National Cancer Institute CARCINOGENESIS Technical Report Series No. 174 1979 BIOASSAY p-PHENYLENEDIAMINE DIHYDROCHLORIDE FOR POSSIBLE CARCINOGENICITY CAS No. 624-18-0 NCI-CG-TR-174 U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE Public Health Service National Institutes of Health
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National Cancer Institute CARCINOGENESIS Technical Report Series No. 174 1979
BIOASSAY p-PHENYLENEDIAMINE DIHYDROCHLORIDE FOR POSSIBLE CARCINOGENICITY
CAS No. 624-18-0
NCI-CG-TR-174
U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE Public Health Service National Institutes of Health
National Cancer Institute CARCINOGENESIS Technical Report Series No. 174 1979
BIOASSAY p-PHENYLENEDIAMINE DIHYDROCHLORIDE FOR POSSIBLE CARCINOGENICITY
CAS No. 624-18-0
NCI-CG-TR-174
U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE Public Health Service National Institutes of Health
BIOASSAY OF
p-PHENYLENEDIAMINE DIHYDROCHLORIDE
FOR POSSIBLE CARCINOGENICITY
Carcinogenesis Testing Program Division of Cancer Cause and Prevention
National Cancer Institute National Institutes of Health
Bethesda, Maryland 20014
U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE Public Health Service
National Institutes of Health
DHEW Publication No. (NIH) 79-1730
REPORT ON THE BIOASSAY OF p-PHENYLENEDIAMINE DHYDROCHLORIDE FOR POSSIBLE CARCINOGENICITY
CARCINOGENESIS TESTING PROGRAM DIVISION OF CANCER CAUSE AND PREVENTION
NATIONAL CANCER INSTITUTE, NATIONAL INSTITUTES OF HEALTH
FOREWORD: This report presents the results of the bioassay of p-phenylenediamine dihydrochloride conducted for the Carcinogenesis Testing Program, Division of Cancer Cause and Prevention, National Cancer Institute (NCI), National Institutes of Health, Bethesda, Maryland. This is one of a series of experiments designed to determine whether selected chemicals have the capacity to produce cancer in animals. Negative results, in which the test animals do not have a significantly greater incidence of cancer than control animals, do not necessarily mean the test chemical is not a carcinogen because the experiments are conducted under a limited set of circumstances. Positive results demonstrate that the test chemical is carcinogenic for animals under the conditions of the test and indicate a potential risk to man. The actual determination of the risk to man from animal carcinogens requires a wider analysis.
CONTRIBUTORS: This bioassay of p-phenylenediamine dihydrochloride was conducted by Litton Bionetics, Inc., Kensington, Maryland, initially under direct contract to the NCI and currently under a subcontract to Tracer Jitco, Inc., prime contractor for the NCI Carcinogenesis Testing Program.
The experimental design was determined by the NCI Project Officers, Dr. N. P. Page (1,2), Dr. E. K. Weisburger (1) and Dr. J. H. Weisburger (1,3). The principal investigators for the contract were Dr. F. M. Garner (4) and Dr. B. M. Ulland (4,5). Mr. S. Johnson (4) was the coprincipal investigator for the contract. Animal treatment and observation were supervised by Mr. R. Cypher (4), Mr. D. S. Howard (4) and Mr. H. D. Thornett (4); Mr. H. Paulin (4) analyzed dosed feed mixtures. Ms. J. Blalock (4) was responsible for data collection and assembly. Chemical analysis was performed by Midwest Research Institute (6) and the analytical results were reviewed by Dr. N. Zimmerman (7).
Histopathologic examinations were performed by Dr. N. J. Wosu (4) at Litton Bionetics, Inc., the pathology narratives were written by Dr. N. J. Wosu (4), and the diagnoses included in this report represent the interpretation of this pathologist. Histopathology findings and reports were reviewed by Dr. R. L. Schueler (8).
iii
Compilation of individual animal survival, pathology, and summary tables was performed by EG&G Mason Research Institute (9); the statistical analysis was performed by Mr. W. W. Belew (7,10), Mr. R. M. Helfand (7) and Dr. J. P. Dirkse, III (11), using methods selected for the Carcinogenesis Testing Program by Dr. J. J. Gart (12).
This report was prepared at METREK, a Division of The MITRE Corporation (7) under the direction of the NCI. Those responsible for this report at METREK are the project coordinator, Dr. L. W. Thomas (7), task leader Ms. P. Walker (7), senior biologist Mr. M. Morse (7), biochemist Mr. S. C. Drill (7), and technical editor Ms. P. A. Miller (7). The final report was reviewed by members of the participating organizations.
The following other scientists at the National Cancer Institute were responsible for evaluating the bioassay experiment, interpreting the results, and reporting the findings: Dr. K. C. Chu (1), Dr. C. Cueto, Jr. (1), Dr. J. F. Douglas (1), Dr. R. A. Griesemer (1), Dr. T. E. Hamm (1), Dr. W. V. Hartwell (1), Dr. M. H. Levitt (1), Dr. H. A. Milman (1), Dr. T. W. Orme (1), Dr. R. A. Squire (1,13), Dr. S. F. Stinson (1), Dr. J. M. Ward (1), and Dr. C. E. Whitmire (1).
1. Carcinogenesis Testing Program, Division of Cancer Cause and Prevention, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
2. Now with the U.S. Environmental Protection Agency, 401 M Street S.W., Washington, D.C.
3. Now with the Naylor Dana Institute for Disease Prevention, American Health Foundation, Hammon House Road, Valhalla, New York.
5. Now with Hazleton Laboratories America, Inc., 9200 Leesburg Turnpike, Vienna, Virginia.
6. Midwest Research Institute, 425 Volker Boulevard, Kansas City, Missouri.
7. The MITRE Corporation, METREK Division, 1820 Dolley Madison Boulevard, McLean, Virginia.
8. Tracer Jitco, Inc., 1776 East Jefferson Street, Rockville, Maryland.
iv
9. EG&G Mason Research Institute, 1530 East Jefferson Street, Rockville, Maryland.
10. Now with the Solar Energy Research Institute, Cole Boulevard, Golden, Colorado.
11. Consultant to The MITRE Corporation, currently a professor in the Department of Statistics at The George Washington University, 2100 Eye Street, N.W., Washington, D.C.
12. Mathematical Statistics and Applied Mathematics Section, Biometry Branch, Field Studies and Statistics Program, Division of Cancer Cause and Prevention, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
13. Now with the Division of Comparative Medicine, Johns Hopkins University, School of Medicine, Traylor Building, Baltimore, Maryland.
SUMMARY
A bioassay of p-phenylenediamine dihydrochloride for possible carcinogenicity was conducted using Fischer 344 rats and B6C3F1 mice. p-Phenylenediamine dihydrochloride was administered in the feed, at either of two concentrations, to groups of 50 male and 50 female animals of each species. The high and low concentrations of p-phenylenediamine dihydrochloride were, respectively, 1250 and 625 ppm for both rats and mice. After a 103-week period of compound administration, there were additional observation periods of 2 weeks for rats and 1 week for mice. Twenty animals of each sex and species were placed on test as controls.
