019 02-F022
 

*UTEEUS (19) (44)
 

IEIOHYCBA 1 (5X)
 

INDOMETPIAL STROMAL POLYP 1 (2X)
 

HEMANGIOMA 1 (2X)
 HEHANGIOSAHCOMA
 

tUTEROS/ENDOMETRIUM (19) (44)
 

PAPILLOMA, NOS 2 (5X)
 

IOVARY (19) (144)
 HEMANGIOMA
 

HFHANGIOSARCOMA
 

NEEVOUS SYSTEM
 

NONE
 

SPECIAL SEkSE ORGANS
 

*HSEDERIAN GLAND (20) (48)
 

ADENOMA, NOS 1 (2%)
 

MUSCOIOSKELEIAL SYSTEM
 

*VERTEBEAL COLUMN (20) (48)
 CSTECSARCOMA 1 (2X)
 

BCDY CAVITIES
 

NONE
 

ALL C1HER SYSTEMS
 

THCEACIC CAVITY
 NEUPOFIEROSARCOMA
 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMEEE OF ANIMALS NECROPSIED
 

B-9
 

HIGH DOSE
 

02-F023
 

(37)
 

1 (3X)
 

1 (3X)
 

(37)
 

(36)
 

1 (3X)
 

1 <3X)
 

(16)
 

(46)
 
TABLE B2 (CONCLUDED) 

CONTROL (VEH) LOW DOSE HIGH DOSE 

02-F019 02-F022 02-F023 

ANIMAL DISPOSITION SUMMARY
 

ANIMAIS INITIALLY IN STUDY 20 50 50
 

NATURAL DEATHS 2 28 37
 

MORIBUND SACRIFICE 5 8
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 18 17 5
 

ANIMAL MISSING
 

3 INCLUDES AUTOLYZFD ANIMALS
 

TDMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 7 42 37
 

TOTAL PRIMARY TUMORS 8 70 51
 

TOTAL ANIMALS WITH BFNIGN TOHORS 2 16 5
 

TOTAL BENIGN TUMORS 2 18 5
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 6 39 37
 

TOTAL MALIGNANT TUMORS 6 52 149
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 1 7 5
 

TOTAL SECONDARY TUMORS 2 8 5
 

TOTAL ANIHALS WITH TUMORS ONCEPTAINBFNIGN OR MALIGNANT
 

TCTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPEIMARY OB METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY IUBOHS: ALL TUMORS EXCEPT SECONDARY TOMORS
 

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

B-IO
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH SULFALLATE
 

TABLE Cl

 

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

CONTROL (VEH) 

01-M001 

LOW

01­

 DOSE 

M016 

HIGH DOSE 

P1-H017 

ANIMALS INITIALLY IN STUDY 50 

ANIMALS BEC8CPSIED U9 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 49 

50 

48 

38 

50 

15 

30 

INTEGUMENTARY SYSTEM
 

*SKIN (49) (1»8) (45)
 

EPIDERMAL INCLUSION CYST 1 (2X) 2 (4X)
 

*SUBCUT TISSUE («9) (48) (45)
 ABSCESS, NOS 1 (2X) 1 (2*)
 

RESPIEATCBY SYSTEM
 

tTPACHEA (1) (26) (21)
 INFLAMMATION, NOS 4 (ICO*)
 

tLUNG/EBONCHUS (49) (28) (21)
 HYPEPPLASIA, LYHPHOID 1 (4X) 3 (14*)
 

tLUNS (49) (28) (21)
 MINEBALIZATION 2 (7*)
 EDEMA, NOS 1 (4*)
 INFLAMMATION, NOS 1 <«*) 2 (10X)
 

INFLAMMATION, NECROTIZISG 2 (7X)
 PNEUMONIA, CHRONIC MURINE 20 (41*) 1 (4X) 1 <5X)
 HYPERPLASIA. LYMPHOID 20 (71X) 13 (62X)
 

HFNATCPCIETIC SYSTEM
 

*BONE HARROW (46) (27) (21) HYPOPLASIA, HEMATOPOIETIC 1 (5X) 

•SPLEEN	 (47) (29) (20) 

FIBBOSIS 1 (2X) 1 (3X) HEMATOPOIESIS 1 (2X) 5 (17X) 2 (10X) 

IHESENTERIC L. NODE (45) (25) (22) CYST. NOS 

t NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMEE R OF ANIMALS NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-3
 
TABLE Cl (CONTINUED) 

CONTROL (VEH ) LOW DOSE 

01-M001 01-M016 

EDEMA, NOS
 1 (1*)
 

CIRCULATCBY SYSTEM 

«HEART 

MINERALIZATION 

(<47) (28) 

1 (1*) 

0MYOCARDIUM 

INFLAMMATION, NOS 

INFLAMMATION WITH

FIBROSIS 

FIBSOSIS, DIFFUSE 

 FIBROSIS 

(147) 

11

1

 (30*) 

 (2X) 

(28) 

10

1

3

6

 (36X) 

 (1*) 

 (11X) 

 (21X) 

*ENDOCARDIUM 

ENDOCARDITIS, BACTERIAL 

METAPLASIA, OSSEOUS 

(17) (28) 

1

1

 (14%) 

 (it*) 

*AOPTA 

PERIARTERITIS 

ARTERIOSCLEROSIS, NOS 

CALCIFICATION, NOS 

(U9) 

H (8*) 

(18) 

2 (<4*1 

DIGESTIVE SYSTEM
 

tSUEMAXILLARY GLAND (1)
 EDEMA, NOS 1 (25*)
 

AESCESS, NOS 1 (25X)
 

FIBROSIS 1 (25X)
 

tLIVER (149) (31)
 CYST, NOS 2 (UX)
 

INFLAMMATION, NOS 3 (6X)
 

NECROSIS, NOS
 

METAMORPHOSIS FATTY 3 (6K) 7 (23X)
 

CYTOPLASMIC VHCUOLIZATION 6 (19*)
 

CYTOIOGIC DEGENERATION 1 (3*)
 

HYPEFPLASIA, NOS 5 (10*)
 

HEMATOPOIESIS
 

*LIVER/CENTRILOBULAE <U9) (31)
 

DEGENERATION, NOS 2 (6*)
 

NECROSIS, NOS 1 (31)
 

ILIVEB/HEPATOCYTES (19) (31)
 NBCROSIS. NOS .
 

