Document GmmvQjL8r2Y3gQ8M5pzbzkeDN

EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. VINYL CHLORIDE PATHOLOGY REPORT Submitted to Manufacturing Chemists Association Washington, D.C. 20009 January 9, 1979 EPL 01992 EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. VINYL CHLORIDE PATHOLOGY REPORT I. Introduction At the request of the Manufacturing Chemists Association, Experimental Pathology Laboratories, Inc. provided a pathologist (Dr. William Busey) to assist in a partial pathology audit of a chronic inhalation study on vinyl chloride conducted at Industrial Bio-Test Laboratories in Decatur, Illinois. This audit was conducted during the week of February 20, 1978 at the IBT Laboratory in Decatur, Illinois. Dr, Busey participated as a member of the audit team and initially his responsibilities were to audit the histopathological findings. Because of the size of the study (2500 animals), and the number of tissues (31 organs per animal) called for to be examined in the protocol, the members of the audit task force were divided into three teams. Dr. Busey was assigned to audit team No. 1 along with Dr. G.K. Hatfield from Diamond Shamrock Corporation, and Drs. Sullivan and Vlaovic from IBT. The details of the tasks performed by audit team No. 1 are presented in the following section of this report. Prior to the initiation of the audit, EPL was provided with a protocol of the study. The following table depicts the experimental design and group designations: EPL 01993 EPL EXPERIMENTAL pathology LABORATORIES, INC. Experimental Design for Vinyl Chloride Study Group Control Tl* T2* T3* T4** Mice MF 100 100 100 100 100 100 100 100 -- Number of Animals Rats MF 100 100 100 100 100 100 100 100 100# Hamsters MF 100 100 100 100 100 100 100 100 -- Specified Exposure Levels* - 50 ppm vinyl chloride 200 ppm vinyl chloride 2500 ppm vinyl chloride 2500 ppm vinyl chloride +Exposure levels have not been verified by audit team *No food, water, or bedding in cages during exposure **Food, water, and bedding to remain in cages during exposure #A11 rats of same sex, either sex permitted The protocol as amended called for exposure of the above groups six hours per day, five days per week. Mice were to be exposed for nine months, rats and hamsters for twelve months. These groups were originally scheduled to be held for equal additional periods of time (nine or twelve months) but subsequently it was agreed they would be kept for their life time or sacrificed if seriously ill. All animals were to be necropsied either at the time of death or sacrifice and a specified set of tissues was to be preserved in 10% neutral buffered formalin. These tissues were to be examined from all animals in the control, T2 and T3 groups. If exposure related lesions were detected in the organs of the T2 or T3 groups, these same organs from the T1 group were to be examined. -201994 EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. II. Audit Tasks Several tasks were assigned to audit team No. 1. These tasks were directed toward: (a) an assessment of the histopathology data developed by the IBT pathologists, (b) an enumeration of the neoplasms diagnosed by IBT, and (c) a verification of the number of tissues examined by IBT microscopically. In addition, EPL was asked to summarize the tissue counts performed by IBT from examination of all of the wet tissue bags, paraffin blocks and microscope slides. This latter task was subsequently performed at EPL. A. Examination of IBT Histopathology Data from Computer Printouts The recorded diagnoses of the IBT pathologists had been input into a computer which in turn generated a series of printouts listing the findings and separating the neoplastic from the non-neoplastic diagnoses. Working with the IBT printout of the neoplastic diagnoses, summary neoplasm tables were generated by EPL for each of the three species. The number of organs indicated as examined by IBT pathologists was tabulated by the audit team, thus providing a denominator for comparative purposes. These neoplasm summaries are found in Tables I, II, and III. In addition to summarizing all of the neoplasms, the audit team extracted all of the diagnoses of angiosarcoma and prepared a separate incidence table for this diagnosis. A summary of the diagnosis of angiosarcoma in any organ, expressed as a percentage of the number of animals examined, is found in Table IV. -3- 01995 EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. B. Microscopic Examination of Selected Neoplastic and Non-neoplastic Lesions Diagnosed by IBT Pathologists Following examination of the computer printout containing the