There were no significant positive associations between the concentrations of p-phenylenediamine dihydrochloride administered and mortality in rats or mice of either sex. Adequate numbers of animals in all groups survived sufficiently long to be at risk from late-developing tumors. Slight dose-related mean body weight depression was observed in female rats and the mean body weights among high dose male rats and dosed female mice were slightly depressed in relation to their respective controls, indicating that the concentrations of p-phenylenediamine dihydrochloride administered to these animals in this bioassay may have approximated the maximum tolerated concentrations. Since no distinct mean body weight depression relative to controls, no significant accelerated mortality, and no other signs of toxicity were associated with administration of p-phenylenediamine dihydrochloride to male mice, it is possible that these animals may have been able to tolerate a higher dietary concentration.
None of the statistical tests for any site in rats or mice of either sex, including time to leukemia or malignant lymphoma analysis in female mice, indicated a significant positive association between compound administration and tumor incidence.
Under the conditions of this bioassay, there was no convincing evidence that dietary administration of p-phenylenediamine dihydrochloride was carcinogenic in Fischer 344 rats or B6C3F1 mice.
vli
TABLE OF CONTENTS
Page
I. INTRODUCTION i
II. MATERIALS AND METHODS 7
A. Chemicals ' B. Dietary Preparation C. Animals D. Animal Maintenance E. Selection of Initial Concentrations "•;: F. Experimental Design I"3
G. Clinical and Histopathologic Examinations H. Data Recording and Statistical Analyses
III. CHRONIC TESTING RESULTS: RATS 23
A. Body Weights and Clinical Observations 23
B. Survival 23
C. Pathology 23
D. Statistical Analyses of Results 2^
33 IV. CHRONIC TESTING RESULTS: MICE
A. Body Weights and Clinical Observations 33
B. Survival 33
C. Pathology 33
D. Statistical Analyses of Results 3"
43 V. DISCUSSION
VI. BIBLIOGRAPHY 44
APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN RATS TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE A-l
APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MICE TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE B~l
APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN RATS TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE C-l
IX
D-l
TABLE OF CONTENTS (Concluded)
APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MICE TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE
LIST OF ILLUSTRATIONS
Figure Number Page
1 CHEMICAL STRUCTURE OF p-PHENYLENEDIAMINE DIHYDROCHLORIDE 2
2 GROWTH CURVES FOR p-PHENYLENEDIAMINEDROCHLORIDE CHRONIC STUDY RATS
DIHY 24
3 SURVIVAL COMPARISONS OF p-PHENYLENEDIAMINE DIHYDROCHLORIDE CHRONIC STUDY RATS 25
4 GROWTH CURVES FOR p-PHENYLENEDIAMINEDROCHLORIDE CHRONIC STUDY MICE
DIHY- 34
5 SURVIVAL COMPARISONS OF p-PHENYLENEDIAMINE DIHYDROCHLORIDE CHRONIC STUDY MICE 35
6 COMPARISONS OF p-PHENYLENEDIAMINE DIHYDROCHLORIDE CHRONIC STUDY FEMALE MICE SURVIVING WITHOUT OBSERVED LEUKEMIAS OR MALIGNANT LYMPHOMAS 42
Treated groups received doses of 625 or 1250 ppm in feed.
"'Number of tumor-bearing animals/number of animals examined at site (proportion). i
"The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability level for the Fisher exact test for the comparison of a treated group with the control group is given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
The 95% confidence interval on the relative risk of the treated group to the control group.
30
TABLE 4
ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC SITES IN FEMALE RATS TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE£
TOPOGRAPHY :MORPHOLOGY
Lung: Alveolar/Bronchiolar Carcinoma or Alveolar/Bronchiolar Adenomab
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
Hematopoietic System: Leukemia or Malignant Lymphoma^
P Values0
Q
Departure from Linear Trend
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
Pituitary: Chromophobe Carcinoma or Chromophobe Adenoma^
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
CONTROL
0/20(0.00)
N.S.
3/20(0.15)
N.S.
P = 0.048
88
7/17(0.41)
N.S.
100
LOW DOSE
1/49(0.02)
N.S.
Infinite 0.023 Infinite
105
15/50(0.30)
N.S.
2.000 0.662 9.943
49
19/44(0.43)
N.S.
1.049 0.545 2.482
82
HIGH DOSE
4/48(0.08)
N.S.
Infinite 0.402 Infinite
105
7/50(0.14)
N.S.
0.933 0.245 5.215
95
25/46(0.54)
N.S.
1.320 0.719 2.991
94
31
TABLE 4 (CONCLUDED)
TOPOGRAPHY : MORPHOLOGY CONTROL LOW DOSE
HIGH DOSE
Mammary Gland:
P Values0
Fibroadenoma 1/20(0.05)
N.S.
5/50(0.10)
N.S.
3/50(0.06)
N.S.
Relative Risk (Control)d
Lower Limit Upper Limit
Weeks to First Observed Tumor
_—
100
2.000 0.249
92.596
98
1.200 0.106
61.724
103
Uterus: Endometrial Stromal Polyp
P Values0
1/20(0.05)
N.S.
4/47(0.09)
N.S.
4/47(0.09)
N.S.
Relative Risk (Control)d
Lower Limit Upper Limit
Weeks to First Observed Tumor
—_
105
1.702 0.186
81.978
105
1.702 0.186
81.978
96
«a
Treated groups received doses of 625 or 1250 ppm in feed.
Number of tumor-bearing animals/number of animals examined at site (proportion). Q
The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability level for the Fisher exact test for the comparison of a treated group with the control group is given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
The 95% confidence interval on the relative risk of the treated group to the control group. £
The probability level of the test for departure from linear trend is given beneath the control group when P < 0.05.
incidence. Based upon these statistical results, there was insuffi
cient evidence that p-phenylenediamine dihydrochloride was a car
cinogen in Fischer 344 rats under the conditions of this bioassay.
For male rats the Cochran-Armitage test did indicate a significant
negative association between dosage and the incidence of chromophobe
adenomas of the pituitary.
To provide additional insight into the possible carcinogenicity
of this compound, 95 percent confidence intervals on the relative
risk have been estimated and entered in the tables based upon the
observed tumor incidence rates. In all of the intervals shown in
Tables 3 and 4, the value one is included; this indicates the absence
of statistically significant results. It should also be noted that
all of the confidence intervals have an upper limit greater than one,
indicating the theoretical possibility of tumor induction in rats by
p-phenylenediamine dihydrochloride that could not be established under
the conditions of this test.
32
IV. CHRONIC TESTING RESULTS: MICE
A. Body Weights and Clinical Observations
There was no evidence of dose-related mean body weight depres
sion in male mice. Mean body weights among dosed female mice were
slightly lower than that among the controls (Figure 4).
