Document Dq2RENxV08LVykobYzwpQgvO

TOXICOLOGY, CARCINOGENICITY, AMI REPRODUCTIVE EFFECTS OF SINGLE AND MULTIPLE EXPOSURES TO VINYL CHLORIDE IN RATS AND MICE (A pre-publication release as of 1 April 1979) This pre-publication release reports a research project sponsored by the Consumer Products Safety Commission under an interagency agreement (1974, 1975-1979) with Chemical Systems Laboratory (formerly Edgewood Arsenal), Aberdeen Proving Ground, Maryland. B.P. McNAMARA, Ph.D. Chemical Systems Laboratory PROJECT OFFICERS: JOSEPH MCLAUGHLIN, JR., Pb.D. Consumer Products Safety Commission t AUTHORS OF RELEASE: ROBERT M. HEHIR, Ph.D., Consumer Products Safetv Commission GEORGE BIERBOWER, D.V.M., Consumer Products Safety Commission DONALD A. WILLIGAN, D.V.M., Ph.D., Donald A. Willigan, Inc., GERALD EOLAJA, MAJ., D.V.M., Ph.D., Chemical Systems Laboratory GLEN E. MARRS, JR., MAJ., D.V.M., Chemical Systems Laboratory DAVID E. HINTON, Ph.D., Univ. of West Virginia Medical School RICHARD L. DIMMICK, Chemical Systems Laboratory JOSEPH S. WILES, Chemical Systems Laboratory ucc 0022^6 SUMMARY REPORT I. INTRODUCTION II, EXPERIMENTAL PROCEDURE A, Exposure Procedures for Toxicology and Carcinogenesis 1. Doses 2, Exposure and Sampling Technique 3, Observation 4. pathology a. Gross and Light Microscopy b. Electron Microscopy Reproduction Study and Carcinogenicity III, RESULTS A, Toxicology (single and multiple exposures} 1, Observations During Exposure 2. Observations After Exposure B, Gross Pathology C, Light Microscopic Pathology 1. Mice (single exposures) 2. Mice (10 exposures) 3. Mice (100 exposures) 4. Rats (single exposure) 5. Rats (multiple exposures) 6. Rats (49 exposures from the reproduction study) 7. Summary of Light Microscopic Studies D, Electron Microscopy Study E, Multtgeneratton Study In Rats UCC 002247 IV. DISCUSSION At Factors to Cqrctnopentctty B. Ytnyl Chloride Carcinogenictty V, CONCLUSIONS VI. Appendix UCC 002248 Toxicology, Carcinogenicity and Reproductive Effects of Single and Multiple Exposures to Vinyl Chloride in Rats and Mice I. INTRODUCTION. The background information on vinyl chloride monomer (_VCM) with technical details Is reviewed In "Scientific and Technical Assess ment Report on Vtnyl Chloride and polyvinyl Chloride", US Environmental 1 Protection Agency, EPA-600/6-76-004, June 1975. VCM studies In rats and mice reported herein were conducted at the Chemical Systems Laboratory (CSL) under Interagency agreement with Consumer Products Safety Comnlsslon (CPSC). II, EXPERIMENTAL PROCEDURES. A, Exposure Procedures for Toxicology and Carcinogenicity. 1, Doses. Male and female rats (Fischer 344) and mice (A/J or ICR) were totally exposed in one of the following ways: a. One single one-fhour exposure at 0, 50, 500, 5,000 or 50,000 ppm. (Fischer 344 rats, ICR Mice) (Appendix, Table I) b. Ten one-hour exposures at 500 ppm (one-hour per day, five days per week for two weeks), (Fischer 344, A/J Mice) (Appendix, Table II) c. One-hundred one-hour exposures at 50 ppm (one-hour per day, five days per week for 20 weeks). (Fischer 344 rats, A/J Mice) 2. Exposure and Sampling Techniques. The exposures were conducted In 1000 liter, stainless steel dynamic flow chambers of the Rochester type. Construction insured laminar flow and uniform exposure of the test animals. During one-hour exposure periods four 0.5 cc gas samples were collected at selected times for concentration analysis, A Hewlett-Packard series 5830A Reporting Gas Chromatograph equipped with a dual-flame ionization detector was used for VCM analysis. 1 UCC 002249 The system is sensitive to 60 parts per billion of vinyl chloride. Following exposure the animals were air-washed in the chamber until less than 1 ppm of VCM was detectable. They were then placed in ; the animal holding area for the remainder of the observation period (up to 24 months). 3. Observation. Animals were observed twice dally for indications of their general health. The following signs were noted: mortality, health, external sores, subcutaneous masses, alertness, and activity. All test groups were weighed weekly for the first eight weeks post exposure and monthly thereafter. No blood chemistry or hematology studies were performed. 4. Pathology. a. Gross and light Microscopy. A complete gross pathological examination was performed on most control and exposed animals which died or were sacrificed. Autolysis precluded such examination in a few cases. Light microscopic examination was made of the following tissues: lung, trachea, heart, liver, stomach, small intestine, large Intestine, spleen, kidneys, bladder, bone marrow (sternum), adrenals, pancreas, duodenum, brain (2 sections), pituitary, spinal cord (cervical), skin, thyroid, uterus-ovaries, eye, ear, nose, muscle and bone (femur). Particular emphasis was placed on examination of brain, lung, liver and the Zymbal gland in the rodent ear. All rodents were to be serially sacrificed at 8, 16, 24 months post exposure. However, the life span of the mice forced some changes in the later times of sacrifice and termination of mouse experiments. For the single exposure the planned 16 and 24 month sacrifices were replaced by 2 ucc 002250 an 18-month sacrifice. The latter is minimum suggested by the National Cancer Institute for cancer bioassays in small rodents. For the multiple dose studies (500 ppm x 10 exposures; 50 ppm x 100 exposures) in mice the final sacrifice was at 20 months rather than 24 months. The change was made in consideration of the risk of animal loss through death and possible cannabollsm. b. Electron Microscopical Studies. For the electron microscopical studies the following animals were selected at random: Groups consisting of five males and five females Fischer 344 rats from each of the single one-hour (50, 500, 5000 and 50,000 ppm) ex posures with equal numbers of their corresponding control group. The groups were sacrificed at 8, 16 and 24 months, respectively, after the exposure. Groups consisting of five males and five female Fischer 344 rats from each of the multiple one-hour exposures (10 x 500 oom, 100 x 50 ppm) with equal numbers of their control group. The groups were sacrificed at the end of 16 and 24 months, respectively, after the final exposure. One group of five male and five female Sprague/Dawley/Wistar rats from the parenteral generation of the reproduction group (49 x 50 ppm, 49 x 500 ppm) with equal numbers of their control group. This group was sacri ficed 24 months after the final exposure. The rats were anesthetized with pentobarbital and their livers were perfused via the vena cava with chilled electron microscopical fixative 2 consisting of 4% formaldehyde, IX glutaraldehyde in 0.1 M phosphate buffer (E.M. McDowell & B.F. Trump, Histological Fixatives Suitable for Diagnostic Light and Electron Microscopy, Arch, Path. Lab. Med. 100:405-415, 1976). After mincing, the tissues were post-fixed In 1% phosphate buffered 3 UCC 002251 osmium tetroxide, dehydrated In graded alcohol solutions, cleared In propylene 3 oxide and embedded in Epon (J.H. Luft, Improvements In Epoxy Resin Embedding Methods, J. Biophys. Blochem. Cytol, 9:409-414, 1961), B. Reproduction Study and Carcinogenicity. Fq male and female parents Sprague/Dawley/Wlstar Rats were exposed to 50 ppm or 500 ppm of VCM one-hour per day, five days per week for 10 weeks (49 exposures) before they were mated. (Appendix, Table III) This assured exposure of all forms of male germ cells. The females were exposed during all phases of the oogenic cycle. The parents were evaluated for numbers of matings, percentages of pregnancies, fertility and lactation indices. The F^, Fg and F3 off-spring were evaluated for litter-size, percent of stillborn pups, post-natal growth, viability, survivability and reproduction anomalies. The parenteral generation of Sprague/Dawley/Wlstar Rats were maintained for 24 months post exposure for carcinogenic evaluation. III. RESULTS. A. Toxicology (single and multiple exposures). 