diagnoses rendered by the IBT pathologists for the various organs examined, selected tissues were examined microscopically. In general, this second microscopic examination correlated well with the original diagnoses and no discrepancies were'noted. The only discrepancies noted were set of slides from apparently two different animals containing the same animal numbers. Since all of the slides were not examined, no assessment of the magnitude of this problem could be made. C. Verification of Number of Tissues Examined Microscopically During the audit selected neoplastic and non-neoplastic lesions were reexamined microscopically, and tissue counts were made from the microscope slides for selected animals. These counts were then checked against the rough pathology reports to assess whether all of the tissues available for microscopic examination were in fact examined. For these selected animals, no discrepancies were noted and the number of tissues available for microscopic examination correlated well with those that were reported. D. Summarization of Counts of Wet Tissue, Paraffin Blocks and Microscope Slides Conducted by IBT Personnel At the request of the vinyl chloride audit task force, IBT examined all of the wet tissues, paraffin blocks and slides from the animals used in the study. This examination consisted of the identi- -4- EPL 01996 EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. fication and counting of the various tissues that were available for possible microscopic evaluation. These counts were submitted on several individual forms and in order to properly evaluate these data, EPL summarized these counts by group, sex, and species. These summaries are presented in Tables V, VI, and VII. III. Summary and Recommendation Inspection of the recorded IBT histopathologic findings clearly indicate that the inhalation exposure of rats, hamsters, and mice to vinyl chloride at 50, 200 and 2500 ppm results in a carcinogenic effect in the liver. This carcinogenic effect was manifest by the development of angiosarcomas, many of which metastasized to regional lymph nodes and the lung. Even though the histopathology is incomplete, the number of angiosarcomas observed clearly indicates the carcinogenicity of vinyl chloride to these three species of laboratory animals. Other" neoplasms were observed and reported but because of the paucity of numbers of specific organs examined, conclusive statements about their relationship to vinyl chloride exposure cannot be made. Tumors of Zymbal's gland were seen in all of the exposure groups in the rats and probably are related to the exposure to vinyl chloride. However, only a small number of these organs were examined in each of the control and exposed groups. Malignant tumors of the mammary gland were seen to a greater degree in the exposed female rats; but, here again, the number of mammary glands examined was small and inconsistent. Therefore, conclusive statements -5- BPL 01997 EPL EXPERIMENTAL PATHOLOGY LABORATORIES. INC. cannot be made. Four tumors of the central nervous system were seen. One of these occurred in a Group II male and three occurred in Group III males. These are important findings in light of recent epidemiological data in man, but their relationship to the exposure to vinyl chloride in this study is undetermined because of the small number of brains examined.. Primary lung tumors occurred with greater frequency in both sexes of vinyl chloride exposed mice at all concentrations. These primary lung tumors were both benign and malignant and were diagnosed as alveologenic adenomas and carcinomas. Inspection of the I8T pathology reports indicates that approximately 50% of the lungs from the mice were examined microscopically. This number of lungs is sufficient to conclude that the inhalation exposure of vinyl chloride to male and female mice had a carcinogenic effect in the lungs. Primary lung tumors were only seen in the male and female hamsters exposed to 2500 ppm of vinyl chloride. Two bronchiolar carcinomas were seen in the males and one in the females, and one bronchiolar adenoma was seen in the males. Primary lung tumors of the hamster are relatively rare; therefore, the presence of these five neoplasms may be meaningful and related to vinyl chloride exposure. Since microscopic examination of a few selected neoplastic and non-neoplastic lesions diagnosed by the IBT pathologists did not reveal any meaningful discrepancies, it would appear that the histopathology conducted on the tissues from these animals was done accurately. Similar -6- EPL 01998 EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. spotchecks