National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 141 

1978 

BIOASSAY OF 

1-PHENYL-3-METHYL-5-PYRAZOLONE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 89-25-8 

NCI-CG-TR-141 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

1-PHENYL-3-METHYL-5-PYRAZOLONE
 

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) 78-1396
 

REPORT ON THE BIOASSAY OF 1-PHENYL-3-METHYL-5-PYRAZOLONE
 

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
 

l-phenyl-3-methyl-5-pyrazolone 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 l-phenyl-3-methyl-5-pyrazolone was
 

conducted by Litton Bionetics, Inc., Bethesda, Maryland, initially
 

under direct contract to the NCI and currently under a subcontract
 

to Tracor Jitco, Inc., prime contractor for the NCI Carcinogenesis
 

Testing Program.
 

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

Weisburger (1,3). The principal investigators for the contract were
 

Dr. S. M. Garner (4,5) and Dr. B. M. Ulland (4,5). Mr. S. Johnson
 

(4) was the coprincipal investigator for the contract. Animal treatment and observation were supervised by Mr. R. Cypher (4), Mr. D. S.
 

Howard (4) and Mr. H. D. Thornett (4); Mr. H. Paulin (4) analyzed
 

dosed feed mixtures. Ms. J. Blalock (4) was responsible for data
 

collection and assembly. Chemical analysis was performed by Midwest
 

Research Institute (6) and the analytical results were reviewed by
 

Dr. N. Zimmerman (7).
 

Histopathologic examinations were performed by Dr. P. K. Hildebrandt (4) at Litton Bionetics, Inc., the pathology narratives were
 

written by Dr. P. K. Hildebrandt (4), and the diagnoses included in
 

this report represent the interpretation of this pathologist. Histopathology findings and reports were reviewed by Dr. R. L. Schueler
 

(8).
 

iii
 
Compilation of individual animal survival, pathology, and summary tables was performed by EG&G Mason Research Institute (9); the
 

statistical analysis was performed by Mr. W. W. Belew (7,10) and
 

Mr. R. M. Helfand (7), using methods selected for the Carcinogenesis
 

Testing Program by Dr. J. J. Gart (11).
 

This report was prepared at METREK, a Division of The MITRE Corporation (7) under the direction of the NCI. Those responsible for
 

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

(7), task leader Ms. P. Walker (7), senior biologist Mr. M. Morse
 

(7), biochemist Mr. S. C. Drill (7), and technical editor Ms. P. A.
 

Miller (7). 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 (1,12), Dr.
 

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

T. W. Orme (1), Dr. R. A. Squire (1,13), 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 U.S. Environmental Protection Agency, 401 M Street
 

S.W., Washington, D.C.
 

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

4.	 Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington, Maryland.
 

5.	 Now with Hazleton Laboratories America, Inc., 9200 Leesburg
 

Turnpike, Vienna, Virginia.
 

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

Missouri.
 

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

Boulevard, McLean, Virginia.
 

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

Maryland.
 

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

Rockville, Maryland.
 

10.	 No wit w h the Solar Energy Research Institute, Cole Boulevard,
 

Golden, Colorado.
 

11.	 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.
 

12.	 No wit w h Clement Associates, Inc., 1010 Wisconsin Avenue, N.W.,
 

Washington, B.C.
 

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of l-phenyl-3-methyl-5-pyrazolone for possible carcinogenicity was conducted using Fischer 344 rats and B6C3F1 mice.
 

l-Phenyl-3-methyl-5-pyrazolone was administered in the feed, at
 

either of two concentrations, to groups of 49 or 50 male and 50
 

female animals of each species. The high and low concentrations of
 

l-phenyl-3-methyl-5-pyrazolone utilized were, respectively, 5000 and
 

2500 ppm for rats and 15,000 and 7500 ppm for mice. Twenty animals
 

of each species and sex were placed on test as controls. After a
 

103-week period of chemical administration, there was an additional
 

observation period of 2 weeks for rats. A 102-week period of chemical administration was followed by an additional 2-week observation
 

period for mice.
 

In both species adequate numbers of animals survived sufficiently long to be at risk from late-developing tumors. Compound-


related mean body weight depression was observed in mice, but not
 

in rats. In addition, no significant accelerated mortality or other
 

signs of toxicity were associated with the dietary administration of
 

l-phenyl-3-methyl-5-pyrazolone to rats; therefore, it is possible
 

that the compound was not administered to rats at the maximum tolerated concentration.
 

There were no tumors in either sex of rats or mice for which a
 

significant positive association could be established between chemical administration and incidence.
 

Under the conditions of this bioassay, there was no evidence for
 

the carcinogenicity of l-phenyl-3-methyl-5-pyrazolone to Fischer 344
 

rats or B6C3F1 mice.
 

viz
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 4
 

A. Chemicals 4
 

B. Dietary Preparation 5
 

C. Animals 6
 

D. Animal Maintenance 6
 

E. Selection of Initial Concentrations 8
 

F. Experimental Design 10
 

G. Clinical and Histopathologic Examinations 13
 

H. Data Recording and Statistical Analyses 14
 

III. CHRONIC TESTING RESULTS: RATS 19
 

A. Body Weights and Clinical Observations 19
 

B. Survival 19
 

C. Pathology 22
 

D. Statistical Analyses of Results 22
 

IV. CHRONIC TESTING RESULTS: MICE 30
 

A. Body Weights and Clinical Observations 30
 

B. Survival 30
 

C. Pathology 30
 

D. Statistical Analyses of Results 33
 

V. DISCUSSION 39
 

VI. BIBLIOGRAPHY 40
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH 1-PHENYL-3-METHYL-5­

PYRAZOLONE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH 1-PHENYL-3-METHYL-5­

PYRAZOLONE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH l-PHENYL-3­

METHYL-5-PYRAZOLONE C-l
 

IX
 
 D-l 

TABLE OF CONTENTS (Concluded)
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH l-PHENYL-3­

METHYL-5-PYRAZOLONE

Figure Number
 

1
 

Table Number
 

LIST OF ILLUSTRATIONS 

Page 

CHEMICAL STRUCTURE OF l-PHENYL-3-METHYL5-PYRAZOLONE 

GROWTH CURVES FOR 1-PHENYL-3-METHYL-5­

PYRAZOLONE CHRONIC STUDY RATS 20 

SURVIVAL COMPARISONS OF l-PHENYL-3-METHYL5-PYRAZOLONE CHRONIC STUDY RATS 21 

GROWTH CURVES FOR 1-PHENYL-3-METHYL-5­

PYRAZOLONE CHRONIC STUDY MICE 31 

SURVIVAL COMPARISONS OF l-PHENYL-3-METHYL5-PYRAZOLONE CHRONIC STUDY MICE 32 

LIST OF TABLES 

Page 

DESIGN SUMMARY FOR FISCHER 344 RATS— 

1-PHENYL-3-METHYL-5-PYRAZOLONE FEEDING 

EXPERIMENT 11 

DESIGN SUMMARY FOR B6C3F1 MICE—l-PHENYL-3­

METHYL-5-PYRAZOLONE FEEDING EXPERIMENT 12 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

1-PHENYL-3-METHYL-5-PYRAZOLONE 23 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE 26 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

1-PHENYL-3-METHYL-5-PYRAZOLONE 34 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE 36 

XI
 
LIST OF TABLES (Concluded) 

Table Number Page 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH 1-PHENYL-3-METHYL-5­

PYRAZOLONE A~3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH l-PHENYL-3-METHYL5-PYRAZOLONE A-7 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH 1-PHENYL-3-METHYL-5­

PYRAZOLONE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH l-PHENYL-3-METHYL5-PYRAZOLONE B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH 1-PHENYL3-METHYL-5-PYRAZOLONE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 1-PHENYL3-METHYL-5-PYRAZOLONE C-8 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH 1-PHENYL3-METHYL-5-PYRAZOLONE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 1-PHENYL3-METHYL-5-PYRAZOLONE D-7 

Xll
 
I. INTRODUCTION
 

l-Phenyl-3-methyl-5-pyrazolone (Figure 1) (NCI No. C03952), an
 

aromatic heterocycle and widely used dye intermediate, was selected
 

for bioassay by the National Cancer Institute because of the increased incidence of bladder cancer observed among workers in the
 

dye manufacturing industry (Anthony and Thomas, 1970; Wynder et al.,
 

1963).
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is 2,4-dihydro-5-methyl-2-phenyl-3H­

*
 

pyrazol-3-one. It is also called 3-methyl-l-phenyl-2-pyrazolin-5­

one; phenyl-3-methylpyrazolone; l-phenyl-3-methyl-5-oxo-2-pyrazoline;
 

l-phenyl-5-(3-methylpyrazolone); Norphenazone; Developer Z; and C.I.
 

(Colour Index) Developer 1.
 

l-Phenyl-3-methyl-5-pyrazolone is an intermediate in the synthesis of at least 36 dyes and pigments, 10 of which are produced in
 

commercially significant quantities in the United States: C.I. Solvent Yellow 16, C.I. Solvent Red 8, C.I. Mordant Yellow 30, C.I. Acid
 

Orange 74, C.I. Solvent Orange 5, C.I. Mordant Red 7, C.I. Pigment
 

Orange 13, C.I. Pigment Red 41, C.I. Acid Yellow 42, and C.I. Acid
 

Orange 56 (Society of Dyers and Colourists, 1956). l-Phenyl-3­

methyl-5-pyrazolone is also used as an intermediate in the synthesis
 

of drugs and is an extremely sensitive reagent for the detection of
 

cyanide (Rose and Rose, 1966).
 

The CAS registry number is 89-25-8.
 
Xon3 

FIGURE 1 

CHEMICAL STRUCTURE OF 1-PHENYL-3-METHYL-5-PYRAZOLONE 
The U.S. produced 17,000 pounds of l-phenyl-3-methyl-5-pyrazolone
 

and sold 14,000 pounds in 1975 (U.S. International Trade Commission,
 

1977). Production data in 1975 are also available for the following
 

dyes and pigments for which l-phenyl-3-methyl-5-pyrazolone is an intermediate: C.I. Acid Orange 74 (20,000 pounds), C.I. Pigment Orange
 

13 (209,000 pounds), and C.I. Acid Yellow 42 (26,000 pounds) (U.S.
 

International Trade Commission, 1977).
 

The potential for exposure to l-phenyl-3-methyl-5-pyrazolone is
 

greatest for laboratory workers and for workers in the dye, pharmaceutical, and chemical manufacturing industries.
 
II. MATERIALS AND METHODS
 

A. Chemicals
 

l-Phenyl-3-methyl-5-pyrazolone, a light yellow powder, was purchased from Aldrich Chemical Company, Milwaukee, Wisconsin. Analysis
 

was performed by Midwest Research Institute, Kansas City, Missouri.
 

The observed melting point (128° to 130°C) conformed with that found
 

in the literature (129° to 130°C) (Jones et al., 1963) and suggested
 

a compound of high purity. Elemental analysis was consistent with
 

C nH1f.N 0, the molecular formula for this compound. However, thin-


layer chromatographic (TLC) plates utilizing two solvent systems
 

(chloroform:methanol and ethyl acetate) indicated five and two impurities, respectively, of lower mobility than the major compound.
 

Each plate was visualized by 254 and 356 nm light, dichromate, and
 

heat. High-pressure liquid chromatography (HPLC) showed one homogeneous peak. Infrared and nuclear magnetic resonance analyses were
 

consistent with the structure of the compound. Ultraviolet (UV)
 

analysis showed a \ of 246 nm with a molar extinction coefficient
 

max
 

(O of 13 x 10 . The literature value was \ = 245 nm with « =
 max
 

18 x 103 (Katritsky and Maine, 1964).
 

A second batch of the compound was purchased five months later
 

from the same supplier. TLC utilizing the same solvent systems described above showed the presence, respectively, of two and one impurities of lower mobility. HPLC again showed one homogeneous peak,
 
and the melting point and elemental analyses were similar to those
 

observed with the first batch. UV analysis (X = 246 with « =
 max
 

1.22 x 10 ) observed in O.ln NaOH was almost identical with that reported in the literature (\ = J
 246 with e = 1.17 x 10 ) (Katritsky
 

max


and Maine, 1964).
 

Throughout this report, the term l-phenyl-3-methyl-5-pyrazolone
 

will be used to refer to this material.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals
 

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

l-Phenyl-3-methyl-5-pyrazolone was administered to the dosed animals
 

as a component of the diet.
 

The chemical was removed from its container and a proper amount
 

was blended with an aliquot of the ground feed using a mortar and
 

pestle. Once visual homogeneity was attained, the mixture was placed
 

in a 6 kg capacity Patterson-Kelley standard model twin-shell stainless steel V-blender along with the remainder of the feed to be prepared. After 20 minutes of blending, the mixtures were placed in
 

double plastic bags and stored in the dark at 4°C. The mixture was
 

prepared once weekly.
 

Dosed feed preparations containing 2500 and 5000 ppm of 1-phenyl3-methyl-5-pyrazolone were analyzed spectrophotometrically. The results immediately after preparation ranged from 92.8 to 97.9 percent
 

with a mean of 95.7 percent of theoretical, including correction for
 
analytical method of recovery used. Data were not corrected for any
 

loss which may have been due to chemical instability or reactivity.
 