1 - Observation During Exposure. Exposures of rats and mice for one-hour to concentrations of 50, 500, 5,000, and 50,000 ppm produced no remarkable signs of toxicity 'except for mice exposed to the highest level. Fifty percent of the males were hyperventilating after 45 minutes of exposure; twitching and possible 4 ucc 002252 ataxia were noted. At 59 minutes tremors were seen. Females showed some hyperactivity at 40 minutes and 25* showed respiratory difficulty and ataxia after 55 minutes. No other effects were noted. There were no remarkable signs of toxicity during the ten repeated exposures of mice and rats at 500 ppm of VCM, nor in rats during the 49 exposures at 500 ppm, nor in either species during the 100 exposures at 50 ppm. 2. Observations After Exposure There were no consistent or dose-related differences between control and exposed (single or multiple) mice or rats In: death rate, (Appendix, Fig I IX, inclusive) toxic signs, gain In body weight. (Appendix, Figs X-XVIII inclusive B. Gross Pathology There was a suggestion of a higher frequency of masses in the lungs and livers of mice and rats exposed once or repeatedly at the higher dose levels. (500, 5,000, 50,000 ppm) C. Light Microscopic Pathology 1. Mice (single exposure) Histological examination at 8 and 18 months in ICR mice exposed unce to concentrations of 50, 500, 5000 or 50000 ppm showed the following changes attributable to VCM. (Appendix, Table IV-VI, inclusive) The development of lung adenomas Increased with exposure to higher dose levels of vinyl chloride: control - 12/120 or IQS 50.000 ppm - 45/137 or 32.8S 5.000 ppm - 24/143 or 16.8% 500 ppm - 18/139 or 12.9% 50 ppm - 14/139 or 10.1% Progression to carcinoma was minimal: 5 ucc 002253 control - 0/120 or OX 50.000 ppm - 3/137 or 2.2X 5.000 ppm - 1/143 or 0.7X 500 ppm - 1/139 or 0.7X 50 ppm - 0/139 or OX Pneumonitis was evident in all animal groups which were exposed to VCM in doses of 500 ppm or more. 2. Mice (10 exposures) Changes attributable to VCM in A/J mice exposed ten times at 500 ppm am examined at 8,16 and 20 months are as follows (Appendix, Tables VII-X, inclusive) The Induction of pulmonary adenomas: controls - 31/90 or 34.4X 500 ppm - 124/166 or 74.7X Progression to malignancy (carcinoma) in the test group was greater than in the controls: controls - 3/90 or 3.3X 500 ppm - 22/166 or 13.3X 3. Mice (100 exposures) Marginal increases in the occurrence of adenomas of the lungs were attributable to VCM In A/J mice exposed 100 times at 50 ppm and evaluated at 8, 16 and 20 months. (Appendix, Tables VII-X, Inclusive) controls - 29/84 or 34.5X 50 ppm - 65/158 or 44.IX Progress to malignancy (carcinoma) was apparent: controls - 2/84 or 2.4X 50 ppm - -7/158 or 4.4X 6 UCC 002254 4. Rats (single exposures) Except for aggravation of latent pulmonary changes, particularly bronchopneumonia, changes attributable to VCM were not apparent in any of the tissues evaluated microscopically at 8, 16 or 24 months from Fischer 344 rats exposed to concentrations of 50, 500, 5000 or 50,000 ppm. (Appendix , Tables XI-XIV, inclusive) 5. Rats (multiple exposures) No changes attributable to VCM were apparent in any of the tissues evaluated microscopically at 8, 16 or 24 months from Fischer 344 rats exposed ten times at 500 ppm or 100 times at 50 ppm. (Appendix, Tables XV-XVII, inclusive) 6. Rats (49 exposures from the reproductive study) The following histological observations were made 24 months post exposure In Sprague-Dawley/Wlstar rats which had been exposed 49 times at 50 ppm or 500 ppm of VCM, (Appendix, Table XVII) Neoplastic and non-neoplastic lesions were observed in approxi mately equal frequency In control and test animals. The only lesions that occurred in higher frequency in the VCM exposed animal than in control rats were eosinophilic cell alterations presented as foci and/or areas. The appearance of these foci was related to dosage. The nature of these lesions is controversial. Some pathologists feel that basophilic lesions have greater significance with respect to neoplasm development than do other cellular alterations. Others feel that all of these cellular alterations may be part of a spectrum capable of progressing to the formation of neoplastic nodules. 7. Summary of Light Microscopy Studies The carcinogenic, or possible related changes (eosinophilic foci) attributable to exposure of VCM in rats and mice are summarized in Table 1. 7 ucc 002255 TABLE 1. VINYL CHLORIDE STUDIES Experimental Group* PPfi ------- DAYS-------- -------- SUMMARY (1 hr/day) (ppm-hrs) SPECTeT/ NUMB'ElTTXPQ'SElj------------ RESULTS A 50,000 1 50,000 ICR Mice 180 33i adenomas Fischer Rats 178 negative c 500 49 24,500 Sprague-Dawley/ 49 eosinophilic Hi star Rats changes A 5,000 1 5,000 ICR Mice 180 16.8% adenomas Fischer Rats 180 negative B 500 10 5,000 AJ Mice 180 74% adenomas Fischer Rats 180 negative B 50 100 5,000 AJ Mice 179 41% adenomas Fischer Rats 180 negative c 50 49 2,450 Sprague-Dawley/ 47 Eosinophilic Hi star changes A 500 1 500 ICR Mice 180 like controls Fischer Rats 190 negative A 50 1 50 ICR Mice 180 like controls Fischer Rats 180 negative TOTAL EXPOSED TOTAL CONTROL COMBINED TOTAL 2263 7TS 305? *A - 170 ICR Mice (82 male and 88 female) and 17* Fischer Rats (92 male and 79 female) served as control animals for the single exposure (50,000; 5,000; 500; 50 PPM) studies. B - 190 AJ Mice (89 male and 101 female) and 200 Fischer Rats (100 male and 100 female) served as control animals for the multiple exposure (10 X 500 PPM and 100 X 50 PPM) studies. gO c(-J k tn VI an c - 45 Sprague-Dawley/Wistar (20 male and 25 female) served as control animals for the multigeneration reproduction study. D. Electron Microscopical Results. 1. Seneral. In general, these studies Indicate that exposure to vinyl chloride Increased organelle turnover (Fig. XIX) as well as loss of volume control (bleb formation) and Increased lysosomal activity In the liver of rats (Figure XIX). These alterations progressively decreased as recovery time after exposure increased. s Hepatocellular carcinoma was seen in one male Fischer rat which had received ten exposures of 500 ppm. Lymphosarcoma was noted in one female Fischer rat which had received a single exposure at 500 ppm. Since these were individual cases, and since no cancers were seen at 50,000 ppm, the lymphosarcoma and the hepatocellular carcinoma are not likely related to the vinyl chloride exposure. Thus, It appears that exposure to vinyl chloride did not produce cancer in rats In any of these single or multiple exposures. General conclusions concerning these various segments of the electron microscopical results follow. 2. General us^gn of 8-Month Recovery to Single Exposure 5o, 500, 5000, 50,000 ppm-hrj of Vinyl Chloride in Fischer Rats. a. Alterations from control (F1q. XVIII) occurred In all treated animals at each concentration, b. Changes were less severe In female animals. c. Hepatocytlc alteration Included lipid accumulation (Fig. XIX), Increased dense bodies (Fig. XX), llpofuschin granules (Fig. XX) and residual bodies. These are an Indication of cytoplasmic sublethal injury. tion. d. Alteration was incremental with Increasing exposure concentra e. Most severe changes involved cellular necrosis with subsequent phagocytosis by Kupffer cells# (Fig XXI) seen in single and multiple exposures. 9 UCC 002257 f. Extruded areas of hepatocyte cytoplasm (in bleb formation)(Fig. xix may indicate a means of removal of altered portions of cells. 3. General Conclusion of 8-Month Recovery to Multiple Exposures (500 ppm X 10 exposures; 50 ppm X 100 exposures) to Vinyl " Chloride in Fischer Rats. a. Changes were seen In male and female treated animals. (Fig. XXI) b. Less alteration was encountered in female rats. c. Cellular changes was greater than that seen in single dose animals. d. Alterations involving hepatocyte nuclei, not seen in single dose animals, were encountered in males of this group. 