1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

NO. 83 

BIOASSAY OF 

DAMINOZIDE 

FOR POSSIBLE CARCINOGEN ICITY 

CAS No. 1596-84-5 

NCI-CG-TR-83 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

DAMINOZIDE
 

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) 78-1333
 

BIOASSAY OF
 

DAMINOZIDE
 

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
 

daminozide 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 environmental chemicals have the
 

capacity to produce cancer in animals. Negative results, in
 

which the test animals do not have a greater incidence of cancer
 

than control animals, do not necessarily mean that the test
 

chemical is not a carcinogen, inasmuch as the experiments are
 

conducted under a limited set of circumstances. Positive results
 

demonstrate that the test chemical is carcinogenic for animals
 

umder the conditions of the test indicate that exposure to
 

the chemical is a potential risk man. The actual determination of the risk to man from anima., c '.rcinogens requires a wider
 

analysis.
 

CONTRIBUTORS: This bioassay of daminozide was conducted by
 

Litton Bionetics, Inc., Keasington, Maryland, initially under
 

direct contract to the NCI and currently under a subcontract to
 

Tracer Jitco, lie., prime contractor for the NCI Carcinogenesis
 

Testing Program.
 

The experimental design and doses were chosen by Drs. E. K.
 

Weisburger1, J. H. Weisburger1'2, N. P. Page1'^ and F. M.
 

Garner^. The administration of the test chemical and the
 

observation of the animals were supervised by Dr. F. M. Garner^,
 

with the technical assistance of Mr. R. Cypher^, Mr. H. D.
 

Thornett^, and Mr. D. J. Howard^.
 

iii
 
Histologic examinations of rats were performed by Drs. H.
 

Seibold^, B. C. Zook , and N. J. Wosu^ and those of mice by Drs.
 

R. J. Montali^, F. M. Garner^, N. J. Wosu^, and H. Seibold^.
 

Histologic sections of all tumors and hyperplasias were
 

reexamined by Dr. Montali, who also reviewed all diagnoses and
 

prepared the initial interpretative pathology summary.
 

Animal pathology tables and survival tables were compiled at EG&G
 

Mason Research Institute^. The statistical analyses were
 

performed by Dr. J. R. Joiner" and Ms. P. L. Yong", using methods
 

selected for the bioassay program by Dr. J. J. Gart^. Chemicals
 

used in this bioassay were analyzed under the direction of Dr. E.
 

Murrlllo

0

, feed samples were analyzed by Mr. H. Paulin AH, and the
 

results of the analyses were reviewed by Dr. S. S. Olin°.
 

This report was prepared at Tracer Jitco^ under the direction of
 

NCI. Those responsible for the report at Tracer Jitco were Dr.
 

L. A. Campbell, Director of the Bioassay Program; Dr. S. S. Olin,
 

Deputy Director for Science; Drs. J. F. Robens and C. H.
 

Williams, toxicologists; Dr. R. L. Schueler, pathologist; Dr. G.
 

L. Miller, Ms. L. A. Waitz, and Mr. W. D. Reichardt, bioscience
 

writers; and Dr. E. W. Gunberg, technical editor, assisted by Ms.
 

Y. E. Presley and Ms. P. J. Graboske.
 

The following scientists at NCI were responsible for evaluating
 

the bioassay, interpreting the results, and reporting the
 

findings: Dr. Kenneth C. Chu, Dr. Cipriano Cueto, Jr., Dr. J.
 

Fielding Douglas, Dr. Dawn G. Goodman, Dr. Richard A. Griesemer,
 

Dr. Harry A. Milman, Dr. Thomas W. Orme, Dr. Robert A. Squire^,
 

Dr. Jerrold M. Ward.
 

1-Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

iv
 
with the National Institute of Occupational Safety and
 

Health, Rockville, Maryland.
 

^Litton Bionetics, Inc., 5516 Nicholson Lane, Kensington,
 

Maryland.
 

5

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

Rockville, Maryland.
 

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

Maryland.
 

^Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute, National
 

Institutes of Health, Bethesda, Maryland.
 

°Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of daminozide, a plant growth regulator, for possible
 

carcinogenicity was conducted by administering the test chemical
 

in the diet to Fischer 344 rats and B6C3F1 mice.
 

Groups of 50 rats and 50 mice of each sex were administered
 

daminozide at one of two doses, either 5,000 or 10,000 ppm, for
 

104 weeks, then observed for an additional week. Matched
 

controls consisted of 20 untreated males and 20 untreated females
 

of each species. All surviving rats and mice were killed at 105
 

weeks.
 

Mean body weights of the high-dose female mice were appreciably
 

lower than those of the corresponding controls, while mean body
 

weights of all other dosed groups of rats and mice were essentially unaffected. No other clinical signs related to administration of daminozide were observed. Sufficient numbers of animals
 

in all groups of rats and mice were at risk for development of
 

late-appearing tumors.
 

In the male rats, no tumors occurred at incidences that were
 

significantly higher in dosed groups than in controls, except for
 

interstitial-cell tumors of the testis (controls 13/20, low-dose
 

49/50, high-dose 47/50). These tumors occurred, however, at a
 

high spontaneous rate (182/220) in historical-control male rats;
 

thus, the association of the interstitial-cell tumors with
 

administration of the chemical is doubtful.
 

In the female rats, adenocarcinomas of the endometrium and
 

leiomyosarcomas of the uterus occurred only in the dosed groups
 

(adenocarcinomas: controls 0/19, low-dose 5/50, high-dose 3/50;
 

leiomyosarcomas: controls 0/19, low-dose 1/50, high-dose 3/50).
 

The incidences in the dosed groups were too low to be statistically significant; however, the low incidence of these tumors in
 

historical—control female rats (2/220 adenocarcinoma and 0/220
 

leiomyosarcoma) indicate that the occurrence of these tumors in
 

vil
 
the dosed female rats was associated with the administration of
 

daminozide.
 

In the male mice, there was a dose-related trend (P = 0.008) in
 

the incidence of hepatocellular carcinomas; also, the incidence
 

in the high-dose group was significant (P = 0.020) compared with
 

that in the controls (controls 0/14, low-dose 7/50, high-dose
 

13/46). The incidence of these tumors in the historical-control
 

male mice was, however, 21/216; thus, the association of the
 

hepatocellular carcinomas with administration of daminozide is
 

not clear. In the female mice, only three such tumors occurred.
 

It is concluded that under the conditions of this bioassay,
 

daminozide was not carcinogenic in the male Fischer 344 rats or
 

in the female B6C3F1 mice. In male B6C3F1 mice, the induction of
 

hepatocellular carcinomas may have been associated with the
 

administration of the test chemical. Daminozide was carcinogenic
 

in female Fischer 344 rats, inducing adenocarcinomas of the
 

endometrium of the uterus and leiomyosarcomas of the uterus.
 

viii
 
TABLE OF CONTENTS
 

Page
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical. 3
 

B. Dietary Preparation 3
 

C. Animals. 4
 

D. Animal Maintenance 5
 

E. Subchronic Studies 6
 

F. Designs of Chronic Studies 8
 

G. Clinical and Pathologic Examinations 8
 

H. Data Recording and Statistical Analyses 11
 

III. Results -Rats 17
 

A. Body Weights and Clinical Signs (Rats) 17
 

B. Survival (Rats) 17
 

C. Pathology (Rats) 20
 

D. Statistical Analyses of Results (Rats) 22
 

IV. Results - Mice 27
 

A. Body Weights and Clinical Signs (Mice) 27
 

B. Survival (Mice) 27
 

C. Pathology (Mice) 30
 

D. Statistical Analyses of Results (Mice) 31
 

V. Discussion 35
 

VI. Bibliography 39
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in
 

Rats Fed Daminozide in the Diet 41
 

Table Al Summary of the Incidence of Neoplasms in
 

Male Rats Fed Daminozide in the Diet 43
 

Table A2 Summary of the Incidence of Neoplasms in Female
 

Rats Fed Daminozide in the Diet 47
 

ix
 
Page 

Appendix B Summary of the Incidence of Neoplasms in 

Mice Fed Daminozide in the Diet. 51 

Table Bl Summary of the Incidence of Neoplasms in 

Male Mice Fed Daminozide in the Diet 53 

Table B2 Summary of the Incidence of Neoplasms in Female 

Mice Fed Daminozide in the Diet 57 

Appendix C Summary of the Incidence of Nonneoplastic Lesions 

in Rats Fed Daminozide in the Diet 61 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Fed Daminozide in the Diet.... 63 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Fed Daminozide 

in the Diet 67 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Fed Daminozide in the Diet 71 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Fed Daminozide in the Diet.... 73 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Fed Daminozide in the Diet.. 80 

Appendix E Analyses of the Incidence of Primary Tumors 

in Rats Fed Daminozide in the Diet 89 

Table El Analyses of the Incidence of Primary Tumors in Male 

Rats Fed Daminozide in the Diet 91 

Table E2 Analyses of the Incidence of Primary Tumors in Female 

Rats Fed Daminozide in the Diet 95 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Fed Daminozide in the Diet 101 

Table Fl Analyses of the Incidence of Primary Tumors in Male 

Mice Fed Daminozide in the Diet 103 

Table F2 Analyses of the Incidence of Primary Tumors in Female 

Mice Fed Daminozide in the Diet 106 
Page 

TABLES 

Table 1 Design of Daminozide Chronic 

Feeding Studies in Rats 9 

Table 2 Design of Daminozide Chronic 

Feeding Studies in Mice 10 

FIGURES 

Figure 1 Growth Curves for Rats Fed Daminozide 

in the Diet 18 

Figure 2 Survival Curves for Rats Fed Daminozide 

in the Diet 19 

Figure 3 Growth Curves for Mice Fed Daminozide 

in the Diet 28 

Figure 4 Survival Curves for Mice Fed Daminozide 

in the Diet 29 

xi
 

I. INTRODUCTION
 

Daminozide (CAS 1596-84-5; NCI C03827) is a hydrazine compound
 

that has been used as a plant growth regulator since 1962
 

(Riddell et al., 1962). It retards stem growth, protects against
 

heat, drought, and frost, and induces the development of multiple
 

flowers (Thomson, 1976). The Environmental Protection Agency
 

(1976) has established residue tolerances of 1-55 ppm on a
 

variety of crops which include cherries, plums, apples,
 

nectarines, peaches, pears, grapes, melons, tomatoes, brussel
 

sprouts, peppers, and peanuts; residues of 0.02-2 ppm are allowed
 

in the meat or milk of animals exposed to the herbicide.
 

Daminozide was selected for testing for carcinogenic activity
 

because other hydrazine compounds had been shown to be carcinogenic, as reviewed by Toth (1975), and because of the potential
 

for long-term human exposure to this chemical during agricultural
 

application or from residues in food products.
 

II. MATERIALS AND METHODS
 

A. Chemical
 

Daminozide, which is the common name for succinic acid mono-2,2­

dimethylhydrazide, was obtained from Aldrich Chemical Co.,
 

Milwaukee, Wisconsin, in three lots (Lot Nos. 101927, 121327, and
 

071637). The identity and purity of each lot was confirmed by
 

analysis. Elemental analyses (C, H, N) were consistent with
 

C

6H12N203> tne molecular formula for daminozide. Titration with
 

sodium hydroxide indicated purities close to 100% (100.0, 100.2,
 

and 99.3, respectively). Thin-layer chromatography showed a
 

single component. Melting point and infrared and nuclear
 

magnetic resonance spectra were consistent with the structure.
 

The chemical was stored at 4°C in the original container.
 

B. Dietary Preparation
 

A 6-kg diet was prepared twice per week for mice and three times
 

per week for rats. To obtain each dietary concentration, the
 

appropriate weight of daminozide was mixed with a small portion
 

of Wayne® Lab Blox animal feed (Allied Mills, Inc., Chicago,
 

111.), using a mortar and pestle. This premix was then added to
 

the remaining weight of feed and mixed in a twin-shell blender
 
for at least 15 minutes. Feed preparations containing the test
 

chemical were stored at 1°C for no longer than 1 week.
 

The stability of the chemical in feed was checked by triplicate
 

analyses at 5,000 and 10,000 ppm. The recovery of test chemical
 

from feed decreased about 20% after the feed was stored for 10
 

days at 25°C; thus, some decomposition of the test chemical
 

appeared to occur in the feed hoppers under the conditions of
 

this bioassay.
 

Ground Wayne® Lab Blox animal meal not containing the test
 

chemical was used as the diet for the control groups of animals.
 

C. Animals
 

Rats and mice of each sex, obtained through contracts of the
 

Division of Cancer Treatment, National Cancer Institute, were
 

used in these bioassays. The rats were of the Fischer 344 strain
 

obtained from A. R. Schmidt/Sprague-Dawley, Madison, Wisconsin,
 

and the mice were B6C3F1 hybrids obtained from Charles River
 

Breeding Laboratories, Inc., Wilmington, Massachusetts. On
 

arrival at the laboratory, the animals were approximately 4 weeks
 

of age. All animals were quarantined for 2 weeks. After the
 

period of quarantine, all animals were weighed individually and
 

segregated into equal weight groups. Cage assignments were made
 
by selecting one animal from each group so that the total weights
 

of animals in each cage were about the same.
 

D. Animal Maintenance
 

All animals were housed in temperature- and humidity-controlled
 

rooms. The temperature was maintained at 21-25°C and the
 

relative humidity at 45-55%. There were 15 changes of room air
 

per hour, and the air was passed through both incoming and
 

exhaust HEPA (High Efficiency Particulate Air) filters. The
 

animal rooms were positively pressurized with respect to the exit
 

hall and negatively pressurized with respect to the entrance
 

hall. Cool white fluorescent lighting was provided 8 hours per
 

day. Test and control diets were available ad libitum and were
 

replenished three times per week. Tap water, acidified with HC1
 

to pH 2.5, also was available ad libitum.
 

Rats were housed four per cage and mice five per cage in solid
 

polycarbonate cages. Each cage was covered with a wire mesh
 

screen and a sheet of filte-r paper. Heat-treated hardwood chip
 

bedding (Absorb-Dri®, Lab Products, Garfield, N.J.) was used in
 

the cages. Cages and water bottles were sanitized twice per week
 

and feed hoppers once per week at approximately 82°C; bedding was
 

replaced twice per week.
 

Rats and mice were housed in separate rooms. Control animals and
 
dosed animals were housed in the same room. Animals administered
 

daminozide were housed in the same room as animals of the same
 

species administered the following chemicals:
 

RATS
 

hydroxytriphenylstannane (CAS 76-87-9)
 

2-nitroethenylbenzene (CAS 102-96-5)
 

N-(aminocarbonyl)-2-bromo-2-ethylbutanamide (CAS 77-65-6)
 

MICE
 

hydroxytriphenylstannane (CAS 76-87-9)
 

4,4'-diisocyanato-3,3'-dimethoxy-l,l'-biphenyl (CAS 91-93-0) 

N-(aminocarbonyl)-2-bromo-2-ethylbutanamide (CAS 77-65-6) 

N.N'-diethylthiourea (CAS 105-55-5) 

iodomethanesulfonic acid, sodium salt (CAS 126-31-8)
 

2,5-cyclohexadiene-l,4-dione, dioxime (CAS 105-11-3)
 

3-hydroxy-(3 alpha, 5 beta) cholan-24-oic acid (CAS 434-13-9)
 

4-amino-2-nitrophenol (CAS 119-34-6)
 

ethylenediaminetetraacetic acid, trisodium salt trihydrate
 

(EDTA) (CAS 150-38-9)
 

All of these compounds were administered in the diet, with the
 

exception of 2-nitroethenylbenzene and 3-hydroxy-(3 alpha, 5
 

beta)cholan-24-oic acid, which were administered by gavage, and
 

iodomethanesulfon-ic acid, sodium salt, which was administered by
 

intraperitoneal injection.
 