C. Animals
 

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

contracts of the Division of Cancer Treatment, National Cancer Institute. All rats were supplied by the Frederick Cancer Research Center,
 

Frederick, Maryland. All mice were supplied by Charles River Breeding
 

Laboratories, Inc., Wilmington, Massachusetts.
 

Rats and mice were approximately 4 weeks old when received. Upon
 

receipt, animals were examined for visible signs of disease or parasites. Obviously ill or runted animals were culled. The remaining
 

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

Animals which did not manifest clinical signs of disease were placed
 

on test at this time. Animals were assigned to groups and distributed
 

among cages so that the average body weight per cage was approximately
 

equal for a given species and sex.
 

D. Animal Maintenance
 

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


controlled rooms. The temperature range was 22° to 26°C and the relative humidity was maintained between 45 and 55 percent. Incoming
 

air was filtered through HEPA filters (Flanders Filters, McLean, Virginia) at a rate of 12 to 15 complete changes of room air per hour.
 
Fluorescent lighting was provided 8 hours per day (9:00 a.m. to 5:00
 

p.m.).
 

All rats were housed four per cage by sex and all mice five per
 

cage by sex. Throughout the study dosed and control animals of both
 

species were housed in polycarbonate cages (Lab Products, Inc., Garfield, New Jersey) suspended from aluminum racks. Racks were fitted
 

with a continuous stainless steel mesh lid over which a sheet of
 

filter paper was firmly secured. Filter paper was changed at 2-week
 

intervals, when the racks were sanitized. Clean cages and bedding
 

®


were provided twice weekly. Ab-sorb-dri hardwood chip bedding
 

(Wilner Wood Products Company, Norway, Maine) was used in polycarbonate cages for the entire bioassay.
 

Acidulated water (pH 2.5) was supplied to animals in water bottles filled by an automated metering device that was checked daily
 

for diluting accuracy. Water bottles were changed twice weekly, and
 

sipper tubes were washed at weekly intervals. During the period of
 

chemical administration, dosed and control animals received treated
 

®
 

or untreated Wayne Lab-Blox meal as appropriate. The feed was
 

supplied in hanging stainless steel hoppers which were refilled three
 

times per week and sanitized weekly. Food and water were available
 

ad libitum for both species.
 

All dosed and control rats were housed in a room with other
 

&
 

rats receiving diets containing N,N'-diethylthiourea (105-55-5) and
 

*
 

CAS registry numbers are given in parentheses.
 
4-nitro-o-phenylenediamine (99-56-9); and other rats intubated with
 

dosed solutions of 3-(chloromethyl)pyridine hydrochloride (3099-31-8).
 

All dosed and control mice were housed in a room with other mice
 

receiving diets containing 2,4-dimethoxyaniline hydrochloride (54150­

69-5); 4'-(chloroacetyl)-acetanilide (140-49-8); p-phenylenediamine
 

dihydrochloride (624-18-0); 4-nitro-o-phenylenediamine (99-56-9); and
 

nithiazide (139-94-6); and other mice intubated with dosed solutions
 

of trimethylphosphate (512-56-1); 2-(chloromethyl) pyridine hydrochloride (6959-47-3); 3-(chloromethyl)pyridine hydrochloride (3099-31-8);
 

and pivalolactone (1955-45-9).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

l-phenyl-3-methyl-5-pyrazolone for administration to dosed animals
 

in the chronic study, subchronic toxicity tests were conducted with
 

both rats and mice. Rats were distributed among nine groups, each
 

consisting of five males and five females. l-Phenyl-3-methyl-5-pyrazolone was incorporated into the basal laboratory diet of seven of
 

the nine groups of rats in concentrations of 2150, 3160, 4600, 6800,
 

10,000, 14,700, and 21,600 ppm. The two remaining rat groups served
 

as control groups, receiving only the basal laboratory diet.
 

Mice were distributed among ten groups, each consisting of five
 

males and five females. l-Phenyl-3-methyl-5-pyrazolone was incorporated into the basal laboratory diet of eight of the ten groups of
 

mice in concentrations of 2160, 3150, 4600, 6800, 10,000, 14,700,
 

8
 
21,500, and 31,600 ppm. The remaining two mouse groups served as
 

control groups, receiving only the basal laboratory diet.
 

The dosed dietary preparations were administered for a period
 

of 7 weeks, followed by a 1-week observation period during which all
 

animals were fed the basal laboratory diet. Individual body weights
 

and food consumption data were recorded twice weekly throughout the
 

study. Upon termination of the observation period, all survivors
 

were sacrificed and necropsied.
 

At the end of the subchronic test, mean body weight gain among
 

male rats receiving a dietary concentration of 4600 ppm was 4 percent
 

less than the mean body weight gain of their controls, while female
 

rats receiving the same concentration displayed a mean body weight
 

gain 4 percent greater than that of their controls. At a dietary
 

concentration of 6800 ppm, the mean body weight gain of male rats
 

was 9 percent less than the mean body weight gain of their controls,
 

while the mean body weight gain of female rats receiving the same
 

concentration was 1 percent less than that of their controls. No
 

deaths occurred in any dosed group; one female control died. The
 

high concentration selected for administration to dosed rats in the
 

chronic bioassay was 5000 ppm.
 

At the end of the subchronic test, mean body weight gain among
 

male mice receiving a dietary concentration of 14,700 ppm was 12 percent less than the mean body weight gain of their controls, while
 

female mice receiving the same concentration displayed a mean body
 
weight gain which was 11 percent less than that of their controls.
 

At a dietary concentration of 21,500 ppm, the mean body weight gain
 

of male mice was 17 percent less than that of their controls, while
 

female mice receiving the same concentration displayed a mean body
 

weight gain 19 percent less than that of their controls. No deaths
 

occurred in any group. The high concentration selected for administration to dosed mice in the chronic bioassay was 15,000 ppm.
 

F. Experimental Design
 

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

treated and untreated observation periods) are summarized in Tables
 

1 and 2.
 

All rats were approximately 6 weeks old at the time the test was
 

initiated and were placed on test simultaneously. The dietary concentrations of l-phenyl-3-methyl-5-pyrazolone utilized were 5000 and
 

2500 ppm. Throughout this report those rats receiving the former
 

concentration are referred to as the high dose groups and those
 

receiving the latter concentration are referred to as the low dose
 

groups. Dosed rats were supplied with feed containing l-phenyl-3­

methyl-5-pyrazolone for 103 weeks followed by an additional 2-week
 

observation period.
 

All mice were approximately 6 weeks old at the time the test was
 

initiated and were placed on test simultaneously. The dietary concentrations of l-phenyl-3-methyl-5-pyrazolone utilized were 15,000
 

10
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

1-PHENYL-3-METHYL-5-PYRAZOLONE FEEDING EXPERIMENT
 

l-PHENYL-3­

INITIAL METHYL-5­ OBSERVATION PERIOD 

GROUP PYRAZOLONE TREATED UNTREATED 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) 

MALE
 

CONTROL 20 0 0 105
 

LOW DOSE 49 2500 103
 

0 2
 

HIGH DOSE 50 5000 103
 

0 2
 

FEMALE
 

CONTROL 20 0 0 105
 

LOW DOSE 50 2500 103
 

0 2
 

HIGH DOSE 50 5000 103
 

0 2
 

Concentrations given in parts per million.
 

11
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

1-PHENYL-3-METHYL-5-PYRAZOLONE FEEDING EXPERIMENT
 

INITIAL

GROUP

SIZE

l-PHENYL-3­

 METHYL-5­

 PYRAZOLONE 

CONCENTRATION3 

OBSERVATION PERIOD 

TREATED UNTREATED 

(WEEKS) (WEEKS) 

MALE 

CONTROL 20 0 0 104 

LOW DOSE 50 7,500 

0 

102 

2 

HIGH DOSE 50 15,000 

0 

102 

2 

FEMALE 

CONTROL 20 0 0 104 

LOW DOSE 50 7,500 

0 

102 

2 

HIGH DOSE 50 15,000 

0 

102 

2 

Concentrations given in parts per million. 

12
 
and 7500 ppm. Throughout this report those mice receiving the former concentration are referred to as the high dose groups and those
 

receiving the latter concentration are referred to as the low dose
 

groups. Dosed mice were supplied with feed containing l-phenyl-3­

methyl-5-pyrazolone for 102 weeks followed by an additional 2-week
 

observation period.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. From the first day, all animals were inspected twice
 

daily for mortality. Food consumption data were collected at monthly
 

intervals from 20 percent of the animals in each group. Body weights
 

were recorded once monthly throughout the bioassay.
 

All moribund animals or animals that developed large, palpable
 

masses that jeopardized their health were sacrificed. A necropsy
 

was performed on each animal regardless of whether it died, was sacrificed when moribund, or was sacrificed at the end of the bioassay.
 

The animals were euthanized by carbon dioxide asphyxiation, and were
 

immediately necropsied. The histopathologic examination consisted of
 

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

gross lesions taken from sacrificed animals and, whenever possible,
 

from animals found dead.
 

Tissues were preserved in a 10 percent neutral buffered formalin
 

solution, embedded in paraffin, sectioned, and stained with hematoxylin and eosin prior to microscopic examination.
 

13
 
Slides were prepared from the following tissues: skin, subcutaneous tissue, lungs and bronchi, trachea, bone marrow, spleen, lymph
 

nodes, thymus, heart, salivary gland, liver, gallbladder (mice), pancreas, esophagus, stomach, small intestine, large intestine, kidney,
 

urinary bladder, pituitary, adrenal, thyroid, parathyroid, testis,
 

prostate, brain, tunica vaginalis, uterus, mammary gland, and ovary.
 

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 recorded in each group at the time that the test
 

was initiated.
 

H. Data Recording and Statistical Analyses
 

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

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

as recommended by the International Union Against Cancer (Berenblum,
 

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

14
 
These data were analyzed using the statistical techniques described in this section. Those analyses of the experimental results
 

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect
 

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

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

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.,
 

15
 
lymphomas), the denominators consist of the numbers of animals necrops led.
 

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

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.
 

16
 
A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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

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 wets calculated from
 

17
 
the exact interval on the odds ratio (Gart, 1971). The relative risk
 

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

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

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

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

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

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

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the relative risk have been included in the tables of statistical analyses.
 

The interpretation of the limits is that in approximately 95 percent
 

of a large number of identical experiments, the true ratio 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 t.he conditions of this test.
 

18
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

No distinct mean body weight depression was associated with compound administration in either male or female rats (Figure 2).
 

No abnormal clinical signs were recorded.
 

B. Survival
 

The estimated probabilities of survival for male and female
 

rats in the control and l-phenyl-3-methyl-5-pyrazolone-dosed groups
 

are shown in Figure 3. The Tarone tests for positive association
 

between dosage and mortality were not significant for either male or
 

female rats. Due to the relatively high mortality of control female
 

rats beginning with week 76, a significant (P = 0.003) negative
 

association between dose and mortality and a significant (P = 0.006)
 

departure from linear trend were indicated by the Tarone test.
 

For male rats, 74 percent (37/50) of the high dose, 59 percent
 

(29/49) of the low dose, and 65 percent (13/20) of the control were
 

alive at the termination of the study. Thus, adequate numbers of
 

males were at risk from late-developing tumors.
 

For female rats, 88 percent (44/50) of the high dose, 88 percent
 

(44/50) of the low dose, and 55 percent (11/20) of the control group
 

survived on test until the termination of the study. Thus, adequate
 

numbers of females survived sufficiently long to be at risk from
 

late-developing tumors.
 

19
 
750­ •750 

MALE RATS CONTROL 

LOW DOSE 

600­ -600 

HIGH DOSE 

< 

oc 

450­ -450 

I 

O 

g300- -300 

CO 

150­ -150 

I \ I I 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

750­ •750 

CO 

600­

FEMALE RATS CONTROL 

LOW DOSE 

HIGH DOSE 

-600 

<r 

H 450­ X 

UJ 

-450 

0300-

m 

-300 

UJ 

5 150­ -150 

15 30 

n 1 r 

45 60 75 

TIME ON TEST (WEEKS) 

90 105 120 

FIGURE 2 

GROWTH CURVES FOR 1-PHENYL-3-METHYL-5-PYRAZOLONE CHRONIC STUDY RATS 

20 
33 

30 

PROBABILITY OF SURVIVAL PROBABILITY OF SURV

o
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a
 
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).
 

A variety of tumors was observed in both the control and dosed
 

groups. The spontaneous occurrence of these lesions, however, is not
 

uncommon in this strain of rats.
 

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

rats (Appendix C).
 

The results of this pathologic examination indicate that under
 

the conditions of this bioassay the administration of l-phenyl-3­

methyl-5-pyrazolone did not induce any toxicologic or neoplastic
 

lesions in Fischer 344 rats.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

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

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

were observed in at least one of the control or l-phenyl-3-methyl5-pyrazolone-dosed groups and where such tumors were observed in at
 

least 5 percent of the group.
 