4. General Conclusion of 16-Month Recovery After Single Exposure T$0, 506, 5000, $0,6(30 ppm x 1 hr) to Vinyl Chloride in Fischer Rats. a. Normal morphology (Fig. XXII) was noted after 16 months in the lower exposure concentrations. When compared to the above alterations at 8 months the 16 months recovery period appeared to be sufficient for return to control morphology. b. When changes in controls were subtracted from those in treated animals, the most significant finding was hepatocyte necrosis in high dose males. 5. General Conclusion of 16-Month Recovery to Multiple Exposures T500 ppm X 10 exposures; 50 ppm X 100 exposures in Fischer Rats. a. Electron microscopic evidence supportive of a diagnosis of hepatocellular carcinoma was seen in one male given 10 doses of 500 ppm vinyl chloride. (Fig. XXIII) b. Other changes included cleft-like spaces in hepatocytes and Kupffer cells. These were interpreted as age-related changes in hemoglobin/ hemosiderin metabolism. I c. Changes were more severe in males. 10 UCC 002258 6. General Cnc1ii5ion to 24-Month Recovery from Single Dose (50, 500. 5000, 56,000 ppm) VinyT Chloride in Fischer Rats. a. Age-related changes including formation of cleft-like spaces (possibly related to hemoglobin/hemosiderin), inflammatory infiltrate (pericholan gitis) and some collagen accumulation were seen in controls (Fig. XXIV) and treated animals in this group. b. In this group hepatocytlc Injury - not encountered in controls - was observed. At 5000 ppm one female showed evidence of lymphosarcoma. 7. General Conclusion to 24-Month Recovery After Multiple Doses (30 ppm X 10) of Vinyl Chloride in Fischer Rats. a. Controls and treated animals showed changes similar to those seen after 24 months recovery to single exposure. b. These changes were regarded as age-related and ncn-specificlFig.XXV 8. General Conclusion to 49 Exposures for 1 Hr at Daily Intervals followed by 24 Month of Recovery (50 and~500 ppmTTsprague/ Pawley/Wistar RatsTT a. Age-related non-specific change was encountered in all groups. b. Most advanced changes were related to 500 ppm group of male rats in which cell swelling and platelet aggregation were seen. E. Multiqeneration Study in Rats (Spraque/Dawley/Wistar). No consistent changes attributable to VCM were found in FQ parents which were exposed to 50 ppm or 500 ppm of the vinyl chloride monomer one hour per day, five days per week for ten weeks before mating and evaluated for numbers of matings, percentages of pregnancies, fertility and lactation indices. The F-j, Fg, and offspring were evaluated for litter size, percent of stillborn pups, post-natal growth, viability, survivability and reproduction anomalies. (Appendix, Table XIX - XXV, inclusive) Electron microscopical examination of the parent rats which were held for 24 months after the 49th exposure revealed age related changes, cell swelling, and platelet aggregation but no tumors,(Fig. XXV): 11 UCC 002259 IV. DISCUSSION. A. Factors In Carcinogenicity. Theoretically, a single molecule of a carcinogenic substance may produce a cancer-if it Is not destroyed in the body before it reaches a suscept ible body cell, if It makes a carcinogenic biochemical combination ("hit") with the cell, If this combination is not repaired, if the cancer cells are not de stroyed by the Immune system, and If other host factors are favorable to carcinogenicity. On the other hand it may be assumed that not every molecule will make a carcinogenic "hit", that some "hits" will be repaired, and some "hits" may not develop Into tumors because of unfavorable host factors, and in some situations the cancer cells may be destroyed by the bodies immune system. The higher the dose rate the greater the number of hits and the greater the freguency of tumor production. The above considerations suggest the existence of "no effect" doses, threshold doses, and dose response curves which have the hockey stick shape 4 described by Bryan and Shimkin. These considerations have been discussed for radiation5 * 6 and chemical carcinogenesis 7 ' 8 by numerous authors. The Food Protection Committee, Food and Nutrition Board of National Q Academy of Sciences - National Research Council noted that dose-response re lationship applied to carcinogens. The higher the dose, the greater is the incidence of response and the shorter the time required to elicit the response. This relationship has been reported for 1,2,5,6,-dibenzoanthracene (DBA)4, 20-methylcholanthrone (MC) , and DMBA plus croton oil , p-dimethylaminoazo- 13 *|4 benzene , and carbon tetrachloride. Dose response curves have been given for ultraviolet light1^ and ionizing radiation There are non-tumorigenic levels of exposure to carcinogens for given experimental conditions. Carcinogens do not produce cancers in all exposed animals. In bioassays, the lower dose levels sometimes do not produce tumors while higher dose do produce tumors.'11,12,13 ijGn 002260 12 The possibility Is recognized that "no effect" doses may exert carcinogenic effects which are too weak to be detected with the numbers of animals used in routine testing. The concept of lifetime accumulative, non-tumorlgenic and tumorlgenic doses of radiation has been adopted. The Federal Radiation Council 18 stated that for occupationally exposed personnel the accumulated dose of radiation to the whole body, head, trunk, active blood forming organs, gonads or lens of the eye shall not exceed: 1. In any calendar quarter, 1.25 roentgen equivalent mammal (rem) 2. Total lifetime dose of 5 (N-18) rem where N equals the present age in years. B. Vinyl Chloride and Carcinogenicity. Maltonl described a dose-response relationship for the carcinogeni effect of vinyl chloride in animals. The neoplastic response was related 19 20 to the length of exposure. ' Lee et al_., 21 noted that the incidence and severity of tumors increased with the concentration of VC and the length of exposure. The above statments indicated that the total dose (concentration X exposure time) may be of importance in the carcinogenicity of vinyl chloride. Total inhaled dose can be approximated by the Haber concept. 22 In its simplest form this concept states that the total inhaled dose, Ct (mg min/cu m) is the product of C (concentration in mg/cu m) X t (time in mi nutes). 13 IJCC 002261 The concentration can be expressed also in part per million (ppn) and the time can be expressed in hours producing Ct in ppm-hr. Factors for breathing rate and detoxication can be included when these data are available. However, the simplified Ct approximation of total inhaled dosage is sometimes useful. A rough calculation of total dosages has been made for some of the data of Maltoni Lee et al_. Viola et al_.,^ Caputo et al. 25 and Keplinger et al_. These Ct calculations are sunmarized in Tables 2 and 3. In the studies of P.L. Viola, A. Bigotti and A. Caputo23 tumors were seen in rats which had been exposed to 30,000 ppm of vinyl chloride four hours per day, five days per week for 12 months. Positive effects were obtained at the total dose (CT) of 28,800,000 ppm-hrs. A. Caputo, P.L. Viola and A. Bigotti 24 exposed rats to vinyl chloride four hours per day, five days per week for 12 months. The con centrations were 20,000; 10,000; 5,000; 2,000; 500; or 50 ppm. The total dose (Ct) for the 50 ppm was 48,000 ppm-hrs. No tumors were produced at this level. The total dose for 500 ppm was 480,000 ppm-hrs. Tumors did occur at the latter dose appearing as early as eight months was 320,000 ppm-hrs. Tumors appeared in the rabbits after nine months at 10,000 ppm. Thus the lowest total dose was 7,200,000 ppm-hrs. M.L. Keplinger et al_., 25 exposed rats, hamsters and mice to vinyl chloride. Only the data on mice was sufficiently complete for examination of total dose effects. The animals were exposed seven hours per day, five days per week for eight months. The lowest Ct was 56,000 ppm-hrs. This Ct and all higher ones did produce tumors in mice. 14 UCC 002262 trtUL-t L MALTONI VINYL CHLORIDE STUDIES Calculations based on reference 19 TEST SPECIES RESULTS (CARCINOGENESIS) - PPM-HRS BT1 RATS BT3 RATS BT6 RATS BT7 RATS BT4 MICE K* *' thousand (000) QUESTIONABLE AT 52K* NEGATIVE AT 17K QUESTIONABLE AT 85K