NUMBER OF ANIMALS WITH TISSUE EXAMINED MICBOSCOPICAILY
 

NUMBFR OF ANIMALS NECROPSIED
 

C-4
 

HIGH DOSE
 

01-B017
 

(21)
 

(21)
 

6 (29X)
 

1 <5X)
 

(21)
 

(45)
 

1 (2%)
 

(22)
 

1 (5*)
 

1 (5X)
 

2 (9X1
 

1 (5*)
 

(22)
 

(22)
 

1 (5*1
 
HYPERTROPHY, HOS
 

*BILE DDC1
 

INFLAMMATION, NOS
 

HYPERPLASIA, NOS
 

tPANCREAS
 

PEHIAHIERITIS
 

ATROPHY. NOS
 

IESOPHAGUS
 

INFLAMMATION, NOS
 

*STOSACH
 

MINERALIZATION
 

THROMBOSIS, NOS
 

INFLAMMATION, NOS
 

ULCEB, NOS
 

REACTION. FOREIGN BODY
 

NFCROSIS, FOCAL
 

HYPEFKERATOSIS
 

ACANTHOSIS
 

iCOLON
 

INFLAMMATION, NOS
 

URINABY SYSTEM
 

*KIDNEY
 

MINERALIZATION
 

CONGESTION, NOS
 

PYELONEPHRITIS, NOS
 

PYONEPHEOSIS
 

ABSCESS, NOS
 

INFLAMMATION, CHPONIC
 

NEPHBOPATHY, TOXIC
 

tKIDNEY/CORTEX
 

CYST, NOS
 

IKIDNEY/PELVIS
 

DILATATION, NOS
 

INFLAMMATION, NOS
 

tURINARY BLADDER
 

INFLAMMATION. NOS
 

TABLE Cl (CONTINUED)
 

CONTROL (VEH)
 

01-H001
 

(49)
 

3 (6%)
 

(46)
 

5 (11*)
 

(1)


1 (1COX)
 

(16)
 

1 (2X)
 

(46)
 

1 (2X)
 

(47)
 

2 (4X)
 

1 (2X)
 

1 (2%)
 

37 (79X)
 

(47)
 

(47)
 

(46)
 

_ - 1 -12XJ.
 

LOW DOSE
 

01-8016
 

7 (23X)
 

(«3)
 

9 (19X)
 

16 (33%)
 

(26)
 

1 (4*)
 

2 <8X)
 

(26)
 

(30)
 

4 (13X)
 

1 (3X)
 

7 (23X)
 

7 (23*)
 

(26)
 

(3D
 

3 (10X)
 

1 (3*)
 

1 (3X)
 

27 (87X)
 

1 (3*)
 

(31)
 

5 (16X)
 

(31)
 

1 <3X)
 

1 (3X)
 

(27)
 

* NUMBER OF ANIMALS SITH TISSUE FXABINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-5
 

HIGH DOSE
 

P1-M017
 

3 (14X)
 

(45)
 

11 (24X)
 

14 (31X)
 

(21)
 

1 <5X)
 3 (14X)
 

(21)
 

(25)
 

1 (4X)
 

4 (16X)
 

1 (4X)
 

6 (24X)
 

7 (28 X)
 

(20)
 

(22)
 

1 (5X)
 

15 (68X)
 

9 (U1%)
 

(22)
 

(22)
 

1 (5X)
 

(21)
 
TABLE Cl (CONTINUED)
 

ENDOCRINF SYSTEM
 

•PITUITARY
 

CYST, NOS
 

HYPERPLASIA, CHROHOPHOBE-CEIl
 

•ADRENAL
 

CYTOLOGIC DEGENERATION
 

ANGIECTASIS
 

*ADRENAI, COBTEX
 

CYTOPLASBIC VACUOLIZATION
 

ATROPHY. NOS
 

HYPEFTHOPHY, NOS
 

•THYROID
 

CYST, NOS
 

CYSTIC FOLLICLES
 

HYPEEPLASIA, C-CELL
 

HYPEFPLASIA, FOLLICULAR-CELL
 

•PARATHYROID
 

HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEM
 

*HABBARY GLAND
 

GALACTOCELE
 

CYST, NOS
 

•PROSTATE
 

INFLAMBATION, NOS
 

*SFBINAL VESICLE
 

INFLAMMATION, NOS
 

•TESTIS
 

BINEPALIZATION
 

GRANULOBA, SPERMATIC
 

ATEOPHY, NOS
 

*EPIDIDYBIS
 

MINERALIZATION
 

CONTROL (VEH)
 

01-M001
 

(41)
 

1 (2%)
 

(46)
 

8 (17X)
 

(1*6)
 

(48)
 

1 (2*)
 

1 (2X)
 

<t (8%)
 

(46)
 

2 (4*)
 

(49)
 

1 (2X)
 

1 (2%)
 

(34)
 

9 (26X)
 