E. Subchronic Studies
 

Subchronic feeding studies were conducted with rats and mice to
 

estimate the maximum tolerated doses of daminozide, on the basis
 

of which low and high concentrations (hereinafter referred to as
 
"low doses" and "high doses") were determined for administration
 

in the chronic studies. In the subchronic studies, daminozide
 

was added to the animal feed in concentrations of 10,000, 20,000,
 

30,000, 40,000, or 50,000 ppm. Feed containing the chemical was
 

provided to groups of five male and female animals of each
 

species for 6 weeks, and the standard commercial diet was
 

similarly provided to groups of five male and female animals of
 

each species. All surviving animals were killed and necropsied
 

at week 8.
 

In dosed male and female rats, soft stools and diarrhea were
 

noted at doses of 20,000 ppm and above. After week 7, there were
 

no effects on mean body weight gain in rats, except for a slight
 

weight depression in the males administered 50,000 ppm. Water
 

consumption also increased in this group after week 3 of the
 

study.
 

There were no effects on mean body weight gain in the mice
 

administered the different doses; however, during week 2, two
 

female mice died that were administered 30,000 ppm. There were
 

no gross pathologic changes noted in any of the dosed animals.
 

The low and high doses for the chronic studies using rats and
 

mice were set at 5,000 and 10,000 ppm.
 
F. Designs of Chronic Studies
 

The designs of the chronic studies are shown in tables 1 and 2.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity, and
 

animals that were moribund were killed and necropsied. Rats and
 

mice were weighed individually at regular intervals. Palpation
 

for masses was carried out at each weighing.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of all major tissues, organs, or gross lesions from
 

killed animals and from animals found dead. The following
 

tissues and organs were routinely subjected to microscopic
 

examination: skin, lymph nodes, mammary gland, salivary gland,
 

bone marrow, trachea, lungs and bronchi, heart, thyroid, parathyroid, esophagus, stomach, small intestine, large intestine,
 

liver, gallbladder (mice), pancreas, spleen, kidney, adrenal,
 

urinary bladder, prostate or uterus, testis or ovary, brain, and
 

pituitary. Occasionally, additional tissues were also examined
 

microscopically. The different tissues were preserved in 10%
 

buffered formalin, embedded in paraffin, sectioned, and stained
 

with hematoxylin and eosin. Special staining techniques were
 

utilized when indicated for more definitive diagnosis.
 
Table 1. Design of Chronic Feeding Studies of Daminozide
 

in Rats
 

Sex and 

Test 

Group 

Initial 

No. of 

Animals3 

Daminozide 

in Diet 

(ppm)b 

Time 

Dosed 

(Weeks) 

on Study 

Observed 

(weeks) 

Male 

Matched-Control 20 0 105 

Low-Dose 50 5,000 104 1 

High-Dose 50 10,000 10.4 1 

Female 

Matched-Control 20 0 105 

Low-Dose 50 5,000 104 1 

High-Dose 50 10,000 104 1 

a

Animals were approximately 42 days of age when placed on study.
 

^Test diets were made available ad libitum, 7 days per week.
 
Table 2. Design of Chronic Feeding Studies of Daminozide
 

in Mice
 

Sex and 

Test 

Group 

Initial 

No. of 

Animals3 

Daminozide 

in Diet 

(ppm)b 

Time 

Dosed 

(Weeks) 

on Study 

Observed 

(weeks) 

Male 

Matched-Control 20 0 105 

Low-Dose 50 5,000 104 1 

High-Dose 50 10,000 104 1 

Female 

Matched-Control 20 0 105 

Low-Dose 50 5,000 104 1 

High-Dose 50 10,000 104 1 

a

Animals were approximately 42 days of age when placed on study.
 

^Test diets were made available ad libitum, 7 days per week.
 

10
 
A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals were
 

missing, cannibalized, or judged to be in such an advanced state
 

of autolysis as to preclude histopathologic evaluation. Thus,
 

the number of animals from which particular organs or tissues
 

were examined microscopically varies, and does not necessarily
 

represent the number of animals that were placed on study in each
 

group.
 

H. Data Recording and Statistical Analyses
 

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

System (Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design,
 

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

Against Cancer (Berenblum, 1969). Data tables were generated for
 

verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear on the possibility of carcinogenicity are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

11
 
procedure of Kaplan and Meier (1958) and are presented in this
 

report in the form of graphs. Animals were statistically
 

censored as of the time that they died of other than natural
 

causes or were found to be missing; animals dying from natural
 

causes were not statistically censored. Statistical analyses for
 

a possible dose-related effect on survival used the method of Cox
 

(1972) for testing two groups for equality and Tarone's (1975)
 

extensions of Cox's methods for testing for 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 is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic examination was required to detect lesions prior to histologic sampling
 

(e.g., skin or mammary tumors), or when lesions could have
 

appeared at multiple sites (e.g., lymphomas), the denominators
 

consist of the numbers of animals necropsied.
 

The purpose of the statistical'analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

12
 
significantly higher proportion of tumors than did the control
 

animals. As a part of these analyses, the one-tailed Fisher
 

exact test (Cox, 1970) was used to compare the tumor incidence of
 

a control group with that of a group of dosed animals at each
 

dose level. When results for a number of dosed 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) 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), was also used. Under the
 

assumption of a linear trend, this test determines 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 is a positive dose relationship. This method also provides a two-tailed test of departure
 

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

13
 
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 an animal died naturally or was 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 with its control was calculated
 

14
 
from the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as Pt/Pc where pt is the true binomial
 

probability of the incidence of a specific type of tumor in a
 

dosed group of animals and pc is the true probability of the
 

spontaneous incidence of the same type of tumor in a control
 

group. The hypothesis of equality between the true proportion of
 

a specific tumor in a dosed 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
 

dosed 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% of a large number of identical experiments, the
 

true ratio of the risk in a dosed 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 (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
 

15
 
indicates that there is a theoretical possibility of the
 

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

detected under the conditions of this test.
 

16
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of all groups of dosed rats were generally
 

comparable with those of the matched controls, except toward the
 

end of the study, when the mean weights of the high-dose males
 

were slightly depressed (figure 1). Fluctuation in the growth
 

curve for the male rats may be due to mortality; as the size of a
 

group diminishes, the mean body weight may be subject to variation. No other clinical signs related to administration of
 

daminozide were reported.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats fed daminozide in the diet at
 

the doses of this bioassay, together with those of the matched
 

controls, are shown in figure 2.
 

The results of the Tarone test for positive dose-related trend in
 

mortality are not significant in either sex. At least 70% of the
 

animals lived to the end of the study (44/50 of the high-dose
 

males, 37/50 of the low-dose males, 14/20 of the control males;
 

39/50 of the high-dose females, 35/50 of the low-dose females,
 

and all 20 of the control females; one female control died in the
 

17
 
sooH 

o 

UJ a 5 

m 

§ a to 

z a UJ 

5 

100-1 MALE RATS 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

40 50 60 70 

TIME ON STUDY (WEEKS) 

8 B! 

n § 

FEMALE RATS 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

—I— —I— —I— 

40 50 60 70 

TIME ON STUDY (WEEKS! 

Figure 1. Growth Curves For Rats Fed Daminozide in the Diet 

18 
PROBABILITY OF SURVIVAL PROBABILITY 

O O O O O 

8 S S 8 3 S S 

"g =•— C o a _ > O D. 

i

r

- CD 

ro § i 

V) g -

CJ 

h

Tl m 

S o 

-n o 

m 

m 4 x -S ^ -j |S»- tfi ° 

S 

S" g­

<D 

q

S' 
last week of the study). Sufficient numbers of rats of each sex
 

were at risk for development of late-appearing tumors. In male
 

rats, the control group had an increased incidence in mortality
 

compared with that of the dosed groups from week 60 to the end of
 

the study.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

Appendix A, tables Al and A2; findings on nonneoplastic lesions
 

exe summarized in Appendix C, tables Cl and C2.
 

A variety of neoplastic and nonneoplastic changes were observed
 

in the low- and high-dose groups of rats administered daminozide
 

and in the control groups.
 

The most significant finding in this bioassay was the increased
 

incidence of malignant tumors of the uterus in the rats administered the test chemical. The incidences were as follows:
 

Matched Low High
 

Control Dose Dose
 

Number of animals with tissues
 

examined microscopically 19 50 50
 

Uterus/Endometrium
 

Endometrial Adenocarcinoma 0 5(10%) 3(6%)
 

Uterus
 

Leiomyosarcoma 0 1(2%) 3(6%)
 

20
 
The endometrial adenocarcinomas were characterized by irregular
 

acini of pleomorphic, hyperchromatic epithelial cells that
 

invaded the myometrium and in many cases extended to the serosa
 

and mesometrium. Most of the adenocarcinomas were scirrhous; two
 

metastasized to the lungs and one to the bladder wall. Several
 

appeared to arise from endometrial stromal polyps, which also
 

occurred in 6/50 (12%) low-dose and 4/50 (8%) high-dose female
 

rats that did not have adenocarcinomas. The leiomyosarcomas were
 

rather poorly differentiated, with the exception of one which
 

occurred in a high-dose female. They consisted of spindle cells,
 

often with abundant eosinophilic cytoplasm and elongated pleomorphic nuclei. The cells formed irregular, interwoven patterns
 

which obliterated the endometrium and extended through the entire
 

uterine wall. Giant spindle cells and bizarre nuclear forms were
 

evident in some areas. In one high-dose animal, a leiomyosarcoma
 

occurred simultaneously with a noninvasive endometrial adenocarc inoma.
 

Other tumors and hyperplasias that occurred in the control and
 

dosed rats were of the expected types and incidences for this age
 

group of Fischer 344 rats. A few more sebaceous gland carcinomas
 

of the ear (Zymbal's gland tumors) and adenomas of the preputial
 

glands and lungs were encountered in the dosed groups when
 

compared with controls. Although these tumors are not common in
 

21
 
aging Fischer 344 rats, the differences are not considered great
 

enough in magnitude to be meaningful.
 

All of the nonneoplastic lesions were considered to be spontaneous changes that occurred with similar frequency and intensity in
 

control and dosed rats.
 

Based on the histopathologic examination, it is concluded that
 

under the conditions of this bioassay administration of
 

daminozide was associated with low numbers of endometrial
 

adenocarcinomas, as well as of leiomyosarcomas, of the uterus.
 

D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

least two animals of one group and at an incidence of at least 5%
 

in one or more than one group.
 

In the male rats, the results of the Cochran-Armitage test for
 

positive dose-related trend in the incidence of interstitial-cell
 

tumors of the testis are significant (P = 0.003), but an indicated departure from linear trend is observed (P = 0.042),
 

because of the relatively steep increase in incidence observed in
 

the dosed groups. The results of the Fisher exact test show that
 

the incidences in both the low- and high-dose groups are signifi22
 
cantly higher than that in the matched controls (P = 0.009 and
 

P = 0.004, respectively). Data compiled to date on the
 

historical-control male rats at this laboratory show, however, an
 

incidence of 182/220 (83%). The matched-control group of male
 

rats had a higher incidence of mortality from week 60 to the end
 

of the study than did the dosed groups, and this may account for
 

the apparent significance of the incidence of interstitial-cell
 

tumors of the testis in the dosed groups.
 

In the female rats, the results of the Cochran-Armitage test for
 

the incidence of lung tumors are significant (P = 0.036), but the
 

results of the Fisher exact test are not significant. A negative
 

linear trend (P = 0.021) is observed in the combined incidence of
 

C-cell adenomas and carcinomas of the thyroid, and the results of
 

the Fisher exact test for the comparison of the incidence in the
 

high-dose group with that in the matched controls is 0.032 in the
 

negative direction; this P value of 0.032 is above that of 0.025
 

required for statistical significance when the Bonferroni
 

criterion for multiple comparisons is considered. While the
 

difference in survivals of the high-dose and control groups is
 

not statistically significant (P > 0.05), some increase in
 

mortality in the dosed group is observed, and this may account
 

for the incidence of the thyroid tumors in the negative
 

direction.
 

23
 
Although the results of the Cochran-Armitage and Fisher exact
 

tests on the incidences of adenocarcinomas of the
 

uterus/endometrium and of leiomyosarcomas of the uterus in female
 

rats were not significant, these tumors are rarely observed in
 

female untreated-control rats. Data compiled to date from all
 

laboratories in the bioassay program show that the incidence of
 

adenocarcinomas of the uterus/endometrium is 4/1,659 (0.24%) and
 

that the incidence of leiomyosarcomas of the uterus is 1/1,659
 

(0.06%). Of the 220 female untreated-control rats observed to
 

date in the present laboratory, no leiomyosarcomas of the uterus
 

and only two adenocarcinomas of the uterus (0.9%) were found.
 

Assuming that these adenocarcinomas (2/220 [0.9%]) follow a
 

binomial distribution, the probability of occurrence of 3 or more
 

tumors in 50 high-dose animals is less than 0.002, while the
 

occurrence of 5 or more tumors in 50 low-dose animals is less
 

than 0.001. Assuming that the leiomyosarcomas with assumed
 

incidence of 1/221 (0.5%) follow a binomial distribution, the
 

probability of the occurrence of 3 or more tumors in 50 high-dose
 

animals is less than 0.0001.
 

In each of the 95% confidence intervals of relative risk, shown
 

in the tables, except that for the incidence of tumors of the
 

testis, a value of one is included; this indicates the absence of
 

significant positive results. It should also be noted that each
 

24
 
of the intervals has an upper limit greater than one, indicating
 

the theoretical possibility of the induction of tumors by
 

daminozide, which could not be detected under the conditions of
 

this test.
 

25
 

IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of the high-dose female mice were lower than
 

those of the matched controls throughout the bioassay, while mean
 

body weights of the other dosed groups were essentially
 

unaffected (figure 3). Fluctuation in the growth curve may be
 

due to mortality; as the size of a group diminishes, the mean
 

body weight may be subject to variation. No other clinical signs
 

related to administration of daminozide were observed.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice fed daminozide in the diet at
 

the doses of this bioassay, together with those of the matched
 

controls, are shown in figure 4.
 

In the male mice, the results of the Tarone test for positive
 

dose-related trend in mortality are not significant. There were
 

32/50 (64%) of the high-dose group, 38/50 (76%) of the low-dose
 

group, and 11/20 (55%) of the controls alive at the end of the
 

bioassay. Five of the male controls and four of the male high-


dose group were reported missing during the study. In the
 

females, the results of the Tarone test are significant (P =
 

27
 
Q D O Q Q A n n a S A A 

3 30-1 

|»H 

CD 

Z 

4 

Ul 15­

5 

MALE MICE 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

1­

10 20 40 50 80 70 100 

TIME ON STUDY (WEEKS) 

880088 0 DQ a o 

o 8 ° D A A AAA 8 

o o n A A 

O D . A A A A B A A A A A 

A A A 

A A 

A A 

z o 25- an X 

o

a 

< 16' 

5 

FEMALE MICE 

G MATCH ED CONTROL 

O LOW DOSE 

A HIGH DOSE 

40 50 60 70 100 110 

TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves For Mice Fed Daminozide in the Diet 

28 
PROBABILITY OF SURVIVAL

 PROBABILITY O

> i — s £ ', *c g gS c

j

9

 a

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0.044). There were 30/50 (60%) of the high-dose group, 32/50
 

(64%) of the low-dose group, and 17/20 (85%) of the controls
 

alive at the end of the bioassay. Sufficient numbers of mice of
 

each sex were at risk for development of late-appearing tumors.
 

C. Pathology (Mice)
 

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.
 

There was a relatively high incidence of hepatocellular carcinomas observed in the dosed mice. Hepatocellular carcinomas were
 

observed in 7/50 (14%) low-dose and 13/46 (28%) high-dose male
 

mice and in 3/40 (8%) low-dose female mice. No hepatocellular
 

carcinomas were observed in the high-dose female mice or in the
 

control mice of either sex. Grossly, the liver tumors were
 

multinodular friable masses that usually measured 1 cm or greater
 

in diameter. Some had necrotic or hemorrhagic areas. Microscopically, the liver tumors had a variety of patterns including
 

disorganized plates, irregular trabeculae, and acinar-like
 

structures of hepatocyte-like cells that were often hyperchromatic and pleomorphic, with basophilic or vacuolated cytoplasm.
 