of the number of tissues reported to have been examined did not reveal any significant discrepancies. Inspection of the tissue counts conducted by the 1ST personnel of the wet tissue, paraffin blocks and microscope slides indicate that significantly fewer tissues were available for microscopic examination than were called for by the protocol. The reason for this discrepancy became apparent when the necropsy sheets and animal observation books were examined. The tissue from a large number of the animals in this study was not obtained. This discrepancy is unfortunate in light of the fact that the inhalation exposure of these three species of laboratory animals to vinyl chloride might have provided further definition of the pathology produced, the relationship to dose and the variation between species. In conclusion, the results of this investigation revealed some rather severe deficiences in the conduct of this study, primarily with regard to the numbers of tissues that were saved. The pathologic evaluation of the limited material that was available appears to have been conducted accurately. The available data indicate that vinyl chloride at 50, 200 and 2500 ppm has a carcinogenic effect on the liver of rats, mice and hamsters and a carcinogenic effect on the Zymbal's gland of the rat. Similarly, there appears to be a carcinogenic effect in the lungs of the mice. These conclusions are based upon the assumption that the concentrations expressed in this report were actually achieved EPL 01999 -7- EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. and maintained throughout the course of the study. Any significant excursions in the low level (50 ppm) may negate the conclusion of the carcinogenic effect of vinyl chloride at this level. Because of the serious deficiency in the number of tissues saved from animals in both the control and exposed groups, it is recommended to MCA that no further pathology work be done on this study. WILLIAM R. BUSEY, 1TV.M., Ph.D. Pathologist WMB/sfh -8- EPL 02000 TABLE I RAT - TUMOR SUMMARY DATA ADRENAL (No. Examined) Cortical Carcinoma Cortical Adenoma Medullary Carcinoma BRAIN (No. Examined) Neuroepithelioma CECUM (No. Examined) Lymphosarcoma DUODENUM (No. Examined) Lymphosarcoma HEART (No. Examined) Angiosarcoma KIDNEYS (No. Examined) Lymphosarcoma Nephroblastoma Reticulum Cell Sarcoma Tubular Adenoma Lipoma Metastatic Angiosarcoma LIVER (No. Examined) Angiosarcoma Heptocellular Carcinoma Lymphosarcoma Control Control T1 Male Female Male T1 Female T2 Male T2 Female (11) (26) (8) (9) (19) (7) 11 1 * Cio) (10) (7) (69) (21) (21) (9) (60) (8) (8) (1) (74) (7) (7) (6) (52) (18) 1 (17) 1 (16) 1 (64) (4) (4) (4) (59) (70) 2 (70) (72) (75) (60) (68) (70) 11 1 11 31 1 1 (73) (77) (62) (69) (72) 11 17 35 47 1 12 EPL Experimental Pathology Laboratories, Inc. EPL 02001 TABLE I RAT - TUMOR SUMMARY DATA ADRENAL (No. Examined) Cortical Carcinoma Cortical Adenoma Medullary Carcinoma BRAIN (No. Examined) Neuroepithelioma CECUM (No. Examined) Lymphosarcoma DUODENUM (No. Examined) Lymphosarcoma HEART (No. Examined) Angiosarcoma KIDNEYS (No. Examined) Lymphosarcoma Nephroblastoma Reticulum Cell Sarcoma Tubular Adenoma Lipoma Metastatic Angiosarcoma LIVER (No. Examined) Angiosarcoma Heptocellular Carcinoma Lymphosarcoma T3 Male (5) T3 Female (5) T4 T4 Female Male __None-- (6) 1 (6) (0) (5) 3 (3) (0) (5) (3) (1) - (5) (.71). (62) (75) (69) 1. (64) 1 (69) (78) (69) (70) 51 63 49 EPL Experimental Pathology Laboratories, Inc. EPL 02002 TABLE I RAT - TUMOR SUMMARY DATA LIVER (continued) Reticulum Cell Sarcoma Metastatic Adreno-cortical Carcinoma Control Male Control Female T1 Male 3 42 T1 Female 1 1 T2 Male LUNG (No. Examined) Alveologenic Adenoma Bronchogenic Adenocarcinoma Bronchogenic Adenoma Bronchiolar-Alveolar Adenoma Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma Metastatic Hepatocellular Carcinoma Metastatic Fibrosarcoma Metastatic Mammary Gland Adenocarcinoma Metastatic Zymbal's Gland Carcinoma Metastatic Adreno-cortical Carcinoma Metastatic Adenocarcinoma N.O.S. (73) 1 2 1 (73) (77) (58) (71) 1 1 11 I1 2 42 1 9 7 24 1 12 1 MESENTERY (No. Examined) (2) (3) (12) (11) (31) Metastatic Hepatocellular Carcinoma 1 T2 Female (68) 2 28 1 (43) EPL Experimental Pathology Laboratories, Inc. EPL 02003 TABLE I RAT - TUMOR SUMMARY DATA LIVER (continued) Reticulum Cell Sarcoma Metastatic Adreno-cortical Carcinoma T3 Male T3 T4 Female Female T4 Male None 1 LUNG (No. Examined) Alveologenic Adenoma Bronchogenic Adenocarcinoma Bronchogenic Adenoma Bronchiolar-Alveolar Adenoma Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma Metastatic