(49)
 

1 (2%)
 

(44)
 

9 (20*)
 

(49)
 

» NUBBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUBBER OF ANIHALS NECHOPSIED
 

C-6
 

LOW DOSE
 

01-M016
 

(30)
 

1 (3X)
 

(28)
 

14 (SOX)
 

(28)
 

1 (4X)
 

1 (4X)
 

14 (SOX)
 

(27)
 

1 (4X)
 

(3)
 

3 (100X)
 

(48)
 

(9)
 

4 (44X)
 

(48)
 

1 (2X)
 

(27)
 

1 <4X)
 

2 (7X)
 

8 (SOX)
 

(48)
 

.,., 1 (?*)
 

HIGH DOSE
 

01-H017
 

(20)
 

1 (5X)
 

2 (inxj
 

(20)
 

8 (KOX)
 

(20)
 

6 (30X)
 

(21)
 

2 (10X)
 

5 (24X)
 

(1)
 

1 (100X)
 

(45)
 

(14)
 

5 (36X)
 

(45)
 

(21)
 

3 (14X)
 

(45)
 
TABLE Cl (CONTINUED) 

CONTROL (VEH) 

01-M001 

LOW DOSE 

01-M016 

HIGH DOSE 

01-M017 

GRANULOMA, SPERMATIC 

ATROPHY, NOS 

1 (2X) 1 (2%) 

NEPVCUS SYSTEM 

•BRAIN 

ABSCESS, NOS 

(47) (27) (22) 

1 (5%) 

SPECIAL SENSE OBGANS 

*EYI 

MINERALIZATION 

INFLAMMATION, NOS 

CATARACT 

PHTHISIS BBLBI 

*EYE/CRYSTALLINE LENS 

DEGENERATION, NOS 

*EYE/LACBIMAL 3LAND 

HYPEBPLASIA. NOS 

*HABDERIAN GLAND 

INFLAMMATION, NOS 

(49) 

1 (2*) 

(49) 

(49) 

(49) 

1 (2X) 

(48) 

1 (2%) 

1 (2X) 

1 (2*) 

(48) 

(48) 

1 (2*) 

(48) 

(45) 

(45) 

1 (2*) 

(45) 

(45) 

MUSCUIOSKELETAL SYSTEM 

*SKELETAL MUSCLE 

EEGENERATION. NOS 

(49) 

1 (2%) 

(48) (45) 

BODY CAVITIES 

*ABDOtlINAL CAVITY 

NECROSIS, FAT 

*PEBICAPDIDM 

INFLAMMATION, NOS 

*MESENTERY 

PERIARTEBITIS 

(49) 

(49) 

5 (10X) 

(49) 

2 (4*) 

(48) 

1 (2X) 

(48) 

(48) 

1 (2X) 

(45) 

(45) 

(45) 

ALL OTHER SYSTEMS 

NONE 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

C-7 
TABLE Cl (CONCLUDED)
 

CONTROL (VEH) LOW DOSE HIGH DOSE
 

01-M001 01-H016 01-M017
 

SPECIAL MORPHOLOGY SUMMARY
 

NECROPSY PERF/NO HISTO PERFORMED 2 12
 AUTO/NECBOPSY/NO HISTO 8 3
 

AUTOLYSIS/NO NECROPSY 1 2 5
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

C-8
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE RATS TREATED WITH SULFALLATE 

CONTROL (VEH) LOU DOSE
 

01-F001 01-F016
 

ANIMALS INITIALLY IN STUDY 50 50
 ANIMALS NECROPSIED 50 50
 ANIMALS EXAMINED HISTOPATHOLOGICALLY ** 50 i»7
 

INTEGUMENTARY SYSTEM
 

*SKIN (50) (50)
 INFLAMMATION, NOS 1 12%)
 

RESPIRATORY SYSTEM 

tTRACHEA 

INFLAMMATION. NOS 

(5) 

5 (100X) 

(44) 

#LDNG/ERONCHUS 

ABSCESS, NOS 

(50) (ft 4) 

1 (2X) 

tLUNG 

CYST, NOS 

INFLAMMATION, NOS 

PNEUMONIA, CHBONIC MURINE 

HYPEPPLASIA, LYHPHOID 

(50) 

15 (3GX) 

(114) 

2

3

32

 (5*) 

 (7X) 

 (73X) 

HEMATOPOIETIC SYSTEM
 

#BONE HAREOH (50) (43)
 

MITAKORPHOSIS FATTY 1 (2X)
 

LEUKEMOID REACTION 1 (2*)
 

tSPLEEN (50) (45)
 

LEUKEMOID REACTION 1 (2X)
 

HEMATOPOIESIS 5 (10X) 8 (18%)
 

•SPLENIC	 CAPSULE (50) (45)
 

FIBROSIS 1 (2X)
 

CIHCHLATORY SYSTEM
 

#HEART (50) (44)
 

MINERALIZATION
 

# NUMBE R OF ANIMALS WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-9 

HIGH DOSE
 

01-F017
 

50
 

48
 

39
 

(48)
 

(28)
 

(29)
 

(29)
 

2 (7X)
 

1 (3X)
 

12 (41X)
 

(28)
 

(28)
 

7 (25X)
 

(28)
 

(28)
 

2 (7X1
 
TABLE C2 (CONTINUED) 

CONTROL(VEH) LOR DOSE HIGH DOSE 01-F001 01-F016 01-F017 

FIBROSIS, DIFFUSE 1 (2X) 1 (4X) 

PERIABTEEITIS 1 (2X) 

NECROSIS, NOS 1 (2X) 

*HYOCARDIUM (50) (14) (28) 