There was a loss of lobular architecture in the neoplastic areas
 

that compressed and invaded normal surrounding hepatic
 

30
 
parenchyma. Cords of neoplastic hepatic cells often traversed
 

blood-filled spaces that lay in some of the affected zones. The
 

hepatocellular carcinomas metastasized to the lungs of one low-


dose and two high-dose male mice and to the adrenal of one
 

high-dose male mouse. An angioma was observed in the liver of a
 

high-dose male mouse, and an angiosarcoma was observed in the
 

liver of a low-dose female.
 

Other tumors and hyperplasias that occurred in the control and
 

dosed groups were of the expected types and incidences for aged
 

B6C3F1 mice.
 

All of the nonneoplastic lesions were considered to be
 

spontaneous changes, and most have been previously documented by
 

long-term experiments in B6C3F1 mice. These changes occurred
 

with similar incidences in control and dosed groups.
 

Based on the histopathologic examination, a relationship may
 

exist between the numbers of hepatocellular carcinomas observed
 

in the dosed mice and the administration of daminozide under the
 

conditions of this bioassay. However, the incidences seen are
 

within the range expected for this strain of mice.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

31
 
of the incidences of those primary tumors that occurred in at
 

least two animals of one group and at an incidence of at least 5%
 

in one or more than one group.
 

In the male mice, the results of the Cochran-Armitage test for
 

positive dose-related trend in the incidence of hepatocellular
 

carcinomas are significant (P = 0.008), and the results of the
 

Fisher exact test show that the incidence in the high-dose group
 

is significantly higher (P = 0.020) than that in the controls.
 

Data compiled to date on the historical-control male mice at this
 

laboratory show, however, an incidence of 21/216 (10%), and data
 

compiled to date on the historical-control male mice of the
 

entire bioassay program show an incidence of 266/2,182 (12.2%).
 

The incidence of hepatocellular carcinomas in the female mice is
 

not significant using either statistical test. When the combined
 

incidence of adenomas and carcinomas of the liver in each sex is
 

analysed, the results of the Fisher exact test are not significant for either the low- or high-dose groups. The results of the
 

Cochran-Armitage test for the combined incidence show a probability level of 0.031 in males, whereas in females it is above the
 

0.05 level.
 

Significant trends in the negative direction are observed in the
 

incidences of lipomas of the peritoneum and of the mesentery in
 

the female mice. This significance in the negative direction may
 

32
 
be accounted for by the increased mortality in the dosed groups
 

of animals when compared with that in the control group. The
 

results of the Fisher exact test are not significant.
 

In each of the 95% confidence intervals of relative risk, shown
 

in the tables (except that for the incidence of hepatocellular
 

carcinoma of the liver in high-dose males), a value of one is
 

included; this indicates the absence of significant positive
 

results. It should also be noted that each of the intervals has
 

an upper limit greater than one, indicating the theoretical
 

possibility of the induction of tumors by daminozide, which could
 

not be detected under the conditions of this test.
 

33
 

V. DISCUSSION
 

Under the conditions of this bioassay, daminozide was only
 

slightly toxic to female B6C3F1 mice. Mean body weights of the
 

high-dose female mice were lower than those of the controls; mean
 

body weights of all other dosed groups of rats and mice were
 

essentially unaffected, and these animals may have been able to
 

tolerate a higher dose. No other clinical signs related to
 

administration of daminozide were recorded, and sufficient
 

numbers of rats and mice of each sex were at risk for development
 

of late-appearing tumors.
 

In the male rats, no tumors occurred at incidences that were
 

significantly higher in dosed groups than in controls, except for
 

interstitial-cell tumors of the testis (controls 13/20, low-dose
 

49/50, high-dose 47/50). These tumors occurred, however, at -a
 

high spontaneous rate (182/220) in historical-control male rats;
 

thus, the association of the interstitial-cell tumors with
 

administration of the chemical is doubtful.
 

In the female rats, adenocarcinomas of the endometrium and
 

leiomyosarcomas of the uterus occurred only in the dosed groups
 

(adenocarcinomas: controls 0/19, low-dose 5/50, high-dose 3/50;
 

leiomyosarcomas: controls 0/19, low-dose 1/50, high-dose 3/50).
 

In historical-control female rats in this laboratory, 2/220
 

35
 
adenocarcinomas and 0/220 leiomyosarcomas of the uterus were
 

found. Assuming binomial distributions for these tumors, the
 

probability of 3/50 leiomyosarcomas in high-dose animals and 5/50
 

adenocarcinomas in low-dose animals is less than 0.001. Thus,
 

the occurrence of these tumors in the dosed animals was
 

associated with the administration of daminozide.
 

In the male mice, there was a dose-related trend (P = 0.008) in
 

the incidence of hepatocellular carcinomas; furthermore, the
 

incidence in the high-dose group was significant (P = 0.020)
 

compared with that in the controls (controls 0/14, low-dose 7/50,
 

high-dose 13/46). The incidence of these tumors in the
 

historical-control male mice was, however, 21/216; thus, the
 

association of the hepatocellular carcinomas with administration
 

of daminozide is not clear. In the female mice, only three such
 

tumors occurred.
 

Alveolar/bronchiolar adenomas or carcinomas occurred at higher
 

incidences in the dosed groups of both male and female mice
 

(males: controls 4/14, low-dose 15/50, high-dose 18/46; females:
 

controls 1/20, low-dose 8/39, high-dose 10/48), but none of these
 

incidences were significantly higher than those in the matched
 

controls. The incidence of alveolar/bronchiolar adenomas and
 

carcinomas in historical-control male mice at the laboratory was
 

39/216 and that of female controls was 12/217. Since these
 

36
 
incidences were also lower than those of the dosed groups, they
 

suggest that, at least in the dosed females, there may have been
 

an increase in the incidence of these tumors of the lung.
 

Daminozide has a low acute toxicity, having an acute oral LD5Q of
 

8,400 mg/kg for rats (Farm Chemicals Handbook, 1977). One long-


term feeding study has been reported (Farm Chemicals Handbook,
 

1977) in which no effects were noted after rats and dogs were fed
 

the compound at doses up to 3,000 ppm for 2 years.
 

Uaminozide was previously reported to induce angiomas and
 

angiosarcomas of the blood vessels, adenomas and adenocarcinomas
 

of the lungs, and adenomas of the kidneys in Swiss albino mice
 

during lifetime administration of the chemical at a concentration
 

of 2% in drinking water (Toth et al., 1977). None of these types
 

of tumors were observed at incidences in the present bioassay
 

which could be clearly associated with administration of the test
 

chemical; however, alveolar/bronchiolar adenomas or carcinomas
 

occurred at increased incidences in both sexes of mice and in the
 

high-dose female rats. The differences in results may have been
 

due to differences in the species and strains of animals used or
 

in the route of administration of the test chemical.
 

1,1-Dimethylhydrazine, produced by hydrolysis of the daminozide,
 

may have contributed to the carcinogenicity of the test chemical,
 

as previously suggested (Toth et al., 1977). 1,1-Dimethylhydra37
 
zine has been shown to induce alveolar/bronchiolar adenomas or
 

adenocarcinomas in Swiss albino mice administered the chemical by
 

gavage (Roe et al., 1967) and also vascular angiosarcomas in the
 

same strain of mice administered the chemical in the drinking
 

water (Toth, 1973).
 

It is concluded that under the conditions of this bioassay,
 

daminozide was not carcinogenic in the male Fischer 344 rats or
 

in the female B6C3F1 mice. In male B6C3F1 mice, the induction of
 

hepatocellular carcinomas may have been associated with the
 

administration of the test chemical. Daminozide was carcinogenic
 

in female Fischer 344 rats, inducing adenocarcinomas of the
 

endometrium of the uterus and leiomyosarcomas of the uterus.
 

38
 
VI. BIBLIOGRAPHY
 

Armitage, P., Statistical Methods in Medical Research, John Wiley
 

& Sons, Inc., New York, 1971, pp. 362-365.
 

Berenblum, I., ed., Carcinogenicity Testing; A_ Report of the
 

Panel on Carcinogenicity of the Cancer Research Commission
 

of the UICC, Vol. _2, International Union Against Cancer,
 

Geneva, 1969.
 

Cox, D. R., Regression models and life tables. J. R. Statist.
 

Soc. B 34;187-220, 1972.
 

Cox, D. R., Analysis of Binary Data, Methuen & Co., Ltd., London,
 

1970, pp. 48-52.
 

Environmental Protection Agency, Title 40-Protection of
 

Environment, Code of Federal Regulations, 180.246, 1976.
 

Farm Chemicals Handbook, Meister Publishing Co., Willoughby,
 

Ohio, 1977, p. D9.
 

Gart, J. J., The comparison of proportions: a review of
 

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Statist. Inst. 39;148-169, 1971.
 

Kaplan, E. L. and Meier, P., Nonparametric estimation from
 

incomplete observations. J. Am. Statist. Assoc. 53;457-481,
 

1958.
 

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

Peters, J. A., Carcinogenesis bioassay data system. Comp.
 

and Biomed. Res. _7:230-248, 1974.
 

Miller, R. G., Jr., Simultaneous Statistical Inference,
 

McGraw-Hill Book Co., New York, 1966, pp. 6-10.
 

Riddell, J. A., Hageman, H. A., J'Anthony, C. M., and Hubbard, W.
 

L. , Retardation of plant growth by a new group of chemicals.
 

Science 136;391. 1962.
 

Roe, F. J. C., Grant, G. A., Millican, D. M., Carcinogenicity of
 

hydrazine and 1,1-dimethylhydrazine for mouse lung, Nature
 

216:375-376, 1967.
 

39
 
Saffiotti, U. , Montesano, R., Sellakumar, A. R., Cefis, F., and
 

Kaufman, D. G., Respiratory tract carcinogenesis in hamsters
 

induced by different numbers of administrations of benzo (a)
 

pyrene and ferric oxide. Cancer Res. 32;1073-1081, 1972.
 

Tarone, R. E., Tests for trend in life table analysis. Biometrika
 

6^:679-682, 1975.
 

Thomson, W. T., Fumigants, growth regulators, repellents and
 

rodenticides. Agricultural Chemicals - Book III;
 

Miscellaneous Chemicals, Thomson Publications, Fresno,
 

Calif., 1976, pp. 46-47.
 

Toth, B., Wallcave, W., Patie, K. , Schmeltz, I., and Hoffmann,
 

D., Induction of tumors in mice with the herbicide succinic
 

acid 2,2-dimethylhydrazine. Cancer Res. 37:3497-3500. 1977.
 

Toth, B., Synthetic and naturally occurring hydrazines as 

possible cancer causative agents. Cancer Res. 35:3693-3697, 

1975. 

Toth, B., 1,1-Dimethylhydrazine (unsymmetrical) carcinogenesis in
 

mice. Light microscopic and utrastructural studies on
 

neoplastic blood vessels, J. Natl. Cancer Inst. 50;181-194,
 

1973.
 

40
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS FED DAMINOZIDE IN THE DIET
 

41
 

TABLE A1. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

FED OAMINOZIDE IN THE DIET 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

ANIMALS INITIALLY IN STUDY 20 50 50
 

AHIHALS NECROPSIED 20 50 50
 

ANIMALS EXAHINSD HISTOPATHOLOGICALLY 20 50 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50) (50)
 

PAPILLOMA, NOS 1 (2%)
 

SQUAMOUS CELL PAPILLOMA 1 (5*)
 

SQUAMOUS CELL CARCINOMA 1 (2X)
 

FIBROMA 1 (2X)
 

*SUBCUT TISSUE (20) (50) (50)
 

FIBROMA 1 (2X)
 

FIBROSARCOMA 2 (H%) 2 (H%)
 

RESPIRATORY SYSTEM
 

tLUNG (20) (50) (50)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (2X)
 

ALVEOLAK/BRONCHIOLAR ADENOMA 2 (U%)
 

ALVEOLAR/BRCNCHIOLAR CARCINOMA 1 (5«) 1 (2%)
 

THYMOMA, METASTATIC 1 (5%)
 

OSTEOSARCOMA, METASTATIC 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS (20) (50) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (2X)
 

LEUKEMIA, NOS 1 (5»)
 

UNDIFFERSNTIATED LEUKEMIA 1 (2%)
 

tTHYMUS (1) (2) (1)
 

THYMOMA, MALIGNANT 1 (100X)
 

CIRCULATORY SYSTEM
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUBBEP OF ANIMALS NECROPSIED
 

43
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

DIGESTIVE SYSTEM
 

*LIP
 

PAPILLOMA, NOS
 

*LIVSR
 

HEPATOCELLULAR CARCINOMA
 

tSTOMACH
 

SQUAMOUS CELL CARCINOMA
 

#SMALL INTESTINE
 

LEIOMYOMA
 

URINARY SYSTEM
 

#KIDNEY
 

TUBULAE-CELL ADENOCARCINOMA
 

THYMOMA, METASTATIC
 

ENDOCRINE SYSTEM
 

•PITUITARY
 

CHROMOPHOBE ADENOMA
 

f ADRENAL
 

PHEOCHROMOCYTOMA
 

PHEOCHEOMOCYTOMA, MALIGNANT
 

•THYROID
 

FOLLICULAR-CELL CARCINOMA
 

C-CELL ADENOMA
 

C-CELL CARCINOMA
 

#PARATHYEOID
 

ADENOMA, NOS
 

•PANCREATIC	 ISLETS
 

ISLET-CELL ADENOMA
 

REPRODUCTIVE SYSTEM
 

*PREPUCE


TSJCHCEPITHELIQHA


MATCHED LOW DOSE HIGH DOSE 

CONTROL 

(20) (50) (50) 

1 (2X) 

(20) (50) (50) 

1 (2X) 

(20) (50) (50) 

1 (2X) 

(19) (50) (50) 

1 (2X) 

(20) (50) (50) 

1 (2X) 

1 (5%) 

(18) (43) (49) 

2 (5%) 2 (4X) 

(20) (50) (50) 

2 (10%) 2 (4%) 1 (2X) 

1 (2X) 

(16) (38) (43) 

1 (6X) 

2 (13X) 3 (8X) 

2 (5X) 

(12) (22) 

1 (5X) 

(22) 

(20) (48) (49) 

1 (5X) 

 (20) (50) (50) 

1_ 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBEB OF ANIMALS NECEOPSIED
 

44
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE 

CONTROL 

*PREPUTIAL GLAND (20) (50) (50) 

CARCINOMA,NO S 1 (5%) 

ADENOHA , NOS 1 (5X) 1 (2%) H (S%) 

*TESTIS (20) (50) (50)
 

INTERSTITIAL-CELL TUHOR 13 (65X) 46 (92X) HI (94X)
 

8ERVOU5 SYSTEM
 

fBRAIN/MENINGES (19) (50) (50)
 

MESOTHELIOMA, BETASTATIC 1 (5X)
 

IBRAIN (19) (50) (50)
 

SQUABOUS CELL CARCINOMA, METASTA 1 (2%)
 

THYWOHA, BETASTATIC 1 (5%)
 

MEDULLOBLASTOHA 1 (2%)
 

SPECIAL SENSE ORGANS
 

*EYELID (20) (50) (50)
 

FIBKOSARCOMA 1 (2%)
 

*ZYNBAL'S GLAND (20) (50) (50)
 

SEBACEOUS ADENOCARCINOHA 1 (2%) 1 (2%)
 

MUSCOLOSKELETAL SYSTEM
 

*VERTEBRA (20) (50) (50)
 

OSTEOSARCOMA 1 (2%)
 

BODY CAVITIES
 

*ABDOHINAL C-AVITY (20) (50) (50)
 

LKIOMYOHA 1 (2X)
 

*PERITONEOB (20) (50) (50)
 

BESOTHELIOMA. NOS 1 (2%) 1 (2X)
 