POSITIVE AT 170K, 850K, 2000K, 3000K POSITIVE AT 24.600K NEGATIVE AT 52K, 260K QUESTIONABLE AT 520K POSITIVE AT 2600K, 6200K, and 10.400K POSITIVE AT 30K, 150K, 300K, 600K, 1500K and 3600K t ' ucc 002263 15 TABLE 3 OTHER Vim (ifiLUHiutAfll *% 9%. * W siuuitsAVlftShaasA AUTHORS VIOLA, BIGOTTI.23' CAPUTO CAPUTO, VIOLA,24 ' BIGOTTI KEPLINGER et al.,25 LEE 21 CONSUMER PRODUCTS 1 SAFETY COMMISSION SPECIES RATS RATS RABBITS MICE (RATS & HAMSTERS) MICE ICR MICE A/J MICE FISCHER RATS RESULTS (CARCINOGENESIS) - PPM-HRS POSITIVE AT 28, 80QK NEGATIVE AT 48K POSITIVE AT 320K, 1280K, 3200K 6400K, 12.800K POSITIVE AT 72Q0K POSITIVE AT 56K ALL TESTS ESSENTIALLY NEGATIVE BELOW AMD POSITIVE ABOVE 48K 50 AND 500 - NEGATIVE 5000 - BORDERLINE POSITIVE 50,000 - POSITIVE 5C00 - POSITIVE 50, 500, 5000 AND 50,000 - NEGATIVE 24,500 - EOSINOPHILIC' LOCI, NO CANCERS ucc 002264 Calculations (Tables 1 and 2 Appendix) on the Consumer Products Safety Commission studies indicate that strongly positive carcinogenic effects may not appear in rats until the total dose (Ct) of VCM reaches or exceeds 50,000 ppm-hrs. The maximum, response in the rats was the ap pearance of eosinophilic cells which might suggest a pre-cancerous change. The CPSC VCM tests in mice indicate that increased frequencies of adenoma may appear in this species at Ct's of 5,000 ppm-hr and above. This total dose for carcinogenicity is in general agreement with Lee et al_,,21 (48,000 ppm-hrs) and Maltoni19,20 (between 30,000 ppm-hrs and 150, 000 ppm-hrs), and the other investigators. 23 ' 24 ' 25 In general, in orders of magnitude,carcinogenic tendencies are seen in some species at Ct's of 5.000 to 50,000 ppm-hrs. Definite carcinogenicity appears in both rats and mice at Ct's of 50,000 to 500,000 ppm-hrs, and high incidences of carcinogensis are noted in mice and rats at Ct's of greater than 500,000 ppm-hrs. The previous studies^9'29,2^*23*24,25 and present cpsq study are in agreement as to the dose-time relationship for carcinogenesis related to vinyl chloride exposure. All of these studies considered collectively may indicate that there may be a life-time total dose for vinyl chloride below which carcinogenicity is not likely to occur. This "no cancer" Ct seems to be below 5000 ppm-hr for mice. For histological confirmed carcinogenesis the "no cancer" Ct in rats appears to be greater than 50.000 ppm/hrs. The results of the light or the electron microscopical study did not reveal carcinogenic responses in rats at Cts from 50-50,000 ppm-hrs. However, Ct's of 2450 ppm-hr produced eosinophilic loci but no ijcr 17 002265 tumors in rats in the C?SC tests. As an additional consideration in the total dose concept, Ct's of 5000 ppm-hr produced increased incidence of adenomas in mice when the exposures were at 5000 ppm for one day, 500 ppm for 10 days or 50 ppm for 100 days. The increased incidence of adenomas produced by the single exposure of mice at 5000 ppm is of borderline significance. How ever, there is the indication that even single exposure of sufficient magnitude may be carcinogenic in sensitive species. The dose-response relationship and the "no effect" dose con cept have been described for other carcinogens. It is difficult to relate these animal studies to man. Data on vinyl chloride exposure in plants have been limited. However, acute dizzyness, headache, nausea and chronic liver damage have been seen in vinyl chloride workers. It is assumed that peak exposure levels of several thousand parts per million were experienced at times. Air monitoring of one group of plants during 1950-59 indicate that tiniaweighted (8-hr) average exposure were 120-385 ppm. (This would give daily Cts of 960-3080 ppm-hrs). Peak exposures possibly exceeded 1 COO ppm. This may not have been typical of all polyvinyl chloride plants.^ Data on vinyl chloride in ambient air are limited also. Atmospheric measurements in the vicinity of production plants indicate that concentrations are below 1 ppm. One peak grab sample of 33 ppm has been reported at 0.5 kilometer from the center of one plant.^ The time-weighted threshold limit value of the American Conference of Government Industrial Hygienest for 1977 was 200 ppm. There is a notice of intended change. The Environmental Protection Agency ' has established UCC 1 8 002266 the following emission limits for vinyl chloride: (1) formation and purification processes is 10 pmm, (2) emissions from equipment preceding and including the stripper in the plant process flow is 10 ppm, (3) emissions from equipment following the stripper are to be controlled by stripping dispersion resins to 2000 ppm and other resins to 400 ppm. Assuming that man is as sensitive as the mouse or rat the carcinogenic effects of vinyl chloride might be expected after an accumulated Ct of about 5000 ppm-hrs or ' epproximately 150,000 ppm-hr;(Maltoni BT3). res;:ct Based on the high pollution values given above (1000 ppm-hr per day) car cinogenic doses could have been accrued In one week (mouse) or 30 weeks (rat). At the TLV of 200 ppm (daily Ct of 1600 ppm-hr) the time to accumulate a carcinogenic dose would be 3 days (mouse) or about 19 weeks (rat). Comparable accumulation times at 10 ppm (80 ppm-hrs per day) would be about 12 weeks (mouse) or 375 weeks (rats). The calculations are based upon a five day working week. IV. CONCLUSIONS. A. Except at the highest concentration, 50,000 ppm, where possibly anesthetic-type effects were seen vinyl chloride produced no pharmacotoxlc (excluding pathology) signs In mice or rats during or after exposure. B. No reproductive or teratogenic changes attributable to vinyl chloride were found. Mating; % pregnancies; fertility, lactation indices, % stlll-borns; litter size; post-natal growth; viability, survivability, anomalies. C. Vinyl chloride seemed to produce pneumonitis in mice and to aggravate bronchopneumonic In rats. D. Vinyl chloride produced eosinophilic changes in rats but no 19 UCC 002267 frank (light microscopy) carcinogenesis. E. Vinyl chloride produced increased in frequency of tumors In mice. F. Electron microscopical studies revealed some hepatocellular changes but no carcinogenesis related to the vinyl chloride exposures at Ct's of 50,000 ppm-hrs or less. 1. The carcinogenic effectiveness of VCM depends upon concentration and exposure time, Ct. 4 * 5 ' 6 2. There were VCM doses which were not carcinogenic and there appeared to be a total accumulated dose above which tumors were produced!3*13-1 Similar effects have been noted with other chemicals. 3. Tumors were not seen In mice at Ct's of 500 ppm-hrs nor in rats at 50,000 ppm-hrs or less.4'5,6,9,10,11 4. Carcinogenic effects of VCM were seen in mice at total doses (Cts) of 5,000 ppm-hrs and above.'11 20 UCC 002268 1. Scientific and Techn-'cal Assessment Report on Vinyl Chloride and Polyvinyl Chloride, US Erironmental Protection Agency, EPA-600/i>-75-004, June 1975. 2. E.M. McDowell & B.F. Trump. Histological Fixatures Suitable for Dignostic Light and Electron Microscopy. Arch. Path. Lab. Med. 100:405-414, 1976. 3. J.H. Luft, Improvements In Epoxy Resfn Embedding Methods, J. Biophys. Blochem. Cytol. 9:409-414, 1961. 4. Bryan, W.R., and M.B. Shlmkin. Quantitative Analysis cf Dose-Response Data Obtained with Three Carcinogenic Hydrocarbons in Strain C3H Male Mice. J. Natl. Cancer Inst., 3:503-531, 1943. 5. J.B. Storer. Radiation Carcinogenesis, Chapter 16, pp 453-433 in Cancer 1. Etiology: A Comprehensive Treajtlse. Editor: F.F. Becker. Plenum Press, N.Y., London. 1975 6. A.C. Upton, J.L. Randolph and J.W. Conklin. Late Effects of Fast Neutrons and Gamma-Rays in Mice as Influenced by the Dose Rats of Irradiation: Induction of Neoplasia, Radiation Res. 41:467,-1970. 7. U. Saffiotti. Identifying and Defini ng Chemical Carcinogens), pp 13111362. Origins of Human Cancer, Book C. H uman Risk Assessr-ivt. Editors: H.H. Hiatt, J.D. Norton, J.A. Wlnsten. Co Id Spring Charter' Confers! css in Cell Proliferation. Vol. 4. 1977. Cold S pring Charter Laboratory. 