Hepatocellular Carcinoma Metastatic Fibrosarcoma Metastatic Mammary Gland Adenocarcinoma Metastatic Zymbal's Gland Carcinoma Metastatic Adreno-cortical Carcinoma Metastatic Adenocarcinoma N.O.S. (78) 2 1 1 32 (70) 2 37 2 (69) 36 1 MESENTERY (No. Examined) Metastatic Hepatocellular Carcinoma (37) (37) (43) EPL Experimental Pathology Laboratories, Inc. RpL 02004 TABLE I RAT - TUMOR SUMMARY DATA MESENTERY (continued) Angiosarcoma Reticulum Cell Sarcoma Control Male Control T1 Female Male T1 Female T2 Male T2 Female 2 2 12 1 29 42 PANCREAS (No. Examined) Islet Cell Adenoma (35) (39) (38) (23) (23) (8) 71 1 PARATHYROID (No. Examined) Adenoma (6) (5) 1 (2) (9) (8) (1) PITUITARY (No. Examined) Chromophobe PLEURA (No. Examined) Angiosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma (9) (18) 8 14 (0) . (0) (9) (5) (12) (5) 43 5 (0) (1) 1 (4) (7) 45 1 PROSTATE (No. Examined) Adenocarcinoma (9) (10) (19) 1 SKIN (No. Examined) Fibroma Fibrosarcoma Lipoma Liposarcoma Sarcoma, N.O.S. Reticulum Cell Sarcoma Squamous Cell Carcinoma (19) (12) (12) (4) (17) (4) 71 51 22 13 1 1 11 2 1 PL Experimental Pathology Laboratories, Inc. gpL 02005 TABLE I RAT - TUMOR SUMMARY DATA MESENTERY (continued) Angiosarcoma Reticulum Cell Sarcoma PANCREAS (No. Examined) Islet Cell Adenoma PARATHYROID (No. Examined) Adenoma PITUITARY (No. Examined) Chromophobe PLEURA (No. Examined) Angiosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma PROSTATE (No. Examined) Adenocarcinoma SKIN (No. Examined) Fibroma Fibrosarcoma Lipoma Liposarcoma Sarcoma, N.O.S. Reticulum Cell Sarcoma Squamous Cell Carcinoma T3 Male T3 Female T4 Female T4 Male None 38 41 42 (4) (2) (4) (3) (2) (0) (4) (0) (4) 2 (4) (5) (4) 3 44 1 1 (3) 3 (3) (1) (3) 1 1 11 EPL Experimental Pathology Laboratories, Inc. EPL 02006 TABLE I RAT - TUMOR SUMMARY DATA Control Male Control ~n-- T1 T2 Female Male Female Male T2 Female MAMMARY GLAND (No. Examined) Fibroadenoma Adenoma Adenocarcinoma (1) I ZYMBAL'S GLAND (No. Examined) Carcinoma (9) (62) 75 23 5 (16) (4) 4 (7) 2 (52) 45 8 24 (3) 4 (7) (17) 1 (19) 10 7 8 (6) 2 SPLEEN (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma (65) 2 (69) 2 (66) 1 1 2 (54) (62) 1 1 (65) SKELETAL MUSCLE (No. Exam.) Metastatic Angioma (9) (15) (6) (7) (T4) (3) STOMACH (No. Examined) Lymphosarcoma (9) (20) (8) (7) (18) (4) 1 TESTIS (No. Examined) Interstitial Cell Tumor Angioma (54) (64) (41) 2 1 THYMUS (No. Examined) Lymphosarcoma (1) (0) (0) (0) (2) (4) 2 THYROID (No. Examined) Lymphosarcoma C-Cell Adenoma (ID (22) (12) (ID (16) (4) 1 1 EPL Experimental Pathology Laboratories, Inc. EPL 02007 TABLE I RAT - TUMOR SUMMARY DATA T3 Male MAMMARY GLAND (No. Examined) Fibroadenoma Adenoma Adenocarcinoma 1 T3 Female T4 Female T4 Male None 35 12 8 14 ZYMBAL'S GLAND (No. Examined' Carcinoma (8) 9 SPLEEN (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma (71) 1 SKELETAL MUSCLE (No. Exam.) Metastatic Angioma (1) (5) 4 (67) (0) 4 (0) (0) STOMACH (No. Examined) Lymphosarcoma (3). (0) (0) TESTIS (No. Examined) Interstitial Cell Tumor Angioma (5) THYMUS (No. Examined) Lymphosarcoma (0) (1) (0) THYROID (No. Examined) Lymphosarcoma C-Cell Adenoma (3) (2) (0) EPL Experimental Pathology Laboratories, Inc. EPL 02008 TABLE I RAT - TUMOR SUMMARY DATA Control Control Male Female T1 T1 Male Female T2 Male T2 Female THYROID (continued) Follicular Adenoma Follicular Adenocarcinoma 1 1 TRACHEA (No. Examined) Lymphosarcoma (14) (27) (17) (15) (23) (9) 1 UTERUS (No. Examined) Reticulum Cell Sarcoma (36) 1 (35) (11) LYMPH NODE (No. Examined) Fibrosarcoma (12) (21) (8) (7) (17) 1 (5) Total Number Neoplasms 44 141 58 134 1?3 155 No. Animals with Angiosarcomas S) 0 PM) 0 NR) 2 T) 2 %) 2.4 0 12 7 3 0 12 13 33 62 0 00 0 4 0 13 20 40 69 0 15.7 26.7 52.6 84.1 Total No. Primary Angiosarcomas 2 0 12 29 69 96 No. Primary Angiosarcomas per Rat 0.0125 0 0.145 0.387 0.908 1.170 S=Sacrificed PM=Died During Study T=Total NR=Newly Read tPL Experimental Pathology Laboratories, Inc. 02009 TABLE I RAT - TUMOR SUMMARY DATA THYROID (continued) Follicular Adenoma Follicular Adenocarcinoma T3 Male T3 T4 Female Female T4 Male None TRACHEA (No. Examined) Lymphosarcoma (4) (3) (0) UTERUS (No. Examined) Reticulum Cell Sarcoma (2) (0) LYMPH NODE (No. Examined) (3) (0) (0) Fibrosarcoma -P* CD Total Number Neoplasms 168 161 No. Animals with Angiosarcomas S) 10 PM) 41 . NR) 2. .................................. T) 53 . %) 63.9 . 