No other clinical signs were recorded.
B. Survival
The estimated probabilities of survival for male and female mice
in the control and p-phenylenediamine dihydrochloride-dosed groups
are shown in Figure 5. For both male and female mice the Tarone test
for association between dosage and mortality was not significant.
There were adequate numbers of male mice at risk from late-
developing uumors as 84 percent (42/50) of the high dose, 76 percent
(38/50) of the low dose, and 80 percent (16/20) of the control group
survived on test until the termination of the study.
For females, with 84 percent (41/49) of the high dose, 88 percent
(44/50) of the low dose, and 85 percent (17/20) of the control group
surviving on test until termination of the study, there were adequate
numbers at risk from late-developing tumors.
C. Pathology
Histopathologic findings on neoplasms in mice are summarized in
Appendix B (Tables Bl and B2); findings on nonneoplastic lesions are
summarized in Appendix D (Tables Dl and D2).
33
FIGURE4 GROWTH CURVES FOR p-PHENYLENEDIAMINE DIHYDROCHLORIDE CHRONIC STUDY MICE
34
TIME ON TEST (WEEKS) FIGURES
SURVIVAL COMPARISONS OF p-PHENYLENEDIAMINE DIHYDROCHLORIDE CHRONIC STUDY MICE
35
A variety of tumors occurred both in the control and dosed
groups. A few neoplasms occurred only in the dosed groups or with
greater frequency in dosed groups as compared with controls. The
neoplasms which were observed have all been reported to occur spon
taneously in this strain of mice. No neoplasms were considered to
be compound-related.
A number of degenerative, proliferative and inflammatory changes
was encountered in dosed and control mice. The incidence and severity
of these nonneoplastic lesions are not unusual for aging B6C3F1 mice.
Based upon the results of this pathology examination, p-pheny
lenediamine dihydrochloride was not carcinogenic to B6C3F1 mice under
the conditions of this bioassay.
D. Statistical Analyses of Results
The results of the statistical analyses of tumor incidence in
mice are summarized in Tables 5 and 6. The analysis is included for
every type of malignant tumor in either sex where at least two such
tumors were observed in at least one of the control or p-phenylene
diamine dihydrochloride-dosed groups and where such tumors were ob
served in at least 5 percent of the group.
The number of weeks to first observed leukemia or malignant lym
phoma in female mice decreased from 98 weeks in the control group to
67 weeks in the low dose group and then to 31 weeks in the high dose
group, while the incidences in the low dose and high dose group,
although not significantly different, were higher than those found in
36
___
TABLE 5
ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC SITES IN MALE MICE TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE*
TOPOGRAPHY :MORPHOLOGY
Lung: Alveolar /Bronchiolar Carcinoma
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
Lung: Alveolar/Bronchiolar Carcinoma or Alveolar/Bronchiolar Adenoma^3
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
Hematopoietic System: Leukemia or Malignant Lymphoma^
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
CONTROL
1/20(0.05)
N.S.
104
4/20(0.20)
N.S.
104
3/20(0.15)
P - 0.032(N)
83
LOW DOSE
6/49(0.12)
N.S.
2.449 0.332
110.166
80
10/49(0.20)
N.S.
1.020 0.346 4.068
70
7/50(0.14)
N.S.
0.933 0.245 5.215
64
HIGH DOSE
6/50(0.12)
N.S.
2.400 0.325
108.021
104
8/50(0.16)
N.S.
0.800 0.250 3.327
1/50(0.02)
N.S.
0.133 0.003 1.568
37
96
104
___
___
38
TABLE 5 (CONCLUDED)
The probability level for the Cochran-Armitage test is given beneath the incidence of tumors in
LOW HIGH TOPOGRAPHY : MORPHOLOGY CONTROL DOSE DOSE
Treated groups received doses of 625 or 1250 ppm in feed.
Number of tumor-bearing animals/number of animals examined at site (proportion) £
the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability level for the Fisher exact test for the comparison of a treated group with the control group is given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
The 95% confidence interval on the relative risk of the treated group to the control group.
TABLE 6
ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT SPECIFIC SITES IN FEMALE MICE TREATED WITH p -PHENYLENEDIAMINE DIHYDROCHLORIDE*
TOPOGRAPHY: MORPHOLOGY
Lung: Alveolar/Bronchiolar Carcinoma or Alveolar/Bronchiolar Adenoma"3
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
Hematopoietic System: Leukemia or Malignant Lymphoma'3
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
Liver: Hepatocellular Carcinoma
P Values0
Relative Risk (Control) Lower Limit Upper Limit
Weeks to First Observed Tumor
LOW CONTROL DOSE
0/20(0.00) 2/48(0.04)
N.S. N.S.
_— Infinite 0.128
Infinite
104
2/20(0.10) 10/50(0.20)
N.S. N.S.
2.000 0.488 17.808
98 67
1/20(0.05) 3/48(0.06)
N.S. N.S.
1.250 0.110 64.251
104 87
HIGH DOSE
3/49(0.06)
N.S.
Infinite 0.255
Infinite
103
10/49(0.20)
N.S.
2.041 0.498 18.154
2/45(0.04)
N.S.
0.889 0.050 51.294
39
31
104
40
TABLE 6 (CONCLUDED)
LOW HIGH TOPOGRAPHY : MORPHOLOGY CONTROL DOSE DOSE
Treated groups received doses of 625 or 1250 ppm in feed.
Number of tumor-bearing animals/number of animals examined at site (proportion). CThe probability level for the Cochran-Armitage test is given beneath the incidence of tumors in the control group when P < 0.05; otherwise, not significant (N.S.) is indicated. The probability level for the Fisher exact test for the comparison of a treated group with the control group is given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than in the control group.
The 95% confidence interval on the relative risk of the treated group to the control group.
the control group. To further examine these results an additional,
life-table analysis was performed. Figure 6 shows the probability of
survival without a known leukemia or malignant lymphoma for female
mice. The Tarone test did not indicate any significant differences
between dosed and control groups.
No other statistical test in mice of either sex indicated a sig
nificant positive association between the administration of p-pheny
lenediamine dihydrochloride and an increased tumor incidence at any
site. Based on these statistical results, there was insufficient
evidence that p-phenylenediamine dihydrochloride was a carcinogen in
B6C3F1 mice under the conditions of this bioassay.
To provide additional insight into the possible carcinogenicity
of this compound, 95 percent confidence intervals on the relative
risk have been estimated and entered in the tables based upon the
observed tumor incidence rates. In all of the intervals shown in
Tables 5 and 6, the value one is included; this indicates the absence
of statistically significant results. It should also be noted that
all of the confidence intervals have an upper limit greater than one,
indicating the theoretical possibility of tumor induction in mice by
p-phenylenediamine dihydrochloride that could not be established under
the conditions of this test.