8. T.H. Maugh II, Chemical Carcinogens: How Dangerous are Lo.v Doses? Science, 202:37-41. Oct 1978. Problems in the Evaluation of Carcinogenic Hazard freui the Use of ood Additives. Publication 749. The Food Protection Ccnr.ittee, Food and Nutrition Board of National Academy of Sciences - National Researlcl i Council December 1959. ! 10 Horton, A.W., and Dorothy T. Denman. Carcinogenesis of the Skin. A Re-examination of Methods for Quantitative Measurement of the Potencies of Complex Materials. Cancer Research 15:f01-709, 1965. 11. Poel, W.C. Effect of Carcinogenic Dosage and Duration of Exposure on Skin-Tumor Induction In Mice. J. Natl. Cancer Inst 19-44, 1959. 22: 12. Graffl, A. Untersuchungen uber den Mechanismus der Carcerogfenese und die Wlrkungsweise cancerogener Relze. Abhandl. deut. Akad, Hiss. Berlin, 53:1-27, 1953. 13. Druckery, H. Pharmacological Approach to Carcinogenesis, in Clba foundation Symposium on Carcinogenesis: Mechanisms of Action. Boston: Little, Brown and Company, 1959. ;pp 110-130. UCC 21 002269 14. Eschenbrenner, A.B., and Eliza Miller. Studies on Hepatomas. I.. Size and Spacing of Multiple Doses in the Induction of Carbon Tetra chloride Hepatomas. J. Natl. Cancer Inst., 4:385-388, 1944. 15. Blum, H.F., On the Mechanism of Cancer Induction by Ultraviolet Radiation. J. Natl. Cancer Inst., 11:463-495, 1950. Ijj. Finkel, Miriam P, Mice, Men and Fallout. (The potential danger of strontium 90 is appraised on the basis of data from animal experi ments). Science 128:637-641, 1958. "17. Mole, R.H. The Dose-Response Relationship in Radiation Carcinogenesis. Brit. Med. Bull. 14:184-189, 1958. '...18. The Federal Radiation Council (Report No. 1, Background Material for the Development of Radiation Protection Standards, 1960, Government Printing Office, Washington, DC). 19. C. Maltoni. The Value of Predictive Experimental Bioassay in 1 Occupational and Environmental Carcinogenesis. An Example: Vinyl Chloride. Ambio. 4:18-23, 1975. 20. C. Maltoni and G. Lefemine. Carcinogenicity Assay of Vinyl Chloride. Ann. NY Acad. Sci. 246:195-218, 1975. 21. C.C. Lee, J.C. Bhandari, J.M. Winston, W.B. House, R.L. Dixon and J.S. Woods. Carcinogenicity of Vinyl Chloride and Vinylidene Chloride, J. Tox. Environ. Health 4:15-30, 1978. 22. F. Haber, "Funf Vortrage aus den Yahren 1920-23": No. 3. Die chemie im Kriege: No. 5. Zur geschichte des gaskarnpes, Juluis Springer, Berlin, 1924 - reference through Prentiss: Chemicals in War, McGraw-Hill Book Company, Inc., New York, 1937. 23. P.L. Viola, A. Bigotti and A. Caputo. Oncogenic Response of Rat Skin, Lungs and Bones to Vinyl Chloride, Cancer Research 31:516-522, 1971. 24. A. Caputo, P.L. Viola and A. Bigotti. Oncogenicity of Vinyl Chloride at Low Concentrations in Rats and Rabbits. 0. Int. Res. Commun. 21:1582, 1974. 25. M.L. Keplinger, J.W. Goode, D.E. Gordon, and J.C. Colandra, Interim Results of Exposure of Rats, Hamsters and Mice to Vinyl Chloride. Ann. NY Acad. Sci. 246:219-224, 1975. 26. Threshold Limit Values for Chemical Substances and Physical Agents in The Workroom Environment with Intended Changes for 1977, American Conference of Governmental Industrial Hygienist. 27. Title 40-Protection of the Environment. Environmental Protection Agency, Part 61-National Environmental Standards for Hazardous Air Pollutants, Stand ard for Vinyl Chloride, Federal Register, Vol 41, No. 205, Thursday, Oct 21, 1976. 002210 22 000 APPENDIX Table I. Single Exposure Schedule of Animals To VCM Species Sex Dose ppm Exposure dote AH PH Fischer Rat H 50 3/4/75 500 3/11/75 5000 3/18/75 50000 3/25/75 F 50 500 5000 50000 3/4/75 3/11/75 3/18/75 3/25/75 M ICR House F 50 500 5000 50000 3/4/75 3/11/75 3/18/75 3/25/75 50 500 5000 50000 3/4/75 3/11/75 3/18/75 3/25/75 Rat H Neg/Cont. F Neg/Cont/ House M Neg/Cont. F Neg/Cont. Exposure group si/e(s) 90 90 85 90 90 100 95 88 90 90 90 90 90 90 90 90 92 79 82 88 Aye at time of exposure weeks 15 16 17 18 15 16 17 18 15 16 17 18 15 16 17 18 15-18 15-18 15-18 15-18 23 t APPENDIX Table II . Exposure Schedule for Animals Exposed Repeatedly to VCM Species Fischer Rat A/J House Fischer Rat A/J llouse Sex Dose ppm U 50 500 F 50 500 U 50 500 F 50 500 U Neg. control F Neg. control U Neg. control F Neg. control Exposure periods days 100 10 100 10 100 10 100 10 100(c) 10(c) 100(c) 10(c) 100(c) 10(c) 100(c) 10(c) Exposure dates From To 8/27/75 7/7/75 8/27/75 7/7/75 7/7/75 8/27/75 7/7/75 8/27/75 -- - 1/26/76 7/18/75 1/26/76 7/18/75 7/18/75 1/26/76 7/18/75 1/26/76 _ -- - NOTE: (c) control for corresponding dose above. Exposure Group size(s) Start End 90 86 90 90 90 87 90 90 90 87 90 90 90 88 90 90 50 50 50 50 50 47 50 50 40 39 50 50 50 50 50 50 Age Start 21 14 21 14 15 8 15 8 21 14 21 14 15 8 15 8 End wks 41 16 41 16 35 10 35 10 41 16 41 16 35 10 35 10 002272 C o o 24 APPENDIX Table III. VCM Multigeneration Study Fg Parents Group I II III Compound Air VCM VCM Dose Control Low Dose (50 ppm) High Dose (500 ppm) Number of Males 25 25 Number of Females 25 25 25 25 ucc 002273 25 100 n Figure I SPONTANEOUS MORTALITY MALE ICR MICE SINGLE DOSE STUDY VINYL CHLORIDE MONOMER 60 PPM CONTROL 50.000 PPM 5.000 PPM 500 PPM % MORTALITY (CUMULATIVE) 00 40 30 30 10 - AGE Or MICE (MONTHS) 26 ucc 002274 Figure II SPONTANEOUS MORTALITY FEMALE ICR MICE SINGLE DOSE STUDY VINYL CHLORIDE MONOMER 50.000 PPr. 5.000 PIT,; 50 PPM CONTRGL 003 PFi,` AGE OF MICE (MONTHS) 27 UCC 002275 % M O T.A LtT',' (CUMULATIVE) 28 Figure IV CONTROL 5.000 PPM 500 PPM 50 PPM 50.000 PPM \ UCC 29 002277 % MORTALITY (CUMULATIVE) Figure V SPONTANEOUS MORTALITY EXPOSED Q EXPOSED CONTROL </CONTROL 30 UCC 002278 Figure VI SPONTANEOUS MORTALITY </E EXPOSED 9 EXPOSED 9CONTROL </cCONTROL ucc 002279 31 ;i Figure VII </E EXPOSED CONTROL </c A OCONTROL 1-12 13 14 IS 16 17 18 19 20 21 22 23 AGE OF MICE (MONTHS) 32 o&o 70 60 - 0 - 40 - OD aO 30 S 20 - 10 - wO oo Figure VIII SPONTANEOUS aJcrTALITY FISCHER RATS 100 DAY STUDY VINYL CHLORIDE MONOMER i 17 18 13 AGE OF HATS (ViOMTKSI 33 ' r 20 70 "I 00 - / .t A\ -uV* > < D 40 - SO3 < sOs 30 20 - 10 - ftj oc MsM O Figure IX SPONTANEOUS MORTALITY FISCHER RATS 100 DAY STUDY VINYL CHLORIDE MONOMER ACE OF RATS (MONTHS) ' 31! CONTROL Figure X GROWTH CURVES OF HALE ICR MICE EXPOSED ONCE TO VCM to < K 13 > H htoLD H L'J , rI /' rrOoo O>x-v 0to3 WEEKS POST EXPOSURE 35 Figure XI GROWTH CURVES OF FEMALE ICR MICE EXPOSED ONCE TO VCM MEAN A N IM A L W E IG H TS I N GRAl mO $ 36 MEAN A N IM A L HEIG HTS I N GRAMS Figure XII GROWTH CURVES OF MALE FISCHER RATS EXPOSED ONCE TO VCM tVEEKS POST EXPOSURE Figure XIII GROWTH CURVES OF FEMALE FISCHER RATS EXPOSED ONCE TO VCM MEAN A N IM A L WEIGHTS I N GRAMS oC glivoort CD CD 38 Figure XV GROWTH CURVES OF A/J MICE EXPOSED TO VCH IN IOO DAY STUDY in x c<c 13 in H X CD H Id < X H2< 2 < Sg ro O mO - CD CD *tO I t J I Figure XVI GROWTH CURVES OF FISCHER RATS EXPOSED TO VCM IN 10 DAT STUDY 500-q MEAN A N IM A L W EIGHTS IN GRAMS 0 20 80 100 Figure XVII GROWTH CURVES OF FISCHER RATS EXPOSED TO VCM IN 100 DAY STUDY MEAN A N IM A L W EIGHTS I N GRAMS oC So foO tao 0 20 EJ oJ 80 100 WEEKS POST EXPOSURE k2 Table IV. Over-all Summary Incidence of Non-neoplastic Changes and Histologically-Proven Neoplasms Within The Liver and Lungs of ICR Swiss Mice Exposed to Vinyl Chloride. Single Inhalation Exposure Tissue/Response Exposure Dose Level (PPM) Sex of Animals Animals Per Group* : Control 0 MF 62 77 Liver :Number Evaluated : 50 Zi - hepatic cell necrosis - hepatic cell vacuolation (lipidosis) - hepatic cell hypertrophy - hepatic cell hyperplasia - angiectasis - sinusoidal reticulosis 2 10 2 11 1 Incidence of Response 50.000 Vinyl chloride 5,000 500 MFM 74 82 76 FM 82 72 F 75 63 78 68 76 61 72 355 74 4 2 12 3 28 84 44 14 5 6 5 13 50 M 81 64 2 1 1 F 80 68 3 4 - hepatic cell adenoma - hepatic cell carcinoma - hemangioma - hetnangiosarcoma 21 12 2 416 19 2 21 11 Lung - pneumonitis :Number Evaluated : 50 1 - bronchio-alveolar adenoma - bronchio-alveolar carcinoma 4 70 6 8 61 76 65 21 10 13 31 14 14 121 78 66 17 19 10 8 73 21 15 4 10 8 1 68 7 6 002291 Total includes animals from scheduled sacrificed (8 and 18 month periods) and spontaneous deaths. o o 43 Table V. Summary of Incidence of Histologicaliy-Proven Neoplasms Within Tissues ICR Mice Exposed to Vinyl Chloride Monomer (Single Inhalation Exposure) 1-hr Inhalation Exposure in ppm 50,000 Spontaneous Deaths 0-6 Months 7-12 Months M F Total H F Total 000 5 11 16 i ' 13"18 Months 1* M F Tota 1 Schedule Sacrifice 8 Months 18 Months r Total. H Total 33 32 65 4 1 5' 7 17 24 Totals -tlaJs____ ____ Fspalfi_____ 49 61 5,000 0 22 4 2 6 22 27 49 2 13 33 | 45 5 13 18 i 500 0 5 5 6 8 14 19 14 33 l 01 5 24 29 31 51 . f 15 1 47 50 0 0 0 4 15 19 9 18 27 0 0 0 2 14 16 ss! i 0 2020 1 1 14 41 00 2 12 14 18 ! 