None of the statistical tests for any site in rats of either sex
 

indicated a significant positive association between chemical administration and an increased tumor incidence. Thus, at the dose levels
 

22
 
_ __ 

___ 

TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN MALE RATS TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE*
 

TOPOGRAPHY : MORPHOLOGY
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma-t0
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

NJ
 

Pituitary: Chromophobe Adenoma
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Cortical Adenoma or
 

Adenoma NOS*5
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

3/20(0.15)
 

N.S.
 

95
 

6/18(0.33)
 

N.S.
 

97
 

1/19(0.05)
 

N.S.
 

105
 

LOW
 

DOSE
 

10/49(0.20)
 

N.S.
 

1.361
 

0.406
 

7.138
 

89
 

16/45(0.36)
 

N.S.
 

1.067
 

0.495
 

2.880
 

80
 

3/48(0.06)
 

N.S.
 

1.187
 

0.105
 

61.031
 

89
 

HIGH
 

DOSE
 

8/50(0.16)
 

N.S.
 

1.067
 

0.295
 

5.813
 

86
 

16/44(0.36)
 

N.S.
 

1.091
 

0.506
 

2.939
 

82
 

0/50(0.00)
 

N.S.
 

0.000
 

0.000
 

7.102
 
___ 

___ 

TOPOGRAPHY: MORPHOLOGY
 

Adrenal: Pheochromocytoma or Pheochromocy toma , Malignant**
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pancreatic Islets: Islet-Cell Adenoma 

ro P Values0 

•PDeparture from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Testis: Interstitial-Cell Tumor
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

CONTROL
 

4/19(0.21)
 

N.S.
 

—_
 

103
 

0/18(0.00)
 

N.S.
 

P = 0.026
 

15/19(0.79)
 

N.S.
 

90
 

LOW
 

DOSE
 

8/48(0.17)
 

N.S.
 

0.792
 

0.250
 

3.278
 

96
 

4/49(0.08)
 

N.S.
 

Infinite
 

0.357
 

Infinite
 

89
 

36/49(0.73)
 

N.S.
 

0.931
 

0.728
 

1.368
 

87
 

HIGH
 

DOSE
 

6/50(0.12)
 

N.S.
 

0.570
 

0.158
 

2.520
 

88
 

0/47(0.00)
 

N.S.
 

43/49(0.88)
 

N.S.
 

1.112
 

0.884
 

1.497
 

86
 
TABLE 3 (CONCLUDED)
 

created groups received doses of 2500 or 5000 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.
 

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


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

group when P < 0.05.
 NJ
 

Ln
 
___ 

___ 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN FEMALE RATS TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE*
 

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar/Bronchiolar Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

Malignant Lymphomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma or
 

Acidophil Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/20(0.00)
 

N.S.
 

3/20(0.15)
 

P = 0.041(N)
 

94
 

11/18(0.61)
 

N.S.
 

76
 

LOW
 

DOSE
 

3/50(0.06)
 

N.S.
 

Infinite
 

0.250
 

Infinite
 

99
 

2/50(0.04)
 

N.S.
 

0.267
 

0.024
 

2.190
 

103
 

23/46(0.50)
 

N.S.
 

0.818
 

0.525
 

1.494
 

83
 

HIGH
 

DOSE
 

3/50(0.06)
 

N.S.
 

Infinite
 

0.250
 

Infinite
 

105
 

1/50(0.02)
 

N.S.
 

0.133
 

0.003
 

1.568
 

103
 

19/45(0.42)
 

N.S.
 

0.691
 

0.429
 

1.304
 

90
 
___ 

__ _ 

___ 

TOPOGRAPHY : MORPHOLOGY
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma0
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polyp
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

CONTROL
 

1/17(0.06)
 

N.S.
 

105
 

1/20(0.05)
 

N.S.
 

100
 

2/19(0.11)
 

N.S.
 

94
 

LOW
 

DOSE
 

4/45(0.09)
 

N.S.
 

1.511
 

0.168
 

72.703
 

105
 

3/50(0.06)
 

N.S.
 

1.200
 

0.106
 

61.724
 

95
 

3/48(0.06)
 

N.S.
 

0.594
 

0.076
 

6.774
 

105
 

HIGH
 

DOSE
 

1/44(0.02)
 

N.S.
 

0.386
 

0.005
 

29.672
 

105
 

1/50(0.02)
 

N.S.
 

0.400
 

0.005
 

30.802
 

105
 

7/50(0.14)
 

N.S.
 

1.330
 

0.289
 

12.469
 

103
 
TABLE 4 (CONCLUDED) 

Treated groups received doses of 2500 or 5000 ppm in feed. 

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

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. 

N5 

CO 
used and under the conditions of the test, there was no evidence that
 

l-phenyl-3-methyl-5-pyrazolone was carcinogenic in Fischer 344 rats.
 

In female rats the Cochran-Armitage test indicated a significant
 

negative association between dose and the incidence of leukemia or
 

malignant lymphoma. The Fisher exact tests, however, were not significant.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative risk
 

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

tumor incidence rates. In all of the intervals shown in Tables 3 and
 

4, the value one is included; this indicates the absence of statistically significant results. It should also be noted that all of the
 

confidence intervals have an upper limit greater than one, indicating
 

the theoretical possibility of tumor induction in rats by l-phenyl-3­

methyl-5-pyrazolone that could not be established under the conditions
 

of this test.
 

29
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

There was mean body weight depression in dosed male and female
 

mice when compared with controls (Figure 4).
 

No abnormal clinical signs were recorded.
 

B.	 Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and l-phenyl-3-methyl-5-pyrazolone-dosed groups are
 

shown in Figure 5. The Tarone tests for positive association between
 

dosage and mortality were not significant for either male or female
 

mice.
 

Adequate numbers of male mice were at risk from late-developing
 

tumors, as 86 percent (43/50) of the high dose, 80 percent (40/50)
 

of the low dose, and 80 percent (16/20) of the control group survived
 

on test until the end of the study. Two control males were missing
 

starting with week 11.
 

For female mice, 68 percent (34/50) of the high dose, 76 percent
 

(38/50) of the low dose, and 90 percent (18/20) of the control group
 

survived on test until the end of the study, thus providing adequate
 

numbers of mice at risk from late-developing tumors. Three high dose
 

females were missing starting with week 8.
 

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).
 

30
 
50 

TIME ON TEST (WEEKS) 

90 

50­ -50 

40­ -40 

C/3 

< 

cc 

55 30 — 

hX 

-30 

>­ 20 — Q 

O 

CD 

Z 

LU 

5 10 — 

CONTROL 

LOW DOSE 

-20 

-10 

FEMALE MICE HIGH DOSE 

I 

15 

I 

30 

I ' I T I 

45 60 75 

TIME ON TEST (WEEKS) 

90 

^ I 

105 120 

FIGURE 4 

GROWTH CURVES FOR 1-PHENYL-3-METHYL-5-PYRAZOLONE CHRONIC STUDY MICE 

31 
PROBABILITY OF SURVIVAL

 PROBABILITY OF SUR

C5 0 0 o 0 <3 O 0 o 

C3 SJ -t^ en oo CD C3 NJ •fe 0> 

1 1 1 1 1 1 1 1 1

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There was an increased incidence of lymphoreticular neoplasms in
 

low dose male mice. These are, however, common spontaneous neoplasms
 

in mice. Lymphoreticular neoplasms occurred with approximately the
 

same frequency in dosed and control female mice.
 

The incidence of follicular cysts of the ovary was slightly
 

elevated in dosed female mice compared to controls. However, this
 

lesion is frequently seen in aged B6C3F1 female mice. A variety of
 

other nonneoplastic lesions was seen and did not appear to be related
 

to compound administration.
 

The results of this pathologic examination indicate that under
 

the conditions of this bioassay, the administration of 1-phenyl3-methyl-5-pyrazolone was not carcinogenic to B6C3F1 mice.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

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

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

were observed in at least one of the control or l-phenyl-3-methyl5-pyrazolone-dosed groups and where such tumors were observed in at
 

least 5 percent of the group.
 

None of the statistical tests for any site in mice of either sex
 

indicated a significant positive association between chemical administration and an increased tumor incidence. Thus, at the dose levels
 

used in this experiment, there was no evidence that l-phenyl-3-methyl5-pyrazolone was carcinogenic in B6C3F1 mice.
 

33
 
___ 

___ 

___ 

TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN MALE MICE TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE*
 

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar/Bronchiolar Adenoma or
 

Alveolar/Bronchiolar Carcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

to
 

Hematopoietic System: Leukemia or
 

Malignant Lymphomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

3/17(0.18)
 

P = 0.007(N)
 

104
 

2/18(0.11)
 

N.S.
 

104
 

2/18(0.11)
 

N.S.
 

104
 

LOW
 

DOSE
 

3/47(0.06)
 

N.S.
 

0.362
 

0.055
 

2.514
 

104
 

11/50(0.22)
 

N.S.
 

1.980
 

0.502
 

17.385
 

70
 

3/48(0.06)
 

N.S.
 

0.563
 

0.072
 

6.411
 

93
 

HIGH
 

DOSE
 

0/49(0.00)
 

P = 0.015(N)
 

0.000
 

0.000
 

0.570
 

4/50(0.08)
 

N.S.
 

0.720
 

0.117
 

7.578
 

68
 

1/49(0.02)
 

N.S.
 

0.184
 

0.003
 

3.372
 

90
 
TABLE 5 (CONCLUDED) 

LOW HIGH 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE DOSE 

Liver: Hepatocellular Adenoma or 

Neoplastic Nodule or Hepatocellular 

Carcinoma*5 8/18(0.44) 8/48(0.17) 6/49(0.12) 

P Values0 P = 0.007(N) P = 0.024(N) P = 0.007(N) 

Relative Risk (Control) 0.375 0.276 

Lower Limit 0.156 0.099 

Upper Limit 0.994 0.791 

Weeks to First Observed Tumor 92 93 90 

Treated groups received doses of 7500 or 15,000 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 signifi­ cant (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.
 
___ 

___ 

TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC
 

SITES IN FEMALE MICE TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE2
 

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar /Bronchiolar Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

u>
 Malignant Lymphoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

1/20(0.05)
 

N.S.
 

104
 

5/20(0.25)
 

N.S.
 

97
 

2/20(0.10)
 

P = 0.046(N)
 

104
 

LOW
 

DOSE
 

3/46(0.07)
 

N.S.
 

1.304
 

0.115
 

66.966
 

104
 

8/49(0.16)
 

N.S.
 

0.653
 

0.222
 

2.293
 

85
 

2/47(0.04)
 

N.S.
 

0.426
 

0.034
 

5.603
 

104
 

HIGH
 

DOSE
 

1/46(0.02)
 

N.S.
 

0.435
 

0.006
 

33.420
 

101
 

12/47(0.26)
 

N.S.
 

1.021
 

0.400
 

3.310
 

0/46(0.00)
 

N.S.
 

0.000
 

0.000
 

1.459
 

53 
TABLE 6 (CONCLUDED)
 

Treated groups received doses of 7500 or 15,000 ppm in feed.
 

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

c
 

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

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

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

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

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

OJ
 
For male mice the possibility of a negative association between
 

dose and the incidence of liver neoplasms and of lung neoplasms was
 

observed. For female mice the Cochran-Armitage test indicated a
 

significant negative association between dose and the incidence of
 

hepatocellular adenomas. The Fisher exact tests, however, were not
 

significant.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

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

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

Tables 5 and 6, the value one is included; this indicates the absence
 

of statistically significant results. It should also be noted that
 

many of the confidence intervals have an upper limit greater than one,
 

indicating the theoretical possibility of tumor induction in mice by
 

l-phenyl-3-methyl-5-pyrazolone that could not be established under
 

the conditions of this test.
 

38
 
V. DISCUSSION
 

Adequate numbers of animals in all groups survived sufficiently
 

long to be at risk from late-developing tumors. A moderate depression of mean body weight gain relative to controls was observed in
 

dosed male and female mice, but not in any rat group. In addition,
 

no significant accelerated mortality or other signs of toxicity were
 

associated with the dietary administration of l-phenyl-3-methyl-5­

pyrazolone to rats; therefore, it is possible that the compound was
 

not administered at the maximum tolerated concentrations.
 

No neoplasms in either sex of either species occurred for which
 

a significant positive association between chemical administration
 

and incidence could be established. All observed neoplasms were of
 

types and incidences known to occur spontaneously in Fischer 344
 

rats or B6C3F1 mice.
 

Under the conditions of this bioassay, there was no evidence for
 

the carcinogenicity of l-phenyl-3-methyl-5-pyrazolone in Fischer 344
 

rats or B6C3F1 mice.
 

39
 
VI. BIBLIOGRAPHY
 

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

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

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

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

J. Wiley & Sons, New York, 1971.
 

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

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

Union Against Cancer, Geneva, 1969.
 

Chemical Abstracts Service, The Chemical Abstracts Service (CAS)
 

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

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

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

Co., Ltd., London, 1970.
 