8 50 8 64 79.0 5 53 3 61 81.3 - Total No. Primary Angiosarcomas No. Primary Angiosarcomas per Rat 93 no 96 1.069 1.358 1.28 S=Sacrificed PM=Died During Study T=Total NK=Newly Read EPL Experimental Pathology Laboratories, Inc. EPIj 02010 TABLE II MICE - TUMOR SUMMARY DATA HEART (No. Examined) Lymphosarcoma Angiosarcoma KIDNEY (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma Control Male (SO) 2 Control Female (45) (52) -- (45) - 32 2 LIVER (No. Examined) Angiosarcoma Hepatocellular Carcinoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Mammary Carcinoma Hepatocellular Adenoma Metastatic Squamous .Cell Carcinoma LUNG (No. Examined) Alveologenic Adenoma Alveologenic Carcinoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Mammary Carcinoma Metastatic Angiosarcoma Metastatic Squamous Cell Carcinoma (52) 3 __ 1 1 .......................... - (51) 8 1 3 2 - (45) 3 1 - (45) 2 2 T1 Male T1 Female (56) 1 . __ (58) -------------- (58)' 3 2 1 (62) 7 2 1 (59) 18 1 2 1 (62) 28 1 - (58) 68 4 2 1 1_ (62) 34. .6 2 7 1 2 EPL Experimental Pathology Laboratories, Inc. EPL 02011 TABLE II MICE - TUMOR SUMMARY DATA HEART (No. Examined) Lymphosarcoma Angiosarcoma KIDNEY (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma T2 Male (64) T2 Female (59) (69) " 1 1 1 (62) 1 1 LIVER (No. Examined) Angiosarcoma Hepatocellular Carcinoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Mammary Carcinoma Hepatocellular Adenoma Metastatic Squamous Cell Carcinoma LUNG (No. Examined) Alveologenic Adenoma Alveologenic Carcinoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Mammary Carcinoma Metastatic Angiosarcoma Metastatic Squamous Cell Carcinoma ' (70) 71 1 1 -------(70) 66 1 1 1 (64) 60 1 - -- (64) 53 3 4 1 EPL Experimental Pathology Laboratories, Inc. T3 Male (46) T3 Female (40) (50) 2 2 (51) 52 (48) 1 (50) 61 1 1 1 ..... _ , . . (51) 92 10 (50) 61 8 1 26 45 EPL 02012 TABLE II MICE - TUMOR SUMMARY DATA LYMPH NODE (No. Examined) Lymphosarcoma Angiosarcoma Reticulum Cell Sarcoma Control Male (3) 1 Control Female (2) 2 1 MESENTERY (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Fibrosarcoma (2) 1 1 - - ----- (2) 1 OVARY (No. Examined) Angiosarcoma (10) 1* - PANCREAS (No. Examined) Angiosarcoma Lymphosarcoma ill (14)_ 1____ ......... SAL IVARY GLAND (No. Examined) (0) (0) Angiosarcoma - ------ -- SKELETAL MUSCLE (No. Examined Lymphosarcoma (0)_ (0) --- ' SKIN (No. Examined) Lymphosarcoma Fibrosarcoma Squamous Cell Carcinoma (0). (3) 1 T1 Male (2) T1 Female (2) 1 -- ---- -- (10) 7 2 1 -. (27) 29 1 1 (5)7 1 on (5) i1 ' (7) - . 0) - ----- - (6) (3) (4) 1 1 1 Diagnosed angioma EPL Experimental Pathology Laboratories, Inc. EPL 02013 TABLE II MICE - TUMOR SUMMARY OATA LYMPH NODE (No. Examined) Lymphosarcoma Angiosarcoma Reticulum Cell Sarcoma T2 Male (0) T2 Female (2) -- -- MESENTERY (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Fibrosarcoma OVARY (No. Examined) Angiosarcoma (12) (10) 11 10 1 -- -- - ...... .... (4) 1 PANCREAS (No. Examined) Angiosarcoma Lymphosarcoma ......(3) SALIVARY GLAND (No. Examined) ~ Angiosarcoma (1) . (4) .. . - (1) 1 SKELETAL MUSCLE (No, Examined ) Lymphosarcoma (1) 1 (2) SKIN (No. Examined) Lymphosarcoma Fibrosarcoma Squamous Cell Carcinoma (1) (2) 2 - T3 Male (0) T3 Female (1) 1 ----- (6) 10 -- (7) 8 0) - (1) 0) _ (2) ' ..... (0) (0) (0). 0) (1) 1 EPL Experimental Pathology Laboratories, Inc. EPL 02014 TABLE II MICE - TUMOR SUMMARY DATA MAMMARY GLAND (No. Examined) Adenosquamous Carcinoma Adenocarcinoma Control Male (0) Control Female 0) 1 T1 Male 0) 1 T1 Female (24) 9 19 SPLEEN (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma Metastatic Mammary Carcinoma (46) (41) 23 11 -- - _ .. STOMACH (No. Examined) Adenocarcinoma Lymphosarcoma Reticulum Cell Sarcoma (8) 07) " 1 1 -- URINARY BLADDER (No. Examined; ' (6) Lymphosarcoma 1 UTERUS (No. Examined) Sarcoma, N.O.S. " -- (8) --- 04) 1 Total Neoplasms 31 19 (57) 2 1 (62) 3 2 - (6) (2) 1 - (5) - - - 0) "(5) - 120 145 No. Animals with Angiosarcoma: S) 0 PM) 0 T) 0 %) 0 Total No. Primary Angiosarcom. s No. Primary Anaiosarcomas/Mou: e 0 0 0 0 1 2.2 1 .02 4 0 20 33.3 30 0.5 3 0 40 61.5 69 1.06 EH Sacrificed, PM=Died During Study, T=Total Experimental Pathology Laboratories, Inc. TABLE II MICE - TUMOR SUMMARY DATA MAMMARY GLAND (No. Examined) Adenosquamous Carcinoma Adenocarcinoma T2 Male (0) T2 Female (9) 6 5 SPLEEN (No. Examined) Angiosarcoma Lymphosarcoma Reticulum Cell Sarcoma Metastatic Angiosarcoma Metastatic Mammary Carcinoma (68) ' 3 1 1 1 (63) 1 1 STOMACH (No. Examined) Adenocarcinoma Lymphosarcoma Reticulum Cell Sarcoma (0) (2) T3 Male (3) 1 2 T3 Female (7) 8 8 (47)" (50) 2 1 1 (0) (1) URINARY BLADDER (No. Examined Lymphosarcoma UTERUS (No. Examined) Sarcoma, N.O.S. (0) 0) -- - - - ... (2) - - ........... Total Neoplasms 152 136 (0) -- - (0) (0) 160 127 No. Animals with Angiosarcoma S) 3 PM) 0 T) 74 0 