41
TIME ON TEST (WEEKS)
FIGURE 6 COMPARISIONS OF p-PHENYLENEDIAMINE DIHYDROCHLORIDE CHRONIC STUDY FEMALE MICE
SURVIVING WITHOUT OBSERVED LEUKEMIASOR MALIGNANT LYMPHOMAS
42
V. DISCUSSION
There were no significant positive associations between the
concentrations of p-phenylenediamine dihydrochloride administered
and mortality in rats or mice of either sex. Adequate numbers of
animals in all groups survived sufficiently long to be at risk from
late-developing tumors. Slight dose-related mean body weight de
pression was observed in female rats and the mean body weights among
high dose male rats and dosed female mice were slightly depressed in
relation to their respective controls, indicating that the concen
trations of p-phenylenediamine dihydrochloride administered to these
animals in this bioassay may have approximated the maximum tolerated
concentrations. Since no distinct mean body weight depression rela
tive to controls, no significant accelerated mortality, and no other
signs of toxicity were associated with administration of p-phenylene
diamine dihydrochloride to male mice, it is possible that these ani
mals may have been able to tolerate a higher dietary concentration.
None of the statistical tests for any site in rats or mice of
either sex, including time to leukemia or malignant lymphoma analysis
in female mice, indicated a significant positive association between
compound administration and tumor incidence.
Under the conditions of this bioassay, there was no convincing
evidence that dietary administration of p-phenylenediamine dihydro
chloride was carcinogenic in Fischer 344 rats or B6C3F1 mice.
43
VI. BIBLIOGRAPHY
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Anthony, H.M., and G.M. Thomas, "Tumors of the Urinary Bladder: An Analysis of the Occupations of 1,030 Patients in Leeds, England." Journal of the Rational Cancer Institute 45:879-895, 1970.
Armitage, P., Statistical Methods in Medical Research, Chapter 14. J. Wiley & Sons, New York, 1971.
Barabashova, G.D., V.A. Izmak'skii, and E.G. Milliarfsi, "Genesis of Absorption Spectra of Benzene Derivatives Containing n-twoacceptor Contra Systems." DOKL. AKAD. - NAUK SSSR 190:600, 1970.
Berenblum, I., editor, Carcinogenicity Testing. International Union Against Cancer, Technical Report Series, Vol. 2. International Union Against Cancer, Geneva, 1969.
Blijleven, W.G.H., "Mutagenicity of Four Hair Dyes in Drosophila Melanogaster." Mutation Research 48:181-186, 1977.
Burnett, C., E.I. Goldenthal, S.B. Harris, F.X. Wazeter, J. Strausburg, R. Kapp, and R. Voelker, "Teratology and Percutaneous Toxicity Studies on Hair Dyes." Journal of Toxicology and Environmental Health 1:1027-1040, 1976.
Burnett, C., B. Lanman, R. Giovacchini, G. Wolcott, and R. Scala, "Long-term Toxicity Studies on Oxidation Hair Dyes." Food and Cosmetics Toxicology 13:353-357, 1975.
Burnett, C. , R. Loehr, and J. Corbett, "Dominant Lethal Mutagenicity Study on Hair Dyes." Journal of Toxicology and Environmental Health 2:657-662, 1977.
Chemical Abstracts Service, The Chemical Abstracts Service (CAS) Ninth Collective Index, Volumes 76-85, 1972-1976. American Chemical Society, Washington, D.C., 1977.
Corbett,'J.P., and J. Menkart, "Hair Coloring." CUTIS 12:190-197, 1973.
44
Cox, D.R., Analysis of Binary Data, Chapters 4 and 5. Methuen and Co., Ltd., London, 1970.
Cox, D.R., "Regression Models and Life-Tables." Journal of the Royal Statistical Society, Series "B" 34:187-220, 1972.
Gart, J.J., "The Comparison of Proportions: A Review of Significance Tests, Confidence Limits, and Adjustments for Stratification." International Statistical Institute Review 39:148-169, 1971.
Hawley, G.G., editor, The Condensed Chemical Dictionary, 8th edition. Van Nostrand Reinhold Company, New York, 1971.
Hossack, J.N., and J.C. Richardson, "Examination of the Potential Mutagenicity of Hair Dye Constituents Using the Micronucleus Test." Experientia 33:377-378, 1977.
Kaplan, E.L. , and P. Meier, "Nonparametric Estimation from Incomplete Observations." Journal of the American Statistical Association 5 :457-481, 1958.
Linhart, M.S., J.A. Cooper, R.L. Martin, N.P. Page, and J.A. Peters, "Carcinogenesis Bioassay Data System." Computers and Biomedical Research 7:230-248, 1974.
Markland, W.R., "Hair Preparations." Kirk-othmer Encyclopedia of Chemical Technology, 2nd edition, Vol. 10. Interscience Publishers, New York, 1966.
Miller, R.G., Simultaneous Statistical Inference. McGraw-Hill Book Co., New York, 1966.
Nishioka, H., "Detection of Carcinogenicity of Color Cosmetics in Bacterial Test Systems." Mutation Research 38:345, 1976.
Pouchert, L.J., The Aldrich Library of Infrared Spectra, 2nd edition. Aldrich Chemical Company, Milwaukee, Wisconsin, 1975.
Saffiotti, U., R. Montesano, A.R. Sellakumar, F. Cefis, and D.G. Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced by Different Numbers of Administration of Benzo (a) Pyrene and Ferric Oxide." Cancer Research 32:1073-1079, 1972.
Sax, N.I., "General Chemicals." Dangerous Properties of Industrial Materials. N.I. Sax, editor. Van Nostrand Reinhold Company, New York, 1975.
45
Society of Dyers and Colourists, Colour Index, 2nd edition, Volume 4( Yorkshire, England, 1956.
Stanford Research Institute, 1977 Directory of Chemical Producers, U.S.A. Menlo Park, California, 1977.
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Tarone, R.E., "Tests for Trend in Life-Table Analysis." Biometrika 62 679-682, 1975.
U.S. International Trade Commission, Imports of Benzenoid Chemicals and Products, 1974. USITC Publication 762, U.S. Government Printing Office, Washington, D.C., 1976.
U.S. International Trade Commission, Synthetic Organic Chemicals: United States Production and Sales, 1975. USITC Publication 804, U.S. Government Printing Office, Washington, D.C., 1977.
Windholz, M., editor, The Merck Index: An Encyclopedia of Chemicals and Drugs, ninth edition. Merck and Co., Inc., Rahway, New Jersey, 1976.
Wynder, E.L., J. Onderdonk, and N. Mantel, "An Epidemiological Investigation of Cancer of the Bladder." Cancer 16:1388-1407, 1963.