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

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

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

Tests, Confidence Limits, and Adjustments for Stratification."
 

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

Jones, R., A.J. Ryan, S. Sternhell, and S.E. Wright, "Structures of
 

5-Pyrazolones and Derived 4-Arylazo-5-pyrazol-ones." Tetrahedron
 

JJ^:1497, 1963.
 

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

Observations." Journal of the American Statistical Association
 

53^:457-481, 1958.
 

Katritsky, A.R. and F.W. Maine, "Tautomers of Heteroaromatic Compounds
 

with Five-membered Rings. IV. 1-Substituted Pyrazolin-5-ones."
 

Tetrahedron 20:299, 1964.
 

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.
 

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

Co., New York, 1966.
 

40
 
Rose, A. and E. Rose, editors, The Condensed Chemical Dictionary, 7th
 

edition. Van Nostrand Reinhold Company, New York, 1966.
 

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

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

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

Society of Dyers and Colourists, Colour Index, 2nd edition, Volume 3.
 

Yorkshire, England, 1956.
 

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

j>2_:679-682, 1975.
 

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

United States Production and Sales, 1975. USITC Publication
 

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

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

41
 

Review of the Bioassay of l-Phenyl-3-Methyl-5-Pyrazolone*
 

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 DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be exposed.
 

The members of the Clearinghouse have been drawn from
 

academia, industry, organized labor, public interest groups,
 

State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 

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

as ad hoc members. The Data Evaluation/Risk Assessment
 

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

NCI-sponsored bioassays of chemicals studied for carcinogenic­ ity. It is In this context that the below critique is given
 

on the bioassay of l-Phenyl-3-Methyl-5-Pyrazolone for
 

carcinogenicity.
 

The reviewer agreed with the conclusion given in the
 

report that l-Phenyl-3-Methyl-5-Pyrazolone was not carcinogenic in rats or mice, under the conditions of test. He
 

said that the study was "straightforward" and moved that
 

the report 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.
 

43
 

•ftU.S. GOVERNMEN T PRINTIN G OFFICE : 1978-260-899/3 1
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE
 

TABLE AI 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH 

1-PHENYL-3-METHYL-5-PYRAZOLONE 

CONTROL (UNTR) LOW DOSE 

11-11*75 11-11*73 

HIGH DOSE 

11-11*71 

ANIMALS INITIALLY IN STUDY 20

ANIMAL S NECBCESIED 20

ANIMAL S EXAMINED HISTOFATHOLOG ICALL7** 19

 S50 

 U9 

 1*9 

50 

50 

50 

INTFGtJMFNTARY SYSTEM
 

*EK!1, (20) (U9) (50)
 

FIBROMA 1 (2%)
 

*SUECUT TISSUE (20) (49) (50)
 UNDIFFERENTIATED CAFCIN01A 1 (2%)
 EASAL-CELL CARCINOMA 1 (5f)
 

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

MYXOMA 1 (2%)
 

FHABDCMYCSARCCMA 1 (2%)
 

HEMANGIOMA 1 (2^)
 

RFSEIFATCFY SYETFP
 

*LUNG (18) (1*7) («9)
 

CARCINOMA, NOS, MEIASTATIC 1 (2%)
 ALVFOLAR/EPONCHIOLAR ADENOM A 1 (6)f) 1 (2X) 1 (2%)
 EHECCHFCMCCYTCMA, METASTA^IC 1 (2%)
 

FHABBOMYCSARCOMA, PFTA ST»TIC 1 (2K)
 

HEfATCECIETIC EYET"?
 

*EBAIN (19) (U3) ,5n)
 

MALIGNAN T EETICULCSIS 1 (2«)
 

*MUITIPLE ORGANS (20) (1*9) (50)
 

MALIGNANT LYMPHOMA, NOS 3 (6%)
 LEUKr HIA,NOS 3 (15X) 9 (18%) 5 (1051)
 

#SPLFEN (19) (1*9) (1*9)
 

SARCOMA, NCS 1 (5%)
 

1EUKFMIAXNOS l_i2SJ
 

* kUMEE R OF ANIMAL S WIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* MJCEE K OF ANIMAL S NECEOPSIF D 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

9 50 ANIMAL S HER E INITIALL Y IN SIUC Y BUT ONE WAS DELETE D WHE N FOUN D TP BE A FEMAL E 

ANIMA L IN A MAL E GROUP . 

A-3 
TABLE Al (CONTINUED) 

CONTPOL (ONTR) LOW DOSE HIGH 

11-1175 11-1U73 11-1 

CIFCIL ATCRY SYSTFC 

ItHZBPT (18) (16) (19) 

ADENCCAECINOCA, NC5 1 (6%) 

DIGESTIVE SYST^I 

»LIVFP (19) (19) (18) 

N3CELASTIC NCDULF 1 

hEEATOCFLLULAP CAPCINOMA 1 (2%) 

UFINAF Y Slfl^K 

NCNE 

ENtCCFIN f SYST!H 

»EIT!!TTAFY (18) (IS) (11) 

CP50POEHCSE PDENCC'A 6 (33%) 16 (36%) 16 

»ADFENA L (19) (IS) (50) ADFNCHA , NOS 1 (5?) 

CCFTICAI AEFNCN A 3 (6%) 

FHFCCHFCPlOCYTOdA U (21%) 6 (13%) 5 tH^CCHFCKOCYTOMA, MALIGNAN T 2 (it*) 1 

dThYHOID (11) (16) (16) 

CAFCINCMH , NCS 1 

FOLLICUL«P-CrLL CA"CINOHA 1 (2%) 

C-C"LL ADENOM A 2 (1%) 2 

IFANCFEATI C ISLETS (18) (19) (17) 

ISLFT-CFLL ADENOM A « (8%) 

BEFFCIUCTTVE SYSTEM 

*KAfMAF Y GLAN D (20) (19) (50) 

AEENC1A , NOS 1 

EIEKCM A 1 

FTBPCJTENOMA 1 (5%) 1 (2%) 1 

*PPFPUTIAl GLAN D (20) (19) (50) 

* NUMB" F OE ANI1ALS MICEOSCOPICALLY 

* UUCBFP OF ANI1AL S NECPOTSIEr 

EOSE
 171
 

(2%)
 

(36%)
 

(10%)
 

(2%)
 

(2%)
 

(1%)
 

(2%)
 

(2%)
 

(2%)
 

A-4
 
TABLE A1 (CONTINUED) 

CONTROL(UNTR )

11-1U75

 LOW DOSS

 11-1173

 HIG H DOSE 

 11-1U71 

ITFSTIS

INTFPSTITIAL-CELL TUMOP

 (19)

 15 (79%)

 (U9)

 36 (13%)

 (1(9) 

 13 (88*) 

*ERAIN

GLIORA, DOS

 (19)

 1 (5%) 

 («8) (50) 

SEFC1A I

NON E 

 SENE F CBGAli S 

BUSCULOSKELETAL SYSTEM 

NCI.E 

BCEY CAVITIES 

*AEDCMINAL CAVITY

fESCTHELIOHA, NOS

*Tt)NICA VAGINALIS

KESOTHELIOBA, NOS

 (20)

 (20)

 (U9)

 1 (2X) 

 (U9)

 (50) 

 (50) 

2 (U)S) 

All CTHEF SYSTEMS 

NONI 

ANIPA L riSfOSITIO N SUBMAR Y 

ANIMAL S INITIALLY IN STUDY 

NATDFAL CEATHa 

MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIBA L MISSING 

ANIMA L DELETFD/WPONG SEX 

20 

13 

SO 

9 

12 

29 

1 

50 

2 

11 

37 

* KUMBER CE ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* MitEEH CF ANIMAL S NFCROPSIEE 

A-5
 
TABLE A1 (CONCLUDED)
 

CONTROL (UNTR) LOU DOSF HIGH DOSE
 

11-1475 11-1U73 11-1t71
 

T UKCF SUMMAR Y
 

TCTAL AMHAL S WITH ERIBAR Y THMCBS* 19 U9 U9
 

10TAI FRTKAR Y TUMOPS 35 88 88
 

TOTAL ANIMAL S WITH EFNIGN TU^OPS 18 US U7
 

TCTAL EENIGN TUMORS 2B 70 72
 

TOTAL fNIBALS WITH MALIGNAN T TDNOPS 7 15 13
 

TC^AI MAIIGNAN T TDHOPS 7 17 13
 

TOTBI ANIMALS KITH SFCONDAFY TUMOES* 1 2
 TOTAL SFCONDARY TUTORS 1 2
 

1CTAI ANIKAL S WITH TUrORS UNCFFTAIN­ EFNICN OB HALIGNANT 1 3
 

TCTAL UNCERTAIN T'lCPRS 1 3
 

TOTAL ANIMAL S WITH TUMOR? UNCHFTAINEFII'ARY CF 1ETASTATIC
 

TCTAI UNCERTAIN THMORS
 

* ERICAFY TUMOFS: ALL TUMOSS FXCEPT SECONDARY TUMORS
 

* SECONDARY TUMORS: MJTAS^ATIC TUMORS OE TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-6
 
TABLE A2 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH 

1-PHENYL-3-METHYL-5-PYRAZOLONE 

CONTR0 1 (UNTR ) LO W DOSE HIG H DOSF 

11-1176 11-1U7U 11-1U72 

ANIMAL S INITIALL Y IN STUDY 20 50 50 

ANIMAL S NECBCESIE D 20 50 50 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 20 50 so 

INTEGUMENTARY SYSTEM
 

*SKIfc (20) (50) (50)
 

KEPATCACANTHOMA 1 (2%)
 

*SUECDT TISSUE (20) (50) (50)
 

SQUAMOUS CELL PAPILLCMA 1 (2JS)
 

ACENOSQUAMOUS CARCINOMA 1 (2%)
 

FIBRCAEENOMA 1 (2%)
 

RESEIFATCRY JYST^P
 

(LUNG (20) (50) (50)
 

ALVEOLAR/BBONCHIOLAR ADENOMA 3 (6%) 3 (6X)
 

GBANDLOSA-CELL CARCINOMA, METAST 1 (2%)
 

CSTICSARCCBA, METASTATIC 1 (5«)
 

HEPATCEOIETIC SYSTEM
 

IE8AI N (20) (1(9) (50) MALIGNAN T R*TICULOSIS 1 (5T) 

*MULTIPLF ORGANS (20) (50) (50)
 

fALIGNANT LYHPHOMA, NOS 1 (5J)
 

LEUKEMIA,NOS 2 (10%) 2 (Uf) 1 (2%)
 

CIFCDLATCFY SYSTEP
 

NONE
 

DIGESTIVE SYSTEM
 

t DDMBE R OF ANIMAL S BITH TISSUE EXAMINE D 1ICFOSCOPICALLY 

* MJMBFP OF ANIMAL S NECROPSIED 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-7 
UPINAR Y SYSTE M 

SCN E 

ENICCFINF £Y£TF M
 

«FITDTTAFY
 

CHROMOEHCBE ADENOMA
 

CHFCPOEHCEE CARCINOM A
 

ACIECFHIl ADENOM A
 

HEMANGIOPA
 

*ADPENAL
 

COPTICAL ADCNCM A
 

FHFCCHFCCCCYTCMA
 

FHFCCHFOMOCYTOMA, MALIGNAN T
 

HT-HYROIT
 

FCL1ICULAR-CFLL CAFCINOMA
 

C-CFLL ADENOM A
 

C-CEIL CAFCINOM A
 

»FANCFFATI C ISLETS
 

TSIET-CFLL ADENOM A
 

HFEFCCUCTIVE SYfl'1
 

+MAMMAF Y GLAN T
 

ACFNCMA , NOS
 

ACTNCCAPCINOBA , NOS
 

CYSTADENCMA, NOS
 

F:BRCADFKO«.A
 

*VAGIN A
 

FIBFCSAECOM A
 

Kl^IFUS
 

ADENCCAECINO"A, NOE
 

LIFCHA
 

ENTCMETFIA L SIPOMAL POLYP
 

CHORICCAFCINOM A
 

tCVAP Y 

,^£li t_£A PC I NO ^A

FR C F ANIBAL S WIT H TISSU *

M'CEER O F ANIMAL S NFCPOPSIE f 

TABLE A2 (CONTINUED) 

CONTRO L (ONTR ) LOW DOSF 

11-1476 1 1-1474 

(18) (46) 

11 (61%) 22 (48%) 

1 (2%) 

1 (2%) 

(20)	 (50)
 

2 (4%)
 

(17)	 (45)
 

1	 (6%) 3 (7%)
 

1 (2%)
 

(20)	 (49)
 

1 (2%)
 

(20)	 (51)
 

1 (2%)
 

1 (5%) 3 (6«)
 

(20)	 (50)
 

1 (2%)
 

(19)	 (49)
 

2 (11%) 3 (6%)
 

(19) (48) 

, 

 EXAMINE D MICROSCOPICALL Y 

HIGH DOSE 

11-1172 

(45)
 

19 (42*)
 

1 (2%)
 

(47)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(44)
 

2 <5X)
 

1 (2%)
 

(50)
 

(50)
 

1 (2%)
 1 (2%)
 