INFLAMMATION, NOS 4 (9X) 2 (7X) 

FIEROSIS, DIFFUSE 1 (2X) 

DEGENERATION. NOS 2 <4X) 

4ENDOCABDIUM (50) (44) (28)
 

HYPERPLASIA, NOS 1 (2X)
 

*AORTA (50) (50) (48)
 

ABTEBIOSCLEROSIS, NOS 1 (2X)
 

DIGESTIVE SYSTEM
 

tSALIVABY GLAND (1) INFLAMMATION, SOPPUBATIVE 1 (100*) 

*LIVER (50) (47) (31) 

INFLAMMATION, NOS 4 (8X) 

GPANULOMA, NOS 1 (2X) 

METAMORPHOSIS FAT1Y 1 (2X) 5 (11X) 3 (10X) 

CYTOPLASMIC VACUOLIZATION 1 (2X) 2 (6X) 

FOCAL CELLULAR CHANGE 1 (2X) « (9X) 1 (3X) 

HYPERTROPHY, NOS 11 (35X) 

HYPERPLAEIA, NOS 2 (6X) 

HEMATOPOIESIS 1 <3X) 

*LIVER/CENT8ILOBULAB (50) (47) (31)

 

NECROSIS, NOS 1 (2<) 1 (3X)

 

ULIVER/PERIPORTAL (50) (47) (31)

 

NECROSIS, NOS 1 <2X)

 

(tLIVER/HEPATOCYTES (50) (17) (31)

 

HYPERTROPHY, NOS 13 (28X)

 

*BILE DOCT (50) (50) (48) 

DILATATION, NOS 1 (2X) 7 (14X) 18 (38X) 

INFLAMMATION, NOS 25 (SOX) 19 (40X) 

HYPERTROPHY, NOS 1 (2X) 

HYPERPLASIA^ NOS 34 (68X) 29 (60X) 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NDMEEE OF ANIMALS KECROPSIED
 

C-IO 
•PANCREAS
 

ATROPHY, NOS
 

ATROPHY, FOCAL
 

*STOHACH
 

ULCER, NOS
 

ULCER. FOCAL
 

CALCIUM DEPOSIT
 

HYPERKERAIOSIS
 

ACANTHOSIS
 

•LARGE INTESTINE
 

NENATODIASIS
 

PARASITISM
 

URINABY SYSTEM
 

tKIDNEY
 

MINERALIZATION
 

HYDRONEPHBOSIS
 

PYELONEPHRITIS, NOS
 

INFLAMMATION, CHROMIC
 

NEEHROPATHY, TOXIC
 

HYPEEPLASIA, NOS
 

tKIDNEY/PELVIS
 

INFLAMMATION, NOS
 

INtOCFINI SYSTEM
 

tPITBITARY
 

CYST, NOS
 

HYPEFPLAEIA. CHBOMOPHOBE-CELL 

ANGIEC1ASIS 

tADRENA L 

CIGENERATION, NOS
 

CYTOLOGIC DEGENERATION
 

HYPERTROPHY, NOS
 

ANGIECTASIS
 

HEMATOPOIESIS
 

tADRESAL CORTEX
 

HYPERTROPHY. NOS
 

TABLE C2 (CONTINUED)
 

CONTROL (VEH)
 

01-F001
 

(50)
 

(50)
 

5 (10X)
 1 (2%)
 

(49)
 

1 (2X)
 

(50)
 

1 (2X)
 

23 (46X)
 

(50)
 

(50)
 

1 (2X)
 

(50)
 

17 (31X)
 

LOW DOSE
 

01-F016
 

(44)
 

1 (2%)
 

1 (2X)
 

(43)
 

2 (5*)
 

2 (5X)
 

1 <2X)
 

1 (2X)
 

(43)
 

1 (2X)
 

(U4)
 

1 (2X)
 

18 (41X)
 

1 (2X)
 

(i»«)
 

1 (2*)
 

(44)
 

2 (5X)
 

1 (2X)
 

(45)
 

21 (47X)
 

9 (20X)
 

* NOMBE8 OF ASIHALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECEOPSIED
 

C-ll
 

HIGH DOSE
 

01-F017
 

(28)
 

1 («X)
 

(35)
 

1* (11X)
 

3 (9X)
 

6 (17X)
 

9 (26X)
 

(28)
 

(28)
 

2 (7*)
 

12 (13X)
 1 (4X)
 

(28)
 

2 (7X)
 

(26)
 

2 <8X)
 

(30)
 

12 (UPX)
 

12 (40X)
 

11 (37X)
 

1 (3X)
 

(3P)
 
TABLE C2 (CONTINUED) 

CONTROL (VEH) LOW DOSE HIGH DOSE
 

01-F001 01-F016 01-F017
 

tTHYROID (50) (11) (28)
 

CYSTIC FOLLICLES 3 (7X) 2 (7X)
 HYPEBPLASIA. NOS 2 (5») 1 (IX)
 

HYPEEPLASIA. PAPILLARY 1 <4X)
 HYPERPLASIA, C-CELL 2 {«») 3 <7X) a (itX)
 HYPEFPLASIA, FOLLICULAR-CELL 2 (»X) 1 (2X)
 

•PANCREATIC ISLETS (50) (11) (28)
 

HYPEFPLASIA, NOS 1 (2X)
 

HEPBCEOCTIVE SYS1EN
 

*HAMf!ARY GLAND (50) (50) (18)
 GALACTOCILE 1 <2X)
 

CYS1. NOS 2 (IX)
 

HYPERPLASIA, DOS 1 (2«)
 