46
Review of the Bioassay of p-Phenylenediamine Dihydrochloride* for Carcinogenicity by the Data Evaluation/Risk Assessment Subgroup
of the Clearinghouse on Environmental Carcinogens
October 25, 1978
The Clearinghouse on Environmental Carcinogens was established in May, 1976, in compliance with DHEW Committee Regulations and the Provisions of the Federal Advisory Committee Act. The purpose of the Clearinghouse is to advise the Director of the National Cancer Institute (NCI) on its bioassay program to identify and to evaluate chemical carcinogens in the environment to which humans may be exposed. The members of the Clearinghouse have been drawn from academia, industry, organized labor, public interest groups, and State health officials. Members have been selected on the basis of their experience in carcinogenesis or related fields and, collectively, provide expertise in chemistry, biochemistry, biostatistics, toxicology, pathology, and epidemiology. Representatives of various Governmental agencies participate as ad hoc members. The Data Evaluation/Risk Assessment Subgroup of the Clearinghouse is charged with the responsibility of providing a peer review of reports prepared on NCI-sponsored bioassays of chemicals studied for carcinogenicity. It is in this context that the below critique is given on the bioassay of p-Phenylenediamine dihydrochloride for carcinogenicity.
The reviewer said that there was no convincing evidence that, under the conditions of test, p-Phenylenediamine dihydrochloride was carcinogenic in either treated rats or mice. After briefly describing the experimental design, he said that there were no unusual highlights or concerns worthy of special comment. Based on the results of the study, the reviewer said that p-Phenylenediamine dihydrochloride would not appear to pose a carcinogenic risk for humans. A motion to accept the report as written was seconded and approved unanimously.
Clearinghouse Members present:
Arnold L. Brown (Chairman), University of Wisconsin Medical School Joseph Highland, Environmental Defense Fund William Li-jinsky, Frederick Cancer Research Center Henry Pitot, University of Wisconsin Medical Center Verne A. Ray, Pfizer Medical Research Laboratory Kenneth Wilcox, Michigan State Health Department
* 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.
OU.S. GOVERNMENT PRINTING OFFICE: 1978 281-217/3295 1-3
APPENDIX A
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN RATS TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE
TABLE A1 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED
WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE
CONTROL (UNTR) LOH DOSE 11-1155 11-11*53
A N I M A L S INITIALLY IN S T U D Y 20 50 ANIMALS NECROPSIED M 20 50 A N I M A L S E X A M I N E D HI STOPATHOLOGICALLY 20 50
INTEGUMENTARY SYSTEM
*SKIN (20) (50) SEBACEOUS A D E N O C A R C I N O M A 1 (2*) K E R A T O A C A N T H O M A 1 (2*) F I B R O M A 1 (5X)
*SUBCUT TISSUE (20) (50) F I B R O M A 1 (5%) 1 (2X) F I B R O S A R C O M A 1 (5*) N E U R O F I B R O K A 1 (2X)
R E S P I R A T O R Y S Y S T E M
tLUNG (18) (47) A L V E O L A R / B R O N C H I O L A R A D E N O M A 1 (6*) 2 (<•«) ALVEOLAR/BRONCHIOLAR CARCINOMA 1 (2X) S E B A C E O U S A D E N O C A R C I N O M A , METAST 1 (2%) FIBROSARCOMA. METASTATIC
H E M A T O P O I E T I C SYSTEM
* M U L T I P L E O R G A N S (20) (50) M A L I G . L Y M P H O M A , U N D I F F E R - T Y P E 1 (5X) M A L I G . L Y M P H O M A , HISTIOCYTIC TYPE L E U K E M I A , NOS 2 (10X) 11 (22X) U N D I F F E R E N T I A T E D L E U K E M I A O R A H U L O C Y T I C L E U K E M I A 1 (55) 1 (2X) M O N O C Y T I C L E U K E M I A
C I R C U L A T O R Y S Y S T E M
NOSE
t N U M B E R OF A N I M A L S M I T H TISSUE E X A M I N E D MICROSCOPICALLY * N U M B E R OF A N I M A L S NECROPSIED **EXCLUDES PARTIALLY AUTOLYZED ANIMALS
HIGH DOSE 11-1151
50 50 18
(50)
150) 1 (25) 1 (2X)
(»6)
1 (2<)
(50)
1 (2«) 9 (18%) 1 (25) 1 (25) 1 (2X)
A-3
TABLE Al (CONTINUED)
CONTROL (UNTR) LOU DOSE HIGH DOSS 11-1455 11-1453 11-1451
DIGESTIVE SYSTEH
•SALIVARY GLAND (19) (50) (45) ADENOCARCINOMA, NOS 1 (2*) 1(2%)
*MULTIPLE ORGANS MALIGNANT LYMPHOMA, NOS MALIG. LYMPHOMA. UNDIFFER-TYPE MALIG. LYMPHOMA, LYMPHOCYTIC TYPE HALIG. LYMPHOMA, HISTIOCYTIC TYPE MALIGNANT LYMPHOMA, MIXED TYPE LEUKEMIA, NOS
•SPLEEN HEMANGIOSARCOMA
tMESENTERIC L. NODE MALIGNANT LYMPHOMA, NOS MALIGNANT LYMPHOMA, MIXED TYPE
tPEYERS PATCH MALIGNANT LYMPHOMA. NOS
CONTROL(OHTH) 22-2455
20 20 20
(20)
(20)
3 (15*) 1 (5X) 1 (5X)
(20) 1 (5X)
1 (5%) 1 (5X)
(20)
(20)
(20)
LOR DOSE 22-2K53
50 50 50
(50) 1 <2X)
(H9) 1 (2«) H (8X) 6 (12*)
(50) 1 (2X) 1 (2«)
1 <2X) 1 (2«) 1 (2«)
(«8)
(U7) 1 (2X) 1 (2*)
(50)
t N U M B E R OF A N I M A L S WITH TISSUE E X A M I N E D MICROSCOPICALLY * N U M B E R OF A N I M A L S NECROPSIED "EXCLUDES PARTIALLY AUTOLYZED ANIMALS
* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OF ANIMALS NECROPSIED
B-4
TABLE Bl (CONTINUED)
CONTROL (ONTH) LOU DOSE HIGH DOSE 22-2455 22-2U53 22-2U51