1 (2%)
 

1 (2*)
 

(50)
 

(50)
 

1 (2%)
 

1 (2%)
 

7 (14%)
 

1 (2%)
 

(50) 

A-8 
TABLE A2 (CONTINUED) 

CONTROL (UNTR)

11-1176

 LOW DOSE

 11-1(4711

 HIGH DOSF 

 11-1472 

NEBVCtE SYSTEM 

NONE 

SPECIAL EEkSE CBGANS 

NONE 

MUECUICSKELETA L SYSTEf 

*EKUIL

CSTEOSABC3MA

 (20)

 1 (5«) 

 (50) (50) 

BOFY CAVITIES 

NON" 

ALI ClhEP SYSTEMS 

NONE 

ANIPA L DISPOSITION SUrPARY 

SNII1ALS INITIALLY IN STUDY 20 50

NATUKAL DEATHS 7 U

CCRIEUND SACRIEICF 2 2 

SCHECULEC SACBIPICZ 

ACCIDENTALLY KILLED 

TERMINAL SACEIPICT 11 U l»

ANIMA L MSSING 

9 INCLUDES AUIOLYZED ANIMALS . 

* KDPBER OF AMBAL E WITH TISSUE EXAMINED MICROSCOPICALLY 

* MJBEER OE ANIMAL S NECROPSTED 

 50 

1 

5 

 t<l 

A-9
 
TABLE A2 (CONCLUDED)
 

CONTROL (UNTR) LOW DOSE HIGH DOSE
 

11-1 «76 1 1-1«71» 11-1«72
 

TDFTP SUMMAR Y
 

TCTAL ANNULS WITH EEIHAEY TUNOFS* 12 36 25
 

TOTAL PBI1AR Y TUPOFS 20 1(7 «6
 

TCTAL ANIMAL S kITH PFNIGN TtjflOBS 11 32 2U
 

TOTAL 5EMGN TD10FS 15 11 38
 

TCT»L AMCAL S KIT" MALIGNANT TUMOBS 5 5 7
 

TCTPL MALIGNAN T TUMORS *• 6 8
 

TOTAL ANIMAI S HITH SECONDARY TUHOFS* 1 1
 

TOTAL SECONDARY TUBOPS 1 1
 

TOTAL ANIMAL S WITH TUMORS UNCrSTAIN-


HFNICN OP MALIGNANT
 

TOTAL UNC'BTAIN TU10FS
 

TOTAL AMMAL S WITH TUPIO=?S UNCER^AINFPIPAFV OF 1FTASTATIC
 

TOTAL UNCERTAIN TUfORS
 

* PRI1AHY TUBOBS: ALL TU10RS EXCEPT SECONDARY TUI10ES
 

* SfCCh'CARY TUMORS: MFTASTATIC TDBOPS OR TUMOFS INVASIVE INTO AN ADJACENT ORGAN
 

A-IO
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE
 

TABLE Bl 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH 

1-PHENYL-3-METHYL-5-PYRAZOLONE 

CONTROL(UNTR ) LOB DOSE HIGH DOSE 

22-2U75 22-21173 22-2171 

ANIMAL S INITIALL Y IN E1UDY 20 50 50 

ANIMAL S HISSING 2 

ANIMAL S NECHCESIED 18 50 50 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* * 18 19 50 

INTEGUMENTAR Y SYSTE M 

NON E 

RESEIFATOBY SYSTEM 

• LUNG

HEPATOCELLULAR CARCINOMA, M*)TAST 

ALVFCLAR/BPONCHIOLAR ADENOMA

ALVECLAR/BBONCHIOLAR CARCINOMA

SAFCCBA, DCS, METASTATIC 

 (17) 

2

 1

 (12%) 

 (6%) 

(17) 

3 (6%) 

(«9) 

1

1

 (2*) 

 (2*) 

HEPATCECIFTIC SYSTEM
 

•MDLTIPLE ORGANS (18) (50) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (2%) 1 <2X)
 

MALIG.LYBPHOHA, HNEIFFER-TYPf 1 (2%)
 

PALIG.LYBPHOMA, HISTIOCYTIC TYPE 3 (6%) 1 (2%)
 

LEUKFMIA.NOS 1 (6%) 1 (2%) 2 (US)
 

GBANOLCCYTIC LEUKEMIA 1 (2*)
 

tSPlEEN (17) (UK) (U8)
 

FIBPOSAKCOHA 1 (2X)
 

MALIG.LYMPHOMA, UNEIFFER -T YPE 1 (2%)
 

#LYMPH NODE (16) (39) (*5)
 

MALIGNANT LYPPHCMA, NOS 1 (3%)
 

tPEEENTFPIC L. NODE (16) (39) (15)
 

MALIGNANT LYMPHOMA, NOS 1 (6%)
 

BALIG.LYPPHOMA, HISTIOCYTTC TYPE 2 (5%)
 

CIECUIATCBY SYSTEM
 

NCNI _
 

* NUMBF R OF ANIMAL S WITH TISSUE EXAMINE E MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S SECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
TABLE Bl (CONTINUED) 

CONTROL (UNTB)

22-2«75

 LOW DOSE

 22-2473

 HIGH DOSE 

 22-2171 

HC*ST.IVt SYSTF" 

H1IVF.R

HCPATOCFILULAR ADENOMA

HEOPIASTIC NODtUF

HEPATOCELLULAP CARCINOMA

SARCOMA, NOS

HIHANGIOCA

HFfANGIOSAFCOMA

 (18)

 5 (2B*|

 1 (6*) 

2 (11X)

 (18)

 6 (13*)

 3 (6*)

 («9) 

5 (10*) 

1 (2*) 

1 (2*) 

1 (2*) 

1 (2*) 

UFIHAS Y SYS1EP 

NCNF 

FNICCFIM SYS1EB 

»ADF"NAI

ADFNCCARCINOMA , SOS

EHFCCHROMCCYTOMA

 (16)

 1 (6*)

 (U3)

 1 (2*) 

1 (2*) 

 (US) 

BFEFCCUCTIVF SYS1EC 

SON11 

NTFVCCS SYST'N 

NCNE 

SPECIAL SENSE OBGANS 

NONS 

flUSCDICSKFIETAI SYSTIP 

8CNF 

BCEY CAVITIES 

# dUPBFK OF ADIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* M'fEFR OF ANIMAL S NFCPOPSIFD 

B-4
 
TABLE Bl (CONCLUDED)
 

CONTROL (DNTR) LOW DOSE HIGH DOSE
 

22-2U75 22-2U73 22-2471
 

A ll CTHFF SYSTEMS
 

NCNE
 

DISECSITICN SUPI1APY
 

ANIHALS INITIALLY IN STUDY 20 50 50
 NATURA L DEATHS 10
 PCRIBOND SACRIFICE
 

SCHEtULEC SACRIFICF
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 16
 AMBA L BISSING 2
 

INCLUDES AUTOLYZED ANIHALS
 

TUPICF SUBBAE J 

TC1AL ANIfALS HITH EBIBARY TUMOES* 11 20 TOTAL PRIMAR Y TUKOFS 1U 25 It 

TOTAL ANIMALS WITH BENIGN TUBOES 9 6
 TOTAL EENIGN TOBOPS 10

 

TOTAL ANIBAL S WIT H BALIGNAN T TIJBOPS 3 15 8 

TOTAL BALIGNANT TUBORS 5 15 e
 

TOTAL ANIMAL S WITH SECONDARY TUMORS* 2
 

TOTAL SECONDASY TUMORS 2
 

TOTAL ANIBAL S WITH TUBORS UNCFPTAINEENIGN OE BALIGNANT 1
 

TCTAL UNCERTAIN TUHORS 1
 

TOTAL ANIMALS KITH TUMORS UNCERTAINEBIBAR Y OR METASTATIC
 

TCTAL UNCERTAIN TUBORS
 

* PRIMARY TUMORS: ALL TDMORS EXCEPT SECONDARY TUMORS
 

» SBCCNEAPY TUMORS: BETASTATIC TUMORS OR TOBORS INVASIVE INTO AN ACJACFNT ORGAN
 

B-5
 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH 

1-PHENYL-3-METHYL-5-PYRAZOLONE 

CONTRO L (UNTR ) LOW DOSE HIGH DOSE 

22-2176 22-2I474 22-2472 

AtHfAI S IKITIAH Y IN STCDY 2 0 50 50 

ANIMAL S MISSIN G 1 3 

ANIMAL S NECECPSIF D 2 0 149 147

 

ANICAL S EXAMINE D HISTOIATHOLOGICALLY* * 2 0 149 146

 

INTBGOMENTABY SYSTEM
 

*5UECDT TISSUE (20) (149) (147)
 

FIBROSAFCOHA 1 (2%)
 

RFSEIFATCBY SYSTEK
 

• LUN G (20) (116) (146) 

ALVECLAR/BEONCHIOIA P ADFNOH A 1 (5?E) 3 (7*) 1 (211) 

FIEROSARCOMA , PIETASTATIC i (2X) 

HF^ATCECI^TI C £YETI K 

*BULTIPLF CPGAHS (20) (149) (U7)
 

PALIGNANI LYMPHOdA. NOS 1 (2X) 1 (2«)
 

BALIG.LYHPHOMA, ONTIFFFR-TYPE 2 ( \C%) 1 (2X) 1 (2X)
 

PHLIG.LYPEHONA, LYBPHOCYTIC TYPE 1 (5») 1 (2%)
 

CALIG.LYKEHONA, HISTIOCYTIC TYPE 1 (5%) 3 (6*) 3 (6X)
 

LEUKEMIA,NOS 2 (US)
 

UNDIFFEBENTTATIE LEUKEMIA 1 (2X)
 

FRYTHPnCYTIC LEUKEMIA 1 (2X)
 

GFANULOCYTIC LEUKFPIIA 1 (2K)
 

#EON5 HABPOS (19) (145) (UO)
 

CALIGNANT LYCPHOKA, NOS 1 (2*)
 

»SPL5EN (19) (145) (142)
 

BEMANGIOM A 1 (2X)
 

MALIG.LYrEHOHA, HISTIOCYTIC TYPE 1 (5%)
 

»LYBP H MOD E (17) (140) (39) 

PDENOCARCINOflA , NOS 1 (3X) 

*BES^NTFPIC L. NODE (17) («0) (39) 

I KUBEE R O F ANIMAL S WIT H TISSU E EXAMINE D MICROSCOPICALLY 

* NUKEE P O F ANIMAL S KECBOPSIE D 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-6 
TABLE B2 (CONTINUED) 

CONTROL (UNTR )

22-2476

 LOW DOSE

 22-217U

 HIGH DOSE 

 22-2172 

KLIVE R

PALICNAN T LYPFHCr<A , NCS

KALIG.LYNFHOMA , UNEIF F EF -TYPE

 (20) (17) (16) 

1 (2%) 

1 (2*) 

CIFCUIATCFY EYSTEP 

KCNE 

EIGTSTIVI SYSTI M 

*IIVER

HEPATOCEILULAS AEENOHA

*STC1ACH

KERATCACANTH01A

 (20)

 2 (1051)

 (20)

 (<47)

 2 (U%) 

 (1*6)

 (l»6) 

 (15) 

1 (2%) 

URINAR Y SYSTEM 

NCNF 

ENCCCKIN T SYSTEM 

• THYFOIt

FCLLICULAP-CELL ADEHCKA

 (6) (30)

 1 (3«) 

 (22) 

PFFFCEUCTIVF SYSTEM 

(K.TERDS

ItlorYCMA

INEOfl^TPIAL STRONA1 POLYP

HFMANGIOSA

• OVAR Y

EAPHLAB Y ADENCPI A

GRANULCSA-CE1 L TUMO R

 (19)

 1 (5%)

 (16)

 1 (6%)

 (16)

 2 (IX)

 1 (2*) 

 (H2)

 1 (2X) 

 (U5) 

1 (2X) 

1 (21) 

 (38) 

1 (3*) 

MFVCDS

NCN E 

 SYSTE f 

SFfCIA L EE^£ I CFGAN E 

t MJKEE R

* liOKEEB

 OF

 OF

 ANIBAL S

 ANIMAL S

 WIT H ^ISSDE

 NECROPSIE D 

 EXAMINE D MICROSCOPICALLY 

B-7 
TABLE B2 (CONCLUDED)
 