0 65 9 12 00 51 63 %) 96.1 91.5 Total No. Primary Angiosarcom is 85 75 No. Primary Angiosarcomas/Mou :e 1 -10 _____ U15 89.5 64 - 1-12 88.7 71 1n EPL S=Sacrificed, PM=Died During Study, T=Tota1 Experimental Pathology Laboratories, Inc. EPL 02016 TABLE III HAMSTERS - TUMOR SUMMARY DATA ADRENAL (No. Examined) Cortical Adenoma Cortical Carcinoma Control Male Control Female (9) (4) HEART (No. Examined) Lymphosarcoma (45)... (32) KIDNEYS (No. Examined) Angiosarcoma Reticulum Cell Sarcoma Tubular Adenoma Metastatic Mammary Adenocarcinoma (53) . (36) --------- -- - LIVER (No. Examined) Angiosarcoma Lymphosarcoma Metastatic Granulosa Cell Metastatic Mammary Adenocarcinoma LUNG (No. Examined) Alveolar Adenoma Angiosarcoma Bronchiolar Carcinoma Bronchiolar Adenoma Metastatic Angiosarcoma Metastatic Melanosarcoma Metastatic Adenocarcinoma . (5.0).. (33). ... .. -- . (49). (32) EPL Experimental Pathology Laboratories, Inc. T1 Male T1 Female (12) ..(4) 1 (41)_ . (30) (43) (33) . 1 ... 1 1 (44)... ... 5 . . (30) .2 . (42) (29). 1 EPL 02017 TABLE III HAMSTERS - TUMOR SUMMARY DATA ADRENAL (No. Examined) Cortical Adenoma Cortical Carcinoma HEART (No. Examined) Lymphosarcoma T2 Male (18) 1 T2 Female (11) 1 (35) . (51). KIDNEYS (No. Examined) Angiosarcoma Reticulum Cell Sarcoma Tubular Adenoma Metastatic Mammary Adeno- carcinoma LIVER (No. Examined) Angiosarcoma Lymphosarcoma Metastatic Granulosa Cell Metastatic Mammary Adenocarcinoma LUNG (No. Examined) Alveolar Adenoma Angiosarcoma Bronchiolar Carcinoma Bronchiolar Adenoma Metastatic Angiosarcoma Metastatic Melanosarcoma Metastatic Adenocarcinoma r o o4* (57) _1 (36). . . 1 . .. (52). . 11 1. ... 1 (36)__ (49) 12 1 .1 __ T3 Male T3 Female (5) (2) 1 (45) _ (23) 1. . (50) (24) . .7 _ (48) _ 38. 1 (26) 18.. ----- - . (44) (26) 1 2 21 1 74 . epl Experimental Pathology Laboratories, Inc. 02018 TABLE III HAMSTERS - TUMOR SUMMARY DATA LUNG (Continued) Metastatic Mammary Adeno carcinoma Metastatic Uterine Adeno carcinoma Control Male Control Female --. MESENTERY (No. Examined) Angiosarcoma Melanosarcoma Adenocarcinoma, N.O.S. ESOPHAGUS (No. Examined) Papilloma (0) (0) (7) . (2)_ ! OVARIES (No. Examined) Granulosa Cell, Malignant Thecal Cell PANCREAS (No. Examined) Islet Cell Adenoma ... .. (14)_. (16) 1 (4) PLEURA (No. Examined) Metastatic Angiosarcoma (0) (0) SKELETAL MUSCLE (No. Examined Lymphosarcoma (3) (3) SKIN (No. Examined) Angiosarcoma - (5) (2) EPL Experimental Pathology Laboratories, Inc. T1 Male T1 Female ._ ,- (5) (3) 11 1 (7). (3) (13) 1 (20) ... ... (11) . .1 (0) (0) (7) . (2) (7) (4) EPL 02019 TABLE III HAMSTERS - TUMOR SUMMARY DATA LUNG (Continued) Metastatic Mammary Adenocarcinoma Metastatic Uterine Adenocarcinoma MESENTERY (No. Examined) Angiosarcoma Melanosarcoma Adenocarcinoma, N.O.S. ESOPHAGUS (No. Examined) Papilloma OVARIES (No. Examined) Granulosa Cell, Malignant Thecal Cell PANCREAS (No. Examined) Islet Cell Adenoma PLEURA (No. Examined) Metastatic Angiosarcoma T2 Male T2 Female 2 1 (ID 2 .(3) 3 1 (14) ,. -- --- (9) J (16) 1 _1 _ (16) ... 1 (12) (0) (0) - SKELETAL MUSCLE (No. Examined Lymphosarcoma (16) . SKIN (No. Examined) Angiosarcoma (16) . 2, (5) - (12) T3 Male T3 Female -(8) (4) 74 (4) (3) (2) (4) (3). 1 0) (0) 1 (5) (3) . 1 (7) (3) EPL Experimental Pathology Laboratories, Inc. EPL 02020 TABLE III HAMSTERS - TUMOR SUMMARY DATA SKIN (Continued) Fibroma Fibrosarcoma Melanosarcoma Metastatic Uterine Adenocarcinoma Control Male Control Female - MAMMARY GLAND (No. Examined) Adenocarcinoma Adenoma (0) (2) SPLEEN (No. Examined) Angiosarcoma Metastatic Mammary Adenocarcinoma Lymphosarcoma (40).. . .. (31)_. STOMACH (No. Examined) Papilloma THYMUS (No. Examined) Lymphosarcoma (3) 1____ (0) (3) (Pi. UTERUS (No. Examined) Adenocarcinoma Angiosarcoma Papilloma - (22) 1* T1 Male T1 Female 1 .1 (0) . (4) _ . (36) 3 (26) 2 -- (8), (3) 3 (0) (0) (15) EPL * Diagnosed as angioma. -29- Experimental Pathology Laboratories, Inc. EPL 02021 table III HAMSTERS - TUMOR SUMMARY DATA SKIN (Continued) Fibroma Fibrosarcoma Melanosarcoma Metastatic Uterine Adenocarcinoma T2 Male MAMMARY GLAND (No. Examined) Adenocarcinoma Adenoma (0) T2 Female 1 2 - (12) 3 1 SPLEEN (No. Examined) Angiosarcoma Metastatic Mammary Adenocarcinoma Lymphosarcoma STOMACH (No. Examined) Papilloma (34) 5 . (47i 11 ................... 1 (16) . .(14) 1 10 THYMUS (No. Examined) Lymphosarcoma UTERUS (No. Examined) Adenocarcinoma Angiosarcoma Papilloma (2) -- - _(3). (33) 2 1 1 T3 Male T3 Female .. (0) (3) 1 - (41) __ (_22) 89 1. ... (7)_ 2 -- - (4). 