MUSCULOSKELETAL SYSTEM
NOHF,
BODY CAVITIES
* P E H I T O N E U M (20) (50) (50) LIPOMA 1 (2X)
AIL OTHEB SYSTEMS
•MULTIPLE O R G A N S SARCOMA, NOS H E M A N G I O S A R C O M A
(20) 1 (5*)
(50)
1 (2*)
(50) 1 (2%)
A N I M A L DISPOSITION S U M f l A R Y
A N I M A L S I N I T I A L L Y I N STUDY 20 50 50 N A T U R A L D E A T H S u 9 8 M O R I B U N D SACRIFICE 3 S C H E D U L E D SACRIFICE A C C I D E N T A L L Y KILLED TERMINAL SACRIFICE 16 38 112 A N I M A L MISSING
8 INCLUDES iUTOIYZED A N I M A L S
I N U M B E R OF A N I M A L S MITH TISSUE E X A M I N E D MICROSCOPICALLY * NUMBER OF A N I M A L S NECROPSIED
B-5
TABLE Bl (CONCLUDED)
CONTROL (UNTR) LOW DOSE HIGH DOSE 22-2455 22-21*53 22-2451
I NUMBER 0? ANIMALS «ITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OF ANIMALS NECROPSIED
B-8
HIGH DOSE 22-2U52
(l»5) 6 (13X) 2 <»*)
(3U) 1 (3*)
(t«) 1 (2X) 1 (2X)
(49)
(18)
1 (2*) 1 (2S)
(48) 1 (2X)
TABLE B2 (CONTINUED)
CONTROL ( O N T R ) LOW DOSE HIGH DOSE 22-2156 22-2454 22-2452
M U S C U L O S K E L E T A L SYSTEM
N O N E
BODY CAVITIES
* P E R I T O N E H M (20) (50) (49) L I P O M A 1 (2*)
ALL O T H E R SYSTEMS
'MULTIPLE O R G A N S (20) (50) (49) SARCOMA, NOS 1 (5*)
A N I M A L DISPOSITION S U M M A R Y
A N I M A L S INITIALLY I N STUDY 20 50 50 N A T U R A L DEATHS 3 6 8 M O R I B U N D SACRIFICE SCHEDULED SACRIFICE A C C I D E N T A L L Y RULED T E R M I N A L SACRIFICE 17 44 41 A N I M A L MISSING
8 INCLUDES A U T O L I f Z E p A j l I M A L S
* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OF ANIMALS NECROPSIED
B-9
TABLE B2 (CONCLUDED)
"-" ——————.—.—-.— _—.________ -T—-_- T .__!—-
CONTBOL (UNTB) LOW DOSE HIGH DOSE 22-2U56 22-2U5U 22-2452
TUMOR SUMMARY
TOTAL ANIMALS WITH PRIMARY TUMORS* a 17 21 TOTAL PRIMARY TUMORS 9 22 28
TOTAL ANIMALS WITH BENIGN TOMORS 2 7 12 TOTAL BSNIGS TUBOES 3 7 14
TOTAL ANIMALS 1ITH MALIGNANT TUMORS 5 14 13 TOTAL MALIGNANT TUMORS 5 15 14
TOTAL ANIMALS BITH SECONDARY TUMORS) t 1 TOTAL SECONDARY TUMORS 1
TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
TOTAL UNCERTAIN TUMORS
TOTAL ANIMALS HITH TUMORS UNCERTAINPBIMARY 08 METASTATIC 1 TOTAL UNCERTAIN TUMORS 1
« PRIMARY TUMOHS: ALL TUMORS EXCEPT SECONDARY TUMORS t SECONDARY TUMORS: METASTATIC TUMORS OR TUMOBS INVASIVE INTO AN ADJACENT ORGAN
B-10
APPENDIX C
SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN RATS TREATED WITH
p-PHENYLENEDIAMINE DIHYDROCHLORIDE
TABLE Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS
TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE
HIGH DOS2 11-1451
50 50 48
(50)
(50) 1 < 2 X )
(46) 2 (4X) 1 (2X) t (9«) 1 (2X) 1 (2X)
21 (USX)
3 (7X)
(45) 1 (2X)
(4«)
(47) 1 12XL
A N I M A L S I N I T I A L L Y I N STUDY A U I H A L S NECROPSIED A N I M A L S E X A M I N E D HISTOPATHOLOGICAI.LY
I N T E G U M E N T A R Y SYSTEM
*SKIN CYST. NOS
*SUBCUT TISSDE D E R M A L INCLUSION CYST
RESPIRATORY SYSTEM
J L U N G ATELECTASIS T H R O M B O S I S , NOS C O N G E S T I O N , NOS H E M O R R H A G E I N F L A M M A T I O N , INTERSTITIAL P N E U M O N I A , CHRONIC M U R I N E FIBROSIS H Y P E R P L A S I A , A D E N O M A T O U S
HEMATOPOIETIC SYSTEM
#BONE MA3ROH H Y P E R P L A S I A , HEMATOPOIETIC
•MEDIASTINAL L .NODE CONGESTION, NOS H E M O R R H A G E
C I R C U L A T O R Y SYSTEM
t H E A R T INFLAMMATION*. CHRONIC
I N U M B E R OF A N I M A L S WITH TISSUE* N U M B E R OF A N I M A L S NECROPSIED "EXCLUDES PARTIALLY AUTOLYZED ANIMALS
CONTROL (UNTR) 11-1455
20 ** 2°
20
(20)
(20)
(18) 2 (11X)
2 (11X)
11 (61X) 1 (6X) 1 (6XJ
(19)
(20) 1 < 5 X ) 1 (5X)
(19)
LOU DOSE 11-1U53
50 50 50
(50) 1 < 2 X )
(50) 1 ( 2 X )
(47)
3 (6X)
1 (2X) 2t (51%)
2 (»X)
(«8) 1 (2X)
(46)
(46)
E X A M I N E D MICROSCOPICALLY
C-3
TABLE Cl (CONTINUED)
CONTROL (UNTB) LOW DOSE HIGH DOSE 11-1155 11-1153 11-1151
IADBENAL M2DULLA (19) (50) (46) HYPEFr>LASIAt NOS 1 i2U
» N U M B E P O F A N I M A L S W I T H T I S S U E E X A M I N E ! ) M I C R O S C O P I C A L L Y * N U M B E R O F A N I M A L S N E C R O P S I E D
C-5
TABLE Cl (CONTINUED)
CONTROL (ONTR) LOW DOSE HIGH DOSE 11-1155 11-1U53 11-1151
t NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY * NUMBEP OF ANIMALS NECROPSIED
C-9
TABLE C2 (CONTINUED)
CONTROL (UNTP) LOW DOSE 11-11156 11-1454
*COLON (19) (U7) PARASITISM 2 (11X) 8 (17X)
U R I N A R Y SYSTEM
tKIDNEY (20) (149) I N F L A M M A T I O N , FOCAL 1 (2%) I N F L A M M A T I O N . CHRONIC 9 (45X) 29 (59X) NEPHROPATHY. TOXIC 2 (»*)
E N D O C R I N E S Y S T E M
f P I T U I T A R Y (17) (44) CYST, NOS 3 (18*) 6 (14%) H E M O R R H A G E H E M O R R H A G I C CYST 1 (6%) 2 (5X)
I A D R E N A L (18) (45) LIPOIDOS1S 1 <6X) CYTOPLASMIC VACUOLIZATION
ITHYROID (20) (48) U L T I M O B R A N C H I A L CYST 1 (2%) CYSTIC FOLLICLES H Y P E R P L A S I A , C-CELL 3 (6X) H Y P E R P L A S I A , FOLLICULAR-CELL 1 (2*)
• T H Y R O I D FOLLICLE (20) (48) HYPERPLASIA, CYSTIC
R E P R O D U C T I V E SYSTEM
*HAMMAHY GLAND (20) (50) DILATATION/DUCTS 2 (»*)
tUTEPUS (20) (47) ABSCESS, NOS POLYPOID HYPERPLASIA 1 <5».)