CONTROL(UNTR) LOU DOSE HIGH DOSE
 22-2176 22-2U7I* 22-2172
 

BUECULOSKELETA L SYSTFB
 

NCNF
 

BCEY CAVITIfE
 

NONE
 

A ll CTFFR SYETFBS
 

NONF
 

ANIFA1 DIStCEITICN SUPFABY
 

ANTPALS INITIALLY IN STUDY 20 50 50
 NATURA L CTA^HS) 2 9 13
 PORIFUND SACRIFICE 3
 

SCHFrULEF. SACRIFICE
 ACCICENTALLY KILLET
 

TERC.INAL SACRIFICE 18 33 3K
 AliIPAl MISSING 1 3
 

a INC1UDES ADTOLYZED ANIMALS
 

TDMOE SUMMAF i
 

TC1AI ANICAL S WITH EEIHAEY TUKORC * 9 19 15
 TOTAL PRIMAR Y TUfCFS 10 20 ie
 

TOTAL ANIMAL S KITH BENIGN TriMOFS S 10 14
 

TOTAI BENIGN 'UMOPS 5 10 5
 

TCTAL ANIMAL S KITH MALIGNANT TUMORS 5 9 13
 TOTAL MALIGNAN T TUMORS 5 9 13
 

TCTAL ANIMALS WITH SECONDARY TDMORS* 1
 TOTAL SECONDARY TUBOPS 1
 

TOTAL ANIBAL S KITH TUMORS UKCIPTAINEENIGN OP MALIGNAN T 1
 TCTAL UNCERTAIN TUMORS 1
 

TOTAL ANIBALS KITH TUMORS USCFFTAIN­ FFIMARY CR META5TATIC
 TOTAI UNCERTAIN TUMORS
 

* PRIMAR Y TUMCRS: ALL TUMOBS "XCEFT SECONDARY TUBORS
 t SECONDARY TUMORS: f^TASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-8
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE
 

TABLEC1

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE

 

(rONTROI (UNTB) 

11-1475 

LOW DOSE

11-1473

 HIGH DOSE 

 11-1471 

ANIBALS INITIALLY IN STUDY 

ANirAl S NECBCFSIED 

ANIBALS EXAMINE D HISTOEATHOLOGICALLY** 

20 

20 

19 

cB50

49

49

 50 

 50 

 50 

INTEGIBFNTAP Y SYSTE M 

liCtlE 

BISEIFATCF Y SYSTE B
 

ILUNG (18) (47) (49)
 

BINFFALIZATICN 1 (6*)
 

ATELFCTASIS 1 (2S)
 THROtlBCSIS, NOS 1 (2%)
 

CCNGESTICN, NOS 7 (15X) 2 (4%)
 

ETEBA, NOS 1 (215)
 HEMOFRHAGE 2 (4%) 1 (2%)
 

INFLABMATION, ACUTF 1 (2%)
 PNEUMONIA, CHRONIC MUHINE 4 (22«) 4 (9S) 5 (10X)
 

HYPEFPIASIA, ACENOMATOUS 1 (6%) 2 (4*) 1 (2%)
 

HISTICCYTOSIS 1 (6%) 1 (2%)
 

HEPATCECIETIC SYSTEM
 

(19) (49) (49)
 

INFARCT, NOS 1 (2%)
 

HEKCSIDFFOSIS 1 (215)
 

HYFEFTHCEHY, NOS 1 (2%)
 

HYFEPFLASIA, CIFFISE 1 (2*)
 

HYPERPLASIA, RITICDLUM CELL 1 (2%)
 

HEMATOFOIFSIE
 2	 (4%)
 

tBESENTFRIC L. NODE (19) (48) (47)
 

LYBPHANGIFCTASIS 1 <5X)
 

HYFEFFLASIA, LYBFHOIE 1 (2%)
 

CIFCUIA1CF Y SYSTE M 

tHFART/ATHIO H (18) (46) (49) 

THBOBB£SISi_NCS 2 jjjjiL 

* NUPBF P CF ANI1ALS.HIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* MOfEE B CF ANIBAI S NECFOESIE D ** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

3 50 ANIBAL S WER E INITIALL Y IS STUD Y BUT ONE KA S DELETED WHF N FOUN D TO BE A FEBALE 

ANIBA L IN A MAL E GROUP . 

C-3 
IMYCCAPEIU M
 

FIPRCSIS
 

•FNCCCAFBIUM
 

THROMBOSIS, NOS
 

*AHTEBY
 

INFLAMMATION , NOS
 

*AORTA
 

MINERALIZATION
 

'PULMONARY APT^RY
 

MldFFALIZA^ICN
 

*M,*EENTF.RIC APTPRY
 MINERALIZATIO N
 

FIBROSIS
 

DIGESTIVE SYSTEM
 

•LIVER
 

CONGESTION, NOS
 

CHCLANRirFIBFOSIS
 

HEPATITIS, TOXIC
 

NECROSIS, FOCAL
 

fITAMORPHOSIS FATTY
 

EASOIHILIC CYTO CHANGE
 

FCCAI CEILULAF CHANGE
 

tLIVtp/CFNTfilLOEULAP
 

N^CPCSIS, NOS
 

•LIVEP/HFPATOCYTFS
 

FYP^FPLAJIA, DIFFUSE
 

*EIIF PUCT
 

HYPEPFLASIA, NOS
 

HYPEFFLASIA, FOCAL
 

•PANCREATIC	 ACINU S
 

ATROPHY, NOS
 

•STC1ACH
 

t MJrEF R O F (NIPIAL S WIT H TI?SU E

* MJPEI F O F ANIBAI S NECPOPSIE D 

TABLECl (CONTINUED) 

CONTROL (UNTR) LOU DOSE
 

11-1475 11-1473
 

(18)	 (46)
 

9 (50%) 10 (22X)
 

(18) (46)
 

(2") (49)
 

1 (2*)
 

(20)	 (49)
 

1 (2*)
 

(20)	 (49)
 

1 (2%)
 

(20)	 (49)
 

1 (2X)
 

1 (2%)
 

(19) (49)
 

4	 (21%)
 

1 (2*)
 

1 <2X>
 

1 (5%) 1 (2%)
 

2 (4%)
 

(19)	 (49)
 

2 (<tX)
 

(19)	 (49)
 

1 (5%)
 

(20)	 (49)
 

1 (5X) 2 (4X)
 

1 (5%)
 

(18)	 (49)
 

1 (6%) 1 (2%)
 

(18) (47)
 

 EXAMINE D MICROSCOPICALL Y 

HIGH COSE
 

11-1471
 

(49)
 

15 (31X)
 

(49)
 

1 <2X)
 

(50)
 

(50)
 

(50)
 

1 (2X)
 

(50)
 

(48)
 

1 (2%)
 

2 (4X)
 

1 (2%)
 

5 (10X)
 

1 (2X)
 

2 (4*)
 

(48)
 

(48)
 

(50)
 

1 (2X)
 

(47)
 

2 (4*)
 

(49)
 

C-4 
TABLE Cl (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH DOSE
 

11-1U75 11-1U73 11-1U71
 

ULCEK, FCCAL 1 (2%)
 

TNFLABBATICN, CHFONIC 1 (2X)
 

HYPEFPLASIA, FOCAL 1 (2X)
 

• LARGE INTESTINE (19) («8) (50)
 

KEBATODIASIS 5 (26%) 18 (38%) 21 (12%)
 

UFINAF Y EYETEB
 

• KIDNEY (19) (149) (50)
 

BltiEFALIZATICN 1 (2X) 1 (2%)
 

CCNGESTICN, NOS 1 (2%) 1 (2%)
 HEMOFRHAGE 1 (2%)
 INFLAMMATION, NOS 1 (2X) 1 (2%)
 PYELONEPHRITIS, ACUTE 1 (2X)
 INFLAMMATION, CHRONIC 1U (7<t%) 21 (13%) 37 (7UX)
 

DEGENERATION, HYALINE 1 (2%) 1 (2X)
 

• KIDNEY/TUEULE (19) (U9) (50)
 

DILATATION, NOS 1 (2%) 1 (2%)
 

• URINARY BLADDER (11) (37) (39)
 

OBSTRUCTION, NOS 1 (3X)
 

INFLAMMATION, ACUTE 1 (3X)
 

*URETHBA (20) (M9) (50)
 

INFLAMMATION, NOS 1 (2%)
 

ENDOCRINE SYSTEM
 

• EITUITAPY (18) (15) <t«>
 HEBOPRHAGE 1 (2*)
 

HEBOFRHAGIC CYST 1 (6%) 1 (2%)
 

ANGIECTASIS 1 (6*)
 

• ADRENAL (19) (it 8) (50)
 

CCNGESTION, NOS 1 (2X)
 

•ADRENAL	 CORTEX (19) (18) (50)
 

DEGENERATION, LIPCID 1 (2%)
 

BETABOEPHOSIS FATTY 1 (2%)
 

tADBFNAL MEDULLA	 (19) (U8) (50)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* KUPEER OF ANIMAL S NFCROPSIED
 

C-5 
#ThYEOir
 

FCIIICULAE CYST, NCS
 

HYEEPPLA^IA, C-CELL
 

•PANCREATIC	 ISLETS
 

hYPEEPLASIA, EOCAL
 

RIEFCEUCTIVE SYSTE!"
 

*EBCSTAT£
 

INFLAMMATION, NOS
 

INFLAMMATION , SDPPURATIVE
 

INFLABKATION, ACUTE
 

INFLAMMATION , CHFONIC
 

(tTFSTIS
 

RINFPA1IZATICN
 

ATROPHY, NOS
 

NEtVCU S SYSTEM
 

tEPAI N
 

HYDROC^PHALOS, INTERNAL
 

SPECIAL SENSE CPGAN S
 

*ChCEOID
 

THROPEOSIS, NO?
 

MUSCUICSPEIETA L	 SYSTEP
 

*SKEIFTJL MDSCLI
 

INFLAMMATION, PYCGPANUL01P TOUS
 

BCTY CAVITIES
 

•PFSENTFRY
 

FERIARTERITIS
 

TABLECl (CONTINUED) 

CONTROL (UNTR) LOW DOSE HIGH
 

11-1475 11-1473 11-1
 

(14) (46) (46)
 

1
 

1 (7%) 2
 

(18)	 (49) (47)
 

1 (2X)
 

(9)	 (42) (40)
 

2 (5% )
 

1 (11%)
 

1	 (2*)
 1
 

(19) (49) (49)
 

1	 (5X) 1 (2%) 1
 

3 (6*) 1
 

(19)	 (43) (50)
 

1 (2*) 1
 

(20)	 (49) (50)
 

1 <2X)
 

(20)	 (49) (50)
 

1 (2*)
 

(20)	 (49) (50)
 

2 (4%) 1
 

DOSE
 

471
 

<2X)
 

(MX)
 

(3X)
 

(2X)
 

(2X)
 

(21)
 

(2X)
 

* MIPEF P CF ANIMAL S WIT H TISSUE EXAMINE D MICFOSCOPICALLY 

* dUPE^ R OF ANIBAL S NECROPSIE D 

C-6 
TABLECl (CONCLUDED)
 

CONTROL (UNTR) LOU DOSE HIGH DOSE
 11-11(75 11-1U73 11-1M71
 

ALL CTHEP SYSTEMS
 

AEIECSE TISSUE
 INFLAMMATION, GEANULONHTOUS
 

NECROSIS, TAT
 

SEICIAL CCFEHCLCGY SOMBAR Y
 

AUTC/NFCROESY/NO HISTO
 

» NDMEEE OF ANIilALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* hUBEEB OF ANIMALS NECROPSIED
 

C-7
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE 

ANIMAL S INITIALLY IN STUDY


ANIfAL S NECFCFSIED


AKICAL S EXAMINE D HI S10PATHOLOGICALL Y


INTIGLMFNTAP Y SYSTEM
 

*SKIN


FPIDFE1AL INCLUSION CYST
 

BF.SFIEATCFY SYSTFP
 

»IUNG


DILATATION, NOS


AIELFCTASIS
 

CCNGFSTICN, NOS


EC^MA, NOS


HEMOFPHAG"
 

INFLAMMATION , INTERSTITIAL
 

ENFUCCNIA , CHRONIC SUPINE
 

CONTROL (UNTR)
 

11-1<t7f
 

 20
 

 20
 

 ** 20
 

 (20)
 

 (20)
 

1 (5%)
 

1 (5%)
 

1 (5%)
 

INFLAMMATION, FOCAL S7! AN OLOMATOU 

FIBFCSIS 

HYFFPFLA£TA , AEINOCATOUS 

HYFFFPLASIA, ALVFCLA P FPT^HEIIUH 

HISTIOCYTOSIS 1 (5%) 

"F.M7CECIETIC SYSTFB 

KECNF. BAPFO W

PYFLCSCL"ECSIS 

 (19) 

tSPLFFN

FIBEOSIS 

FJMCSIEEPOSIE

 (20) 

1 (5%) 

tCEPVICAL LYMPH NODE

ANGI'CTASIS 

 (19) 

•FESFNTFPIC L. NODS

INFI.AMMATICN.I CHRONIC 

 (19) 

LOW POSE 

11-1«7(t 

50 

50 

50 

HIGH DOSE 

11-1172 

50 

50 

50 

(50) 

(50) 

1 

a 

5 

1 

6 

1 

1 

2 

(2%) 

(8%) 

(10%) 

(2X) 

(12%) 

(2%) 

(2%) 

(«*) 

(50) 

1 

(50) 

1 

1 

3 

1 

1 

1 

(2%) 

(2%) 

(2%) 

(6%) 

(2%) 

(2%) 

(2%) 

(US) 

1 

(U9) 

1 

1 

(KB) 

(D8) 1 

(2%) 

(2*) 

<2«) 

.1251 _ . 