*VAGINA (50) (50) (18)
 PIBFOSIS 1 (2X)
 

tOTERUS (19) (16) (28)
 

HYDFOHETRA 9 (18%) 1 (2X)
 

INFLAMMATION, NOS 2 {»«)
 

PYOHETRA 3 (7X) 2 (7X)
 

tUTERtJS/ENDOHBTRIUM (19) (16) (28)
 

INFLAMMATION, NOS 2 (IX)
 

HYPERPLASIA, NOS 7 (15X)
 

HYPEHPLASIA, CYSIIC 3 (6X) 8 (17X) 5 (18X)
 

tOVARY (19) (11) (28)
 

CYST, NOS 2 (IX) 1 (2X) 1 (UX)
 

FOLLICULAF CYST, NOS 2 (5X) 1 (IX)
 

NERVOUS SYSTEM
 

*PINEAL BODY (50) (50) (18)
 HEHOFRHAGE 1 (2X)
 

SPECIAL SENSE ORGANS
 

*EYE (50) (50) (18)
 

INFLAMMATION. NOS
 

* NUHBEF OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-12
 
TABLE C2 (CONCLUDED)
 

CONTROL (VEH) LOU DOSE HIGH DOSE
 

01-F001 01-F016 01-F017
 

SYNECHIA. ANTIRIOP 1 (2%)
 

CA1APACT 3 (6%) 1 (2X)
 

DYSFLASIA, DOS 1 (2%)
 

•EYE/COBNEA	 (50) (50) (18)
 

IHFLANMATION, NOS 1 (2X)
 

*EYE/IPIS (50) (50) (48)
 

INFLAMMATION, NOS 1 (2*) 1 (2%)
 

MUSCUtOSKFLETAL SYSTEH
 

NONE
 

BCEY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 1 1
 

NECROPSY PERF/NO HISTO PERFORMED 3 6
 

AUTO/NECROPSY/HISTO PERF 1
 

AUTO/NECROPSY/NO HISTO 3
 

AUTOLYSIS/NO NECROPSY 2
 

» NUMBER OF ANIMALS WITH TISSUE EXAHINED MICROSCOPICALLY
 

* NUBBEH OP ANIBALS NECPOPSIED
 

C-13
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH SULFALLATE
 

TABLEDI

 

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

(CONTROL (VEH) LOW DOSE HIGH DOSE 

02-H019 02-B020 02-B021 

ANIMAL S INITIALLY IN STUDY 20 50 50 

ANIMAL S MISSING 1 1 

ANIMAL S NECPOPSIED 20 46 49 

ANIMAL S EXAMINE D HI STOPATHOLOGICALLY** 20 46 149 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (46) (49)
 

EPIDERMAL INCLUSION CYST 3 (7)1)
 ULCER. NOS <5X)
 ABSCESS, NOS 1 (2X)
 

*SUECUT TISSUE (20) (46) (49)
 

EPIDERMAL INCLUSION CYST 1 (2X)
 

HEKOERHAGIC CYST 1 (2X)
 

INFLAMMATION, NOS 1 (2%)
 

ULCER, NOS 1 (2%)
 ABSCESS, NOS 2 (1CX)
 

HESPIBA1CRI SYSTEM
 

*NASAL MUCOSA (20) (46)
 

INFLAMMATION, SUPPURATIVE 1 (2»)
 

*TPACHEA (18) (38) (41) 

HYPEPPLASIA, PAPILLARY 

fLUNG (20) (43) (45)
 CONGESTION, NOS 4 (9X)
 

HEMORRHAGE 1 (2X) 2 (4X)
 INFLAMMATION, NOS 1 (2X) 3 (7X)
 

PNEUMONIA, CHRONIC MURINE 13 (30X)
 INFLAMMATION, CHRONIC FOCAL 1 (2X>
 HYPEFPLASIA, ADENOPATOUS 1 (9X)
 

METAPLASIA, SQUAMOOS 1 (2X)
 HYPEFPLASIA, LYMPHOID 5 (11X)
 

HEMAIOPOIETIC SYSTEM
 

fSPLEEN (44) 

AMYLOIDOSIS 

i NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NOMEE R OF ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AHTOLYZED ANIMALS 

D-3
 
TABLE Dl (CONTINUED)
 

CONTROL (VEH) LOU DOSE HIGH DOSE
 

02-M019 02-M020 "2-M021
 

HYPEFPLASIA, LYSPHOID 2 (6X)
 HEBATOPOIESIS 5 (15X) 9 (20 X)
 

tLYBPH NODE (16) (41) (12)
 INFLAHHATION, NOS 1 (2X)
 HYPEPPLASIA, NOS 3 (7X)
 ANGIECTASIS 1 <2X)
 HYPERPLAEIA, LYMPHOID 2 (5X)
 

•CERVICAL LYMPH NODE (16) (11) (12)
 HYPERPLASIA, NOS 1 (2X)
 HYPERPLASIA, LYMPHOID 1 (2X)
 

tHESENTERIC L. NODE (16) (11) (12)
 CONGESTION, NOS 1 (2X)
 INFLAMMATION, NOS 2 (13X) 3 <7X)
 

ABYLOIDOS1S 1 (2X)
 ATROPHY, NOS 1 (2X)
 HYPERPLASIA, NOS 6 (15X)
 HYPFRPLASIA, LYBPHOID 3 (7X) 1 (2X)
 

tTHYHUS (11) (10) (13)
 AHYLOIDOSIS 1 <3X)
 ATROPHY, NOS 1 (3X)
 HYPEPTROPHY, NOS 1 (3X)
 

CIRCULATORY SYSTEM
 

tHFART (20) (10) (13)
 