((UTERUS/ENDOMETRIUM (20) (47) CONGESTION, SOS 1 12%l
* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OF ANIMALS NECROPSIED
HIGH DOSE 11-1452
(47) 2 (4X)
(48)
21 (441) 6 (13X)
(46) 3 (7*) 1 (2X)
(46)
1 (2*)
(43)
1 (2%)
(43) 1 (2*)
(59)
(47) 1 (2%)
(47) _
C-IO
TABLE C2 (CONTINUED)
CONTROL (ONTB) LOM DOSE HIGH DOSE 11-11(56 11-1l»5<t 11-U52
FIBROSIS 1 (2X) H Y P E R P L A S I A , N O S 1 (5*) 3 <6«) 1 (2*) H Y P E R P L A S I A , CYSTIC 1 (9*) » (9*)
* O V A R Y CYST, NOS
(20) (18) (U7) 2 («*)
FOLLICULAR CYST, NOS 1 (5X) 3 (6«)
t S E S O V A R I U H (20) (18) (147) P A R O V A R I A N CYST 1 (5X)
N E R V O U S S Y S T E M
NOSE
SPECIAL SENSE O R G A N S
N O N E
HUSCULOSKELETAL SYSTEM
N O N E
BODY CAVITIES
* P E K I T O N E U f l (20) (50) (50) I N F L A M M A T I O N , N O S 1 (2«)
* M E S E N T E 8 Y (20) (50) (50) STEATITIS 1 (2*)
ALL OTHER SYSTEMS
N O N E
SPECIAL M O R P H O L O G Y S U M M A R Y
NO L E S I O N R E P O R T E D 2 5
t N U B 3 E R OF A N I M A L S K I T H TISSUE E X A M I N E D MICROSCOPICALLY * N U M B E R O F A N I M A L S N E C R O P S I E D
C-ll
TABLE C2 (CONCLUDED)
t*
AUTO/NBCROPSY/HISTO PERF AOTO/NECROPSY/NO HISTO
RUflBBR OF A N I M A L S KITH TISSUE KORBER OF AHtHALS NECROPSIED
CONTROL (UMTS) 11-1*56
2
LOU DOSE 11-1U54
1
E X A M I N E D MICROSCOPICALLY
HIGH DOSE 11-1452
2
C-12
APPENDIX D
SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MICE TREATED WITH
p-PHENYLENEDIAMINE DIHYDROCHLORIDE
TABLED! SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE
TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE
CONTROL (UNTR) LOB DOSE 22-2155 22-2153
A N I M A L S INITIALLY IN STUDY 20 50 ANIHALS NECROPSIED ** 20 50 A N I M A L S E X A M I N E D HISTOPATHOLOGICALLY 20 50
I N T E G U M E N T A R Y SYSTEM
*SKIN (20) (50) ULCER, NOS 1 (5X) G R A N U L O M A , NOS
• SUBCUT TISSUE (20) (50) ABSCESS, NOS 1 (2X)
R E S P I R A T O R Y SYSTEM
f t U N G (20) (149) I N F L A M M A T I O N , MOLTIFOCAL P N 5 U M O N I A , ASPIRATION P N E U M O N I A , CHRONIC M U 8 I N E 1 (20X) « (8X) I N F L A M M A T I O N , CHRONIC 2 (10X) 1 (2X) I N F L A M M A T I O N , CHRONIC FOCAL 3 (15X) 3 (6X) H Y P E R P L A S I A , A D E N O M A T O U S
I L U N G / A L V E O L I (20) (19) HISTIOCYTOSIS
HEMATOPOIETIC SYSTEM
IBONE M A R R O W (19) (15) H Y P E R P L A S I A , HEMATOPOIETIC 1 (5X)
*SPLEEN (20) (U8) H Y P E R P L A S I A , R E T I C U L U M CELL H Y P E R P L A S I A , L Y M P H O I D 1 (2%)
•MESENTERIC L. NODE' (20) (U7) L Y B P H A N G I E C T A S I S .
* N U M B E R OF A N I M A L S WITH TISSUE E X A M I N E D MICROSCOPICALLY * N U M B E R OF A N I M A L S NECROPSIED **EXCLUDES PARTIALLY AUTOLYZED ANIMALS
H E M O R R H A G E H Y P E R P L A S I A , L Y M P H O I D
1 (5X) 1 (2X)
1 (2X)
C I R C U L A T O R Y SYSTEM
• H E A R T ENDOCARDIOSIS
(20) (48) 1 < 2 X )
(50)
• M Y O C A R D I U M INFLAMMATION, FOCAL
(20) 1 (5*)
(18) 1 (2X)
(50)
* A O R T A I N F L A M M A T I O N , FOCAL
(20) (50) (50) 1 (2X)
DIGESTIVE S Y S T E M
ILIVER I N F L A M M A T I O N , MULTIFOCAL L I P O G R A N U L O M A I N F L A M M A T I O N , FOCAL G 8 A N U L O M ATOU NECROSIS, H E M O H R H A G I C M E T A M O R P H O S I S FATTY C Y T O P L A S M I C V A C U O L I Z A T I O N FOCAL C E L L U L A R C H A N G E A T Y P I A , NOS M U L T I N U C L E A T E G I A N T - C E L L H Y P E R P L A S I A , FOCAL H Y P E R P L A S I A , DIFFUSE
tLIVER/PEBIPOP.TAL I N F L A M M A T I O N , NOS I N F L A M M A T I O N , C H R O N I C
(20) (19)
1 (2X)
(50) 11
(2X) (2X)
IBILE DUCT CYST, NOS
(20) 1 (5X)
(19) (50)
I P A N C S E A S CYST, NOS CYSTIC DUCTS
(20) (U9)
1 ( 2 X )
(48) 1 (2X)
• P A N C R E A T I C ACINUS A T R O P H Y , NOS
(20) (19) 1 (2X)
(18)
•STOMACH I N F L A M M A T I O N ^ FOCAL
(20) .
(50) 1 Jt2Sl_
(18) 112%) __ _.