HESOTHELIOHA, BALIGNANT 1 (5X)
 

ALL OTHER SYSTEMS
 

NONE
 

t NOBBER OF ANIMALS WITH TISSUE EXAMINBD MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

45
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY


NATURAL DEATHS


MORIBUND SACRIFICE


SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 TERHINAL SACRIFICE


ANIMAL MISSING
 

S INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 

TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TOMORSt
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT
 

TOTAL UNCERTAIN. TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUHORS
 

 20


 1
 

2


 14


16
 

26
 

13
 

20
 

6
 

6
 

2
 

4
 

 50 50 

3 3 

 10 3 

 37 44 

50 49 

70 69 

48 47 

60 58 

8 10 

9 10 

2 

3 

1 1 

1 1 

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

46
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

FED DAMINOZIDE IN THE DIET 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

ANIHALS INITIALLY IN STUDY 20 50 50
 

ANIHALS NECROPSIED 20 50 50
 

ANIHALS EXAMINED HISTOPATHOLOGICALLY 20 50 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50) (50)
 

SQUAMOUS CELL CARCINOMA 1 (2%)
 

TEICHOEPITHELIOMA 1 (2%)
 

SEBACEOUS ADENOCARCINOBA 1 (5%)
 

*SUBCUT TISSUE (20) (50) (50)
 

LIPOMA 2 (4X)
 —____ _ — «—«._______*._.__..__.__._.______________«*«»•_______ — *- — — •— >.••—•*_______•••«
 

RESPIRATORY SYSTEM
 

tLDNG (20) (50) (48)
 

ADENOCARCINOMA, NOS, METASTATIC 3 (6%)
 

ALVEOLAH/BRONCHIOLAR ADENOMA U (8X)
 

SEBACEOUS ADENOCARCINOMA, METAST 1 (5X)
 _ __ _ _ _ —— _ — —___—____• _ _ _ — »__»«.__-.«._.._ — — — -.•..-._ — _ — —.— — — —»— — — — — — — — — — — •— —— — »—••••••.— — — — •«
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS (20) (50) (50)
 

LEUKEMIA,NOS 1 (2«)
 

UNDIFFERENTIATED LEUKEMIA 1 (2%)
 

LYMPHOCYTIC LEUKEMIA 1 (2X)
 

GRANULOCYTIC LEUKEMIA 2 (4X)
 

tLYMPH NODE (20) («9) (49)
 

ADENOCARCINOBA, NOS 1 (2%)
 

CIRCULATORY SYSTEM
 

__NONE
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS HECROPSIED
 

47
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

"~mr
 ""'" '" "~ •— — -• —. — - — •— i-J- TM. -M _ in—. _. n -._.m i -. 1_ -._•_! « — ~m -_—
 

MATCHED LOW DOSE
 

CONTROL
 

DIGESTIVE SYSTEM
 

tSALIVARY GLAND (20) {« 8)
 

ADENOCARCINOHA, NOS 1 (2X)
 

*LIVER (19) (50)
 

HEPATOCELLtlLAR ADEHOHA 1 (2X)
 

URINARY SYSTEM
 

#OHINARY BLADDER (19) (43)
 

PAPILLCMATOSIS 1 (2X)
 

ADENOCARCINOMA, NOS, METASTATIC 1 (2X)
 

ENDOCRINE SYSTEM
 

fPITOITARY (19) (« 5)
 

CHROHOPHOBE ADENOMA 3 (16X) 7 (16X)
 

CHROMOPHOBE CARCINOMA 3 (7X)
 

•ADRENAL	 (20) (48)
 

CORTICAL ADENOMA 1 <5«)
 

PHEOCHROMOCYTOMA. MALIGNANT 1 (5X)
 

tTHYROID (15) (38)
 

C-C5LL ADENOMA 2 (13X) 2 (5X)
 

C-CELL CARCINOMA 2 (13X) 1 (3X)
 

REPRODUCTIVE SYSTEM
 

*MAHMARY GLAND (20) (50)
 

ADENOMA, NOS 1 (2X)
 

ADENOCARCINOMA, NOS
 

PAPILLARY ADENOCARCINOMA 1 (2X)
 

ACINAR-CELL ADENOMA 1 (2S)
 

PIBROADENOMA 3 (15X) 9 (18X)
 

*MAMMARY DUCT (20) (50)
 

ADENOMA, NOS
 

*PREPOTIAL GLAND (20) (50)
 

_ _ACIl!QHAx HOS , 1 JI281
 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

48
 

HIGH DOSE
 

(48)
 

(50)
 

1 (2X)
 

(43)
 

(43)
 

8 (19X)
 

(48)
 

(44)
 

2 (5X)
 

(50)
 

1 (2X)
 

2 (4X)
 

(50)
 

1 (2X)
 

(50)
 

1 I2S1__
 
TABLE A2. FEMALE RATS: NEOPLASMS

tUTERUS


LEIOMYOSARCOBA


ENDOMETRIAL STROMAL POLYP


HEHANGIOHA


#UTERUS/ENDOMETRIUM


ADENOCARCINOHA, NOS


#OVARY


GRANULOSA-CELL TUMOR


NERVOUS SYSTEH
 

IBKAIN


 (CONTINUED) 

MATCHED

CONTROL 

(19)


 (19)


 (19)


 (20)


CHROMOPHOBE CARCINOMA, INVASIVE


SPECIAL SENSE ORGANS
 

*ZYMBAL'S GLAND (20)


SEBACEOUS ADENOCARCINOMA


 LOW DOSE

 (50)


 1 (2%)


 6 (12*)


 (50)


 5 (10*)


 (50)


 1 (2%)
 

(49)


 1 (2*)
 

(50)


 HIGH DOSE 

 (50)
 

3 (6*)
 

4 (8X)
 

1 (2%)
 

 (50)
 

3 (6*)
 

 (49)
 

 (49)
 

 (50)
 

1 (2X)
 _ _ — ^ «____ _ v~^_^__^^__~^__^«__^________ _^^__^__..^^_«^___^.^___^^.,B^^»«»i— — — ^— — ^^«
 

MUSCOLOSKELETAL SYSTEM
 

NONE
 — — — — — — — — —— ~~___^___~_4^_~____~«.«^4_*^.^_4.^^_^^^^_^._«,^^.»^^^^^^_^^^«t^.B^^a>^^«^'>^^«
 

BODY CAVITIES 

*MESENTERY

LIPOMA

HEMANGIOSARCOMA

 (20) (50) (50) 

2 (4%) 

1 (2X) 

ALL OTHER SYSTEMS
 

NONE
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

49
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY

NATURAL DEATHS

MORIBUND SACRIFICE

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE

ANIMAL MISSING 

 20

 1 

 19

 50

 7

8 

 35

 50 

6 

5 

 39 

a INCLUDES AUTOLYZED ANIMALS 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 11 32 28
 

TOTAL PRIMARY TUMORS 13 51 36
 

TOTAL ANIMALS HITH BENIGN TUMORS 8 24 19
 

TOTAL BENIGN TUMORS 9 32 26
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 3 15 9
 

TOTAL MALIGNANT TUMORS H 18 10
 

TOTAL ANIMALS HITH SECONDARY TUMORSf 1 4
 

TOTAL SECONDARY TUMORS 1 5
 

TOTAL ANIMALS HITH TUMORS UNCERTAINBENIGN OR MALIGNANT 1
 

TOTAL UNCERTAIN TUMORS 1
 

TOTAL ANIMALS HITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUHORS
 

# SECONDARY TUMORS: HETASTATIC TUHORS OR TUHORS INVASIVE INTO AN ADJACENT ORGAN
 

50
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE FED DAMINOZIDE IN THE DIET
 

51
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

FED OAMINOZIDE IN THE DIET 

MATCHED LOW DOSE HIGH DOSE 

CONTROL 

ANIMALS INITIALLY IN STUDY 20 50 50 

ANIMALS HISSING 6 4 

ANIMALS NECROPSIED 14 50 46 

ANIHALS EXAMINE D HIS! [^PATHOLOGICALLY 14 50 46 

INTEGUMENTARY SYSTEM
 

*SKIN (14) (50) (46)
 

FIBROMA 1 (7%)
 

NEUROFIBROSARC08A 1 (7X)
 

RESPIRATORY SYSTEM
 

• LUNG (14) (50) (46)
 

HEPATOCELLULAR CARCINOMA, BETAST 1 (2X) 2 (4X)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 2 (14X) 7 (14*) 6 (13X)
 

ALVEOLAR/BRONCHIOLAR CARCINOMA 2 (14X) 9 (18X) 12 (26%)
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS (14) (50) (46)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (2X)
 

MALIGNANT LYMPHOMA, MIXED TYPE 5 (11X)
 

LEUKEMIA,NOS 2 (4X)
 

LYBPHOCYTIC LEUKEMIA 1 (2X)
 

#SPLEEN (13) (40) (43)
 

FIBROSARCOMA 1 (2%)
 

HEMANGIOMA 1 (8X) 1 (3X)
 

ANGIOMA 1 (8%)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (8X)
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2X)
 

#HESENTERIC L. NODE (9) (42) (43)
 

BALIG.LYMPHOHA, LYMPHOCYTIC TYPE 1 (2Jt) 1 (2«)
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2X) 1 (2X)
 

tSMALL INTESTINE (13) (48) (44)
 

NALIGJ.LYHPHQMAJI_LYM.PHQCYT!C_TY.PE.
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

53
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL 

LOW DOSE HIGH DOSE 

IPEYEBS PATCH

MALIG.LYMPHOMA, LYHPHOCYTIC TYPE

 (13) 

1 (8X) 

(48) 

1 (2X) 

(44) 

CIRCULATORY SYSTEM 

NONE 

DIGESTIVE SYSTEM 

*LIVER 

HEPATOCELLULAR ADENOMA 

HEPATOCELLOLAfi CARCINOMA 

SARCOMA, NOS 

ANGIOMA 

*BILS DUCT 

BILE DUCT CARCINOMA 

(14) 

1

(14) 

 (7%) 

(50) 

2 (4X) 

7 (14X) 

(50) 

1 (2%) 

(U6) 

1 (2X) 

13 (28X) 

1 (2X) 

1 (2%) 

(46) 

#PANCREAS

BILE DUCT CARCINOMA, METASTATIC

 (12) (47)

 1 (2%) 

 (42) 

URINARY SYSTEM 

NONE 

ENDOCRINE SYSTEM 

#ADRENAL

HEPATOCELLULAR CARCINOMA, METAST

PHEOCHBOMOCYTOMA

 (8) (35)

 1 (3%) 

 (38) 

1 (3X) 

#THYROID

FOLLICOLAF.-CELL ADENOMA'

 (12) (39)

 1 (3X)

 (35) 

1 (3X) 

tPANCREATIC ISLETS

ISLET-CELL ADENOMA

 (12) (47) (42) 

1 (2%) 

REPRODUCTIVE SYSTEM 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

54
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE 

CONTROL 

NERVOUS SYSTEM 

NON E 

SPECIAL SENSE ORGANS 

NONE 

MDSCULOSKELETAL SYSTEM
 

*SKBLETAL MUSCLE (14) (50)

 

FIBROSARCOMA
 

BODY CAVITIES
 

*ABDOMINAL CAVITY (14) (50)
 

NEOPLASM, NOS
 

LIPOMA 1 (2X)
 

*PERITONEUM (1*0 (50)
 

LIPOMA 1 (2X)
 

^MESENTERY (1t) (50)
 

NEOPLASM, NOS 1 (2X)
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 20 50
 

NATURAL DEATHS 2 9
 

MORIBUND SACRIFICE 1 3
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 11 38
 

ANIMAL MISSING 6
 

i_IS£LOJJE.S AUlO.L.Y.ZSB.AJISiiS
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

55
 

HIGH DOSE 

(46) 

<2X)
 

(46)
 

1 <2X)
 

1 (2X)
 

(46)
 

(46)
 

50
 

10
 

4
 

32
 

4
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 8 28 37
 

TOTAL PRIMARY TUMORS 11 38 51
 

TOTAL ANIMALS WITH BEHIGN TUMORS 5 10 11
 

TOTAL BENIGN TUMORS 6 14 11
 

TOTAL ANIMALS WITH MALIGNANT TUMORS H 22 33
 

TOTAL MALIGNANT TUMORS 5 23 39
 

TOTAL ANIMALS WITH SECONDARY TUMOHS# 2 2
 

TOTAL SECONDARY TUMORS 2 3
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN OR MALIGNANT 1 1
 

TOTAL UNCERTAIN TUHORS 1 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPBIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUMORS OR TUHORS INVASIVE INTO AN ADJACENT OBGAH
 

56
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

FED 0AMINOZIDE IN THE DIET 

MATCHED LOW DOSE 

CONTROL 

ANIMALS INITIALLY IN STUDY 20 50 

ANIMALS HISSING 9 

ANIMALS SECROPSIED 20 41 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 20 41 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE (20) (41)
 

HEBANGIOMA 1 (2X)
 

RESPIRATORY SYSTEM
 

*LUNG (20) (39)
 

CARCINOMA, NOS, MSTASTATIC 1 (3X)
 

ALVEOLAR/BRONCHIOLAR ADENOMA 4 (10X)
 

ALVEOLAH/BRONCHIOLAR CARCINOMA 1 (5X) 4 (10X)
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS (20) (41)
 

MALIGNANT LYMPHOMA, NOS 1 (2X)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (5X) 5 (12X)
 

MALIG.LYBPHOMA, HISTIOCYTIC TYPE
 

MALIGNANT LYMPHOMA, MIXED TYPE 5 (12X)
 

PLASMA-CELL TUMOR
 

LEUKEMIA, NOS 1 (5X)
 

LYMPHOCYTIC LEUKEMIA 1 (5X) 1 (2X)
 

PLASHACYTIC LEUKEMIA 1 (2X)
 

GRANULOCYTIC LEUKEMIA
 

*ABDOMINAL CAVITY (20) (41)
 

MALIGNANT LYMPHOMA, NOS 1 (2X)
 

*SPLEEN (17) Uti
 

HSBAHGIOMA T (3X)
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 1 (6X)
 

ERYTHROCYTIC LEUKEMIA 1 (3X)
 

#MANDIBULAR L. NODE (20) (37)
 

f NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

57
 

HIGH DOSE
 

50
 

2
 

48
 

48
 

(48)
 

(48)
 

8 (17X)
 

2 (4X)
 

(48)
 

2 (4X)
 

4 (8X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

(48)
 

(42)
 

1 (2X)
 

(tO)
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

#MEDIASTINAL L.NODE (20) (37) (40)
 

MALIGNANT LYBPHOBA, NOS 1 (3%)
 

*BESENTERIC L. NODE (20) (37) (40)
 

HEMANGIOMA 1 (3%)
 

BALIG.LYMPHOMA, LYBPHOCYTIC TYPE 1 (5*)
 

HALIGNANT LYMPHOMA, MIXED TYPE 1 (3X)
 

#SNALL INTESTINE (20) (38) (46)
 

MALIG.LYMPUOBA, LYBPHOCYTIC TYPE 1 (2X)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

#LIVER (20) (UO) (48)
 

HEPATOCELLULAR ADENOHA 1 (5X) 1 (3X)
 

HEPATOCELLULAR CARCINOMA 3 (8X)
 

ANGIOSARCOMA 1 (3%)
 

HABARTOMA 1 (3X)
 ___•____.___ ^——•—.— _^_»_^^«___ ••_..__•_ _._..____•__ «._.*— —-^•_—_ — ^—-. — •.^••*— •—•• — »——.
 