(46) 

(50) 

(50) 

2 

(50) 

(IX) 

* NuraF p O F ANIMAL S WIT H TISSU E EXAMINE D MICROSCOPICALL Y * hUfPE F O F ANIMAL S SECFOFSIE D 

** EXCLUDES PARTIALLY AUTOLYZED ANIMALS 
TABLE C2 (CONTINUED)
 

CONTROL (UNTR) 

11-1476 

LOW DOSE 

11-1474 

HIGH DOSE 

11-1472 

INFLAMMATION, GR ANULO M ATCUS 

HYPERPLASIA, LYMPHOID 

1 

1 (2*) 

CIECUIATCRY SYSTEM 

•MYOCARDIUM 

INFLAMMATION, FOCAL 

FIEPOSIS 

(20) 

1 (5%) 

(48) 

1 

9 

(2X) 

(19*) 

(50) 

7 (14*) 

DIGESTIVE SYSTEM 

IIIVFR 

INFLAMMATION, NOS 

INFLAMMATION, FOCAL GR AN ULOMATOU 

SFCRCSIS, FOCAL 

METAMORPHOSIS "ATTY 

BASOFHILIC CYTO CHANGS 

FCCAI CELLULAR CHANGE 

(20) 

1 

1 

5 

(5%) 

(5%) 

(25*) 

(49) 

1 

2 

1 

21 

4 

(2*) 

(4%) 

(2*) 

(43*) 

(8%) 

(50) 

5 (10*) 

14 (28*) 2 (4*) 

*EILE DUCT 

HYPEEPLASIA, NOS 

(20) (50) 

1 (2*) 

(50) 

•PANCREAS 

NECROSIS, FAT 

(20) 

1 (5*) 

(49) (50) 

•PANCREATIC ACINUS 

ATROPHY, NOS 

ATROPHY, FOCAL 

(20) 

1 (5*) 

(49) 

1 <2*> 

(50) 

1 (2*) 

•STOMACH 

ULCER, FCCAL 

HYPERFLASIA, EPITHELIAL 

(20) 

1 (5*) 

(50) (49) 

1 (2*) 

•GASTRIC SUE10COSA 

FCFMA, NCS 

FIEROSIS 

(20) 

1 

1 

(5*) 

(5%) 

(50) 

1 (2*) 

(49) 

•SMALL INTESTINE 

HYPEFPLASIA, LYMPHOID 

(20) (50) 

1 (2*) 

(50) 

•LARGE INTESTINE 

NEMATODIASIS 

(20) 

8 (4Mt) 

(50) 

27 (54*) 

(50) 

24 (48*) 

UEK.ABY SYSTIB 

•KIDNEY 

BINFFALIZATICN 

(20) (50) 

4 1B£L .

(50) 

 2_J£4J 

# NUMEFR OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY 

* KOHEER OF ANIMAL S NECROPSIED 

C-9
 
TABLE C2 (CONTINUED) 

CONTROL(UNTR) LOW DOSE HIGH DOSE
 

11-1176 11-1«7« 11-1472
 

INFLAMMATION, CHRONIC 3 (15«) 7 (1I»X) 4 (8X)
 

l^ECRCSIS, MEDULLAR Y 1 (5*)
 

HYPERPIASIA, TUBULAR CELL 1 (2X)
 

HKICNEY/PFLVTS (20) (50) (50)
 INFLAHBATICN, ACUTE 1 (5X)
 

tUPINARY BLADDER (17) (UU) (37)
 HEMORRHAG E 1 ((,%)
 

INFLAMMATICN, FOCAL 1 (6%)
 

FNCCCFIN! SYSTEB
 

IFITUITHRY (18) (16) (45)
 

CYST, NOS 2 (4%)
 

HEMORRHAGIC CYST 6 (13X) 2 (4X)
 

ANGIECTASIS 1 (6X) 1 <2X) 1 (2%)
 

•ADRENAL	 CORTFX (20) (50) (47)
 

NECEOSIS, FOCAL 1 (5X)
 METAMORPHOSIS FATTY 1 (2%)
 CYTCLOGIC DEGENERATION 1 <5X)
 HYEERFLAJIA, FOCAL 1 (2%)
 

iTHYROID (17) (t5) (44)
 

HYEEFPLAEIA , C-CELL 1 (2%)
 

RFEFCEUCTIVF SYSTEM
 

*FAFKASY GLAND (20) (50) (50)
 

CILATATICN/DUCTS 1 (2X)
 

*VAGINA (20) (50) (50)
 

INFLAMNATICN, ACUTE 1 (2X)
 

fCTFSUS (19) (48) (50)
 

CYST, NOS 1 (2X)
 

INFLAMMATION, FOCAL GRANULOMATOU 1 (2X)
 FIBROSIS 1 (2X)
 

*CERVIX UTF.BI (19) (48) (50)
 

INFLAMMATION, SUEPURATIVE 1 (2X)
 

tCTIRUS/ENDOMETPIUM (19) (48) (50)
 

QYSTi NOS _r _ r 2.J4S1 .2...14X,)
 

* NUBEE R OF ANINALS WITH TISSDE EXAMINED MICROSCOPICALLY
 

* NUBEER OF ANIMAL S NECROPSIEC
 

C-IO
 
INFLAHHATICN, NOS


•OVABY


CYST, NOS


PARCVARIA N CYST


NEFVCUS SYSTEM
 

*EBAIN


HYDHCCEPHALUS, INTERNAL


SPECIAL SEkSE CSGANS
 

NONE
 

HUECDICSFELETAI SYSTEB
 

*STEHNU«


OSTFOSCLEBOSLS


EOE Y CAVITIE S 

*MEEIASTINU«


INFLAHMATICN, GFANULOHATOUS


*rESENTFEY


BINFFALIZATTON


TNFLAHflATICN, GRANULCMATOU S


NECPOSIS, FAT


A LL CTHEF EYSTEB S
 

NONE
 

SPECIAL KCPEHCLCGY EUMMAP Y
 

NC LFSICN FEEORTED


AUTO/NECROPSY/HISTO P?BF


TABLE C2 (CONCLUDED) 

CONTROL (UNTR)


11-1U76


 (19)


 (20)


 (20)


 (20)


 (20)


 2 (10%)


 1


 LOW DOSE HIGH DOSE
 

 11-1!I71» 11-1472
 

1	 (2*)
 

 (US) (50)
 

2 (IS) 2 (14%)
 

1 (2*) 2 (1»X)
 

 (H9) (50)
 

2 (U%)
 

 (50)	 (50)
 

1 (21)
 

 (50)	 (50)
 

1 (2%)
 

 (50)	 (50)
 

1 (2%)
 

1 (2%)
 

2 (H%)
 

t
 

1
 

*	 NUMEEB OF ANIMALS WITH TISSUE EXABINED MICBCSCOPICALLY
 

* MJfEFB OF ANIMALS N-CROPSIED
 

C-ll
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE 

. „ . CONTROL (DNTR) LOU DOSE 

22-2475 22-2473 

AKIFPIS INITIALLY IN STUDY 20 50 ANIMALS KISSING 2 ANIRAL S NECBCFSIED 18 50 ANIMAL S EXAMINED HISTOEATHOLOGICALLY** 18 49 

INTEGUMENTAR Y SYSTEM
 

*SKTN (18) (50) 

FIEROSIS , FOCAL 1 (6*) 

RESFIFATCEY SYSTEM
 

*LUNG/EFCNCHIOLF (17) (H7)
 

INFLAMMATICN, BOS 2 (4«)
 

ILUNG (17) (47)
 

ATELECTASIS
 

THROMBUS, ORGANIZED 1 (2*)
 CCNGFSTICN, NOS 1 (2*)
 

HEMOFPHAGE 2 (4*)
 

INFLAfiMATICN, INTERSTITIAL 3 (18*) 3 (6*)
 

IKFLAKMATICN, SUFFDPATIVE 1 (2X)
 INFLAMMATION, ACUTE FOCAL
 

EEBIVASCDLAR CUFFING 1 (2X)
 CYTOKFGALY 1 (2*)
 FCAM-CELL 2 (UX)
 HYPEPPLASIA, ADENCMATOUS 1 (2X)
 

HEPATCECIETIC SYSTFC
 

»EONE HABHO N (18) (48)
 

HYPEPPLASIA, GRANULOCYTIC
 

FYFLCPOIESIS 1 (2*)
 

#SPLEEN (17) (44)
 

HYPFEFLASIA, BF.TICUL0K CFLL
 

HYEFPPLAEIA, LYMFHOIC 1 (6*) 1 (2X)
 

IC PEC PULP (17) (44)
 

t kUCE" P CF ANIMAL S WIT H TISSU E EXAMINE D (IICFOSCOPICA LL Y 

* liUREE E O F ANIMAL S NECPCPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

HIGH COSE
 

22-2471
 

50
 

50
 

50
 

(50)
 

(49)
 

(49)
 

2 (4«)
 

8 (16X)
 

4 (8X)
 

10 (20X)
 

1 (2«)
 

(47)
 

1 (2*)
 

<U8)
 

1 (2%)
 

2 (4X)
 

(48)
 

D-3
 
TABLE Dl (CONTINUED)
 

HIGH DOSE
 

22-2171
 

(15)
 

1 (2*)
 

(15)
 

1 (2X)
 

1 (2X)
 

1(2%)
 

1 <2X)
 

(18)
 

1 (2X)
 

(18)
 

1 <2X)
 

(50)
 

(19)
 

1 (21.)


1 (2X)
 

1(2%)
 

1 (2X)
 

1 <2X)
 

2 (IX)
 

(19)
 

1 (2X)
 

(19)
 

1 (2X1
 

tMANEIEDLA R L. NCDE


HYFEP.PIASIA, BETICULUH CPLL
 

tMFSENTERIC I. NODE


CONGESTION, NOS
 

CCNGfSTICN, CHRONIC
 

HEMORRHAGE


INFLAMMATION, CHRONIC
 

DEGENERATION, CYSTIC
 

HYFFRPLASIA, PLASMA CELL


HYPERPLAEIA, RETICULOH CELL
 

HYPFFPLAEIA, LYMFHOID
 

HEHATOFOIEEIS
 

CIECtllATCEY SYSTEM
 

IHEAPT/ATRIUM


INFLAMMATION, CHEONIC
 

tMYCCARrlUH


INFLAMMATION, CHEONIC FOCAL
 

*TESTICl)LAR ARTERY


SCLEROSIS
 

DIGESTIVE SYSTEM
 

OLIVER


FIBROSIS, FOCAL
 

CIGFNERATION, NOS
 

KECROSIS, NOS
 

KFCROSIS, FOCAL
 

METAMORPHOSIS FATTY
 

HFFATOCYTOM'GALY
 

HYPEFPLASIA, DIFFDSE
 

POLYPOID HYPERPLASIA
 

tLIVER/PESIPORTAL


INFLAMMATION ACUTE AND CHRONIC
 

INFLAMMATION, CKfiONIC
 

fLIVER/HEPATOCYTES


UYPEJPiiS^ii.DIFFU^E _
 

CONTROL (UNTR)
 

22-2175
 

 (16)
 

 (16)
 

1 (6X)
 

1 (6%)
 

 (17)
 

 (17)
 

 (18)
 

 (19)
 

 (18)
 

 (18)
 

LOW DOSE
 

22-2173
 

(39)
 

(39)
 

1 (3X)
 

1 (3*)
 

1 (3*)
 

1 <3X)
 

(16)
 

(16)
 

(50)
 

1(2%)
 

(US)
 

1 (2%)
 

1 (2*)
 

1 (2%)
 

<«8)
 

1 (2X)
 

(18)
 

J_J2Si__
 

t -KOMEEP OF ANIMALS UITH TISSDF EXAMINED MICROSCOPICALLY
 

* NUfEER OF ANIMALS NECROPSIED
 

D-4
 
• PANCREAS


BILATATICN/DUCTS
 

INFLAtlHATION, ACUTE
 

ATRCPHY, NOS
 

IIAFGE INTESTINE


NFMATODIASIS
 

UR1NAF Y SYST5P
 

*KICNEY


HYDRCNEPHHCSIS
 

CONGESTION, NOS
 

INFLACHATICN, CHRONIC


EERIVASCULAR CUFFING
 

IN?AFCT, HEALEC
 

HYFERFLASIA, TUBULAR CELL
 

#UFINAHY ELADEER


INFLAMMATION , CHECNIC
 

NCCULE
 

HYEEPFLAEIA, EPITHELIAL
 

ENtCCBIN E SYSTEM
 

HACPENA L CORTTX


FYPFFPIASIA, FOCAL
 

*1f-YROID


FYtERPLASIA, FOLLICULA P-CFLL


IFANCPEATIC ISLETS


FYPFRTEOPHY, NOS
 

HYPEPPLAEIA, NOS
 

BEEFCCUCTIVE SYSTEM
 

*SEMINAL VFSICLE


INFLAMMATION, SUPPUP, ATTV
 

tTESTIS/TUBULE


II£ENEBAJICN^_JJ2£


* tvUflEEF , O F ANIMAL S WIT H TISSU E

* (.DrEFR CF ANIMAL S NECfCPSIE D 

TABLE Dl (CONTINUED) 

CONTROL (UNTR) LOW DOSE
 

22-2175 22-2K73
 

 (16)	 (18)
 

1 (2%)
 

1	 (251)
 

 (17) (19)
 

 (18)	 (18)
 

1 (2*)
 

1 (25S)
 

3	 (17X) 3 (61)
 

1 (2*)
 

2 (1)i)
 

1 (2%)
 

 (13) (38)
 

1 (3%)
 

1 (3*)
 

 (16)	 (13)
 

1 <2X)
 

 (7)	 (31)
 

1 (1U%)
 

 (16) (18)
 

1	 (2*)
 

 (18)	 (50)
 

1 (2*)
 

 (17) (17)
 

. JL12SJ
 

 EXAMINE D MICROSCOPICALL Y 

HIGH DOSE
 

22-2171
 

(18)
 

1 (2%)
 

(18)
 

5 (10%)
 

(50)
 

1 (2%)
 

(37)
 

1 (3%)
 

(15)
 

(23)
 

(18)
 

1 (2*)
 

(50)
 

(18)
 

D-5 
TABLE Dl (CONCLUDED) 

CONTROL (UNTR)

22-2175

 LOH DOSE

 22-2173

 HIGH DOSE 

 22-2171 

NFEVCU E SYSTEM 

*ERAIN

KINFEAIIZATICN

 (18)

 2 (11%)

 (19)

 17 (35%)

 (51) 

7 (11*) 

SEICIAL SENSE CBGANS 

NONE 

BUECUIOSKE1ETAI SYSTEB 

NONE 

BCIY CAVITIES 

*ELFURA

INFLAMMATION, FOCAL

INFLAMMATION ACTIVE CHRONIC

GRANULOMA, NO?