I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OF ANIMALS NECROPSIED
D-4
TABLE Dl (CONTINUED)
CONTROL (UNTS) LOU DOSE 22-2155 22-2153
I N F L A M M A T I O N , A C U T E FOCAL
( L A R G E INTESTINE NEM4TODIASIS
(20) (18) 2 (IX)
(COLON (20) (18) I N F L A M M A T I O N ,P A R A S I T I S M
FOCAL 12
(5X) (10%) 9 (19X)
U R I N A R Y SYSTEM
t K I D N E Y (20) (18) M I N E R A L I Z A T I O N 2 (1%) I N F L A M M A T I O N , ACUTE 1 (2X) I N F L A M M A T I O N , CHRONIC 3 (15)5) 3 (6X) N E P H R O P A T H Y 1 (5X) 1 (2%)
( U R I N A R Y BLADDER (15) (10) ATROPHY, NOS
E N D O C R I N E SYSTEM
tTHYROID (20) (16) CYTOPLASMIC V A C U O L I Z A T I O N
(PANCREATIC ISLETS (20) (19) H Y P E R P L A S I A , NOS 2 (IX)
REPRODUCTIVE SYSTEM
(PROSTATE (20) (18) INPLAMMATION, ACUTE
* S E M I N A L VESICLE (20) (50) I N F L A M M A T I O N , N O S
N E R V O U S S Y S T E M
(BRAIN (19) (50) H Y D R O C E P H A L U S , NOS CORPORA AMYLACEA _ JLJ37XJL 15 J30X)L
t N U M B E R OF A N I M A L S WITH TISSUE E X A M I N E D MICROSCOPICALLY * N U M B E R OF A N I M A L S NECROPSIED
HIGH DOSE 22-2151
1 (2X)
(50) 1 (2X)
(50)
8 (16*)
(50)2 ( IX)
(13) 1 (2X>
(18) 1 (2X)
(18)
(18) 1 (2%)
(50) 1 (2%)
(50) 1 (2X)
. _13_126XL _ _.
D-5
TABLE Dl (CONCLUDED)
CONTROL (ONTR) LOR DOSE HIGH DOSE 22-2U55 22-2453 22-2451
t NUMBER OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY * NUMBER OP ANIMALS NECROPSIED
D-6
TABLE D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE
TREATED WITH p-PHENYLENEDIAMINE DIHYDROCHLORIDE
CONTROL (UNTR) LOH DOSE 22-2456 22-2451
A N I M A L S I N I T I A L L Y IN STUDY 20 50 A N I M A L S N E C R O P S I E D 20 50 ANIHALS E X A M I N E D HISTOPATHOLOGICAL!?** 20 50
I N T E G U M E N T A R Y S Y S T E M
NODE
R E S P I R A T O B Y S Y S T E M
*LUNG (20) (48) I N F L A M M A T I O N , INTERSTITIAL 1 (2%) B R O N C H O P N E U M O N I A , A C U T E 1 (2%) P N E U M O N I A , CHRONIC M U R I N S 1 (5X) 10 (21)1) I N F L A M M A T I O N , C H R O N I C 1 (2«) I N F L A M M A T I O N , C H R O N I C FOCAL 1 (5X) 1 (21) PERIARTERITIS 1 (2%)
H E M A T O P O I E T I C S Y S T E M
tSPLEEN (20) (47) H Y P E R P L A S I A , H E M A T O P O I E T I C HYPERPLASIA, LYMPHOID 1 (2X) H E M A T O P O I E S I S 1 ( 2 X )
* L Y M P H NODE (19) (16) I N F L A M M A T I O N , N O S
(MESENTERIC L. N O D E (19) (16) C Y S T , NOS 1 (2X) I N F L A M M A T I O N , N O S 1 (2X)
C I S C U L A T O B Y S Y S T E M
* H E A R T / A T R I U M (20) (17) THPOMBUS, M U R A L _1 15 XI
t N U K B E R O F A N I M A L S W I T H TISSUE E X A M I N E D MICROSCOPICALLY * NUMBER 0F ANIMALS NECROPSIED "EXCLUDES PARTIALLY AUTOLYZED ANIMALS
HIGH DOSE 22-2152
50 49 19
(49)
2 ( IX) 2 (4*) 1 (2X)
(46) 1 (2%) 1 (2X) 1 (2X)
(11) 1 (2X)
(11)
(48) 1 12*1 _ _ _ _ _
D-7
TABLE D2 (CONTINUED)
CONTROL (ONTR) LOU DOSE 22-2456 22-2l»5U
*COROMABY ABTEBY (20) (50) FIBROSIS 1 (2*1
•PULMONARY ARTERY (20) (50) H Y P E R T R O P H Y . NOS 2 <4X)
DIGESTIVE SYSTEM
tLIVER THROMBOSIS, NOS INFLAMMATION, CHRONIC FOCAL NECROSIS, FOCAL I N F A R C T , NOS M E T A M O R P H O S I S FATTY CYTOPLASMIC VACUOLIZATION HYPERPLASIA, POCAL ANGIECTASIS HEMATOPOIESIS
(20)
1 (5X)
(48) 1
1
2
1
12%)
< 2 X )
(4X)
(2X)
ILIVER/PERIPORTAL LYMPHOCYTIC I N F L A M M A T O R Y INFILTR
(20) <«8) 1 (2*)
t P A N C R E A S CYSTIC DDCTS
(20) (47) 2 (4X)
ISTOMACH OLCEH, NOS I N F L A M M A T I O N .I N F L A M M A T I O N ,
FOCAL CHRONIC
(20) (49) 1
2
(2%)
(4X)
iCOLON PARASITISM
(20) (49) 1 (2%)
URINARY SYSTEM
I K I D N E Y (20) (49) H Y D R O N E P H R O S I S 1 ( 2 X ) LYMPHOCYTIC I N F L A M M A T O R Y INFILTR 1 (2X) INFLAMMATION, CHRONIC
I U R I N A R Y BLADDER (15) (43) INFLAaMATIQ!lx_CHRONIC
t N U M B E R OF A N I M A L S W I T H TISSUS E X A M I N E D MICROSCOPICALLY * N U M B E R OF A N I M A L S N E C R O P S I E D
HIGH DOSE 22-2452
(49)
(49)
(4-5)
1 (2%)
1 (2*) 1 (2%)
1 (2*)
1 (2%)
(45)
(45) 2 (4%)
(47)
1 (2%) 1 < 2 X >
(46)
(46) 2 (4T.) 1 (2%) 1 (2X)
(36) 1 J3S)_ _ .
0-8
TABLE D2 (CONTINUED)
HIGH DOSE 22-2452
1 (3X)
(34) 1 (3X)
(44)
(««) 1 (2X)
(41) 1 (2%)
(18)
1 (2%)
(48) 7 (15*)
1 (2X)
(40) 2 (5*) 1 (3X)
(47)
8 (17X) 3 (6*)
PERIARTERITIS
ENDOCRINE SYSTEM
tPITUITARY HEM0RRHAGIC CYST
tADRENAL AMYLOIDOSIS
tADRENAL MEDULLA ATROPHY, FOCAL
•THYROID LIPOIDOSIS
REPRODOCTIVE SYSTEM
tUTERUS HYDROMETRA CYST, NOS EDEMA, NOS
t UTERUS/ENDOMETRIUM CYST, SOS INFLAMMATION, SUPPORATIVE HYPERPLASIA, DIFFUSE HYPERPLASIA, CYSTIC