DRINARY SYSTEM
 

fKIDNEY (20) (41) (48)
 

CARCINOMA, NOS, BETASTATIC 1 (2X)
 

ENDOCRINE SYSTEM
 

IPITUITARY (14) (19) (14)
 

CHROBOPHOBE ADENOBA 2 (14X) 1 (5X)
 

tADRENAL (17) (31) (38)
 

CORTICAL CARCINOMA 1 (3X)
 

*THYROID (13) (28) (38)
 

FOLLICULAR-CELL ADENOBA 1 (8X)
 

tPANCREATIC ISLETS (16) (36) (44)
 

ISiET-CELL_AfiENQMi .
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

58
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

REPRODUCTIVE SYSTEB
 

tUTERUS


ADENOCA/SQUABOUS BETAPLASIA


ENDOMETRIAL STROHAL POLYP


HEHANGIOHA


NEORILEBOBA


tUTERUS/ENDOBETRIUB


CARCINOMA,DOS


tOVARY


LUTEOBA


GRAHOLOSA-CELL TUBOR


HERVOOS SYSTEB
 

HONE
 

SPECIAL SENSE ORGANS
 

NONE
 

.BBSCOLOSKELETAL SISTEB
 

NONE
 

BODY CAVITIES
 

*ABDOMINAL CAVITY


LEIOBYOBA


*PERITONEDM


LIPOBA


*MESENTERY


LIPOBA


ALL OTHER SYSTEHS
 

NOJJ
 

MATCHED LOW DOSE HIGH DOSE 

CONTROL 

 (20) (37) (45)
 

1 (2X)
 

3 (8X)
 

1 (3X)
 

1 (5X)
 

 (20)	 (37) (l»5)
 

1 (3X)
 

 (13)	 (14) (23)
 

1 (8X)
 

1 (8X)
 

 (20)	 (41) (48)
 

1 (2X)
 

 (20)	 (41) (48)
 

2 (10X)
 

 (20)	 (41) (48)
 

2 (10X) 1 (2X)
 

t	 NUBBER OP ANIMALS WITH TISSUE BXABIIED MICROSCOPICALLY
 * NUMBER OP ANIBALS NECROPSIED
 

59
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

ANIMAL DISPOSITION SUMMARY
 

ANIHALS INITIALLY IN STUDY


NATURAL DEATH8


MORIBUND SACRIFICE


SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE


ANIMAL MISSING


9 INCLUDES AUTOLYZED ANIHALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*


TOTAL PRIMARY TUHOBS


TOTAL ANIMALS WITH BENIGN TUMOES


TOTAL BENIGN TUMORS


TOTAL ANIMALS WITH MALIGNANT TUMORS


TOTAL MALIGNANT TUMORS


TOTAL ANIMALS WITH SECONDARY TUMORSt


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT


TOTAL UNCERTAIN TUMORS


 20 50 50
 

3 8 13
 

1 5
 

 17 32 30
 

9 2
 

 13 27 25
 

 17 41 28
 

8 13 10
 

 10 15 10
 

6 19 15
 

6 26 17
 

1
 

2
 

1 1
 

1 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN'TUMORS
 

* PRIMARY TUMORS: ALL TOHORS EXCEPT SECONDARY TOHORS
 * SECONDARY TUMORS: METASTATIC TUMORS OB TUMORS INVASIVE INTO AH ADJACENT ORGAN
 

60
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS FED DAMINOZIDE IN THE DIET
 

61
 

TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

FEDBAMINOZIDEINTHEDIET 

-1-TM "-1 —— —-— — •»•- "- ———••••——»— •———»———.————i
 

MATCHED LOW DOSE
 

CONTROL
 

AHIHALS INITIALLY IN STUDY 20 50
 

ANIHALS NECROPSIED 20 50
 

ANIHALS EXAMINED HISTOPATHOLOGICALLY 20 50
 

INTEGDHENTARY SYSTEH
 

*SKIN (20) (50)
 

EPIDERHAL INCLUSION CYST 2 (4-X)
 

ABSCESS, NOS 1 (2X)
 

*SOBCUT TISSUE (20) (50)
 

HEHOHHHAGIC CYST 1 <5X)
 

ABSCESS, NOS 1 (2X)
 

RESPIRATORY SYSTEH
 

fLUNG/BRONCHUS (20) (50)
 

BRONCHIECTASIS 1 (2X)
 

#LUNG (20) (50)
 

PNEUHONIA, ASPIRATION 1 (5X) 1 (2X)
 

PNEUBONIA, CHRONIC HURINE 7 (35X) 7 (14X)
 

PNEUHONIA INTERSTITIAL CHRONIC 1 (2X)
 

HYPERPLASIA, ADENOBATOUS 2 C*X)
 

HEBATOPOIETIC SYSTEH
 

#SPLEEN (20) (48)
 

HEHOSIDEROSIS 1 (5X)
 

HYPERPLASIA, RETICULUB CELL 1 (2X)
 

HEHATOPOIESIS 1 (2X)
 

tTHYHUS (D (2)
 

HYPERPLASIA, EPITHELIAL
 

CIRCULATORY SYSTEH
 

IHYOCARDIUH (18) (50)
 

# NUHBER OF ANIHALS WITH TISSUE EXAHINED HICROSCOPICALLY
 * NUHBER OF ANIHALS NECROPSIED
 

63
 

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

(50)
 

(50)
 

(50)
 

1 (2X)
 

16 (32X)
 

3 (6X)
 

(48)
 

1 (2X)
 

(D


1 (100X)
 

(49)
 
TABLE C1. MALE RATS: NONNEPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE
 

CONTROL
 

FIBROSIS 2 MIX) 10 (20X)
 

PIBEOSIS, FOCAL « (8X)
 

DIGESTIVE SYSTEM
 

fSALIVARY GLAND (20) (49)
 

ATROPHY, NOS 1 (2X)
 

fLIVER (20) (50)
 

ABSCESS, HOS
 

METAMORPHOSIS FATTI 1 (2X)
 

BASOPHILIC CYTO CHANGE 3 (15X) 5 (10X)
 

HENATOPOIESIS 1 (2X)
 

*BILE DUCT (20) (50)
 

INFLAMMATION, FOCAL
 

•PANCREAS	 (20) (48)
 

FIBROSIS 3 (6X)
 

FIBROSIS, FOCAL 5 (10X)
 

fPANCREATIC ACINUS (20) (48)
 

ATROPHY, NOS
 

ATROPHY, FOCAL
 

fLARGE INTESTINE (16) (49)
 

NEMATODIASIS 5 (31X) 17 (35X)
 

ICOLONIC MUCOUS HEMBR (16) (49)
 

ULCER, NOS 1 (2X)
 

URINARY SYSTEM
 

tKIDNEY (20) (50)
 

INFLAMMATION, CHRONIC 11 (55X) 30 (60X)
 

HYPERTROPHY, NOS
 

IKIDNEY/PELVIS (20) (50)
 

INFLAMMATION, SUPPUBATIVE
 

tURINABY BLADDER (19) (45)
 

INFLAMMATION,. ACUTE_HEMQRRHAGIC__
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECEOPSIED
 

64
 

HIGH DOSE
 

12 (24X)
 

4 (8X)
 

(46)
 

(50)
 

1 (2X)
 

4 (8X)
 

(50)
 

1 (2X)
 

(49)
 

3 (6X)
 

(49)
 

3 (6X)
 

1 (2X)
 

(48)
 

13 (27X)
 

(48)
 

(50)
 

32	 (64X)
 

1 (2X)
 

(50)
 

1 (2X)
 

(42)
 

_ _ 1 J2S1
 
TABLE C1. MALE RATS: NONNEPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL 

LOW DOSE HIGH DOSE 

ENDOCRINE SYSTEM 

•PITUITARY 

CYST, NOS 

HEHORRHAGIC CYST 

(18) 

1 (6X) 

(1*3) (49) 

1 (2*) 

•ADRENAL 

DEGENERATION, LIPOID 

NECROSIS, NOS 

METAMORPHOSIS FATTY 

(20) 

1 (5X) 

(50) (50) 

1 (2X) 

1 (2X) 

•ADRENAL MEDULLA 

DEGENERATION, HYALINE 

(20) (50) (50) 

1 (2X) 

•THYROID 

FOLLICOLAR CYST, NOS 

HYPERPLASIA, C-CELL 

(16) 

1

1

 (6X) 

 (6%) 

(38) 

2 (5X) 

(43) 

7 (16*) 

•PARATHYROID 

HYPERPLASIA, NOS 

(12) (22) (22) 

2 (9X) 

•PANCREATIC ISLETS 

HYPERPLASIA, NOS 

(20) (1*8) 

3 (6X) 

(49) 

REPRODUCTIVE SYSTEM 

*HA«HART GLAND 

GYNECOMASTIA 

(20) (50) 

<2X) 

(50) 

•PROSTATE	 (19) (HO) (47)
 

INFLAMMATION, NOS 1 (2X)
 

HYPERPLASIA, HOS 1 (2%)
 

*SEHIHAL VESICLE (20) (50) (50)
 

INFLAMMATION, SUPPURATIVE 1 (5X) 1 (2X) 1 (2%)
 

•TESTIS	 (20) (50) (50)
 

ATROPHY, NOS 3 <6X)
 

HYPERPLASIA, INTERSTITIAL CELL 1 (2X)
 

NERVOUS SYSTEM
 

•BRAIN	 (19) (50) (50)
 

J2SL
 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

65
 
TABLE C1. MALE RATS: NONNEPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

SPECIAL SENSE OBGANS
 

NONE
 

HUSC0LOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

*HULTIPLE OBGANS (20) (50) (50)
 

PEHIARTERITIS 1 (2%)
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 2
 

t NOPBER OF ANIMALS KITH TISSUE EIAHINED MICROSCOPICALLY
 

* ROBBER OF ANIHALS NECROPSIED
 

66
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

FEDDAMINOZIDEINTHEDIET 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

AHIHALS INITIALLY IN STUDY 20 50 50
 

ANIBALS NECROPSIED 20 50 50
 

ANIHALS EXAMINED HISTOPATHOLOGICALLY 20 50 50
 

INTEGUHEHTARY SYSTEM
 

*SUBCUT TISSUE (20) (50) (50)
 

INFLAMMATION, GRANULOMATOUS 1 (2%)
 

RESPIRATORY SYSTEM
 

fLDNG (20) (50) (U8)
 

CONGESTION, NOS 1 (2%)
 

EDEMA, NOS 1 (2*)
 

PNEUMONIA, LIPID 2 (H%)
 

PNEUMONIA, ASPIRATION 1 (5X)
 

PNEUMONIA, CHRONIC MURINE 5 (25X) 13 (26%) 9 (19S)
 

HEMATOPOIETIC SYSTEM
 

*BONE MARROW (19) (U8)
 

PIBBOSIS, FOCAL 1 (5%)
 

tSPLEEN (19) (50) (48)
 

HEMOSIDEHOSIS 1 (2%)
 

CYTOHEGALY 1 (2%)
 

HYPERPLASIA, LYMPHOID 1 (2X)
 

HEHATOPOIESIS 1 (2X) 1 (2%)
 

CIRCULATORY SYSTEM
 

*MYOCARDIUM (19) (50) (50)
 

FIBROSIS U (8X) <• (8X)
 

FIBROSIS, FOCAL 1 (2X)
 

CALCIFICATIQN^FQCAL
 

# NUMBER OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECEOPSIED
 

67
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

— T,.-_ TM " - • ' - - — ' -'1 —"- - — --"- ->'
 

DIGESTIVE SYSTEM
 

*GOH OF HAHDIBLE
 

ABSCESS, HOS
 

tLIVEE
 

INFLAMMATION, CHRONIC
 

NECROSIS, NOS
 

NECROSIS, FOCAL
 

NECROSIS, CENTRAL
 

METAMORPHOSIS FATTT
 

BASOPHILIC CYTO CHANGE
 

HEHATOPOIESIS
 

*BILE DUCT
 

HYPERPLASIA, NOS
 

tPANCREAS
 

PIBROSIS, FOCAL
 

NECROSIS, NOS
 

tESOPHAGOS
 

INFLAMMATION, NECROTIZING
 

tLARGE INTESTINE
 

NEHATODIASIS
 

ICOLON
 

NEHATODIASIS
 

URINARY SYSTEH
 

•KIDNEY
 

HYDRONEPHROSIS
 

PYELONEPHRITIS, NOS
 

INFLAMMATION, CHRONIC
 

NEPHROSIS, CHOLEHIC
 

INFAHCT, NOS
 

INFARCT, HEALED
 

CALCIFICATION, FOCAL
 

tURINABY BLADDER
 

_ lULAJIATIfilx-SlIEEJUAXIU
 

MATCHED
 

CONTROL
 

(20)
 

(19)
 

2 (11%)
 

5 (26%)
 

(20)
 

1 (5X)
 

(19)
 

1 (5X)
 

(20)
 

(17)
 

8 (47*)
 

(17)
 

(20)
 

2 (10%)
 

1 (5%)
 

(19)
 

LOW DOSE
 

.
 

(50)
 

(50)
 

1 (2%)
 

3 (6X)
 

7 (1UX)
 

(50)
 

(<*8)
 

2 (<»X)
 

1 (2X)
 

(*9)
 

("»«•)
 

8 (18)1)
 

(«4)
 

(50)
 

2 (<»X)
 

9 (18X)
 

1 (2X)
 

1 (2X)
 

(<»3)
 

HIGH DOSE
 

(50)
 

1 (2X)
 

(50)
 

2 (UX)
 

1 (2X)
 

3 (6X)
 

1 (2X)
 

2 (<«X)
 

11 (22X)
 

1 (2X)
 

(50)
 

(H9)
 

(«7)
 

1 (2X)
 

(US)
 

12 (25X)
 

(48)
 

1 (2X)
 

(48)
 

1 (2X)
 

5 (10X)
 

1 (2X)
 

1 (2X)
 

(43)
 

i_i2Si
 

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

68
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

(43)
 

(43)
 

(48)
 

1 (2X)
 

(48)
 

(44)
 

3 (7%)
 

(49)
 

1 (2X)
 

(50)
 

1 (2X)
 

(50)
 

(50)
 

3 (6%)


1 (2X)
 

1 (2X)
 

2 (4X)
 

(50)
 

1 mil
 

*D. BLADDER/MUCOSA
 

ATYPIA, NOS
 

ENDOCRINE SYSTEH
 

•PITUITARY
 

CYST, NOS
 

HYPBRPLASIA, CHROHOPHOBE-CELL
 

•ADRENAL
 

DEGENERATION, LIPOID
 

NECROSIS, FOCAL
 

•ADRENAL CORTEX
 

LIPOIDOSIS
 

•THYROID
 

HYPERPLASIA, C-CELL
 

•PANCREATIC ISLETS
 

HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEH
 

*HAHHARY GLAND
 

GALACTOCELE
 

CYST, NOS
 

HYPERPLASIA, CYSTIC
 

*VAGINA
 

INFLAHHATIOB, SUPPURATIVE
 

•OTERUS
 

CYST, NOS
 

PYOMETRA
 

ABSCESS, NOS
 

HYPERPLASIA, EPITHELIAL
 

POLYP, INFLASHATORY
 

fUTEHUS/ENDOHETRIOH
 

INFLAHHATIOM, NOS
 

lHIiABfliIJ2B*_59£E5IBilUfit NUMBER OF ANIHALS HITB TZSSQB

* NUHBER OF ANIHALS RBCROPSXBO 

-,
 

MATCHED
 

CONTROL
 

(19)
 

(19)
 

(20)
 

(20)
 

1 (5X)
 

(15)
 

(19)
 

(20)
 

(20)
 

(19)
 

3 (16X)
 

2 (11X)
 

(19)
 

1 (5X)
 » (21JI
 

LOW DOSE
 

(«3)
 

1 (2X)
 

(45)
 

1 (2%)
 

1 (2%)
 

(48)
 

1 (2X)
 

(48)
 

(38)
 

(48)
 

(50)
 

1 (2X)
 

1 (2X)
 

(50)
 

1 (2%)
 

(50)
 

1 (2X)
 •» (8X)
 

1 (2X)
 

1 (2X)
 

(50)
 

9 'W\
 

 KXABIIBD HICIOSCOtlCAUf 

69 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED

CONTROL 

 LOW DOSE HIGH DOSE 

INFLAMMATION, CHBONIC

HYPERPLASIA, CYSTIC

HYPERPLASIA, STEOHAL

 2 (4X) 