FCAP-CELL

 (18) (50)

 1 (2%) 

1 (2%) 

 (50) 

1 (2%) 

2 (1!!) 

All CTHFE SYETEBS 

*BDITIPLE ORGANS

EEEIVASCULAR CUFFING

AMYLOIDOSIS

 (18) (50)

 1 (2X) 

1 (2%) 

 (50) 

SEFCIAl rCBtHCIOGY SOPPABY 

NC LESION FFEORTED 6

AfilPAL MISSING/NO NECROPSY 2 

AUTC/NFCEOESY/HISTC BEEF

«OTC/NECFOESY/!)0 HISTO

* MJBEEF OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUBEE R OF ANIMALS NFCRCESIFD 

9

 1 

1 

 13 

D-6
 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH 1-PHENYL-3-METHYL-5-PYRAZOLONE 

CONTROL(UNTR ) 

22-2176 

LOS

22­

 DOS E 

2171* 

ANICAL S INITIALLY IN STUDY

ANIMAL S PISSING 

ANIMAL S NECECPSIED

 20 

 20 

50 

1 

«9 

ANIMALS EXAMINED HISTOPATHOLnGICALLY ** 20 U9
 

INTEGIMINTAR Y SYSTEM
 

NCN"
 

F.ESFIEATCEY SYSTEP
 

*IUNG (20) (16)
 ATFLECTASIS 2 (10*)
 CCNGESTICN, NO? 2 (1C*) 1 (2*)
 HYPEfiEMIA 1 (5%)
 HEMORRHAG E 2 (1C*) 2 («*)
 EFCNCHCFNFUNONIA, NOS 1 (2*)
 INFLAMMATION , INTERSTITIAL 8 («C*) 10 (22*)
 FNFUMCNIA, CHRONIC HURINE 1 (2*)
 GRANULOMA , NOS 1 (2*)
 fESIVASCULAR CUFFING 1 (5%) 3 (7*)
 HYPEFPLASIA, AEENOKATODS 1 (5«)
 

HrfATCfCIFlIC SYSTEM
 

*ECN" MAPEO W (19) (15) 

PEHOSIDE50SI S 1 (5%) 1 (2*) 

hYFEPPLA=IA , GRANULOCYT C 2 (1%) 

FN d") (15) 

HEMOFBHAG E 1 (2*) 

EERIVASCULITI? 

^£C!>CSIF, NOS 

HYPERPLASIA, LYMFHOID 1 (5*) 1 (2%) 

HIMATOPOISSIS 

*1"ESENTEFIC L. NODE (17) (10) 

CONGESTION, NOS 

1 J1S1 

* MirEFR CF ANIBAL S WIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* NUfFF F CF ANI1AL S NFCECPSIE L "EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

HIGH DOSE 

22-2172 

50
 

3
 

17
 

46
 

(16)
 

2 (IX)
 

1 (2*)
 

6 (13*)
 

1 (2*)
 

(10) 

1 <3X ) 

(12) 

1 (2*) 

1 (2%) 

1 (2*) 

1 (2%) 

(39) 

1 (3*) 

D-7 
TABLE D2 (CONTINUED)
 

HIGH DOSE
 

22-2U72
 

1 (3*)
 

1 (3%)
 

(12)
 

1 (2%)
 

<U7)
 

(U6)
 

1 (2X)
 

1 (2%)
 

1 <2X)
 

1 (2%)
 

1 (2%)
 

1 (2«)
 

1 (2%)
 

1 (2%)
 

(M6)
 

1 (2%)
 

1 (2%)
 

(K6)
 

(15)
 

hYPERPLASIA
 

HYFERFLAEIA
 

HYPERPLASIA
 

, NOS
 

, RETICULUM CELL
 

, LYMPHOID
 

CIFCUIATCRY SYSTIf
 

IMYCCARDIUt
 

INFLAfCATICI ACTIVE CHFONIC
 

*PULBONARY ABTEBY
 

HYFEFTROEHY, NOS
 

DIGESTIVE SYSTEM
 

ILIVEH


HEKOERHAGE


INFLAMMATION, FOCAL


LYrpHCCYTIC INFLACMATOPY INFILTP
 

INFLAMRATION , ACUT E FOCPL

INFLAMBATION, ACU1E NFCPOTIZING
 

INFLAMMATION ACUTE AND CHRONIC
 

INFLAMTATICN, CHFONIC FOCAL


ABSCESS, CHRONIC
 

GRANULCMA, NOS
 

FIBPCSIS


EERIVAECULITIS
 

FFRIVASCULAR CUFFING


NECROSIS, NOS


NECROSIS, FOCAL


NECROSIS COASULATIV E 

IKFAFCT , NOS 

HIPATOCYTOMEGALY
 

HYPEPPLASIA, NOS


tLIVER/EERIPORTAL


INFLAMMATION, CHRONIC
 

GBANULOMA, NOS
 

ULIVIP/HF-PATOCYTES


EEGENIEATION, NOS


HYPEPPLASIA, KOS
 

HYEEPFLASIA, DIFFUSE
 

IPANCREAS


CONTROL (UNTR)


22-2t76


(18)
 

(20)
 

 (20)
 

1 (5%)
 

1 (5%)
 

2 (1CS) 

1 (5X)
 

1 (5%)
 

2 (10X)
 

1 (S%)
 

1 (5S)
 

1 (^»)
 

 (20)
 

 (20)
 

1 (5»)
 

 (1S)
 

 LOW DOSE
 

 22-2174
 

1 (3%)
 

2 (5%)
 

1 (2X)
 

(U7)
 

1 (2«)
 

1 (2%)
 

1 <2X)
 

6 (13%)
 

2 (««)
 

1 (2%)
 

(17)
 

(1*7)
 

2 (U«)
 

1 (2%)
 

* NUMBER OF ANIMALS WITH TISSUF EXAMINFD MICROSCOPICALLY
 

* 1.UMEER OF ANIMAL S
 

D-8
 
NICRCSIS, NOS
 

ATROEHY , NOS
 

»FEYEPS PATCH
 

HYPIfpLPSH, lYMPhTID
 

(•LARGE IN'T'STIN'
 

HECATODIASIS
 

UFINAF Y SfSllf
 

KKITN^Y
 

HYDPCN'EhEOSIE
 

INFLAMMATION , CHPOMC
 

FERIVASCULA P COF^H*^
 

INFARCI, HFALE D
 

HYEE^PIAEIA , TUBULA R CELL
 

• KIENFY/flEE'JLLA
 

CYST, NCS
 

INUCCFINF EYE1I.1
 

•ADF1NAL
 

A C Y10IOOSIS
 

"FEFCEUCTIV1' EYETFT
 

ItTFPUS
 

HEMOF^HAGF
 

PF10PPHAGIC CY?^
 

TNFLA "BATICN, NO°
 EYOfETFA
 

INFLA M"ATICN, NECFOTIZINO
 

A"fCFSS, NOS
 

INFLAn^ATTCN , CHRONIC
 

dCEPVI X OTFRI
 

INFLAMMATICN , NOS
 

»01FRDS/IUDCB"TEIl"l
 

CYST, NOS
 

* MlfEr P OF ANI1AL S H1H TI^EU F

* kUCEF R CF ANIHAL S NECPOFSIF P 

TABLE D2 (CONTINUED) 

CONT»OL (UNTR) LOW DOSE
 

22-2176 22-2171
 

1 (5%)
 

(20)	 (15)
 

1 (2%)
 

(20)	 (15)
 

1 (5%) 1 (2?,)
 

(19) (16)
 

5 (26«) 7 (15%)
 

1 (5%) 1 (9*)
 

1 (^1% )
 

1 (2%)
 

(19) (16)
 

(19)	 (12)
 

1 (2«)
 

(19) (16)
 

1 (2%)
 

1 (2%)
 

1 (2H)
 

(19) (16)
 

(19) (16)
 

 EXAMINF D MICROSCOPICALL Y 

HIGH DOSE
 

22-2172
 

1 (2%)
 

(13)
 

(13)
 

1 (2%)
 

(11)
 

1 (2%)
 

1 (9*)
 

(11)
 

1 (2%)
 

(11)
 

(15)
 

1 (2*)
 1 (2%)
 

1 (2%)
 

1 (2%)
 

(15)
 

1 (2%)
 

(15)
 

2 (IX)
 

D-9 
TABLE D2 (CONTINUED) 

CONTRO L (UNTP ) L OW DOSE HIG H COSE 

22-2176 22-247U 22-2U7 2 

INFLAMMATION , S'JFFUPATIVE 

INFLAMFATICN , ACOTE 

1

2

 (5%) 

 (11%) 

1

2

 (2%) 

<<*% ) 1

2

 (2%) 

 (i»%) 

EIE"(CSIS 1 (2%) 

HYPFKPIASIA, NOS 1 (2%) 1 (2%) 

HYPFFELAJTA , CYSTIC 8 (12%) 15 (33%) 19 (U2% ) 

HYPEFPLAflA . STRCMAL 1 (2% ) 

*CVA°Y (16) (U2) (38) 

CYST, NOS 1 (2*) 1 (3%) 

FC1IIC01AR CYST, KC 5 1 (6%) 3 (71!) 6 (16*) 

CCPFUS LCTE'IH CY£: 1 (2*) 

EARCVASIA N CYST 1 (3«) 

THFCKBOSIS, NOS 1 (3%) 

HEdOFEHAG F 1 (3%) 

HEMOFRHAGI C CYST 1 (3%) 

ABSCFSS, NOS 1 (2%) 

NFFVCU f SY=TEf; 

«EPAI>:/MFNISGE S (20) (U7 ) (13) 

INFLABHATICN , NCS 1 (2% ) 

*F.PAI N (20) (U7 ) (<43 ) 

CINFPAIIZATIC N K (20% ) 17 (36%) 14 (33%) 

SEICIM EFNS; racAss 

NCN11 

MUSCUIOSKFIFTA I SYSTEi" 

DON? 

BCIY CAVITIE S 

*AEDCMINAI CAVITY (20) («9) (U7) 

INFLAMflATICN , NFCROTIZIN G 1 (2% ) 

*FLFDSA (20) (U9) (17) 

NCDUL" 1 (2%) 

*CESENTrPY (20) (U9) C*7) 

# NnrEF P O F ANIMAL S WIT H TISSU E FXAKINE D dlCBOSCOPICALL Y 

* NUCBE F OF ANIMAL S NFCFOFSI^C 

D-IO
 
TABLE D2 (CONCLUDED) 

CONTPO t (UNTR )

22-?U7 6

 LO W DOSE

 22-2U7 U

 HIG H DOSE 

 22-2U7 2 

A ll CThEF SYS1FCE 

*BUITIPIF CFGAN f (20) (<49) (K7) FEPIVASCdl«P CUFFING 1 (=;%) 1 (2*) 

A("YLPI->CSIS 1 (2X) 

SFECIAI PCFEF-CLCHY ''U'-CATY 

NC LFSIC N fECFTE T 5 14 

PMPA L riSSISG/N O N^CKODS Y 1 3 

SUTC/NPCFCESY/N C rlS^ T 1 

» NUMBE R O F ANI1AI S HIT h T ISSU E EXAflNE P fICBOSCOPTCALIY 

* NDCEE P O F AM1AI S N'CEOPSIE T 

D-ll 


DHEW Publication No. (NIH) 78-1396