54 (Control] 1 r rTOTALS jVCM 0 7 7 19 36 55 83 31 _ 1______ 1 / j. 2 [9 ^ * ,_ 1 19 63 >. , = ... 37 1 002292 O o 1*4 Table VI. Incidence of Histologically-Proven Meopjasms Within Tissues From ICR Mice Exposed to Vinyl Chloride Monomer (Single Inhalation Exposure) iacrlfice Organ Period Tissue ichedulec Lung 8 months Liver Kidney 50,000 ppm 5, 000 ppm H F Tota 1 M 41 00 00 5l 01 00 F Total l2 01 00 500 ppm M F Total M 0 0 00 1 0 10 0 0 00 50 ppm 0 ppm (Control) Fotal 00 00 00 M 0 0 0 F Total 00 00 00 Stomach 0 0 Other* 0 0 Totals 4 1 18 months Lunq 56 1 Iver 7 3 Kidney 0 0 Stomach 0 0 Other* (1 8 Totals 7 14 Spontaneous Deaths 0-6 men 7-12 mon Lung 00 Liver 0 0 Kidney S tomach 0 0 0 0. Other* Totals Luna ft _ .0 90 }3 Liver Q 2 K1dnev I .2 itarcadi. _Q__ 0 Other* 1 4 Totals 13-18 mon Lunq Liver Kidney Stomach 5 22 5 6 0 11 16 .5 0 0 Other* 2 11 Total 33 32 00 00 52 11 ) * 2 00 00 89 24 0 0 0 0 0 0 6 ? 3- 9 5 16 ?8 10 4 0 13 5 0 0 0 0 0 0 3 0 1 9 0 4 12 5 l 0 4 65 22 00 0 a 0 fl ... n n n . n-- n 00 13 0 1 0 00 0 10 0 0_ 0 00 0 00 00 7 10 4 12 16 1 45 1 ___ Z_ 8 02 1 1 20 11 1 2} 11 0 3 30 22 0 00 00 0 1 11 01 0 00 55 0 7 70 77 0 33 n 18 _ s .. .2.4. -29._ 2 ___1.4-- 16___ 2 12 14 1 '1 00 0 0 1 l 0 ____0-- 0 JB___ ____L_ i 10 00 0 00 00 00 0 0 1 l0 0 00 00 00 0 0 0 fl 0 fl 11 0 22 0 1 4-- i 00 2 2 20 5 "5 0 i ii 1 31 00 0 fl 24 0) 20 2 ------ 0"" 2 0 60 0 00 0l I 0 _Q . 0 11 2 1 20 0 -0. 0 4 61 22 11 10 n 9 0 0 00 00 l1 26 6 8 14 4 15 19 0 11 5 l? 5 5 10 ... 8 34 1 00 0 14 18 ? 27 49 19 2 7? 5 10 4 9 13 4 12 4 2 6 5 7 12 2 30 0 00 1 1 11 0 0 IA 56 T)' 3 6 11 J2___ --JO-- 10____ jj___ 2Q... 24 14 33 9 18 27 |l4 41 55 45 002293 Table VII . Over-all Summary Incidence of Non-neoplastic Ghanges and Histologically-Proven Neoplasms Within The Liver and Lungs of AJ Mice Exposed to Vinyl Chloride (Multiple Inhalation Exposure) Tissue/Response Incidence of Response Exposure Control Vinyl chloride Dose Level (PPM) 0 500 Hours Exposure 10 x 1 Ton Sex of Animal :M F W~ T Animals Per Group* 46 48 78 92 Liver : Number Evaluated : 45 48 zi 89 - hepatic cell necrosis - lymphoid cell infiltrate - hepatic cell lipidosis - neutrophil infiltrate - bile duct hyperplasia - granulomatous foci - sinusoidal reticulosis - hepatocyst - amyloidosis - angiectasls 46 1 2 2 1 6 13 1 2 1 1 1 1 1 1 - hepatic cell adenoma - cholangiocarcinoraa Lung : Number Evaluated : 1 tr 43 ~T 47 1 tr 76 - edema - pneumonitis - bronchio-alveolar hyperplasia 2 21 - bronchio-alveolar adenoma - bronchio-alveolar carcinoma 15 16 3 17 22 56 12 68 11 Total includes animals from scheduled sacrificed (8, 16, 20 month pa riodsj and spontaneous deaths rr 90 1 2 68 10 81 ucc 002294 Table VIII. Over-all Summary Incidence of Nan-neoplastic Changes and Histologically-Proven Neoplasms Within The Lungs of AJ Mice Exposed to Vinyl Chloride. Multiple Inhalation Exposure Tissue/Response Lung - edema - congestion - focal hemorrhage - pneumonitis - bronchio-alveolar hyperplasia - osseous metaplasia - bronchio-alveolar adenoma - bronchio-alveolar carcinoma - reticulum cell sarcoma Exposure Dose Level (PPM) Hours Exposure Sex of Animal Animals Per Group* : Number Evaluated Control G 100 x 1 M 39 Incidence of Response Vinyl chloride 50 100 x 1 F MF 47 81 83 39 45 2 41 2 53 3 77 1 1 2 45 3 1 11 18 27 38 2 34 1 13 TT 30 43 * Total includes animals from scheduled sacrifice (8, 16, 20 month periods) and spontaneous deaths. Uble IX. Summary of Incidence of HistologicaIIy-Proven Neoplasms Within Tissues From AJ Mice Exposed to Vinyl Chloride Monomer (Multiple Inhalation Exposures) Sacrifice Period Spontaneous Deaths 0-6 Months 1-hr Inha la tion Exposures to Vinyl Chloride Monomer 50 p pm x 1 00 CONTROL 500 ppm x 10. j CONTROL M F TOTAt H F TOTAL H F TOTAL M F TOTAL 0 00 0 000 0000 0 TOTALS VCM H M FH 0 2| CONTROLS M 0 F 0 7~I2 Months 8 3 II 0 0 0 2 1 3 0 0 0 10 400 13-20 Months 23 20 43 3 5 8 36 27 63 2 3 5 59 Scheduled Sacrifice 8 Months 3 1 4 0 1 1 6 6 12 0 0 0 9 16 Months 10 6 16 2 3 5 14 8 22 1 1 2 24 20 Months 19 44 63 16 11 27 29 41 70 17 21 38 48 47 5 8 701 14 3 4 85 33 32 Totals 63 74 137 21 20 41 87 85 172 20 25 45 150 159 41 45 oo N<afO>o) iO-i 48 (<tute k. hk.iuonce ox Hii>lxilOjiCAlly-I'roven Ihoplesms Within Tissues From AJ Nice Sacrifice Feriod Scheduled 8 Honths Organ Tissue Long Liver Kidney Stomach Other* Totals Lung Liver Scheduled Si crifice and Soontaneous Deaths 4 SO com x 100 Control ___ 500 own x 100 Control H F Total M . _ F Tamil H F Total H F Tor at 11 2 0 1 _ 1 6 A 17 n A A 00 00 00000a 000000 0 9 0 0_ 00 0 0 00 000o00 00 0 0 00 31 5i. 60 0 0 0 o 1 0 0 . .0 0 0 0 40 l li 6 6 12 0 0 0 9 2 3 5 ___9 _ 9 __17. 1 1 2 6 0 0 0 ft 0 0 00 0 0 16 Honths Kidney Stomach Other* Totals 00 01 90 10 6 0 1 0L 16 t 0 0 .0.. 2 0 o !| 0 0 `i a _0 : 3 1 5 j. _0_ 0. .0. o ..... 9- ... 0 5 o._ s Ik 8 22 9 n 0 1 -o 0 .0 1 _ 0 O_ 0 ... 2 20 Honths Lunq Liver i Il.ty i S.omach Other* ( fotcls w> 28 41 i if>___ n t _2i_! !_2IL----40.-, --66 JA_ 17 JJ________ 23 5) 2 3 ? ii o 0 0i 0 _J____ ft i i; u 1 i ft ! ft ft. * ..ft________ ~~0~ 0 ( 0 1 to p n 0 4 o - v_- > 0 0 1 l< > 3 0 | .0 n^ Q_ ._ . 0 2. 4 0 _6 '9 63 * L> 11 Tf 23 M . 70 17 21 . .L38___ Spontaneous Lunn Deaths Liver TfTHi'ev 0-6 Honths Stomach Other* Totals 0o 00 9 o--. 09 00 00 0. 0 o1 1 Of 0 ot ______________, ------------- -- - -- 000 000 0 0 r0 - 000 of f. n 1* 0' 0 tt o 0 fl ft__ 0 0 0 o 0 1 0 0 0 1 2 1 0 _._G 0 1 2 _Q__ --0 _----------- -ft________ 0 0 _fl________ .0ft _ ft- 000 000 0 0 0_ Lunq Liver 2I 20 } 0 0 o ii i 0 2 0 0 000 10 0 0 00 0 0 7.10 Unnfk Kidney 10 1 0 0 01 0 0 00 0 0 Stomach Other* 00 32 0 5 0 0 0 o 0 0 | ! 0 i 0 1 00 0 0 20 0 0 Totals a 3 11 0 0 o : 1 2 1 3 9 .-.CL- _fl_________ Luna 13 Ifi > 7 C _ i--28-- 23 - -ST- -ft---=---- i ---- --1-- Liver 23 50 1 i1 2 0 20 1 _I_________ 13-20 Kidnev *0 n4 0 ol ft._ ft -JL- -J-ft _________ \________ _________ Honths Stomach 00 0 0 0 o 0 L_ SL _JL_ Q -JJ_______ _________ _________ _______ _________ C O Other* 47 n 1 1 2! 6 H 10 I i 2 o Total 23 20 43 3 5 * UL. 27 83 2 3 5 49 002297 Table XI. Over-all Summary Incidence of Non-neoplastic Changes Within The Lungs of Fischer Rats Exposed To Vinyl Chloride. Single Inhalation Exposure Tissue/Response Exposure Dose Level (PPM) Sex of Animal Animals Per Grouo* : Control :0 :H F : 89 74 Incidence of Response Vinyl chloride 50,000 5,000 500 MF MF HF 86 87 83 93 87 100 50 M r~ 90 91 Lung :Number Evaluated - bronchopneumonia : 85 26 67 10 80 77 45 13 80 87 13 10 85 95 80 15 3 28 74 3 % incidence : 30.6 14.9 56.2 16.9 16.3 11.5 17.6 3 35.0 4.1 * Total includes animals from scheduled sacrifice {8, 16, and 24 month periods) and spontaneous deaths. ucc 002298 50 Table XII. Summary of Incidence of Histologically-Proven Neoplasms Within Tissues From Fischer Rats Exposed to Vinyl Chloride Monomer (Slnlge inhalation Exposure) 1-hr inhalation Exposure in ppm Spontaneous Deaths i 0-6 Months 7-12 Months 13-18 Months 19-24 Months | 8 Non th s MF T* M F T* H F T* M T* ! FM F T* Schedule Sac rifice 16 Months 24 Months M F T* M F T* 50,000 0 0 0 0 4 4 16 20 36 86 47 133 2 0 2 4 2 6 21 34 55 Totals Male Female 129 107 5,000 0 0 0 2 2 4 20 9 29 71 37 108 0 0 0 5 1 6 37 52 89 135 500 0 0 0 0 2 2 18 18 36 57 64 121 0 0 0 13 0 13 82 37 119 170 / 50 0 0 0 0 2 2 18 15 33 89 28 117 0 0 0 7 0 7 64 27 91 178 0 (Control) 0 0 0 0 0 0 13 3 16 102 56 158 0 0 0 11 3 14 60 27 87 I 186 1. 