1 (2X)

 1 (2X) 

1 (2X) 

tOVARY/OVIDDCT

INFLAHBATION, SOPPORATIVE

ABSCESS, NOS

 (19)

 1 (5X)

 (50)

 1 (2X)

 1 (2X) 

(50) 

3 (6X) 

#OVARY

CYST, NOS

ABSCESS, NOS

 (19)

 4 (21X)

 1 (5X)

 (50)

9 (18X)

 1 (2X) 

(49) 

8 (16X) 

NERVOUS SYSTEM
 

tBRAIN (20) (49) (49)
 

COMPRESSION 1 (2X)
 

HYDROCEPHALUS, NOS 1 (2X)
 

SPECIAL SENSE ORGANS
 

NONE
 

MOSCOLOSKELETAL SYSTEM
 

NONE
 — ~_____«_—.~______~~____— ~_— _____*._____*W._AA.W_«.VA.4.W__>«»__W.«.._«».••.*•_•««»<
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED 1 3
 

* NUMBER OP ANIMALS WITH TISSUE EXAMIMED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

70
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE FED DAMINOZIDE IN THE DIET
 

71
 

TABLE D1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTJC LESIONS IN MALE MICE 

FED DAMINOZIDE IN THE DIET 

MATCHED LOW DOSE
 

CONTROL
 

ANIMALS INITIALLY IN STUDY 20 50
 

ANIMALS HISSING 6
 

ANIMALS NECROPSIED 14 50
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 1I» 50
 

INTEGUMENTARY SYSTEM
 

• SKIN (14) (50)
 

CYST, NOS 2 (14X) <2X)
 

*SUBCUT TISSUE (14) (50)
 

GRANULOMA, NOS
 

HISTIQCJTQSIS 

RESPIRATORY SYSTEM 

fLONG/BRONCHUS (14) (50) 

INFLAMMATION, NOS 1 (7X) 1 (2%) 

INFLAMMATION ACTIVE CHRONIC 1 (2X) 

INFLAMMATION, CHRONIC 4 (29X) 2 (4%) 

INFLAMMATION, CHRONIC FOCAL 1 (2%) 

tLUNG (14) (50) 

CONGESTION, NOS 1 (7X) 3 (6%) 

HEMORRHAGE 2 (4%) 

INFLAMMATION, NOS 

INFLAMMATION, FOCAL 

INFLAMMATION, ACUTE 3 (6X) 

INFLAMMATION, CHRONIC 1 <2X) 

INFLAMMATION, CHRONIC FOCAL 1 (2X) 

INFLAMMATION, CHRONIC SUPPURATIV 

GRANULOMA, NOS 1 (7X) 

NODULE 

PERIVASCOLITIS 5 (36X) 

1 (2X) 

4 (8X) 

HYPERPLASIA, ADENOHATOUS 3 (6X) 

LEOKEHOID REACTION 1 (7X) 

tLUNG/ALVEOLI (14) (50) 

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

73
 

HIGH DOSE
 

50
 

4
 

46
 

46
 

(46)
 

1 (2X)
 

(46)
 

(46)
 

1 (2X)
 

(46)
 

3 (7X)
 

1 <2X)
 

1 (2X)
 

1 (2%)
 

7 (15X)
 

1 (2X)
 

(46)
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

— —"- ————"-—— — •--"—-——-—"
 MATCHED LOW DOSE
 

CONTROL
 

HEHATOPOIETIC SYSTEM
 

^MULTIPLE ORGANS (14) (50)
 

HYELOPROLIFERATIVE DISORDER
 

IBONE MARROW (14) (45)
 

HYPERPLASIA, HEHATOPOIETIC
 

tSPLEEN (13) (HO)


INFLAMMATION, FOCAL GRANULOHATOU 1 (3*)
 

HYPERPLASIA, RETICULUM CELL
 

HYPBRPLASIA, LYHPHOID 3 (8X)
 

HEHATOPOIESIS 1 (8*) 5 (13X)
 

tLYMPH NODE (9) (42)
 

HYPERPLASIA, RETICULUH CELL
 

tMANDIBULAR L. NODE (9) (12)
 

HYPERPLASIA, NOS 2 (5X)
 

HISTIOCYTOSIS 1 (2X)
 

PLASMACYTOSIS 1 (2X)
 

HYPERPLASIA, RETICULUH CELL
 

HYPERPLASIA, LYHPHOID 1 (2X)
 

tBRONCHIAL LYHPH NODE (9) (42)
 

HYPERPLASIA, LYHPHOID 1 (2X)
 

tMESENTERIC L. NODE (9) (42)
 

DILATATION, NOS
 

LYHPHANGIECTASIS 7 (17X)
 

CONGESTION, CHRONIC
 

HEMORRHAGE 1 (11X) 3 (7X)
 

INFLAHMATION, GRANULOMATOUS
 

HYPERPLASIA, NOS 2 (5X)
 

HYPEEPLASIA, FOCAL 1 (2*)
 

HISTIOCYTOSIS 1 (11*) 1 (2*)
 

PLASMACYTOSIS
 

HEGAKARYOCYTOSIS 1 (11X)
 

HYPERPLASIA, RETICULUM CELL 1 (2X)
 

HEMATOPOIESIS 1 (2X)
 

CIRCULATORY SYSTEM
 

#HEART (14) (48)
 

ENDQC.ARP.ITIS^BACTE.R.IAL.
 

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

74
 

HIGH DOSE
 

(46)
 

1 (2X)
 

(43)
 

1 (2X)
 

(43)
 

1 (2X)
 

1 (2X)
 

1 (2*)
 

(43)
 

1 (2X)
 

(43)
 

5 (12X)
 

2 (5X)
 

1 (2*)
 

(43)
 

(43)
 

1 (2X)
 

2 (5X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 <2X)
 

(45)
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

HIGH DOSE
 

1 (2X)
 

(45)
 

1 (2X)
 

(46)
 

(46)
 

(43)
 

2 (5X)
 

(46)
 

1 (2X)
 

5 (11X)
 

1 (2X)
 

2 (4X)
 

1 (?*)
 

1 (i 1)
 

2 (4X)
 

4 (9X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

(46)
 

1 (2X)
 

(46)
 

CALCIFICATION, FOCAL
 

#HYOCARDIUH
 

INFLAHHATION, ACUTE
 

INFLABHATION, CHRONIC
 

*CORONARY ARTERY
 

ARTERIOSCLEROSIS, NOS
 

* INGUINAL ARTERY
 

INFLAHHATION, NOS
 

ARTERIOSCLEROSIS, NOS
 

DIGESTIVE SYSTEH
 

fSALIVARY GLAND
 

PERIVASCULITIS
 

#LIVER
 

HULTILOCULAR CYST
 

THROHBUS, ORGANIZED
 

CONGESTION, CHRONIC PASSIVE
 

INFLAHHATION, NECROTIZING
 

INFLAHHATION ACTIVE CHRONIC
 

INFLAHHATION, CHRONIC FOCAL
 

CIRRHOSIS, NOS
 

PERIVASCULITIS
 

DEGENERATION, HYDROPIC
 

NECROSIS, FOCAL
 

INFARCT, NOS
 

HETAHORPHOSIS FATTY
 

HEHOSIDEROSIS
 

CYTOPLASHIC VACUOLIZATION
 

BASOPHILIC CYTO CHANGE
 

HYPERPLASIA, NOS
 

ANGIECTASIS
 

HEHATOPOIESIS
 

^GALLBLADDER
 

CALCULUS, NOS
 

*BILE DUCT
 

DILATATION, NOS
 

MATCHED LOW DOSE
 

CONTROL
 

(14) (48)
 

1 (2X)
 

(14)	 (50)
 

1 (2X)
 

(14)	 (50)
 

1 (2X)
 

1 (2X)
 

(12)	 (47)
 

1 (8X) 6 (13X)
 

(14)	 (50)
 

3 (6X)
 

1 (2X)
 

1 (7X) 2 (4X)
 

3 (6X)
 

1 (7X) 1 (2X)
 

2 (4X)
 

1 (2X)
 

1 (7X)
 

1 (7X)
 

1 (2X)
 

1 (2X)
 

(14) (50)
 

(14)	 (50)
 

1 (2X)
 

t	 NUHBER OF ANIMALS WITH TISSUE EXAHINED MICROSCOPICALLY
 * NUHBER OF ANIHALS NECROPSIED
 

75
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL 

LOW DOSE HIGH DOSE 

HYPERPLASIA, NOS 2 (4X) 

•PANCREAS 

CYST, NOS 

CYSTIC DUCTS 

FIBHOSIS, FOCAL 

PERIVASCULITIS 

BETAHOBPHOSIS FATTY 

ATROPHY, HOS 

(12) (47) 

1 (2X) 

1 (2X) 

1 (2X) 

(42) 

1 (2X) 

1 (2X) 

1 (2X) 

IPANCREATIC DUCT 

INFLAHHATION ACTIVE CHRONIC 

PARASITISH 

(12) 

1 (8X) 

(47) 

1 (2X) 

(42) 

•PANCREATIC ACINUS 

ATROPHY, NOS 

(12) 0*7) 

2 (4X) 

(42) 

•STOHACH

INFLAHHATION, NOS

INFLAHHATION, ACUTE/CHRONIC

HEHOSIDEROSIS

HYP2RPLASIA, FOCAL

 (13) (48) (42) 

1 (2X) 

1 (2X) 

1 (2X) 

1 (2X) 

#PEYERS PATCH (13) (48) (44)
 

HYPERPLASIA, NOS 4 (31X) 9 (19X) 9 (20X)
 

HYPERPLASIA, LYHPHOID 1 (2X)
 

tCOLON (13) (39) (41)
 

NEHATODIASIS 1 (2X)
 

PARASITISH 3 (23X) 7 (18X) 16 (39X)
 

tCOLONIC SOBHUCOSA (13) (39) (41)
 

INFLAHHATION, ACUTE 1 (3X) 1 (2X)
 

INFLAHHATION, CHRONIC 1 (3X)
 

INFLAHHATION, GRAHULOHATOUS 1 (3X)
 

URINARY SYSTEH
 

tKIDNEY (14) (50) (46)
 

IHFLAHHATION, CHRONIC 2 (14X) 1 (2X) 3 (7*)
 

INFLABHATION, CHRONIC FOCAL 1 (2X) 1 (2X)
 

PEHITASCnLITIS 2 (14X) 7 (14X) 6 (13X)
 

INFARCT, HOS 1 (2X)
 

t NOHBEB OF AHIHALS HITQ TISSUE EIAMINED HICROSCOPICALLY
 

* NOBBER OF ANIBALS NECROPSIED
 

76
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

CYTOPLASHIC VACUOLIZATION 1 <7X)
 

HYPERPLASIA, TUBULAR CELL 2 (14X) 2 (4X) 1 (2X)
 

tKIDNEY/CORTEX (14) (50) (46)
 

CYST, NOS 1 (2X)
 

SCAR 1 (2X)
 

tKIDNEY/TUBULE (14) (50) (46)
 

DEGENERATION, NOS 1 (2X)
 

LIPOIDOSIS 3 (21X) 1 (2X) 2 (4X)
 

tKIDNEY/PELVIS (14) - (50) (46)
 

INFLAMMATION, CHRONIC 2 (14X)
 

INFLAMMATION, CHRONIC FOCAL 1 (2X)
 

tURINARY BLADDER (10) (47) (39)
 

LYMPHOCYTIC INFLAMMATORY INFILTH 1 (10X) 1 (2X) 1 (3X)
 

INFLAMMATION, CHRONIC FOCAL 1 (3X)
 

ENDOCRINE SYSTEM
 

IADRENAL (8) (35) (38)
 

CONGESTION, NOS 1 (3X)
 

FIBROSIS, FOCAL 1 (3X)
 

tTHYROID (12) (39) (35)
 

FOLLICULAR CYST, NOS 1 <3X)
 

PEHIVASCULITIS 1 OX)
 

tTHYROID FOLLICLE (12) (39) (35)
 

ECTOPIA 1 (3X)
 

tPANCREATIC ISLETS (12) (47) (42)
 

HYPERPLASIA, NOS 1 (2X)
 

REPRODUCTIVE SYSTEM
 

*SEMINAL VESICLE (14) (50) (46)
 

INFLAMMATION, NOS 1 (7X)
 

INFLAMMATION, CHRONIC 1 (2X)
 

CHOLESTEROL DEPOSIT 1 (7X)
 

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

77
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED

CONTROL 

 LOW DOSE HIGH DOSE 

NERVOUS SYSTEM 

fBRAIH

HEMORRHAGE

CORPORA AMYLACEA

CALCIFICATION, NOS

CALCIFICATION, FOCAL

 (14)

 5 (36%)

 (49)

 1 (2%) 

1 (2X)

 15 (31X)

 (<»5) 

1 (2X) 

1 (2%) 

 13 (29%) 

SPECIAL SENSE ORGANS 

NONE 

MOSCOLOSKELETAL SYSTEH 

*BONE

FIBROUS OSTEODYSTROPHY

 (14) (50)

 1 (2%) 

 (46) 

BODY CAVITIES 

*ABDOHINAL CAVITY

NECROSIS, NOS

 (14) (50) (46) 

1 (2%) 

*PERITONEUM

NECROSIS, NOS

 (14) (50)

 1 (2%) 

 (46) 

*MESENTERY

NECROSIS, FOCAL

 (14) (50) (46) 

1 (2%) 

ALL OTHER SYSTEMS 

THORAX 

NODULE 1 

DIAPHRAGM 

INFLAMMATION, ACUTE NECROTIZING 1 

ADIPOSE TISSUE 

INFLAMMATION, CHRONIC 1 

SPECIAL MORPHOLOGY SUMMARY 

NO_LESIQN_£EPORT£D <j 

* NUMBER OF ANIMALS UITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

78
 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

ANIMAL HISSING/NO NECROPSY 6 4
 

ADTO/NECROPSY/HISTO PEBP 1 1 1
 

* NUMBER OF ANIHALS KITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIHALS NBCBOPSIED
 

79
 
TABLE 02. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

FED DAMINOZIDE IN THE DIET 

MATCHED LOW DOSE 

CONTROL 

ANIMALS INITIALLY IN STUDY 20 50 

ANIMALS MISSING 9 

ANIMALS NECHOPSIED 20 <»1 

ANIMALS EXAHINED HISTOPATHOLOGICALLY 20 41 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

ITRACHEA (16) (31)
 

INFLAMMATION, CHRONIC FOCAL
 

tLUNG/BRONCHUS (20) (39)
 

INFLAMMATION, NOS 2 (10X) 3 (8X)
 

INFLAMMATION, FOCAL 1 (3X)
 

INFLAMMATION ACTIVE CHRONIC
 

INFLAMMATION, CHRONIC 2 (10X) 3 (8X)
 

• LUNG (20) (39)
 

EMPHYSEMA, NOS 1 <3X)
 

CONGESTION, NOS
 

HEMORRHAGE 1 (5X)
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL 1 (5X) 2 (5X)
 

INFLAMMATION, ACUTE
 

PNEUMONIA, CHRONIC MURINE 1 <5X)
 

INFLAMMATION, CHRONIC 1 (3X)
 

SCLEROSIS
 

NODDLE 1 (3X)
 

PERI7ASCDLITIS 8 (40X) 12 (31X)
 

HYPERTROPHY, NOS 1 (3X)
 

HYPERPLASIA, ADENOMATOUS 2 (5X)
 

LEUKEMOID REACTION
 

RETICULOENDOTHBLIOSIS 1 (5X)
 

•LUNG/ALVEOLI (20) (39)
 

mi
 • HUHBBR OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

80
 

HIGH DOSE
 

50
 

2
 

48
 

48
 

(43)
 

1 (2X)
 

(48)
 

6 (13X)
 

1 (2X)
 

4 (8X)
 

(48)
 

6 (13X)
 

1 (2X)
 