PERIAfiTEFITIS 1 (5X) 1 <2X)
 

IBYOCARDIUM (20) (10) (13)
 FIBROSIS 2 (5X)
 

((ENDOCARDIUM (20) (10) (13)
 INFLAMMATION, NOS 1 (3X)
 

DIGESTIVE SYSTEH
 

tSALIVARY GLAND (20) (38) (36)
 INFLAMMATION, NOS 3 (8X)
 

#LIVER (20) (15) (19)
 THROMBOSIS, NOS 12 (24%)
 

* DDMBER OF BNIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NOHEFR OF ANIMALS NECROPSIED
 

D-4
 
CONGESTION. NOS
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

INFLAMMATION, ACDTE
 

INFARCT, NOS
 

AMYLOIDOSIS
 METAMORPHOSIS FATTY
 PIGMENTATION, NOS
 HYPERTROPHY, NOS
 

HYPERTBOPHY, FOCAL
 

HYPERPLAEIA, NODULAR
 

ANGIECTASIS
 

HEHATOPOIESIS
 

tLIVER/CENTRILOBULAR
 

HYPERTROPHY, NOS
 

iLIVtR/KEPATOCYTES
 HYPEKTROEHY, NOS
 

*GALLELADDER
 INFLAMMATION, NOS
 

HYPEBPLASIA, PAPILLARY
 

*BILE DUCT
 

DILATATION, NOS
 

INFLAMMATION, NOS
 

tPANCREAS
 

INFLAMMATION, NOS
 

FIBROSIS
 

FIBROSIS, FOCAL
 

FERIAB1EBITIS
 AMYLOIDOSIS
 AIBOPHY, NOS
 

#STOMACH
 INFLAMMATION, NOS
 

ULCER, NOS
 HYPERPLASIA, EPITHELIAL
 

HYPERKEHATOSIS
 

ACANTHOSIS
 

4PEYEHS PATCH
 HYPEBPLASIA, NOS
 

TABLE Dl (CONTINUED)
 

CONTROL (TEH)


02-M019


2 (10*)
 

(20)


(20)


(20)


(20)


(18)


1 (6X)
 

(19)


(19)


 LOU DOSE
 02-M020
 

3 (7*)
 

2 (4X)
 

1 (2X)
 

2 (4X)
 

1 (2X)
 

2 (»«)
 U (9X)
 

2 (4X)
 

 (45)
 

 (15)
 

 (46)
 

1 (2X)
 

 («6)
 

3 (7X)
 

 (10)
 

2 (5X)
 

1 OX)
 

1 (3X)
 

1 (3X)
 

1 (3X)
 

 (<»0)
 

1 (3X)
 

1 (3X)
 

3 (8X)
 

1 (3X)
 

1 (3X)
 

 (UO)
 

1 J3XJ.
 

* NUMBER OF ANIMALS lilTH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECSOPSISD
 

D-5
 

HIGH DOSE
 

02-H021
 

3 (6X)
 

5 (10X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

26 (53X)
 

1 (2X)
 

(49)
 

1 (2X)
 

(49)
 

3 (6X)
 

(49)
 

1 (2X)
 

(49)
 

1 (2X)
 

(42)
 

(43)
 

1 (2X)
 

2 (5X)
 

2 (5X)
 

(42)
 
tLARlJE INTESTINE
 

NEMATODIASIS
 

*BECTOfl
 

PROLAPSE
 

*ANUS
 

ULCFP. NCS
 

URINARY SYSTEM
 

tKIDNEY
 

HYDRONEPHROSIS
 

PYELONEPHRITIS, NOS
 

INFLAMMATION, NOS
 

INFLAMMATION, CHPONIC
 

NEPHROPA1HY, TOXIC
 

AMYLOIDOSIS
 

IKIDNFY/PELVIS
 

INFLAMMATION, NOS
 

#BRINARY BLADDER
 

CYST, NOS
 

INFLAMMATION, NOS
 

ENDOCRINE SYSTEM
 

• PITUITARY
 

CYS1, NOS
 

tADRENAL
 

HYPBPPLASiA. FOCAL
 

fTllYEOID
 

INFLAMMATION, NOS
 

REPRODUCTIVE SYS1EB
 

*PREPUTIAL GLAND
 

ABSCESS, NOS _
 

t NIIMEE P OF ANIMAL S WITH TISSUE

* NtlBtEE OF ANIHALS NECROPSIFD 

TABLED! (CONTINUED) 

CONTROL (VEH) LOU DOSE
 

02-B019 02-M020
 

(19)	 (40)
 

1 (5X)
 

(20)	 ("6)
 

5 (25*) 5 (11X)
 

(20)	 («6)
 

1 (2%)
 

(20) (»2)
 

4 (10X)
 

6 (1UX)
 

5 (2SX) 14 (33%)
 

26 (62%)
 

14 (20*) 1 <2X)
 

(20)	 C*2)
 

3 (15X) 3 (7X)
 

(18)	 (10)
 

1 OX)
 

5 (13X)
 

(16)	 (39)
 

1 (6X)
 

(16)	 (40)
 

1 (3K)
 

(17) (38)
 

(20)	 (46)
 

1 (5X)
 

 EXAMINE D MICROSCOPICALLY 

D-6 

HIGH DOSE
 

02-M021
 

(43)
 

(49)
 

(49)
 

(43)
 

4 (9X)
 

3 (7X)
 

35 (SIX)
 

(43)
 

2 <5X)
 

(41)
 

(36)
 

(41)
 

(41)
 

1 (2X)
 

(49)
 