1 Totals VCM 0 0 0 2 10 12 72 62 134 303 176 i79 0 2 29 3 32 204 150 354 t____ 101 121 72 89 , Table XIII. Incidence of H i s I o log i c.i l I y-P rovcti asms Within 1 issuer \ rom Fischer Rats Exposed to Vinyl Chicride hcnomei (Single Inhalation Exposure) Sacrifici Organ Period Tissue Scheduled Lunq 8 months Liver Kidney Stomach 0 the r* Total 16 months Lung Liver Kidney S tomach Other* Total 24 months Lunq Liver Kidney Stomach Other* Total 50,000 ppm H F Total 0 00 0 00 0 00 0 00 2 02 2 02 2 02 0 00 0 00 0 00 2 2k 4 26 0 22 ) 3 <1 0 00 0 11 20 28 h8 21 Ik 55 5. 000 ppm K F Total M c0 0 0 0 00 0 0 00 C 0 00 0 0 00 0 0 00 0 0 00 1 0 00 0 0 00 0 0 00 0 51 51 00 3 __2 00 6 12 6 In o3 58 01 0 1 10 3k 49 83 70 37 52 89 82 500 ppm F Total H 00 0 00 0 00 0 00 0 00 0 00 0 011 00 0 00 0 00 0 0 12 6 0 13 36 k 12 01 7 1 9 0 00 30 100 37 119 0 5* 6k 50 ppm 0 ppm (control) F Total M F 00 0 0 00 0 0 00 0 0 00 0 0 00 0 0 00 0 0 0 10 0 00 0 0 00 0 0 00 0 0 0 6 II _ 3 0 7 11 3 2 3. _ 3_ 12 00 3 ? i 2 1 0 000 1 22 76 **7 23 27 91 60 27 Total 0 0 0 0 0 0 0 0 0 0 14 14 5 10 1 1 70 87 002300 O O 52 Table XIV. Incidence of Hislologically-Proven Ntopiasms Within Tissues From Fischer Rats Exposed to Vinyl Chloride Monomer (Single Inhalation Exposure) ' Sacrificc Organ Period Tissue 50,000 ppm M F Total 5 ,000 ppm M F Total M 500 ppm ir , F (Total M Sponlanecus Deaths 0-6 Months Lung Liver Kidney Stomach 0 0 0 0 0 0 0 0 0 0 0 00 0 0 0 00 0 0 0 o0 0 0 0 00 0 00 0 00 0 00 0 00 Other* 0 0 0 0 0 00 0 00 7-12 Months Totals Lung Liver Kideny 0 0 0 0 0 0 0 0 0 0 0 00 o' 0 0 0 0 0 0 0 00 0 10 0 0 0 0 00 0 00 1 10 0 00 Stomach Other* 0 0 0 4 0 0 0 00 4 2* ` 2 4 0 0 00 1 10 Totals 0 4 4 22 40 1 10 13-18 Lung. 1 It Months Liver 2 3 5 5 l 63 5 4 2 62 5 83 4 62 Kidney 0 1 1 0 1 10 0 02 Stomach 0 Other* 13 0 12 00 25 11 0 0 h!r- 0 0 0 5 16 9 22 11 Totals 16 20 36 20 9 29 f!8 13 36 18 19-24 Months Lung 4 Liver 12 Kidney j 2 Stomach j 0 4 8 It 6 18 9 1 3. 3 2 j2 o 5 *6. - 4 4 13 6 i *0 00 Other* | 68 34 102 43 ?7 lb 1 l0 8 12 11 10 16 9 1 f 3" 3 30 4 2 '19 bo Totals f 86 47 | 133 71 37 108 37 64 ... J2' ... 89 50 ppm 0 ppm (control) F Total M 000 000 000 000 000 000 00 0 000 000 000 220 220 1 20 040 020 1 10 13 24 13 15 33 13 0 11 8 0 9 14 033 003 28 ; 34 ! 74 28 M'_7__ 102 F Total 00 00 0. 0 00 00 00 00 00 00 00 00 00 00 00 00 00 3 16 3 16 08 8 22 14 14 46 120 56 158 002301 53 Table XV. Summary of incidence of ri istologic.il ly-Proven Neoplasms Within Tissues From Fischer ^Ts Exposed to Vinyl Chloride Monomer (Multiple Inhalation Exposures) SacrificA __sa. 1pom x 100 Period i M F Total Spontaneous Deaths 000 0-6 Months M 0 Control F Total 00 500 ppm x 100 M F Jfotal 000 Cont ro) M F Total 000 Totals VCM H1 F 00 Control HF 00 7-12 Months 0 0 0 3 0 3 0 0 0 0 0 0 0 0 30 13"18 Month 14 ' 6 20 5 0 5 10 6 16 3 0 3 24 12 80 19-22* Month Ml 51 162 52 29 81 52 29 81 33 34 67 Scheduled Sacrifice 0 0 0 0 0 0 0 1 l 0 0 0 8 Months 16 Months 8 10 18 4 1 5 10 1 11 000 163 0 18 80 85 63 1 00 11 4 1 24 Months 43 43 86 32 29 61 a 71 113 69 27 96 85 Totals 176 no 166 96 59 j 105 j&l 145 114 108 222 166 290 114 101 218 .. 201 56 * 120 od So o !S> 5** Table XVI. Incidence of His to logical I y-Proven Ihiopl -<w. Within Tissues From Fischer Rats Exposed to Vinyl Chloride Monomer vllu'tiple Inhalation Exposure) Spontaneous L'eaihs Sacrlfice Organ Tertod Tissue 0-6 Months 7"I2 Months Luna Liver Kidney Stoaiach Other* Totals . Lunq Liver 50 ppm x 100 H Total 0 0. ... 0 o I0 0 o 10 0 o 10 00o 000 0. 0 00 0 0 Control . II 500 ppen x 100 Control M F Ifotal ;J M F (Total ` M p .... 0 . 00 S0 o _ ! 0 * 0 0___ 10 0 0 0 o i o i! o o 10 0 F Total --0-- . 0 00 0 __ G___ 0 0 0i0 0 0 0 0 0 0 o' 1 0 0 0 0 0 0 0 0 otio 0 0 0 0 0 9 -0- ___| .. -Q _il___ n --___ -JO___ 0 0 o !o 0 , 9 , ,, _JL_____ 0-- -J)___ Kidnev 0000 00 0 0 000 0 Stoaiach o 0 0 .... 9 0 0 ... 0 9 0 9 0 0 Other* 000 3 03 0 0 00 0 0 Totals 0 00 3 03 00000 0 13-18 Months * Luna Liver Kidnev 30}2 }030 ,9 ,,Q 0 0 02 1. 0 k 1 0 1 00 202 1 0 1 0 0 10 0 0 0 0 0 Stoaach Other* 9000 8 6 14 3 00 03 00000 0 k 6 10 1 0 1 I9-24 Months Totals Lung Liver 6 20 5 ii 5 16 9 15 7 22 u 0 5 J 10 k >3 | 9 k *3 ! 9 6 16 k 13 h !*3 30 3 8 5 13 5 8 13 Kidnev S toraach i i 1 2 I2 i 2 1. A 2 lA LI..2 2 10 2 1! *2 1 2 (1 2 13 h 2 Other* 33 37 [120 '32 17 '.j : 3?__. 17 11<9 ! IB 17 oCo Totals [m 51 1 162 ill 29 {52 J2 i si________ L3l 3*1 iiZ____1 55 002303 Table XVtl. Incidence of Histologically-Proven Ikoplasms Within Tissues From Fischer Rats Exposed to Vinyl Chloride Monomer (Multiple Inhalation Exposures) Sacrifice Organ Period Tissue 8 Honths Lung Liver Kidney Stomach Other* Total 16 Months , Lunq Liver Kidney Stomach Other* Total 26 jHonths Lung Liver K i dney Stomach Other* Total Scheduled Sacrifice 50 ppm x 100 Control H F Total H F Total 500 ppm x 100 M F Total Control MF Total 00 0 --o 00 00 00 00 00 00 00 00 00 00 0 . 0__ 0 0 o 0 0 00 .0 .. _ 0.. ....0 0 00 0 00 0 01 0 01 0 __0 0 0 1 1 0 0 __ 0 0 0 0 0 0 0 0 0 0 0 0-- 0 0 0 0 0 0 00 0Q 0 0 0 0 00 0 0 00 68 10 18 8 to 18 j 6 ,1 0 1 1 l -7 2 Cl i . G 0 t. J 0 ' 0 . 0 ... 0 -0 f) 0 0p 1 0 . _0. _--.0___ .-ft.. .. 0- ft- n 0 0 ii 2 0 2 0 00 o o i! o 0 1 0 0 00 1I 5 i i 8 i9 0 00 l ` ! to 5 1 in 0 l !2 i k 2 i6 3 > ; !> 1 ii 5 0 Lo 01 3 6!9 C c . 2 0 (j 0 00 j 35 ill 76 63 86 ! 00 0 : * ^ *25 i 29 jol j j 62 0 t 0 11 68 i 56 6G 23 83 71 l> k. 27 L* g ur\>j Q0 $ 56 Table XVIII. Incidence of Eosinophilic Changes Within Livers of EUA Colony (Sprague-Oawley) Rats-Hu11iple Inhalation Exposures to Vinyl Chloride (UC} Exposure Dose Level (PPM) Hours Exposure Group* Control 0 A __jB_ T._ Hales: Animals per group Eosinophi1ic Focus/foci % Eosinophilic Area/Areas % 9 --179" ii.i 0/9 0.0 18 7-7 1/13 7-7 25 1 "2722 9.1 1/22 4.5 Eosinophilic Alterations** % 1/9 11.1 1/13 7.7 2/22 9.1 Females: Animal per group Eosinophilic Focus/Foci % Eosinophilic Area/Areas % Eoslnlphllic Alterations** % 18 0/18 0.0 4/18 22.2 4/18 22.2 7 0/7 0.0 0/7 0.0 0/7 0.0 25 0/25 0.0 4/25 16.0 4/25 16.0 ,_A . 13 i6/13*** 76.9 1/13 7.7 10/13 76.9 11 4/11 36.4 2/11 13.2 5/M 45.4 VC __ 49. 12 3/11 27.3 I/ll 9.1 4/11 36.4 14 1/13 7.7 0/13 0.0 1/13 7.7 -X. -A_ ___ VC 50 49 _JL_ 25 13/24*** 54.2 2/24 8.3 11 4/io 40.0 0/10 0.0 14 4/11* 36.4 3/H 27.3 14/24**** 4/10 58.3 40.0 5/11** 45.4 25 5/24** 20.8 2/24 8.3 6/24 25-0 19 2/19 10.5 S/9 5.3 3/19 15-8 6 1/6 16.7 1/6 16.7 1/6 16.7 T 25 8/21** 38.1 3/21 14.3 9/21** 42.8 25 3/25* 12.0 2/25 8.0 4/25 16.0 + Group: A Scheduled sacrifice, B = Spontaneous deaths, T = Total +* Eosinophilic fee: and/or areas In the same animal are tabulated one time as an eosinophilic alteration * Different from control at 5-0 - >2.5% level (P = .05 - .025) ** = Different from control at 2.5 - >0.52 level (P = 0.25 - .005) *** * Different from control at 0.5 - >0.05% level (P = .005 - .0005) **** a Different from control at0.05% level (P = .0005) 57 ucc 002305 Table XIX. Summary of Matings and Percentage Pregnancy Observed in the I/CM Three-Generation Study. Fo Gon. F. Gen. F2 Gen. Control* 50 ppm* 500 ppm* Total Females Mated 25 20 3 Total Females Pregnant % Pregnant Females j Total j Females Hated ! 22 S i 68 ! 25 19 95 19 100 " 18 Total Females PregnL i.l I Z Pregnant ' Females Total Feina 1 es Mated Total Females I Pregnant ! 