2 (2) . ..Jo) 1 (4) 1 EPL Experimental Pathology Laboratories, Inc. -30- EPL 02022 TABLE III HAMSTERS - TUMOR SUMMARY DATA EPL Experimental Pathology Laboratories, Inc. EPL 02023 TABLE III HAMSTERS - TUMOR SUMMARY DATA Total Neoplasms T2 Male T2 Female 17 . 61. No. Animals with Angiosarcoma i S) 3 PH) 5 .. __. _ , _ T) 8 . 21.6 2 15. . , 17 26.98 Total No. Primary Angiosarcom is No. Primary Angiosarcomas/ Hamster 12 . 1.5Q 25 1.47 . T3 Male T3 Female 75. _ 41 84 31____ 16__ 39 _ 20. 75.0 ,, ...64.5 - 55 32 1.40 1.60 < -- ............ -- ........-- -- S=$acrificed PM=Died During Study TaTotal -- EPL Experimental Pathology Laboratories, Inc. -32- EPL 02024 TABLE IV PERCENTAGE OF ANIMALS WITH PRIMARY ANGIOSARCOMAS RATS Male Female MICE Male Female HAMSTERS Male Female Untreated Control 2.4 o 0 2.2 0 2.7 I]____ 15.7 26.7 T2 52.6 84.1 33.3 61.5 96.1 91.5 15.2 12.9 21.6 27.0 *Allowed feed and bedding during exposure T3 63.9 89.5 88.7 75.0 64.5 T4* 79.081.3 -33- EPL 02025 TABLE V TISSUE COUNTS - MALE RATS Number of Animals ADRENAL BONE BRAIN EPIDIDYMIS.. ESOPHAGUS EXT. & MID. EAR EYES HEART KIDNEYS............................. LIVER _ . ... LUNG & BRONCHI . .... MUSCLE OVARIES PANCREAS Untreated T-l Control T-2 T-3 81 82 75 86 18 . 15 22 22 18 15 23 24 19 15 23 23 79____ .. 79 _ 73 81 59____ . 69 _ _ .63 .. 10 6 19 73 .0 - 19__ . _ 15___ 22 24 . 81_____ 80 .74 86 . 81_____ . 80._ .. .. _ 73 .. . .86 .. -- ................... . 81 .80 ... 74 86 . 81. 81. .. 74 19.. 17 23 1 -- 69 65 65 86 22 -- 71 - EPL Experimental Pathology Laboratories, Inc. EPL 02026 TABLE V TISSUE COUNTS - MALE RATS PITUITARY PROSTATE SALIVARY GLAND SEMINAL VESICLE SKIN SPINAL CORD SPLEEN STOMACH TESTIS THYMUS THYROID TISSUE MASS TRACHEA URINARY BLADDER UTERUS Untreated Control T-l T-2 T-3 14 13 16 21 11 15 23 8 17 15 23 22 8 5 14 0 24 14 25 22 17 13 21 20 81 78 73 85 19 15 25 20 78 79 73 81 4 318 26 27 30 36 21 24 37 62 49 48 49 52 18 15 25 20 1 EPL Experimental Pathology Laboratories, Inc. -35- EPL 02027 TABLE V TISSUE COUNTS - MALE RATS AORTA Untreated Control T-l 18 13 T-2 19 OPTIC NERVE 16 13 15 . INTESTINES .21 15 24 LYMPH NODE 32 32 38 PERIPHERAL NERVE 18____ . 13 21. . STERNUM TONGUE 15 ... 18 14... . . 23 15 22 NOSE 9___ 12. . 23 DIAPHRAGM 1 00 FAT . 1___ .0 . . .0 UNKNOWN.................................. . 40. 47 .. 39 ZYMBAL'S GLAND 0 00 SKULL 0 ... 13 22 T-3 17 20 .. 23 . 54 16 8 .. 22 16 _ 2 0 ... 35 0 0 EPL Experimental Pathology Laboratories, Inc. -36- KpL 02 028 TABLE V TISSUE COUNTS - FEMALE RATS Untreated Control T-l T-2 T-3 T-4 Number of Animals 92 76 79 79 74 ADRENAL . 30 12 10. 15 9 BONE 28 13 . 10 16 . 10. BRAIN 28 14 10 17 11 EPIDIDYMIS.. ESOPHAGUS . ... 6Q_.. 54 ... 71 75 67 EXT. & MID... EAR .... 18. _ 7. _ 5 .3 .1 EYES ................. ... . 28. .. . 13. _ 11 18. 11 HEART . . . 79........ 65. . 73 _ . 78 . 73 KIDNEYS..................... . 78 .... . 65 . . 74 .... . 77 . . 72. LIVER . . - 77 65. . 75 79 72. LUNG & BRONCHI ,,.. MUSCLE 78 63 74 78 73 30. . 15 . 12 11 10 OVARIES PANCREAS 72 . 62 71 69 67 69 56 67 61 50 EPL Experimental Pathology Laboratories, Inc. EPL 02029 TABLE V TISSUE COUNTS - FEMALE RATS PITUITARY PROSTATE SALIVARY GLAND SEMINAL VESICLE SKIN SPINAL CORD SPLEEN ______ STOMACH TESTIS THYMUS THYROID TISSUE MASS TRACHEA URINARY BLADDER UTERUS Untreated Control T-l 24 9 T-2 11 T-3 12 33 12 13 17 T-4 .8 ... 10 58 29 19 13 . . 12 26 13 8 14 9 78 64 74 79 70 34 14 11 12 13 - .2____ .. 1 . ..2 . 31 21. . 29 60 61 . 69 55 . 47 56 28 . 14. 11 75 64 71 .3 .. ... .. 2_. 32 22 67 67 19 44 13 14 73 66 EFL Experimental Pathology Laboratories, Inc. EPL 02030 AORTA OPTIC NERVE INTESTINES LYMPH NODE PERIPHERAL NERVE STERNUM TONGUENOSE DIAPHRAGM FAT UNKNOWN ZYMBAL'S GLAND SKULL TABLE V TISSUE COUNTS - FEMALE RATS Untreated Control T-l T-2 T-3 27 9 9 15 T-4 5 26 10. 7 15 9 31 17 10 15 . . J3 41 28 38 62 51 21 8 79 9 26 . _ 10 9 5. . 5 24 , . .11 11 10 ,9 16__. J3 .. ,, _,, ____ , 0____ 0 IQ 0. 8 . ... 6 0 .0 . -Q____ 0. Q 0 ... .. 0. . 65 59. 48 38 .53 7 1 0 0 _0 14 9 10 10 8 EPL Experimental Pathology Laboratories, Inc. EPL 02031 TABLE VI TISSUE COUNTS - MALE MICE Number of Animals ADRENAL BONE BRAIN EPIDIDYMIS ESOPHAGUS EXT. & MID. EAR EYES GALL BLADDER HEART KIDNEYS LIVER LUNG MUSCLE NASAL CAVITY j Untreated Control T-l 61 61 14 27 6 13 5 13 56 57 23 10 .6 7 86 60 60 61 60 60 61 60 61 6 13 00 T-2 T-3 82 71 51 52 7 16 3 14 81 66 13 22 16 2 13 5 16 82 71 82 70 82 71 82 71 7 13 1 12 EPL Experimental Pathology Laboratories, Inc. -40- EPL 02032 TABLE VI TISSUE COUNTS MALE MICE OVARIES PANCREAS PITUITARY PROSTATE SALIVARY GLAND SEMINAL VESICLE SKIN SPINAL CORD SPLEEN STOMACH TESTIS THYMUS THYROID TISSUE MASS TRACHEA Untreated Control T-l T-2 T-3 26 17 36. 69 0 0 3 12 . 8 800 3 9 5 14 6____ 5___ . 