1 _(2X)_ _ _
 
IPRCSTiTE
 

CYS1, NOS
 INFLAMMATION, NOS
 

INFLAMMATION, VESICULAR
 HYPKRPLASIA, CYSTIC
 

•SEMINAL	 VESICLE
 

INFLAMMATION, NOS
 

tTESTIS
 

CALCIFICATION, NOS
 

ATROPHY, NOS
 

*EPIDIDYMIS
 GRANULOMA, SPERMATIC
 

NF.RVCUS SYSIEB
 

#EEAIN
 CALCIFICATION, FOCAL
 

SPECIAL SEKSE ORGANS
 

NONE
 

MUSCDICSKELEIAL SYSTEM
 

*SKELETAL MUSCLE
 

INFLAMMATION, NOS
 

FIBFOSIS
 

BCDY CAVITIES
 

NONE
 

ALL CTHIB SYSTEMS
 

NONE
 

TABLE Dl (CONTINUED)
 

CONTROL (VEH)


02-H019


(16)


(20)


(19)


(20)


1 (5%)


(20)


(20)


 LOW DOSE HIGH DOSE
 

 02-BP20 02-B021
 

 (37)	 («1)
 

2 (5*)
 

2 (5X)
 

1 (3«)
 

2 (SX)
 

 (46)	 (49)
 

1 (2%)
 

 (UO) (43)
 

3 (8X)
 

1 (3%) 1 (2%)
 

 (U6) (49)
 

1 (2*) 1 (2%)
 

 (10)	 (12)
 

1 (3*)
 

 (U6) (49)
 

1 (2X)
 

1 (2*)
 

t	 NOMIER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NOMEER OF ANIBALS NFCHOPSIED
 

D-7
 
TABLE Dl (CONCLUDED) 

CONTROL (VEHJ LOM DOSE HIGH DOSE 

02-B019 02-H020 02-H021 

SPECIAL BOREHOLOGY SUMMARY
 

NO LESION BEPOB1ED 2
 

ANIMAL HISSING/NO NECROPSY 1
 

AOTO/NECHOPSY/HISIO PERF 2 1
 

AUTOLYSIS/NO NECROPSY 3
 

* NUSEER OF ANIMALS ilTH TISSUE EXAMINED MICROSCOPICALLY
 

* NUBEEE OF ANIMALS SECROPSIED
 

D-8
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE MICE TREATED WITH SULFALLATE 

CONTROL (VEH) LOW DOSE 

02-F019 02-F022 

ANIMAI S INITIALLY IN STUDY 20 50 

ANIMAL S NFCROPSIF.D 20 48 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY** 19 US 

INTEGUMENTAR Y SYSTEM 

*SKIN (20) (18) 

HEMATOMA . NOS 

*SOECUT TISSUE (20) (18) 

INFLAMMATION , NOS 

NHCROSIS, NOS 1 (2X) 

RESE1EATOPY SYSTEM
 

*LONG (19) (45)
 

CONGESTION, NOS
 

INFLAMMATION. NOS 4 (9X)
 

PNEUMONIA, CHRONIC MDRINE 22 (19%)
 HYPEPPLASIA, ADBNOMATODS
 

HYPEPPLASIA. LYMPHOID
 

HFMATOPOIETIC SYSTEM
 

tSPLEEN (19) (45) HEMOKRHAGIC CYST 1 (2X) INFLAMMATION, NOS 1 (2X) AMYLOIDOSIS 1 (5X) 1 (2X) 

LEUKFMOID REACTION 2 (11X) HYPERPLASIA, LYBPHOID 9 (20X) HEMATOFOIESIS 22 (49X) 

*LYMPH NODE (18) (42) 

HYPEPPLASIA, NOS 2 (5X) 

HYPEEPLASIA, LYMPHOID 1 (2%) 

ICERVICAL LYMPH NODE (18) (42) 

INFLAMMATION. NOS 

* NUMBE R OF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUHBE P OF ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-9 

HIGH DOSE 

02-F023 

50
 

16
 

46
 

(16)

1 (2X) 

(46) 

(37)
 

7 (19X)
 

1 <3X)
 

3 (8X)
 

1 <3X)
 

(42) 

15 (36X) 

(35) 

(35) 
TABLE D2 (CONTINUED) 

CONTROL (VEH ) LCD DOSE HIGH DOSE 

02-F019 02-7022 02-F023 

CALCIFICATION, METASTATIC 1 <2X) 

*MESENTERIC I.' MODE (18) (12) (35) 

CONGESTION, NOS 1 (3X) 

INFLAMMATION , NOS 3 <7X) 

A BYLCI DOSIS 1 (2X) 

HYPERPLASIA , LYMPHOI 0 3 (7X) 

ITHYMUS (U) (U3)
 HYPEFPLASIA. NOS 4 (9*)
 HYPERPLASIA, LYMPHOID 1 (2X)
 

CIRCULATORY SYSTEM
 

•MYOCARDIUM	 (19) (36)
 INFLAMMATION, NOS 2 (5X)
 INFLAMMATION, CHRONIC 1 (2«)
 INFLAMMATION KITH FIBROSIS 1 (2*)
 

DIGESJIVE SYSTEM
 

tSALIVAFY GLAND (17) (38) (29)
 

INFLAMMATION, NOS 1(3%)
 

tLIVER (19) (U7) (46)
 THROMBOSIS, NOS (9X)
 THFOMBUS, ORGANIZED 1 (2X)
 INFLAMMATION, NOS 3 <6X) 14 (9X)
 INFLAMMATION, FOCAL 1 12%)
 ABSCESS, NOS 1 (2X)
 NECROSIS, NOS 1 (2*)
 HYPEETROPHY, NOS <<**)
 HYPERTROPHY, FOCAL (2X)
 HYPERPLASIA, NODULAR (6*) 3 (7X)
 ANGIECTASIS 12 (.26%) 14 (30X)
 

tLIVER/CENTRILOBULAR (19) (17) (46)
 DEGENERATION, NOS 1 (2X)
 NECROSIS. NOS 1 (2*)
 

tLIVER/HEPATOCYTJSS (19) (U7) (46) 

HYPEPTROPHY, NOS 2 <4X) 

*GALLBLADDER (20) (U 8) (46) 

CYST^IJOS 1118). 