21 i ' I i '25 k>w i i l _______ .... '________________ J. _............. !t 1 18 j 9^-7 21 21 i | 18 ! | 100 21 21 % Pregnant Females 92 100 100 Here and in all similar tables. the exposure level refers to that of the Fq generation. I ucc 002306 58 Table XXI. Summary of Average Litter Size for Each Treatment Group and Generation. Control 50 ppm VCM 500 ppm VCM Fj Generation Fg Generation F3 Generation * Significantly different, p = 0.05 Average Litter SizeSTD DEV 11.10 11.32 10.92 8.83* 12.57 11.93 0.38 0.39 0.36 0.36 0.39 0.39 59 UCC 002307 Table XX. Average Litter Size for the Three-Generat ion VCM Study. Control 50 ppm I 500 ppm Total Pups Born F1 Generat ion 195 F2 Generat ion 239 F3 Genera--Li2Q___ 22$ Total Number L i tters 22 19 19 Average Litter Size STD Dev 8.9 0.63 12.6 0.67 11.8 0.67 Total Pups Born 202 234 204 Total j Average Number J Litter Litters 1 Size 21 9-6 18 13.0 18 11.3 Total ST0 Pups Dev Born 0.64 184 0.69 255 0.69 285 Total Number Litters 23 21 21 Average Litter Size STD Dev 8.0 0.61 12.1 0.64 12.6 0.64 002308 oCo 60 Table XXII. Percentage of Stillborn Opus in the VCM Three-Generation Study F1 Generation F2 Generation F3 Generation Control Total Pups Born Total Stillborn 195 1 239 0 225 2 % Stillborn Total Pups Born 0.51 202 0.00 234 0.89 j 204 50 ppm Total % Stillborn Stillborn 5 2.48 2 0.85 5 2.45 500 oom Total Pups Born Total Stillborn % Stillborn 184 3 1.63 255 *> ** 0.78 265 3 1.13 ucc 002309 6i Table XXIII. Numbers, Sex, and Weights of Fj Generation Age Total Number of Pups of Pups Control 50 ppm Days Male Female Male ,, Female 1 1 102 92 105 93 500 ppm Male Female 93 88 Average Weight (Grams) Control Male Female 6.88 6.60 50 ppm Male Female ' 6.98 6.69 500 ppm Male . Female i 7.05 6.76 4 102 7 102 92 105 93 92 88 10.50 10.03 10.50 10.15 10.72 10.37 89 105 93 92 88 15.29 14.71 15.30 14.87 15.76 15.41 14 102 . 21 102 89 105 92 89 105 92 _____ . 4 92 88 .. i 92 j 28 27.8 26.8 -------j 44.5 f 42.4 !( 28.6 ; 44.3 27.2 28.6 42.8 45.4 1 23.4 44.1 002310 oc o 62 Table XXIV. Numbers, Sex, and Weights of F^ Generation Age Total Number of Pups of Pups Control 50 ppm Days Hale Female Hale Female 1 120 119 114 118 4 118 119 114 7 118 119 113 117 116 14 118 119 113 116 21 118 119 113 116 500 ppm Hale Female 123 130 Average Weight (Grams) Control 50 ppm Male____ Female___ Hale . Female 6.38 6.13 6.55 6.16 500 ppm Hale . Female 6.52 6.18 122 130 9.70 9.13 9.82 9.21 10.00 9.27 122 130 14.38 13.85 14.57 13.45 14.72 13.97 122 130 27.77 27.26 28.10 26.15 28.80 27.44 122 130 40.89 40.57 41.54 38.67 42.93 41.20 ogCo So 63 Table XXV. Numbers, Sex, and heights cf Generation Age Total Number of Pups of Pups Control 50 ppm Days Hale Female Hale Female 1 112 111 91 108 4 112 111 91 108 500 ppm Hale Female Average Weight (Grams) Control Male Female 50 ppm Male Female 139 123 6.82 6.38 6.84 6.61 139 123 10.18 9.47 10.35 9.76 500 ppm Hale Female 6.66 6.30 9.87 9.07 7 111 111 91 108 139 123 14.63 13.90 14.81 14.18 13.89 13.25 14 111 111 . 90 108 135 121 26.41 25.39 27.46 25.81 25.53 23.95 i 1 21 111 Ill 90 108 134 121 34.77 33.85 36.75 35.14 35.11 33.40 002312 oC o 64 it t Table XXVI. Viability, Survival, and Lactation Indexes in a Three- Generation Study of Reproductive Performance After of the Fq Parents to VCM Gas. Low Dose High Dose Generation Control 50 ppm 500 ppm F1 IOC 100 99.4 Viability Index F2 99.2 99.7 99.6 F3 100 100 100 Survival Index Day 21 Lactation Index j F1 j 98.5 | F2 | 99.2 F3 j 99.6 i F1 98.5 F2 100 F3 99.6 99.5 99.4 98.7 99.6 99.5 97.3 99.5 99.4 99.1 100 99.5 97.3 Viability Index Day 21 Survival Index Lactation Index No. o:: pros alive at Day 4 x 100 No. 6f pups born alive No. of imps alive at Day 21 -jqq No. c.' pups born alive No. of pups alive at Day 21 x inn No. cf jJ'Vps alive at Day 5" Banerjee, B.N., Course Director, Teratology-Principles and Procedures Related to Fetal Development. Tne Center for Professional Advancement, Sommerville, New Jersey. September 1974. Course Notes. 65 Ucc 002313 Figure XVIII Control - 8 months FIGURE LEGEND Control hepatocyte morphology with normal appearing endothelium adjacent. Xo,000. Negative #193,033 UCC 002314 65 Figure XVIII ' Control - S njonths - . . *' r't * ' r*^ **v^ -'..v.** '- * f?1**' * '*-. w\S##*K'- vv',-^V [, / \ ' ` -*v' --*^" v-' ',:v' >* v.-*-V> " '!*'*? 'w Yiv i . ;.:Y . ?*-'\Y .. f. *-1 "' *' ' .- . -<**' ' *' -;r . , ''.vA'.V-C"i -.->? . ; V. " '* * f' ' ...' \ ** . .*..- i- " ` ~T / 'v,-, ,<- t*wv. !'>-;.>. , -- - , - s * *\ i -S.c* '-*.\/ .v k*_*j #* * ' ^ *' '* - * 1 ,, v-y .a- ,. 1. * -> /*' ' J< /'/ if' u.r. :Y,,";_>:>'ij ; ucc 002315 Figure XIX Single Exposure - 8 months post-exposure FICL"r; TJ'GEMP Ilepatocj'tes show lipid droplet accimulation. At center of "lela, cytoplasm devoid of recognizable organelles is in shape of bleb. Adjacent belb is apparently free in space of Disse. This may represent a means whereby cytoplasmic material may be shed, y.5,000. ;.'.f j -e ^32301 ucc 002316 67 ucc 002317 Figure XX Single Exposure - 8 months post-exposure FIGURE LEGEND Figure shows portion of tv,o ilocyNote numbsrs of snail dense todies with one or i.o+e o'x</tr;-;2-iucent regions. These are lipofuchsin granules. X4.000. Negative #A54,04S 8 l ucc 002318 Figure XXI Multiple Exposure - 8 months post-exposure FIGURE LEGEUD Kupffer cell shows evidence of active phagcieytic function. Large residual body suggests prior engulfraont of entire cell (perhaps a white blood cell). X5.000. negative "198,063 ucc 002320 Figure XXII VINYu aixniDE/MLTIPI DOSE ie cities post exposure controls F&riaTes VP76 136-138 Changes -ve crc.hr:.d in tills control group. Tn hepntccytes, cleft-like spaces sv'n in dense bodies. These were suggestive of a crystalline neatericl eanrcctcd during processing. Kupffer cells were large and contained large residual bodies. Vesicular endopl asniic reticulun and bizarre shaped ni.tochondria were occasional findings. 4 70 UCC 002322 Figure XXII Controls - Multiple Exposure - 16 months post-exposure FIGURE Hepatocyiie eontains numerous dense bodies and lipofuohsin granules. Clef v.hite spaces In dense bodies may represent crystalline irutorial extracted during processing. X5.000. Negative ^107, lf-7 4 ucc 002323 Figure XXII Control - Hu_1_t 1plf_jxp|ure_ -. 16 mont h post - exposure r .c'-' " /'S.<':. -a'- v > : '/ f: ' * , y 'r-f*- r\* . V. ii . ' . - '. v - t- * -s; ", ! , , . ' > n- :>V', - V.! :<: 1 * V /. X;. r.i ' `V >S'::V' '< " XI x <J * * . v Vi v' '-"v t * . <. r?' . * > p' 4'. ,* > >..x* - :* -- v',f: .:n v c . >.v-?d \\ ',* * . .". - r*** 'i ;v *' */ . -v-;i. - > .;.. . > ... /^ .- v V -X't\,i - . , . ` S *. r * "* fc -r* . - . .'7r 1 'V. * f i,- 'v l i;.-... . * Z.Vi . .. . ; t ss im" .v. . : .. "V`- 'vw - . ff. * ^ .> > . * . t -v" V'c^vSr* i r,* # V'.-' . ;/ - . . ' >. !. ...'j, .. J-/ V-- . ^ ' . .. - . . _ 'f ` r-A- ,. .. . . ` .s'.\ y<K -iflVl''.!! *Tjr >s ,, ... ^ \/% .v ; . . . }. . ;.. i* > '* t > 1 ' v'"'*S. " > '`.'/l ' .f E,' ,1, ** ' i.xtr . ' 1 / /i. ':C ,, * l *' I `l, ,.4 ' , s-'y.fV-f".-. C&-- ?>*. .... v- f..:' h *% j. t ,rr:*v,^y-> v.'.y"*;v. ; -., /*'v *>* ;'vj>2i ( , . it *."** ^ 'YY.rvv. v.^ 4 ucc 002324 Figure XXIII Multiple Exposure - 16 months post exposure FIGURE USGEND i Indented nucleus at center of Held consistent with ai ienocarcinocna. Cell at upper left if; necrotic and show's mitochondria with high amplitude swelling and f'lecculunt densities. X5,000. Negative #107,434 f 71 ICC 002325 Figure XXIV Controls - 24 months FIGURE IEGENT) Normal appearance of hepatocyte in this control group. X5.000. Negative *202,293 4 72 occ 002321 X. Figure XXIV Controls - 24 months hr V,?;r t> 'j\- - -> - '/>-y,1 - \ * i ' *,-7li : ! \ t-V-' "'* *, , v 1 * -s ;**;.t*:J*?/,Nv * ' x t i-. ,, ; . *> v.j." - fv`^ ' . ' * -' % s . * 4 . \V "' ' t* ft/, . j- > 3 >iA ' ^*v. /; >vA 1 ,; :/;v' 'V>< , " v _. * -* * * {*\* .*r*-,,;.V' ', * p'-# * \-tV' "* i>, . "``k * ''-!"V **ivi'# '-.',< v ' *;^V *'>-: *'*i.?r* ' *5* w" ' *<! ','**/. V***,* ^'"t * * - -r .y '!* . , i* V .h , ^ - Sk XvZ-.-C Vv\ ; ---. . -- A--. . .'. y' - ", Vjj *'.s'1A"T** 1{ * '1 ' .-;...sJ*: /"v. ' | Vr's,",' h>, /.', . *.*.*. ' * , * \'V ' -t i v* w'r;. ;h.5^sa. ;:.-.v; ' "'di :;t. rr** A i v4. \ 0 u-L .-v1 -jk*:'1 ' -^.v.. ^.L>*-''% **K.-S, rS\ , ,, ._,** - * /- -` 7 , k ,* ^ j ^ vtn* lr 1 ^ ,t |T *- \ Figure XXV VP77 67-71 KALES 10 WEEKLY EXPOSURES TO PPM VI.3YL OiLORIDE POST EXPOSURE 106 WLEKS General Sunmary of Findings: Changes within hepatocytes included increased 1ipofuchsin granules and endoplasmic reticulum dilatation wit): a material of low electron density filling endoplasmic reticulum cisternae. in audition, bile canaliculi frequently showed distended lumens with the supplication of the plasma membrane at this site. Non-hepatccytic changes inuluied enlarged Kupffer cells with abundant lysosanen and bodies end a mononuclear infiltrate into hectic parenchyrv. ' ti 73 ucc 002329 ucc 002330 ].'p 7 7 '7 < 9 A j?