3 (6X)
 

1 (2X)
 

1 (2X)
 

13 (27X)
 

3 (6X)
 

2 (4X)
 

(48)
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

HEBATOPOIETIC SYSTEM
 

tBONE MARROW


HYPERPLASIA, HEMATOPOIETIC


•SPLEEN


INFLAMMATION, GRANULOMATOUS


AMYLOID, NOS


AMYLOIDOSIS


HYPERPLASIA, NOS


LEUKEMOID REACTION


HYPERPLASIA, LYHPHOID


HEMATOPOIESIS


tLYMPH NODE


INFLAMMATION, NOS


HYPERPLASIA, NOS


HYPERPLASIA, DIFFUSE


#BANDIBULAR L. NODE


HYPERPLASIA, NOS


HISTIOCYTOSIS


fBRONCHIAL LYMPH NODE


HYPERPLASIA, NOS


HISTIOCYTOSIS


PLASBACYTOSIS


fBESENTERIC L. NODE


DILATATION, NOS


LYBPHANGIECTASIS


HEMORRHAGE


INFLAMMATION, NOS


INFLAMMATION ACUTE AND CHRONIC


HEMOSIDEROSIS


HYPERPLASIA, NOS


HYPEHPLASIA, FOCAL


HISTIOCYTOSIS


PLASHACYTOSIS


BEGAKARYOCYTOSIS


HEHATOPOIESIS


fRENAL LYBPH NODE


MATCHED LOW DOSE HIGH DOSE 

CONTROL 

 (18) (34) (41) 

1 (3X) 1 (2X) 

 (17) (34) (42) 

1 (2X) 

1 (2X) 

1 (2X) 

1 (2X) . 1 (2X) 

2 (12X) 6 (18X) 

1 (6X) 3 (9X) 2 (5X) 

 (20) (37) (40) 

1 (3X) 

1 (3X) 

1 (3X) 

 (20) (37) (40) 

4 (11X) 5 (13X) 

1 (3X) 

 (20) (37) (HO) 

1 (5X) 1 (3X) 1 (3X) 

1 (3X) 

1 (3X) 

 (20) (37) (40) 

1 (3X) 

1 (3X) 1 (3X) 

1 (3X) 1 (3X) 

1 (3X) 

1 (3X) 

1 (3X) 

2 (5X) 

1 (3X) 

1 (3X) 

1 (3X) 

1 (3X) 

1 (3X) 

 (20) (37) («0) 

U3.S1 

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

81
 
TABLE 01 FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL 

LOW DOSE HIGH DOSE 

HYPERPLASIA, LYHPHOID 

CIRCULATORY SYSTEM 

tHEART 

INFARCT, HEALED 

CALCIFICATION, FOCAL 

(2C) (41) (47) 

1 (2X) 

1 (2*) 

tHYOCARDinn (20) (41) (47)
 

INFLAHHATION, CHRONIC FOCAL 1 (2X)
 

FIBROSIS, FOCAL 2 (5X)
 

*CORONAHY ARTERY (20) (41) (48)
 

ARTERIOSCLEROSIS, NOS 1 (2X)
 

*POLHONARY ARTERY (20) (41) (48)
 

HYPERTROPHY, NOS 1 (5X) 1 (2X) 1 (2X)
 

HYPERPLASIA, NOS 1 (5X) 1 (2X) 1 (2X)
 

*ANTERIOF HENINGEAL A (20) (41) (48)
 

ARTERIOSCLEROSIS, NOS 1 (2X)
 

DIGESTIVE SYSTEH
 

tSALIVARY GLAND (18) (37) (42)
 

INFLAHHATION. CHRONIC FOCAL 1 (6%)
 

PERIVASCULITIS 1 (6X) 3 (8X) 4 (10X)
 

HETAHORPHOSIS FATTY 1 (2X)
 

tLIVER (20) (40) (48)
 

CYST, NOS 1 (5X)
 

CONGESTION, NOS 1 (3%)
 

CONGESTION, CHRONIC PASSIVE 1 (2X)
 

INFLAHHATION, FOCAL 1 (2X)
 

INFLAHHATION, CHRONIC FOCAL 1 OX)
 

FIBROSIS, FOCAL 1 (2%)
 

PERIVASCULITIS 4 (8X)
 

DEGENERATION, HYDROPIC 5 (25X)
 

NECROSIS, COAGULATIVE 1 (3X)
 

INFARCT, NOS 1 (3X) 1 (2X)
 

AHYLOIDOSIS 1 (2X)
 

3ETAHORPHOSIS FATTY 1 (3%) 3 (6X)
 

.___4MSIKlASis
 

f NUHBER OF ANIHALS WITH TISSUE EXAMINED HICROSCOPICALLY
 * NUHBER OF ANIHALS NECROPSIED
 

82
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

HYPERPIASIA, LYMPHOID	 1 (3X)
 

HEMATOPOIESIS	 1 (3X)
 

#LIVER/CENTRILOBULAR (20) (40) (48)
 

CONGESTION, NOS 1 (5X)
 

METAMORPHOSIS FATTY 1 (5X)
 

*GALLBLADDER (20)	 ( U 1 i
 (U8)
 

INFLAMMATION, ACUTE	 1 (2X)
 

*BILE DUCT	 (20) i U 11
 (48)
 

INFLAMMATION ACTIVE CHRONIC 1 (2X)
 

INFLAMMATION, CHRONIC 1 (2X) 2 (UX)
 

INFLAMMATION, CHRONIC FOCAL 1 (5X) 1 (2X) 1 (2X)
 

GRANULOHA, NOS 1 (2X)
 

FIBROSIS 1 (2X)
 

LYMPHOID DEPLETION 1 (5X)
 

tPANCREAS (16) (36) (44)
 

INFLAMMATION, CHRONIC 1 (2X)
 

INFLAMMATION, CHRONIC FOCAL 1 (2X)
 

ATROPHY, FOCAL 1 (2X)
 

fESOPHAGUS (19) (36) (US)
 

INFLAMMATION ACTIVE CHRONIC 1 (3X)
 

•STOMACH	 (20) (39) (45)
 

ULCER, NOS 1 (2X)
 

INFLAMMATION, HEMORRHAGIC 1 (2%)
 

INFLAMMATION, ACUTE FOCAL 1 (3X)
 

CALCIFICATION, NOS 1 <3X)
 

tSMALL INTESTINE (20) (38) (46)
 

INFLAHMATION, CHRONIC FOCAL 1 (2X)
 

•PEYERS PATCH (20) (38) (46)
 

HYPERPLASIA, NOS 2 (10X) 4 (11X] I 6 (13X)
 

HYPERPLASIA, RETICULOB CELL 1 (2X)
 

HYPERPLASIA, LYMPHOID 1 (2»)
 

•DUODENUM	 (20) (38) (46)
 

HYPERPLASIA, LYMPHOID 1 (3X)
 

•LARGE INTESTINE (17) (37) (31)
 

CONGESTION, NOS___ ] 12X1
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

83
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE
 

CONTROL
 

#COLON (17) (37)
 

PARASITISM 1 (6X) 3 (8X)
 

tCOLONIC SUBMUCOSA (17) (37)
 

INFLAMMATION, GRANULOHATOOS 1 (6X)
 

URINARY SYSTEH
 

tKIDNEY (20) (41)
 

HYDRONEPHROSIS 1 (5X)
 

INFLAHHATION, INTERSTITIAL
 

INFLAMMATION ACTIVE CHRONIC 1 (2X)
 

INFLAHHATION, CHRONIC
 

GLOMEKOLONEPHRITIS, CHRONIC 1 (2X)
 

INFLAHHATION, CHRONIC FOCAL 3 (15X)
 

PERIVASCULITIS 5 (25X) 5 (12X)
 

NECROSIS, MEDOLLARY
 

AMYLOID, NOS
 

AMYLOIDOSIS
 

HYPERPLASIA, TOBULAR CELL
 

PLASHACYTOSIS 1 (2%)
 

tKIDNEY/TUBULE (20) (41)
 

LIPOIDOSIS
 

tKIDNEY/PELVIS (20) (41)
 

INFLABMATION ACTIVE CHRONIC 1 (2X)
 

INFLAHHATION, GRANULOHATOUS
 

*ORETER (20) (41)
 

PERIARTERITIS
 

tURINARY BLADDER (15) (31)
 

LYHPHOCYTIC INFLAHHATORY ISFILTR 7 (47X) 7 (23X)
 

INFLAHHATION, CHRONIC 1 (3X)
 

PLASMA-CELL INFILTRATE 1 (3X)
 

NODULE 1 (3X)
 

PERIVASCULITIS 1 (3X)
 

ANGIECTASIS 1 (3X)
 

tO. BLADDEH/HUCOSA
 

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

84
 

HIGH DOSE
 

(31)
 

1 (3X)
 

(31)
 

(48)
 

1 (2X)
 

2 (4X)
 

5 (10X)
 

1 (2X)
 

1 (2X)
 

3 <6X)
 

1 (2X)
 

(48)
 

1 (2X)
 

(48)
 

1 (2X)
 

(48)
 

1 (2X)
 

(33)
 

9 (27X)
 

(33)
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE
 

CONTROL
 

ENDOCRINE SYSTEM
 

•ADRENAL	 (17) (31)
 

CONGESTION, NOS 1 (6%)
 

INFLAHHATION, CHRONIC 1 (3*)
 

DEGENERATION, NOS 1 (3X)
 

AMYLOIDOSIS
 

CYTOLOGIC DEGENERATION
 

•ADRENAL	 MEDULLA (17) (31)
 

CONGESTION, NOS 1 (3X)
 

•THYROID	 (13) (28)
 

INFLAMMATION ACTIVE CHRONIC 1 (<»X)
 

HYPERPLASIA, AD3NOMATOOS 1 C»X)
 

REPRODUCTIVE SYSTEM
 

•UTERUS	 (20) (37)
 

HYDROMETRA 1 (3X)
 

CYST, NOS 2 (5X)
 

HEMORRHAGIC CYST
 

INFLAMMATION, NOS 1 (3X)
 

LYHPHOCYTIC INFLAMMATORY INFILTR 1 (5X)
 

INFLAMMATION, VESICULAR
 

INFLAMMATION ACTIVE CHRONIC 1 (3X)
 

SCLEROSIS
 

METAPLASIA, SQUAMOOS
 

•CERVIX UTERI (20) (37)
 

INFLAMMATION, ACUTE 2 (5X)
 

•UTSRUS/ENDOMETRIUM	 (20) (37)
 

CYST, NOS 5 (25X) 6 (16X)
 

INFLAMMATION, NOS
 

LYMPHOCYTIC INFLAMMATORY INFILTR
 

INFLAMMATION, VESICULAR 2 (10X) 1 (3X)
 

ABSCESS, NOS
 

HYPERPLASIA, CYSTIC 5 (25X) 12 (32X)
 

METAPLASIA, SQUAMOUS
 

•OTERUS/MYOMETRIUM	 (20) (37)
 

INFLAMMATION,. ACU.TE _ _
 

# NOHBER OF A-NIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

85
 

HIGH DOSE
 

(38)
 

1 (3X)
 

1 <3X)
 

(38)
 

(38)
 

(45)
 

4 <9X)
 

1 (2X)
 

1 (2X)
 

2 (4X)
 

1 (2X)
 

3 (7X)
 

(45)
 

5 (11X)
 

(45)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1	 (2%)
 

22 (49X)
 

1 (2X)
 

(45)
 

2-J4S1—
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE
 

CONTROL
 

INFLAMMATION, ACUTE VESICULAR 

#OVAHY/OVIDUCT 

INFLAMMATION ACTIVE CHRONIC 

(20) (37) 

<3X) 

#OVARY 

CYST, NOS 

PAROVARIAN CYST 

HEMORRHAGIC CYST 

ATROPHY, NOS 

(13) 

6 (46X) 

(14) 

3

1

1

 (21X) 

 (7X) 

 (7X) 

NERVOUS SYSTEM
 

tBRAIN/MENINGES (19) (40)
 

INFLAMMATION, CHRONIC 1 (5X)
 

PERIARTERITIS 1 (5X)
 

tBRAIN (19) (40)
 

LYHPHOCYTIC INFLAMMATORY INFILTR 1 (5X)
 

PERIVASCULITIS 1 <5X) 1 (3X)
 

CORPORA AMYLACEA 1 (3X)
 

CALCIFICATION, FOCA;, 4 (21X) 12 (30%)
 

CYTOPLASMIC VACUOLIZATION
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

*BONE (20) (41)
 

FIBROUS OSTEODYSTROPHY 13- (65X) 18 (44X)
 

OSTSOSCLEROSIS 1 (2X)
 

*SKELETAL MUSCLE (20) (41)
 

PARASITISM 1 <2X)
 

BODY CAVITIES
 

*MEDIASTINUM (20) (41)
 

THRQMBUSi_QRGANIZED.
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

86
 

HIGH DOSE
 

<2X)
 

(45)
 

(23)
 

6 (26X)
 

1 C*X)
 

(46)
 

(46)
 

10 (22X)
 

1 (2X)
 

(48)
 

21 (44X)
 

(48)
 

(48)
 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED LOW DOSE HIGH DOSE
 

CONTROL
 

PERIARTERITIS 1 (2X)
 

'PERITONEUM (20) (41) (48)
 

INFLAMMATION, FOCAL 1 12%)
 

'MESENTERY (20) (41) (48)
 

STEATITIS 1 <2X)
 

NECROSIS, FAT 2 (5X)
 

ALL OTHER SYSTEMS
 

'MULTIPLE ORGANS (20) (41) (48)
 

RUSSELL BODY 1 <2X)
 

SPECIAL MORPHOLOGY SUMMARY
 

ANIMAL MISSING/NO NECROPSY 9 2
 

AUTO/NECROPSY/HISTO PERF 2
 

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

87
 

APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS FED DAMINOZIDE IN THE DIET
 

89
 

__ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Topography; Morphology


Lung: Alveolar /Bronchiolar Adenoma
 

or Carcinomab


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Pituitary: Chromophobe Adenomab


P Valuesc>d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Daminozide in the Diet3
 

Matched Low
 

 Control Dose
 

 1/20 (5)	 3/50 (6)
 

 N.S.	 N.S.
 

1.200
 

0.106
 

61.724
 

 105 105
 

 0/18 (0) 2/43 (5)
 

 N.S.	 N.S.
 

Infinite
 

0.129
 

Infinite
 

—	 96
 

High
 

Dose
 

0/50 (0)
 

N.S.
 

0.000
 

0.000
 

7.475
 

2/49 (4)
 

N.S.
 

Infinite
 

0.113
 

Infinite
 

105 
__ 

Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

(continued)
 

Tomography: Morphology


Adrenal: Pheochromocytoma or
 

Pheochromocytoma, Malignantb


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


VO
 

NJ Thyroid: C-cell Carcinomab


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Daminozide in the Dieta
 

Matched
 

 Control
 

 2/20 (10)
 

 N.S.
 

 105
 

 0/16 (0)
 

 N.S.
 

—
 

Low High
 

Dose Dose
 

2/50 (4) 2/50 (4)
 

N.S.	 N.S.
 

0.400	 0.400
 

0.032	 0.032
 

5.277	 5.277
 

105 48
 

0/38 (0) 2/43 (5)
 

N.S.	 N.S.
 

Infinite
 

0.116
 —
 Infinite
 —
 

105
 ——
 
Table El.


(continued)
 

Topography: Morphology
 

Thyroid: C-cell Adenoma or
 

Carcinoma13
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

VO
 

CO
 Preputial Gland: Adenoma
 

or Carcinoma, NOS*5
 

P Value sc»d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Daminozide in the Dieta
 

Matched Low
 

Control Dose
 

2/16 (13) 3/38 (8)
 

N.S. N.S.
 

0.632
 

0.082
 

7.118
 

104 105
 

2/20 (10) 1/50 (2)
 

N.S. N.S.
 

0.200
 

0.004
 

3.681
 

67 99
 

High
 

Dose
 

2/43 (5)
 

N.S.
 