0 _o. 7 : .11 _ 1.0. _ 13. 2_ 9. 4 12 . 60 58 79 71 . _. -- 51_____ 60 _ ,, ._ 82___ __67 _ 52 57 81 64 -- 0 000 0. 3 1 1 4 4 19 12 . 12 8 9 22 EPL Experimental Pathology Laboratories, Inc. -41- EPL 02033 TABLE VI TISSUE COUNTS 7 MALE MICE URINARY BLADDER UTERUS AORTA INTESTINES LYMPH NODE PERIPHERAL NERVE STERNUM TONGUE SKULL UNKNOWN OPTIC NERVE FEET LEGS LIMBS ZYMBAL'S GLAND Untreated Control T-l T-2 T-3 46 47 77 65 2 111 53 59 82 70 11 6 33 33 0 515 0 700 2 1 3 11 3 649 3 11 20 10 6 700 0 000 0 000 0 010 0 006 EPL Experimental Pathology Laboratories, Inc. -42- EPL 02034 TABLE VI TISSUE COUNTS - FEMALE MICE Untreated Control T-l T-2 T-3 Number of Animals 45 76 77 72 ADRENAL 20 .. 29 . 77 64 .. BONE BRAIN 6 .. 9 9 16. . - 2 6 8 13 . EPIDIDYMIS ---- ESOPHAGUS. 7 ..... .2 __ EXT. & MID....EAR.......................... . . 0 _ ____2. 48 .3 . 39. 3 -- -- EYES . . JL__. . ___5___ . . 8 14 ... GALL BLADDER HEART 16____ 43____ 6... . 38 . 75._ . 75 19 . 71 ._ - KIDNEYS LIVER .44 . 75 ,, 77 72 - 44 . 76 77 72 LUNG 44 75 76 72 MUSCLE 9 11 5 14 .. NASAL CAVITY 0 0 5 12 EPL Experimental Pathology Laboratories, Inc. EPL 02035 TABLE VI TISSUE COUNTS - FEMALE MICE OVARIES PANCREAS . PITUITARY PROSTATE SALIVARY GLAND SEMINAL VESICLE SKIN SPINAL CORD SPLEEN . .. STOMACH . TESTIS THYMUS THYROID TISSUE MASS TRACHEA Untreated Control T-l 28 53 20 10 1 .. . 1.. T-2 68 71 5 T-3 64 69 , 10 . --- 0 ... . 2 8 15 - - 3 18 11 . J. .3.. . _ 8 43__ 75 . 75 . 44 74 76 . .. 19 13. 71 70 - - - 0. 0 2 2 4 165 3 52 21 27 12 3 37 33 EPL Experimental Pathology Laboratories, Inc. -44- EPL 02036 TABLE VI TISSUE COUNTS - FEMALE MICE URINARY BLADDER UTERUS AORTA INTESTINES LYMPH NODE PERIPHERAL NERVE STERNUM TONGUE SKULL UNKNOWN OPTIC NERVE FEET LEGS LIMBS ZYMBAL'S GLAND Untreated Control T-l T-2 T-3 23 30 64 60 38 48 75 69 1 045 43 73 74 72 11 2 53 50 0 218 2 3 10 1. 0 8 8 1. 10 5 8 3 45. 19 19 1 570 1 001 0 200 0 000 0 008 EPL Experimental Pathology Laboratories, Inc. EpL 02037 TABLE VII TISSUE COUNTS - MALE HAMSTERS Total Number of Animals ADRENAL BONE BRAIN EPIDIDYMIS... ESOPHAGUS EXT. & MID. EAR EYES GALL BLADDER HEART KIDNEYS LIVER LUNG & BRONCHI MUSCLE NASAL CAVITY (NOSE) Untreated Control T-l T-2 T-3 57 45 _ 46 51 13 12 21.... 9 5 7 17 9 5 7 17 9 56 41 45 56 10.. 9 17 11. 2 .Z 15 4 .4 .... 7___ . 17 9 24 ... 26 25 21 . 57 ._ .. 45 46 . . 61 57 45_ _. 46 61 57 45 46 61 . 57 . 45 46 61 3 8 19 9 2 - - EPL Experimental Pathology Laboratories, Inc. EPL 02038 TABLE VII TISSUE COUNTS - MALE HAMSTERS OVARIES PANCREAS PITUITARY PROSTATE RIB (JUNCTION) SALIVARY GLAND SEMINAL VESICLE. SKIN ......................... SPINAL CORD SPLEEN STOMACH TESTIS THYMUS THYROID TISSUE MASS Untreated Control T-l T-2 T-3 24 28 0. 5 13 12 1 38 .7 .10 5 6. 4....... 8 52. 41 10 14 55 41 .1 2 69 32 27 7 20 17 .19 17 , .16 43 31 44 2 11 7 - 21 _ 0 11 8 11 8 4. --- 59 . 41 -- 54 1 4 4 EPL Experimental Pathology Laboratories, Inc. EPL 02039 TABLE VII TISSUE COUNTS - MALE HAMSTERS TRACHEA Untreated Control T-l 18 17 URINARY BLADDER 12 19 UTERUS AORTA 22 20 OPTIC NERVE 25 INTESTINES LYMPH NODE 16 _ I3 3 .7 PERIPHERAL NERVE STERNUM 1__ 4. 3 3 TONGUE SKULL 3. .6 1 .5 UNKNOWN 13 14 PAPILLARY PROJECTION 2 FEET CYST - 1 EPL Experimental Pathology Laboratories, Inc. T-2 23 30 10 9 32 .15 7 15 16 15 10 3 2 T-3 30 36 .. . -_ 4 46 2 1 7 - ., 8 2 16 - 6 EPL 02040 TABLE VII TISSUE COUNTS - FEMALE HAMSTERS Total Number of Animals ADRENAL _ BONE BRAIN EPIDIDYMIS ESOPHAGUS EXT. & MID... EAR _ eyes ... GALL BLADDER HEART KIDNEYS LIVER LUNG & BRONCHI . . MUSCLE NASAL CAVITY (NOSE) Untreated Control T-l T-2 T-3 35 36 61 31 6 779 3 . 5 8 7 ---- . 3 588 - 7 . ... l 4 8 35 35 35 35 3 6.. 10 5 ,8 57 If 23 36 60 36 . . 60 36 60 36 59 47 8 3 7 12 31 31 31 31 4 EPL Experimental Pathology Laboratories, Inc. -49- EPL 02041 OVARIES PANCREAS PITUITARY PROSTATE RIB (JUNCTION) SALIVARY GLAND SEMINAL VESICLE SKIN SPINAL CORD SPLEEN STOMACH TESTIS THYMUS THYROID TISSUE MASS TABLE VII TISSUE COUNTS - FEMALE HAMSTERS Untreated Control T-l T-2 T-3 18 22 34 18 17 20 27 13 0 .2 2 1 3_ _. 5 . .. 12 ........ 3.. . .. .3 12 3___ 4. . . 7. . 34 _ . . 34 57 15 14 . 3Q 5 5 4-- 30 26 4 0 2 1 3 372 3 343 FPL Experimental Pathology Laboratories, Inc. 020^2 BT'1j table VII TISSUE COUNTS - FEMALE HAMSTERS TRACHEA URINARY BLADDER UTERUS AORTA OPTIC NERVE INTESTINES LYMPH NODE PERIPHERAL NERVE STERNUM TONGUE SKULL UNKNOWN PAPILLARY PROJECTION FEET CYST Untreated Control T-l 12 14 T-2 18 T-3 19 16 12 21 22 32 34 50 28 -- 2 6 20 6 3 26 18 18 30 28 3 38 3 2 33 1 2 47 4 2 . ... 3 6 4 2 43 3 3 2 20 6 1 13 2 - 1 EPL Experiment*! Pathology Laboratories, Inc. -51- EPL 02043