* NUMEEP. OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANI.1ALS NECEOPSIED
 

D-IO
 
*BIIE EUCT
 

INFLAMMATION. DOS
 

tSTOHACH
 

INFLAMMATION. NOS
 

ULCER. NOS
 

*LARGE INTESTINE
 

INFLAMMATION, HEMOREHAGIC
 

NEMATODIASIS
 

*COLON
 

PARASITISM
 

URINAFY SYSTEM
 

tKIDNEY
 

HYDRONEPHROSIS
 

CONGESTION, SOS
 

PYELONEPHRITIS, NOS
 

INFLAMMATION, NOS
 

INFLABHATION, CHRONIC
 

NEPHPOPA1HY, TOXIC
 

•KIDNEY/PELVIS
 

INFLAMMATION, NOS
 

tURINARY BLADDER
 

INFLAMMATION, NOS
 

INFLAMMATION, HFMORRHAGIC
 

INDCCEINE SYSTEM
 

•PITUITARY
 

CONGESTION, NOS
 

HEMORRHAGE
 

INFLAMMATION, NOS
 

tADRENAL
 

CONGESTION, NOS
 

INFLAMMATION, NOS
 

ITHYROID
 

HYPERPLASIA, FOLLICULAR-CELL
 

TABLE D2 (CONTINUED)
 

CONTROL (VEH)
 

02-F019
 

(20)
 

(19)
 

(19)
 

(19)
 

(19)
 

2 (11*)
 

(19)
 

(16)
 

(15)
 

(19)
 

(18)
 

1_I62I
 

LOS DOSE HIGH DOSE
 

02-F022 02-F023
 

(1*8) (16)
 

2 (<IX)
 

(13)	 (38)
 

4 (9S)
 

1 (2%) 1 (3*)
 

(U1) (37)
 

1 (3X)
 

1 (3*)
 

(K1) (37)
 

1 (3X)
 

(4 H) (37)
 

1 (2%) 1 (3*)
 

1 (2X)
 

3 (7»)
 5 (11*)
 

7 (16*) 2 (5X)
 

10 (23X) 26 (70X)
 

(HI) (37)
 

i» (9*)
 

(13)	 (31)
 

8	 (19X)
 

1 (3*)
 

(39)	 (26)
 

1 (3*)
 

1 (3X)
 

1	 (3X)
 

(35)
 

1 (2*)
 

5 (11*)
 

("»«)
 

(U1)	 (32)
 

I	 NUMBER OF ANIMALS SITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECPOPSIED
 

D-ll
 
BEPFCCUCTIVE SYSIEB
 

•VAGINA
 

INFLAMMATION, NOS
 

IUTEBUS
 

HYDBOBETHA
 

INFLAMMATION, NOS
 INFLAMMATION, SUPPUBATIVE
 

FYOBETBA
 

IUTERDS/ENDOBETBIDM
 INFLAMMATION, NOS
 

ISFLABBATION, SOPPOBATIVF
 

INFLAMMATION, VESICULAR
 

HYPEBPLASIA, NOS
 

HTPEPPLASIA, CYSTIC
 

*OVABY
 CYST, NOS
 

FOLLICULAB CYST, NOS
 

FAHCVABIAN CYST
 

HEMOKBHAGIC CYST
 

INFLAMMATION, SUPPURATIVE
 

NESVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

BUSCULCSKELETAL SYSTEB
 

*SKE1ETAL BUSCLE
 

INFLAMMATION, NOS
 

ABSCESS, NOS
 PARA SI II SB
 CALCIFICATION. NOS
 

TABLE D2 (CONTINUED) 

CONTROL (VEH) LOW DOSE HIGH DOSE 

02-F019 02-F022 02-F023 

(20) (16) 

2 (IX) 

(19) (37) 

2 (5X) 1 (3X) 

(5X) 

(5X) 

(19) 

1 (5X) 

(11) (37) 

1 (3X) 

1 (5X) 1 <2X) 

1 (2X) 11 (58X) 17 (39X) 3 (8X) 

(19) (11) 

1 (2X) 

(36) 

« (11*) 

1 (21X) 2 (5X) 

1 <5X) 

1 (5X) 

3 (16X) 

(20) (18) (16) 3 (6X) 1 (2X) 

1 (5*) 

t NUMBER OF ANIBALS KITH TISSUE EXAMINED BICKOSCOPICALLY
 * NUBEEB OF ANIMALS NFCEOPSIED
 

D-12
 
TABLE D2 (CONCLUDED)
 

CONTROL (VEH) LOW DOSE HIGH DOSE
 

02-F019 02-P022 02-F023
 

BODY CAVITIES
 

•PERITONEUM	 (20) (18)
 INFLAMMATION, NOS 2 (IX)
 

ALL C1HIE SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 AUTO/NECROPSY/HIS10 PEEF
 A'JTO/NECBOPSY/NO HISTO
 

AOTOLYSIS/NO NECROPSY
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECHOPSIED
 

D-13
 


DHEW Publication No. (NIH) 78-1370