0.372
 

0.030
 

4.864
 

105
 

4/50 (8)
 

N.S.
 

0.800
 

0.128
 

8.436
 

85
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Daminozide in the Diet3
 

(continued)
 

Matched Low High
 

Topography; Morphology Control Dose Dose
 

Testis: Interstitial-cell Tumorb 13/20 (65) 46/50 (92) 47/50 (94)
 

P Values0 »d P = 0.003 P - 0.009 P = 0.004
 

Departure from Linear Trend6 P = 0.042
 

Relative Risfcf 1.415 1.446
 

Lower Limit 1.043 1.074
 

Upper Limit 1.873 1.830
 

Weeks to First Observed Tumor 101 78 85
 

a

Dosed groups received 5,000 or 10,000 ppm.
 

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

c

Beneath the incidence of tumors in the matched-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

d

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

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

*The 95% confidence interval of the relative risk between each dosed group and the control group.
 
__ 

Table E2. Analyses of the Incidence of Primary tumors In Female Rats
 

Topography: Morphology


Lung: Alveolar /Bronchiolar
 

Adenomab


P Values0 »d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 vo
 

Hematopoietic System: Leukemiab


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Daminozide in the Dieta
 

Matched
 

 Control
 

 0/20 (0)
 

P = 0.036
 

—
 

 0/20 (0)
 

 N.S.
 

Low
 

Dose
 

0/50 (0)
 

N.S.
 

_«.
 

—
 

4/50 (8)
 

N.S.
 

Infinite
 

0.386
 

Infinite
 

83
 

High
 

Dose
 

4/48 (8)
 

N.S.
 

Infinite
 

0.402
 

Infinite
 

102
 

1/50 (2)
 

N.S.
 

Infinite
 

0.022
 

Infinite
 

102
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Daminozide in the Diet3
 

(continued)
 

Matched Low High
 

Topography: Morphology Control Dose Dose
 

Pituitary: Chromophobe Carcinomab 0/19 (0) 3/45 (7) 0/43 (0)
 

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

Departure from Linear Trend6 P = 0.046
 

Relative Riskf Infinite
 _-»
 

Lower Limit 0.265
 _—


Upper Limit Infinite
 

vo Weeks to First Observed Tumor 96
 ——
 

Pituitary: Chromophobe Adenoma
 

or Carcinoma15 3/19 (16) 10/45 (22) 8/43 (19)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Riskf 1.407 1.178
 

Lower Limit 0.424 0.330
 

Upper Limit 7.336 6.361
 

Weeks to First Observed Tumor 105 89 92
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

(continued)
 

Topography; Morphology
 

Thyroid: C-cell Carcinoma'3
 

P Values0 >d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-cell Adenoma or
 

Carcinoma^
 

P Values0>d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Daminozide in the Dieta
 

Matched


Control


2/15 (13)


P = 0.026 (N)


105


4/15 (27)


P = 0.021 (N)


105


 Low
 

 Dose
 

 1/38 (3)
 

 N.S.
 

0.197
 

0.004
 

3.581
 

 99
 

 3/38 (8)
 

 N.S.
 

0.296
 

0.051
 

1.578
 

 99
 

High
 

Dose
 

0/44 (0)
 

N.S.
 

0.000
 

0.000
 

1.140
 

105
 

2/44 (5)
 

P = 0.032 (N)
 

0.170
 

0.018
 

1.085
 

105
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

(continued)
 

Topography; Morphology
 

Mammary Gland: Fibro adenoma^
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

vo Uterus: Leiomyosarcoma'5
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Daminozide in the Diet3 

Matched Low High 

Control Dose Dose 

3/20 (15) 9/50 (18) 2/50 (4) 

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

1.200 0.267 

0.346 0.024 

6.408 2.190 

105 89 105 

0/19 (0) 1/50 (2) 3/50 (6) 

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

Infinite Infinite 

0.021 0.238 

Infinite Infinite 

98 96 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Daminozide in the Dieta 

(continued) 

Matched 

Topography; Morphology Control 

Uterus: Endometrial Stromal Polypb 0/19 (0) 

P Values0 >d N.S. 

Relative Riskf 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Uterus /Endometrium: Adenocarcinoma, 

NOSb 0/19 (0) 

P Valuesc » d N.S. 

Relative Risk^ 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Low
 

Dose
 

6/50 (12)
 

N.S.
 

Infinite
 

0.636
 

Infinite
 

105
 

5/50 (10)
 

N.S.
 

Infinite
 

0.501
 

Infinite
 

83
 

High
 

Dose
 

4/50 (8)
 

N.S.
 

Infinite
 

0.368
 

Infinite
 

105
 

3/50 (6)
 

N.S.
 

Infinite
 

0.238
 

Infinite
 

105
 
__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Daminozide in the Dieta
 

(continued)
 

Matched Low High
 

Topography: Morphology Control Dose Dose
 

Mesentery: Lipoma*5 0/20 (0) 0/50 (0) 2/50 (4)
 

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

Relative Riskf Infinite
 

Lower Limit 0.123
 

Upper Limit —
 Infinite
 —
 

Weeks to First Observed Tumor 92
 —
 

a

Dosed groups received 5,000 or 10,000 ppm.
 

—
 

o
 

o
 

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

c

Beneath the incidence of tumors in the matched-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

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

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

95% confidence interval of the relative risk between each dosed group and the control group.
 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN MICE FED DAMINOZIDE IN THE DIET
 

101
 

Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Topography: Morphology


Lung: Alveolar /Br onchiolar Carcinoma*3


P Values0 »d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar /Bronchiolar Adenoma
 

o
 

CO or Carcinomab


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Daminozide in the Diet3
 

Matched Low 

 Control Dose 

 2/14 (14) 9/50 (18) 

 N.S. N.S. 

1.260 

0.314 

11.287 

 66 89 

 4/14 (29) 15/50 (30)
 

 N.S. N.S.
 

1.050
 

0.425
 

3.858
 

 66 83
 

High
 

Dose
 

12/46 (26)
 

N.S.
 

1.826
 

0.493
 

15.708
 

92
 

18/46 (39)
 

N.S.
 

1.370
 

0.576
 

4.875
 

92
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

(continued)
 

Topography: Morphology


Hematopoietic System: Lymphoma
 

or Leukemiab


P Valuesc'd


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Liver: Hepatocellular Carcinoma*5


P Values0'd


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Daminozide in the Diet3 

Matched Low High 

 Control Dose Dose 

 2/14 (14) 6/50 (12) 11/46 (24) 

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

0.840 1.674 

0.179 0.442 

8.038 14.549 

 103 90 83 

 0/14 (0) 7/50 (14) 13/46 (28) 

P = 0.008 N.S. P = 0.020 

Infinite Infinite 

0.583 1.314 

Infinite Infinite 

86 65 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed Daminozide in the Diet3
 

(continued)
 

Matched Low High
 

Topography: Morphology Control Dose Dose
 

Liver: Hepatocellular Adenoma
 

or Carcinoma^ 1/14 (7) 9/50 (18) 14/46 (30)
 

P Value sc»d P - 0.031 N.S. N.S.
 

Relative Riskf 2.520 4.261
 

Lower Limit 0.411 0.770
 

Upper Limit 107.898 174.781
 

Weeks to First Observed Tumor 105 86 65
 

o
 

Ln a

Dosed groups received 5,000 or 10,000 ppm.
 

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

c

Beneath the incidence of tumors in the matched-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

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

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

95% confidence interval of the relative risk between each dosed group and the control group.
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Fed Daminozide in the Diet3
 

Topography: Morphology
 

Lung: Alveolar /Bronchiolar Carcinoma^
 

P Values0 >d
 

Relative Risk^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar /Brochiolar Adenoma
 

or Carcinoma*5
 

P Value sc»d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

1/20 (5)
 

N.S.
 

105
 

1/20 (5)
 

N.S.
 

105
 

Low
 

Dose
 

4/39 (10)
 

N.S.
 

2.051
 

0.225
 

98.244
 

105
 

8/39 (21)
 

N.S.
 

4.103
 

0.621
 

176.419
 

105
 

High
 

Dose
 

2/48 (4)
 

N.S.
 

0.833
 

0.047
 

48.155
 

72
 

10/48 (21)
 

N.S.
 

4.167
 

0.669
 

176.294
 

72
 
Table F2. Analyses of the Incidence of Primary Tumors in Female. Mice
 

Fed Daminozide in the Diet3 

(continued) 

Topography: Morphology

Matched 

 Control 

Low 

Dose 

High 

Dose 

Hematopoietic System: All Neoplasms^ 5/20 (25) 16/41 (39) 14/48 (29) 

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

Relative Riskf 

Lower Limit 

Upper Limit 

1.561 

0.661 

4.762 

1.167 

0.476 

3.693 

Weeks to First Observed Tumor 97 75 72 

Hematopoietic System: Malignant 

Lymphoma, Lymphocytic Leukemia, or 

Leukemia, NOS° 5/20 (25) 14/41 (34) 13/48 (27) 

P Valuesc'd N.S. N.S. N.S. 

Relative Riskf 

Lower Limit 

Upper Limit 

1.336 

0.560 

4.262 

1.083 

0.434 

3.470 

Weeks to First Observed Tumor 97 75 72 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

(continued)
 

Topography: Morphology


All Sites: Hemangiomab


P Values0 »d


Departure from Linear Trend*3


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 o
 

oo
 

Liver: Hepatocellular Carcinomab


P Valuesc»d


Departure from Linear Trend6


Relative Risk^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Daminozide in the Diet3 

Matched 

 Control 

Low

Dose

 High 

 Dose 

 0/20 (0) 4/41 (10) 0/48 (0) 

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

P = 0.012 

Infinite 

0.471 

Infinite 

— 105 

 0/20 (0) 3/40 (8) 0/48 (0) 

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

P = 0.027 

Infinite 

0.313 

Infinite 

— 105 
__ 

__ 

Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

(continued)
 

Topography: Morphology


Liver: Hepatocellular Adenoma or
 

Carcinomab


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Pituitary: Chromophobe Adenomab


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Daminozide in the Dieta
 

Matched Low
 

 Control Dose
 

 1/20 (5) 4/40 (10)
 

 N.S. N.S.
 

2.000
 

0.220
 

95.888
 

 105 105
 

 2/14 (14) 1/19 (5)
 

 N.S. N.S.
 

0.368
 

0.007
 

6.421
 

 62 105
 

High
 

Dose
 

0/48 (0)
 

N.S.
 

0.000
 

0.000
 

7.780
 

0/14 (0)
 

N.S.
 

0.000
 

0.000
 

3.151
 
_ _ 

Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

(continued)
 

Topography: Morphology


Uterus: Endometrial Stromal Polyp'5


P Values0 »d


Departure from Linear Trend6


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Peritoneum: Lipomab


P Valuesc>d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Daminozide in the Diet3
 

Matched
 

 Control
 

 0/20 (0)
 

 N.S.
 

P = 0.024
 

—
 

 2/20 (10)
 

P = 0.028 (N)
 

 105
 

Low High
 

Dose Dose
 

3/37 (8) 0/45 (0)
 

N.S. N.S.
 

Infinite
 

0.338
 —
 Infinite
 —
 

105
 —
 

0/41 (0) 0/48 (0)
 

N.S. N.S.
 

0.000 0.000
 

0.000 0.000
 

1.632 1.400
 

—
 —

__ 

Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Fed Daminozide in the Diet3
 

(continued) 

Matched Low High 

Topography; Morphology Control Dose Dose 

Mesentery: Lipoma^ 2/20 (10) 1/41 (2) 0/48 (0)
 

P Valuesc»d P = 0.038 (N) N.S. N.S.
 

Relative Riskf 0.244 0.000
 

Lower Limit 0.004 0.000
 

Upper Limit 4.462 1.400
 

Weeks to First Observed Tumor 105 105
 

a

Dosed groups received 5,000 or 10,000 ppm.
 

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

c

Beneath the incidence of tumors in the matched-control group is the probability level for the
 

Cochran-Armitage test when P < 0.05, otherwise, not significant (N.S.) is indicated. Beneath the
 

incidence of tumors in each dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P < 0.05; otherwise,
 

not significant (N.S.) is indicated.
 

d

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

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

95% confidence interval of the relative risk between each dosed group and the control group.
 

Review of tne Bioassay of Daminozide* for Carcinogenlcity
 

by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmenal Carcinogens
 

March 7, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DREW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be
 

exposed. The members of the Clearinghouse have been drawn
 

from academia, industry, organized labor, public -interest
 

groups, State health officials, and quasi-public health and
 

research organizations. Members have been selected on the
 

basis of their experience in carcinogenesis or related fields
 

and, collectively, provide expertise in chemistry, biochemistry,
 

biostatistics, toxicology, pathology, and epidemiology.
 

Representatives of various Governmental agencies participate
 

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

of the Clearinghouse is charged with the responsibility of
 

providing a peer review of reports prepared on NCI-sponsored
 

bioassays of chemicals studied for carcinogenicity. It is in
 

this context that the below critique is given on the bioassay
 

of Daminozide for carcinogenicity.
 

The primary reviewer said that Daminozide was, at most,
 

a borderline carcinogen in female rats, inducing adenocarcinomas
 

of the endometrium and leiomyosarcomas of the uterus. In
 

male mice, Daminozide induced a dose-related incidence of
 

hepatocellular carcinomas. He briefly described the experimental
 

design and conditions under which Daminozide was tested. In
 

his critique, the primary reviewer made the following points:
 

the number of matched controls was too small; the degradation
 

products of Daminozide and their concentrations were not
 

determined; other chemicals were tested in the same room
 

as Daminozide; and the use of historical controls diminished
 

the significance of certain tumors observed in treated animals.
 

The secondary reviewer agreed with the conclusion In
 

the report that Daminozide was carcinogenic in female
 

rats and was associated with the induction of hepatocellular
 

carcinomas in male mice. He pointed out the increased
 

incidence of alveolar/bronchiolar carcinomas and malignant
 

113
 
lymphomas in treated mice. With respect to rats, the secondary
 

reviewer noted the incidence of C-cell hyperplasia of the
 

thyroid in treated females. Given the carcinogenicity of
 

the drug in rats and mice, he concluded that Daminozide
 

poses a potential carcinogenic risk to humans.
 

A motion was made that the report on the bioassay of
 

Daminozide be accepted as written. The motion was seconded
 

and approved unanimously.
 

In further discussion, the inconsistency was noted
 

between the primary reviewer's evaluation that Daminozide
 

is, at most, a borderline carcinogen and the secondary
 

reviewer's statement that it poses a potential carcinogenic
 

risk to humans. A Program staff pathologist said that the
 

finding of relatively rare tumors in the treated animals
 

should take on greater significance, despite their equivocal
 

statistical significance. A Subgroup member added that it
 

was valid to use historical control animals to assess the
 

significance of rare tumors. It was noted that from among
 

318 historical controls, not a single adenocarcinoma of the
 

endometrium or leiomyosarcoma of the uterus was observed.
 The probability of getting eight such tumors by chance in
 

the treated female rats would be remote.
 

Based on the additional discussion, it was moved that
 

the report on the bioassay of Daminozide be accepted as
 

written. The motion was seconded and approved unanimously.
 Another motion was made that the data available were
 

insufficient to assess the potential carcinogenic risk of
 

Daminozide to humans. All the members were in favor of
 

the motion except Dr. Highland, who opposed it.
 

Members present were:
 

Gerald N. Wogan (Chairman), Massachusetts Institute of
 

Technology
 

Arnold Brown, Mayo Clinic
 

E. Cuyler Hammond, American Cancer Society
 

Joseph Highland, Environmental Defense Fund
 

Henry Pitot, University of Wisconsin Medical Center
 

George Roush, Jr., Monsanto Company
 

Michael Shimkin, University of California at San Diego
 

*	 Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

114
 

4U.S. GOVERNMENT PRINTING OFFICE: 1978 260-899/3110 1-3 



OHEW Publication No. (NIH) 78-1333